ABSTRACT BOOK - Final

4th International Conference of the European College of Veterinary Microbiology, 15-17 September 2022, Bari,
Italy
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Organizing Committee
NICOLA DECARO
DVM, PHD, Full Professor of Infectious Diseases of Animals Ebvs®
European Specialist in Veterinary Microbiology Member of European College of Veterinary
MARIALAURA CORRENTE
DVM, Associate Professor, Department of
Veterinary Medicine, University of Bari “Aldo Moro”
GABRIELLA ELIA
Full Professor of Infectious Diseases of Animals, Department of
Veterinary Medicine, University of Bari, Italy
PROF. TOBIAS EISENBERG
EBVS® European Specialist in Veterinary Microbiology Member of European
College of Veterinary Microbiology
DR. LIBBY GRAHAM

MVB MVM PHD DIPL. ECVM MRCVS
PATRÍCIA ALEXANDRA CURADO QUINTAS DINIS POETA
Full Professor EBVS® European Specialist in Veterinary
Microbiology, ESGVM Executive Committee of ESCMID, Head of
Microart- Microbiology and Antibiotic Resistance Team, Head of
Medical Microbiology Laboratory-UTAD, AQV– REQUIMTE, Lisbon
Scientific Committee
PROF. BRYAN MARKEY
BRYAN MARKEY IS ASSOCIATE PROFESSOR OF VETERINARY MICROBIOLOGY AT THE
DUBLIN SCHOOL OF VETERINARY MEDICINE. HE IS HEAD OF THE SECTION OF
VETERINARY PATHOBIOLOGY AND A MEMBER OF THE SCHOOL EXECUTIVE
PROF. CANIO BUONAVOGLIA

DVM, FULL PROFESSOR OF INFECTIOUS DISEASES OF ANIMALS
PROF. DR. PATRICK BUTAYE

DVM, PHD, DIPL. ECVM
DEPARTMENT OF PATHOBIOLOGY, PHARMACOLOGY AND ZOOLOGICAL MEDICINE
LUCA GUARDABASSI
DVM, PHD, PROFESSOR IN ONE HEALTH AMR AT THE UNIVERSITY OF COPENHAGEN
DR. ANTONIO MARTÍNEZ-MURCIA

PROFESSOR OF MICROBIOLOGY, MIGUEL HERNÁNDEZ UNIVERSITY
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Invited Speaker
THE ASF VACCINE IS CLOSER THAN EVER
J. M. Sanchez- Vizcaino
Complutense University of Madrid, Spain
J. M. Sanchez- Vizcaino
ORIGIN AND STUDIES:

Born in Murcia, Spain. He received a bachelor's degree and a doctorate (DVM and Ph.D) from the Faculty of Veterinary Medicine of the Complutense
University of Madrid (UCM), subsequently completing postgraduate studies in animal immunology and virology at Cornell University in New York.
PROFESSIONAL EXPERIENCE:
Upon his return to Spain, he joined the National Institute for Agricultural and Food Research (INIA), where he entered as a researcher in 1978 and
developed a large part of his professional career as a researcher and director of important departments and centers, such as: Director of the
Department of Animal Virology (ASF eradication program in Spain), Director of the Department of Animal Health (control and eradication of ASF
and African horse sickness) and Founding Director of the High Biological Safety Center (BSL3 and BSL3+) for Animal Health Research (CISA) Fight
against classical and African swine fever, horse sickness, PRRS, BSE, etc. until 2002 when he joined the UCM as Professor.
Since 2002 he has been Full Professor of Animal Health at the Complutense University of Madrid, carrying out his teaching and research work in
the Department of Animal Health of the Faculty of Veterinary Medicine and in the VISAVET Center of the UCM, as well as director of the reference
laboratory of the World Organization for Animal Health (OIE) for African Swine Fever and African Horse Sickness.
The scientific contributions of Professor Sánchez-Vizcaíno have contributed significantly to the control and eradication of several animal diseases
on four continents, including African swine fever, African horse sickness and classical swine fever. All this, thanks to the development of new rapid
and sensitive diagnostic methods and reagents, new strategies and epidemiological models, and the development of vaccines. He is currently de
coordinator of the EU VACDIVA project: A safe, DIVA and protected vaccine for ASF in wild boar and domestic pigs.
He has given different training courses and seminars on the early detection of infectious diseases, diagnosis of viral diseases and their control and
eradication, in several countries on the five continents.
He being appointed since the year 2021 as Emeritus Professor of Animal Health at the UCM.
ACHIEVEMENTS AND PUBLICATIONS:
He has more than 260 scientific publications in high-impact international journals, as well as being the author of 47 chapters in internationally
prestigious books (African swine fever in Diseases of swine, African horse sickness in the OIE manual, Trends in Emerging Viral Infections of Swine.
) and digital courses dedicated to immunology, infectious diseases, and health simulations.
He has directed more than 150 national and international research projects and 37 doctoral theses on animal infectious diseases, has participated
in 80 R&D contracts, in 10 patents and has participated as a speaker in more than 500 congresses, conferences, courses and interviews. (TV, radio,
press, video, web) all over the world.
AWARDS AND SCIENTIFIC DISTINCTIONS:
He has several decorations and appointments, among which we highlight: the Commendation of Number of the Order of Agrarian Merit (03/12/99)
and of Food Merit (05/05/03), the Cross of Military Merit with white badge (03 /01/03), all for his contributions to animal health and the control of
infectious diseases. He received the ANAPORC 1990 national pig research award, the "PORCO BRAVO 1999" International Pork Award, the 2000
First Prize in Pork Health and Production, the 2007 Animal Health Award "The Best of La Verdad 2007" and the 2012 Albéitar Awards in the Scientific
Category, as recognition for his contribution to the scientific development of research in Veterinary Science, SEPOR 2019 RESEARCH Award for his
lifelong dedication to research in Animal Health and the achievements made.
He has been awarded the Medal of Merit from the World Organization for Animal Health (OIE), in international recognition of his exceptional
services to veterinary science (Paris 05/24/09), and has been awarded Doctor Honoris Causa by the University of Murcia (Murcia 04/22/10), as well
as named 2013 George C. Poppensiek Visiting Professor in Global Animal Health by Cornell University (Ithaca 09/2013) and Honorary Professor
(Adjunct Professor) of the University of Minnesota (since September of 2018).
CURRENT SCIENTIFIC INTEREST
His scientific interest focuses on the study of the epidemiology and preventive medicine of animal infectious diseases, as well as the development
of new diagnostic techniques, new generation vaccines and models and new strategies for their control. He is currently PI of several research
projects, among which we highlight the European VACDIVA, "H2020-SFS-2019-1, theme: A vaccine against African Swine Fever" endowed with 10
million Euros and the project, financed by the Institute National Health Department, titled the role of COVID 19 in pets.
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Invited Speaker
MALASSEZIA YEAST: AN EMERGING PROBLEM IN MEDICAL
AND VETERINARY MEDICINE
C. Cafarchia
Department of Veterinary Medicine, University of Bari Aldo Moro, Bari, Italy
Malassezia spp. organisms are lipid-dependent yeasts, inhabiting the skin and mucosa of
humans and animals. They are involved in a variety of skin disorders in humans and
animals and may cause bloodstream infections in severely immunocompromised patients.
Despite a tremendous increase in scientific knowledge of these yeasts during the last two
decades, the epidemiology of Malassezia spp. infections and fungemia in animals and
humans remains largely underestimated. In addition, the pathogenic role of these yeasts
species have been suggested only for strains causing skin infections. Since multiple
Malassezia spp. and/or genotypes with varying antifungal profiles may cause unique or
similar pathologies, serious concern about the diagnostic procedures and antifungal
treatment have been raised. Despite the attempt to treat these fungal infections, a trend
toward the recurrence is often observed in humans and animals with dermatitis. Moreover,
the clinical evidence of treatment failure with terbinafine in patients with Pityriasis versicolor
or with itraconazole in dogs with dermatitis as well as with fluconazole or posaconazole in
preventing M. furfur fungemia in humans suggested a probable occurrence of drug
resistance phenomena in these species. However, in vitro susceptibility testing for
Malassezia spp. has not yet been standardized but the low activity of fluconazole,
voriconazole and echinocandins against Malassezia spp. seems to be a common finding
regardless the testing methods. Herein, the most recent literature on Malassezia spp. skin
infection and fungemia, pathogenetic mechanisms of Malassezia spp, diagnostic
procedures, in vitro susceptibility testing and therapeutic approaches will be summarizes
and discussed.
Claudia Cafarchia
D.BSc, Phd, Associate Professor at the Department of Veterinary Medicine, University of Bari
Aldo Moro. She attended the course of Mycology Medical Mycology at The Centraalbureau voor
Schimmelcultures (CBS Fungal Biodiversity Centre), Utrecht, The Netherlands. She is a
member of the ISHAM-Veterinary Mycology Working Group and of the ISHAM-Malassezia
epidemiology and Working Group. Supervisor of Mycology laboratory and Member of the
Scientific Committee of Specialization Courses in "Infectious Diseases, Prophylaxis and
Veterinary Police" and of the Scientific Committee of the PhD course in “Animal Health and
Zoonosis” University of Bari, Italy.
In charge of the courses of Veterinary Mycology at Department of Veterinary Medicine and

Supervisor for degree, Phd and specializations theses. Associate Editor of the Medical
Mycology, Academic Editor of Antibiotics. Responsible and Component of research projects
admitted for funding on the basis of competitive call. The professional and scientific activity is
aimed in studying fungi of medical and veterinary concern. Scientific activity consists of an
overall production of 202 scientific articles in international and national journals; 90 contributions
in international / national conference; 6 book chapters. She is one of two co-editors of a
comprehensive "Micologia veterinaria e comparata” Textbook with ISBN published by Aracne
Editore pp.1-378.
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Invited Speaker
IMPACT OF ANIMAL ROTAVIRUSES ON HUMAN HEALTH
V. Martella
Unversity of Bari Aldo Moro, Department of Veterinary Medicine, Valenzano, Italy
Rotaviruses are important enteric pathogens of humans and animals. Group A rotaviruses
(RVAs) account for up to 1 million children deaths each year, chiefly in developing
countries. Both mono-valent or poly-valent vaccines targeting the main VP7 and VP4
antigens (G and P type, respectively) are now available for prevention of acute gastro-
enteritis (AGE) in children. RVA-associated enteritis is also a major problem in young
calves, piglets and foals. Early in the epidemiological studies for RVAs in humans, either
sporadic cases or epidemics by atypical, animal-like RVA strains were described.
Complete genome sequencing of human and animal RVA strains has revealed a massive
genetic heterogeneity in the 11 double stranded RNA segments across different RVA
strains and has provided evidence for frequent intersections between the evolution of
human and animal RVAs, as a result of multiple, repeated events of interspecies
transmission and subsequent adaptation. After 2015, a pandemic horse-like G3 strain
emerged in human population worldwide. This strain was predominant in several
epidemiological studies. The virus differed in the VP7 gene and in the genome
constellation from classical G3 human viruses, demonstrating the potential impact of this
phenomenon on a global scale.
Vito Martella
Vito Martella received his veterinary degree in 1996 from the Faculty of Veterinary Medicine in
Bari, Italy. Between 1997 and 2000, he completed a PhD on infectious diseases of small
animals, and from 2000 he was a researcher working on various aspects of virology and
diagnostics. From 2001 to 2017, he was Associate Professor and since 2017 he is Full
Professor at the Department of Veterinary Medicine of Bari, Italy. From 2014 to 2018
Coordinator of the PhD program in Animal Health and Zoonoses. From 2018 to 2022,
coordinator of the Master course in Safety of food of animal origin and health (LM86). From
2019 to present, head of Section IV of the Italian Supreme Health Council. His current research
focus is primarily on enteric viruses at the interface between animals and humans, virus
discovery and characterization and optimization of diagnostic tests. He is member of the RCWG
(rotavirus classification working group) and of the ICVT (International Committee for Virus
Taxonomy) calicivirus study group. He has more than 400 publications in peer-reviewed
scientific journal, with more than 15000 citations and a H index of 61 in Scopus.
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Invited Speaker
BROADENING THE UNDERSTANDING OF THE MOLECULAR
BASIS FOR SHIGA TOXIN-PRODUCING ESCHERICHIA COLI
COLONIZATION IN CATTLE
S.A. Barth, C. Berens, M. Weber, C. Menge
Friedrich-Loeffler-Institut, Institute of Molecular Pathogenesis, Jena, Germany
Many cattle are colonized by Shiga toxin (Stx)-producing Escherichia coli (STEC), making
them a major source of human enterohemorrhagic E. coli (EHEC) infections. The highly
virulent E. coli O104:H4 strain having caused the largest outbreak of hemolytic uremic
syndrome in 2011 possessed a blended virulence profile of human adapted
enteroaggregative (EAEC) and EHEC. Experimental infection of calves proved that cattle
can even carry such unusual EHEC strains at least transiently. Harbouring STEC in their
intestinal tract without clinical symptoms, cattle were long believed to resist the detrimental
effects of the Stxs, the principal virulence factors in the pathogenesis of human EHEC-
associated diseases. However, different types of target cells for Stxs, primarily belonging
to the adaptive arm of the immune system, exist in cattle and depressed cellular immune
responses foster intestinal STEC colonization, an effect that can be mitigated by
Shigatoxoid vaccination. Nevertheless, even classical STEC strains differ in their ability to
colonize cattle more persistently (STECper) or only sporadically (STECspo). This difference
is primarily encoded in the accessory rather than the core genome but phenotypic traits
beyond classical virulence factors also correlate with the realization of a specific
colonization pattern. STECper have abandoned some metabolic traits, while STECspo
conserved properties leveraging survival in the environment. The ability to produce Stx,
therefore, seems to be sufficient to qualify pathogenic E. coli strains for both, being
successful colonizers in ruminants as well as emerging human pathogens while a range
of STEC/EHEC strains exist that have further adopted to cattle by metabolic adjustment.
Christian Menge
Prof. Dr. med. vet. Christian Menge
Institute of Molecular Pathogenesis, Friedrich-Loeffler-Institut -
Bundesforschungsinstitut für Tiergesundheit /, Federal Research Institute for
Animal Health
As a licensed DVM, Prof. Menge possesses broad knowledge and expertise in
surveillance, diagnosis, control and eradication of epizootic diseases in accordance
with rules laid down in national and international animal health legislation. He has
particular expertise in the analysis of bacterial host-interactions at the intestinal
mucosal surface of livestock. Using in vitro culture systems and experimental
infection studies in the target species his group elucidated the mechanisms by which
enterohaemorrhagic Escherichia coli (EHEC) manipulates its host´s immune
system. He also has substantial expertise in clinical immunology and the
immunobiology of other bacterial pathogens, e.g., tuberculous and non-tuberculous
Mycobacteria and Coxiella, viruses and parasites in large and companion animal
species. His recent research focusses on drivers and dynamics of host-to-host and
cross-species transmission of bacteria exhibiting antimicrobial resistance (AMR),
also deploying innovative sampling strategies from a One Health perspective.
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Invited Speaker
HEPATITIS E VIRUS AS A ZOONOSIS WITH DOMESTIC
AND WILD RESERVOIRS
J. R. Mesquita
Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto,
Epidemiology Research Unit (EPIUnit), Instituto de Saúde Pública da Universidade do
Porto, Porto,Laboratório para a Investigação Integrativa e Translacional em Saúde
Populacional (ITR), Porto, Portugal
Hepatitis E virus (HEV) is a member of the Hepeviridae family, Orthohepevirus genus,

which can be further subdivided into four species: Orthohepevirus A that can infect humans
and a wide variety of different animals, Orthohepevirus B that circulates in birds,
Orthohepevirus C found in rodents and ferrets, and Orthohepevirus D in bats. The
Orthohepevirus A species has 8 different genotypes (HEV 1-8), with types 1 and 2 infecting
exclusively humans; 3, 4, 7 and 8 with zoonotic transmission; and 5 and 6 infecting
exclusively animals. Among those infecting humans, genotype 3 is treated as an emerging
zoonosis of public health importance, being found in industrialized countries, including
those in Europe. In these countries, consumption of undercooked or raw infected pork, wild
boar and deer meat are major sources of exposure to HEV genotype 3. This presentation
will provide an overview of the current knowledge on zoonotic hepatitis E virus, providing
information on domestic and wild reservoirs for infection.
João Mesquita
João Mesquita (Mesquita JR) holds a bachelor in Veterinary Medicine, a MSc and
a PhD in Virology. He is a Diplomate of the European College of Veterinary
Microbiology since 2019. He has been teaching since 2006 and is currently affiliated
to Instituto de Ciências Biomédicas Abel Salazar – Universidade do Porto, Portugal.
Since 2006 he has been involved in several projects related to infectious diseases
epidemiology. His research interests are focused particularly on human and animal
infectious diseases, particularly on zoonotic agents, with special emphasis on
surveillance and epidemiological tools
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Invited Speaker
CURRENT AND FUTURE POSSIBILITIES OF MALDI-TOF MS AND
NANOPORE SEQUENCING FOR BACTERIAL IDENTIFICATION
AND SUSCEPTIBILITY TESTING
F. Boyen
Department of Pathobiology, Pharmacology and Wildlife Medicine, Faculty of Veterinary
Medicine, Ghent University, 9820 Merelbeke, Belgium.
MALDI-TOF MS has revolutionized bacterial identification in both human and veterinary

medicine. It is a simple, rapid, cost-effective and robust tool for the identification of most
bacteria and fungi to the species level and sometimes even beyond. Even though MALDI-
TOF MS is mainly used for identification, it can also be used to detect antimicrobial
resistance, using different approaches. First of all, specific proteins (peaks) can be
identified that are linked with a certain resistant subpopulation. This peak profiling often
has no direct link with the resistance mechanism, so may only predict resistance in specific
subgroups of a bacterial species in regions where these subgroups are highly prevalent.
Secondly, specific kits allow the detection of strains able to enzymatically degrade
antibiotics (for example ESBL or carbapenemase producing Enterobacteriaceae) in a +/-
2hour protocol. Finally, protocols have been described able to discriminate susceptible
from resistant strains for various antibiotics in various bacterial species after short
incubation with the respective antimicrobial agents using 1 to 6 hour protocols (Idelevich
and Becker, 2021). Increasing quantity and quality of database entries will ensure that
identification of bacteria will ameliorate continuously. In addition, machine learning may
increase possibilities related to identification, AMR detection and, to some extent, strain
typing and detection of host biomarkers (Mortier et al., 2021, Weis et al., 2022). Third
generation sequencing (3GS) allows high-throughput native DNA or RNA sequencing that
can be processed in real-time, providing an important opportunity to reduce sample-to-
result time. Even though there are various providers on 3GS, Oxford Nanopore
Technologies currently are most probable to break through in veterinary medicine. Apart
from identification of pathogens, 3GS also holds possibilities for DNA-based antimicrobial
resistance detection and strain typing. Even though 3GS might lead to the most
comprehensive and most reliable results ever in veterinary microbiology, this technique
also holds several risks. First of all, due to the fact that all observed microbial agents that
are present in the sample are reported, there is a risk of over-interpretation of obtained
results: which of the reported microbial agents are clinically relevant? This is especially
important in samples where a microbiome can be expected to be present. In addition, this
technique may hold a risk of putting too much focus on the pathogen, while especially in
multifactorial diseases also predisposing factors, non-infectious primary problems etc
should not be neglected. Considering this technique is still in its infancy, there is still a lot
of work to be done on setting up and validating workflows. A possible bottleneck may be
the bio-informatics expertise for setting up and validating sequence analysis pipelines. In
addition, using DNA-based detection of antimicrobial resistance determinants to predict in
vivo susceptibility will also require further research and validation. Both hardware and
software-related progress will result in decreasing both price and sample-to-result time of
3GS, provided that general laboratory equipment and 3GS specific consumable prices
would not increase significantly. Automated and species-specific bioinformatics protocols
will be able to increase quality of (deep) sequencing data and shorten processing and
interpretation time (Vereecke et al., 2020). In addition, in near future, it might become
possible for diagnostic labs and maybe even veterinary clinics to have their own
sequencing station without the need for unrealistic investments. While sample preparation
and sequencing can be performed on-site immediately after sampling, online access to the
bioinformatics protocols of specialized providers with appropriate computational power
may allow high quality and even real-time processing and interpretation of the results. Such
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a work flow might result in a sequencing report, possibly within hours after sampling
(Marcolungo et al., 2022). Validation of such point-of-care set-ups and clinically relevant
interpretation will however be of major importance.
REFERENCE LIST
Idelevich EA, Becker K (2021). Matrix-Assisted Laser Desorption Ionization-Time of Flight
Mass Spectrometry for Antimicrobial Susceptibility Testing. J Clin Microbiol.
59(12):e0181419.
Marcolungo L, Passera A, Maestri S, Segala E, Alfano M, Gaffuri F, Marturano G, Casati
P, Bianco P.A., Delledonne M (2022). Real-Time On-Site Diagnosis of Quarantine
Pathogens in Plant Tissues by Nanopore-Based Sequencing Pathogens 11 no. 2: 199.
Mortier T, Wieme AD, Vandamme P, Waegeman W (2021). Bacterial species identification
using MALDI-TOF mass spectrometry and machine learning techniques: A large-scale
benchmarking study. Comput Struct Biotechnol J. 19:6157-6168.
Vereecke N, Bokma J, Haesebrouck F, Nauwynck H, Boyen F, Pardon B, Theuns S (2020).
High quality genome assemblies of Mycoplasma bovis using a taxon-specific Bonito
basecaller for MinION and Flongle long-read nanopore sequencing. BMC Bioinformatics
21(1): 517.
Weis C, Cuénod A, Rieck B, Dubuis O, Graf S, Lang C, Oberle M, Brackmann M, Søgaard
KK, Osthoff M, Borgwardt K, Egli A. (2022). Direct antimicrobial resistance prediction from
clinical MALDI-TOF mass spectra using machine learning. Nat Med. 28(1):164-174.
Filip Boyen
Filip Boyen graduated at the Faculty of Veterinary Medicine, Ghent University,
Belgium in 2002. In that year, he started his PhD research on Salmonella
pathogenesis in pigs at the Laboratory of Veterinary Bacteriology and Mycology
at the same Faculty. He finalized his PhD in 2007, and worked at the Division of
Poultry, Exotic Companion Animals, Wildlife and Experimental Animals for 1 year
as assistant. In 2008, he joined the Laboratory of Veterinary Bacteriology and
Mycology again as laboratory coordinator, a position with diagnostic, teaching and
research responsibilities he still holds today.
His research addresses different bacterial diseases in various animals species,
ranging from an occasional clinical case report, over pathogenesis research and
susceptibility testing, to experimental trials. He considers himself to be a
“generalist”, but his main topics of interest are respiratory and intestinal infections
and antimicrobial resistance in bacterial pathogens of veterinary and zoonotic
importance. The last few years his research focusses also on the use of MALDI-
TOF in diagnostics and innovative applications of this technique.
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Invited Speaker
EVOLUTION OF ZOONOTIC SARS-COV-2 DATABASE AND ITS
INFLUENCE IN ASSIGNING THE PHYLOGENETIC VALUE OF
MUTATIONS FOR LINEAGE-SPECIFIC DETECTION
A. Martínez-Murcia
Department of Microbiology, University Miguel Hernández, and 2Genetic PCR
Solutions™, 03300-Orihuela, Alicante, Spain
One of the pending tasks in the research of SARS-CoV-2 is the determination of a possible
intermediate host in transmission from the natural reservoir. The development of genetic
detection methods is undoubtedly essential to achieve this goal. However, qPCR designs
are based only on the sequences that we are describing and depositing in databases that
do not necessarily represent all natural diversity. Early on this pandemic, our laboratory
has contributed with qPCR kits for SARS-CoV-2 detection and kits specific for some of its
variants of concern (VOC). Post-marketing surveillance plan included a rigorous check of
the new sequences determined and available at the GISAID database (Global Initiative on
Sharing All Influenza Data). In our view, due to the criteria recommended to select samples
for sequencing (i.e., the Spike Gene Target Failure methodology) and the geographically
uneven sequencing capacity, sequences described so far may have been biased to enrich
only a homogenous part of diversity, particularly within the Alpha, Delta and, lately, the
Omicron variants. Phenotypic relevance of mutations, or their possible clinical impact,
cannot always being taken as phylogenetic markers of SARS-CoV-2 variants.
Considerable number of mutations should be determined to properly ascertain the
phylogenetic identification of lineages. A simple sequencing strategy, directed to a 952 bp
S-gene fragment comprising 29 informative mutations, was developed in our laboratory for
affordable determination of most variants of concern.
Antonio Martinez-Murcia
Dr Antonio Martinez-Murcia (6th March 1964) Professor of
Microbiology, Miguel Hernández University, Alicante, since 1993),
B.Sc. in Biochemistry (Univ. Valencia 1987). Ph.D. in Microbiology
(Univ. Reading, UK, 1993). He is an active member of the
“Aeromonas Working-group” of the “International Sub-Committee
on Vibrionaceae, Aeromonadaceae, and related genera”
(International Committee on Systematics of Prokaryotes). Executive
member of Association of Veterinary Laboratory Diagnosis
Specialists (AVEDILA) and board member of The European
Association of Veterinary Laboratory Diagnosticians (EAVLD).
Secretary of the board of the Association of Biotech Companies of
Alicante (AEBA). Member of Spanish Society of Microbiology since
1989 (SEM). He is co-author of > 90 papers in peer-reviewed
journals and book chapters, a patented PCR method (1994), co-
director of 8 doctoral theses, and has described 16 new bacterial
species. Counts with pioneering experience in sequencing
techniques, bacterial phylogeny/evolution/taxonomy, genetic
identification/detection/typing. He has participated in >30 R&D
projects, including several from the European Commission.
Founder and Director of GPS™ laboratory, GENETIC ANALYSIS
STRATEGIES SL, awarded in 2002, 2012 and 2015.
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Invited Speaker
EUROPEAN NETWORK FOR OPTIMIZATION OF VETERINARY
ANTIMICROBIAL TREATMENT
P. Damborg
Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg,
Denmark
The European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT)

is a COST Action launched in 2019 and ending in 2023. The aim of ENOVAT is to optimize
veterinary antimicrobial use with special emphasis on the development of antimicrobial
treatment guidelines and refinement of microbiological diagnostic procedures. In this talk,
the structure of ENOVAT, including its five working groups (WGs), will be briefly introduced
followed by a presentation of selected results. Focus will primarily be on a WG1 survey
from 2021 on practices in veterinary microbiological diagnostic laboratories. Almost 300
respondents from 34 countries took the survey. Results showed large differences between
the practices and standards used. For example, the reported average turnaround time for
antimicrobial susceptibility testing (AST) of fast-growing organisms ranged from 1-2 days
to 6-8 days. Also, quality assurance, use of breakpoints for AST, the level of guidance
provided to veterinary clinicians, and other practices varied substantially between
laboratories. These results indicate a need to heighten standardization of diagnostic
microbiology practices, for example by development of consensus guidelines describing
recommended approaches from sample arrival until diagnostic answer.
Peter Damborg
Peter Damborg received his veterinary degree in 2004 from the
Royal Veterinary and Agricultural College in Copenhagen,
Denmark. Between 2005 and 2008, he completed a PhD on
zoonotic enteric bacteria in dogs, and from 2008 to 2013 he was a
postdoctoral researcher working on various aspects of antimicrobial
resistance and diagnostics. Since 2013, he has been Associate
Professor at the Department of Veterinary and Animal Sciences at
the University of Copenhagen. His current research focus is
primarily on surveillance of AMR in animals and rationalization of
veterinary antimicrobial treatment through development of antibiotic
treatment guidelines and research into novel treatment regimes,
alternative antimicrobial agents (e.g. peptides), and optimization of
diagnostic tests. He is head of a local veterinary diagnostic
laboratory forming the basis of the ECVM Training Centre at the
University of Copenhagen. Since 2018, he has been Chair of the
EUCAST subcommittee VetCAST, and since December 2019 he
has been Chair of the COST Action ENOVAT (European Network
for Optimization of Veterinary Antimicrobial Treatment). Over his
career, he has published more than 70 articles in international
scientific journals with peer-review (https://orcid.org/0000-0001-
9932-5996).
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Invited Speaker
ANTIMICROBIAL STEWARDSHIP AND PETS
N.E.M. Hopman1, I.M. van Geijlswijk2, L. Portengen, J.A. Wagenaar, D.J.J. Heederik,
E.M. Broens
1
Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht
University, Utrecht, the Netherlands
2
Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht
University, Utrecht, the Netherlands
To curb increasing resistance rates, responsible antimicrobial use (AMU) is needed, both
in human and veterinary medicine. In human healthcare, antimicrobial stewardship
programs (ASPs) have been implemented worldwide to improve appropriate AMU. No
ASPs have been developed for and implemented in companion animal clinics yet. The
objective of the present study was to implement and evaluate the effectiveness of an ASP
in 44 Dutch companion animal clinics. The objectives of the ASP were to increase
awareness on AMU, to decrease total AMU whenever possible and to shift AMU towards
1st choice antimicrobials, according to Dutch guidelines on veterinary AMU. The study was
designed as a prospective, stepped-wedge, intervention study, which was performed from
March 2016 until March 2018. The multifaceted intervention was developed using previous
qualitative and quantitative research on current prescribing behavior in Dutch companion
animal clinics. The number of Defined Daily Doses for Animal (DDDAs) per clinic (total,
1st, 2nd and 3rd choice AMU) was used to quantify systemic AMU. A statistically significant
decrease of 15% (7%-22%) in total AMU, 15% (5%-24%) in 1st choice AMU and 26%
(17%-34%) in 2nd choice AMU was attributed to participation in the ASP, on top of the
already ongoing time trends. Use of 3rd choice AMs did not significantly decrease by
participation in the ASP. This study shows that, although AMU in Dutch companion animal
clinics was already decreasing and changing, AMU could be further optimized by
participation in an antimicrobial stewardship program.
Els Broens
Els Broens was trained as veterinarian at the Faculty of Veterinary Medicine, Utrecht
University. After a few years work in farm animal practice, she had several positions as
lecturer and researcher at the Faculty of Veterinary Medicine and the Central Veterinary
Institute of Wageningen University and Research. In 2011, she finished a PhD on
livestock-associated MRSA in pigs at Wageningen University and the National Institute
for Public Health and the Environment. In the same year she became Dutch specialist
in Veterinary Microbiology. After the completion of her PhD and residency, she returned
to the Faculty of Veterinary Medicine in her current position as Director of the Veterinary
Microbiological Diagnostic Centre (VMDC) that processes over 20.000 samples per
year. Besides directing the VMDC, she is deputy chair and principal investigator within
the Division Clinical Infectiology. She co-authored multiple grant proposals and is
involved in several research projects on zoonoses and antimicrobial resistance in
companion animals. She is co-promotor of two PhD-students and supervisor of residents
in Veterinary Microbiology. First for the Dutch specialization, but in 2019 she became
Diplomate for the recently established European College for Veterinary Microbiology
(ECVM) and supervisor of residents for this college. Els is member of several
(inter)national committees and working groups on veterinary microbiology, public health,
zoonoses, antimicrobial resistance and veterinary antibiotic policies. She is co-chair of
the COST Action 18217: European Network for Optimization of Veterinary Antimicrobial
Therapy (ENOVAT) and chair of the ESCMID Study Group for Veterinary Microbiology
(ESGVM).
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Invited Speaker
ANTIMICROBIAL AND BIOCIDE RESISTANCE AMONG FELINE
AND CANINE PATHOGENS FROM DIAGNOSTIC SUBMISSIONS
A.R. Schug 1,2, A.D. Scholtzek 1,2, A.T. Feßler 1,2, B. Kohn 2,3, C. Weingart 2,3, A.-K.
Schink 1,2, A. Bethe 1,2, A. Lübke-Becker 1,2, D. Hanke 1,2, S. Schwarz 1,2
1
Institute of Microbiology and Epizootics, Centre for Infection Medicine, Department of
Veterinary Medicine, Freie Universität Berlin, Berlin, Germany
2
Veterinary Centre for Resistance Research (TZR), Freie Universität Berlin, Berlin,
Germany
4
Small Animal Clinic, Department of Veterinary Medicine, Freie Universität Berlin, Berlin,
Germany
Objectives: Bacterial resistance to antimicrobials and biocides is an increasing threat.

Therefore, the aim of this study was to investigate the resistance properties of canine and
feline bacterial pathogens. Methods: A collection of 62 Staphylococcus aureus, 52
Staphylococcus pseudintermedius, 49 Enterococcus faecalis, 37 Enterococcus faecium,
59 Escherichia coli, 56 Pseudomonas aeruginosa, and 14 Acinetobacter baumannii, was
tested for their susceptibility to antimicrobial agents according to the Clinical and
Laboratory Standards Institute (CLSI) and to biocides (benzalkonium chloride,
chlorhexidine, polyhexanide, and octenidine) using a recently developed broth
microdilution method. Results: Among staphylococci, penicillin resistance was the
dominant resistance property (including 29 methicillin-resistant isolates), followed by
macrolide, fluoroquinolone and tetracycline resistance. E. faecalis showed no expanded
resistance properties, in contrast to E. faecium showing resistance to penicillins,
macrolides, tetracyclines, and fluoroquinolones. A single vancomycin-resistant isolate
carrying vanA was detected. E. coli displayed expanded multiresistance phenotypes,
including a single carbapenem-resistant, blaOXA-48-positive isolate. In addition, three
multiresistant A. baumannii isolates were identified. The minimal inhibitory concentrations
(MICs) of the biocides were unimodally distributed, but differed with respect to the biocide
and the bacterial species investigated. No signs of resistance development were observed
for the bacterial species-biocide combinations tested, but due to solubility limits, some P.
aeruginosa isolates exhibited benzalkonium MICs higher than the highest test
concentration. Conclusions: During this study (multi)resistant isolates were seen among
most bacterial species investigated. Therefore, antimicrobial susceptibility testing is highly
recommended before starting antimicrobial chemotherapy. However, a development of
biocide resistance was not detected among the bacterial species investigated.
Andrea T. Feßler
Dr. med vet. Andrea T. Feßler, PhD
2003 to 2009 study of veterinary medicine at the der Ludwig-Maximilins-Universität in Munich
2012 PhD degree Title of thesis: "Comparative molecular analysis of methicillin-resistant isolates of
Staphylococcus aureus and coagulase-negative Staphylococcus ssp. from cases of mastitis among
dairy cattle" 2016 doctors‘ degree (Dr. med. vet.) "Studien zu Qualitätskontrollbereichen und klinischen
Grenzwerten für Cefoperazon zur Bekämpfung boviner Mastitiden" („Studies on quality control ranges
and clinical breakpoints for cefoperazone for the treatment of bovine mastitis “) 2009-2016 scientist in
the working group „Molecular microbiology and antimicrobial resistance“ from Prof. Dr. Stefan Schwarz
at the Institute for farm Animal Genetics of the Friedrich-Loeffler-Institute, Mariensee since 2017
scientist at the Institute for Microbiology and Epizootics at the Freie Universität Berlin in the working
group of Prof. Dr. Stefan Schwarz
Main research topics
• Antimicrobial Resistance from staphylococci and other bacteria
• Development of quality control ranges and clinical breakpoints for antimicrobial susceptibility
testing
• Development of testing methods and quality control ranges for biocide susceptibility testing
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Invited Speaker
DEVELOPMENT OF RAPID DIAGNOSTICS FOR THE DETECTION
OF PATHOGENS AND AMR
R. M. La Ragione
School of Biosciences and Medicine and School of Veterinary Medicine, University of
Surrey, UK
Culture remains the preferred method for the diagnosis of many human and animal
pathogens. However, it is labour intensive and expensive. Thus, there is an urgent
requirement for rapid, sensitive, specific and economic tests that can be used to diagnose
infections in animals and humans. Moreover, the rapid diagnosis of infections can assist
with the pragmatic and targeted treatment of infections, thus reducing the emergence of
antimicrobial resistance (AMR).
With the advent of Next Generation Sequencing (NGS) technology, and the public
availability of many full pathogen genome sequences, it is now possible to use comparative
genomics to develop robust molecular tests for the rapid and sensitive diagnosis of many
pathogens and AMR genes. One such test, is Loop Mediated Isothermal Amplification
(LAMP). LAMP can be used to identify pathogens and AMR in less than 15 minutes,
directly from a swab, with high specificity and sensitivity. LAMP can be performed with
minimal expertise, simple equipment and the cost is significantly less than culture or PCR.
LAMP can be utilised as both a laboratory based test or in Point of Care (PoC) applications,
and the advent of Artificial Intelligence (AI) algorithms has revolutionised how the test
results can be read and interpreted. This presentation will provide an overview of the
currently available rapid diagnostic tests for pathogen and AMR detection, with a focus on
the development and validation of LAMP assays for use as PoC tests for livestock, poultry,
companion animals and environmental samples.
Roberto La Ragione
Professor Roberto La Ragione – Professor of Veterinary Microbiology and Pathology, School
of Veterinary Medicine and Head of the School of Biosciences and Medicine, University of
Surrey
Roberto graduated in 1995 and then went on to study for a post graduate degree in veterinary
microbiology at the RVC. In 1996 he moved to the government Veterinary Laboratories Agency
to undertake a PhD on the pathogenesis of E. coli in poultry. In 2005 Roberto was appointed
head of pathogenesis and control at the APHA and in 2010 he was appointed Professor of
Veterinary Microbiology and Pathology at the University of Surrey.
Roberto gained the FRCPath in 2010 and in 2012 was appointed the Associate Dean for
Veterinary Strategy in the School of Veterinary Medicine. In 2014 he was appointed Head of
the Department of Pathology and Infectious Diseases and 2019 Deputy Head of School. In
2021 he was appointed Head of the School of Biosciences and Medicine. Roberto is the Chair
of the Royal College of Pathologists Veterinary Pathology Specialty Advisory Committee, Chair
of the Humanimal Trust, a Trustee of the Houghton Trust a member of the APHA Science
Advisory Board, and the past president of the Med-Vet-Net Association and the Veterinary
Research Club. Roberto is an Associate member of the European College of Veterinary
Microbiology (AECVM). In 2020 he was awarded Honorary Associateship of the Royal College
of Veterinary Surgeons (HonAssocRCVS).
Roberto’s current research interests focus on AMR and understanding the pathogenesis of
zoonotic bacterial pathogens. Roberto has a particular interest in the development of control
and intervention strategies, including rapid diagnostics, vaccination, and probiotics for the
control of pathogens such as Salmonella, Brachyspira and E. coli in food producing animals.
Roberto has published over 180 peer reviewed papers in the area of microbiology.
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ORAL, AULA A
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OA1
DIFFERENT GENOTYPES OF MALASSEZIA PACHYDERMATIS ISOLATED FROM DOGS

I. Brodard, A. Thomann, V. Perreten, S. Kittl
Institute of Veterinary Bacteriology, University of Bern, Bern, Switzerland
Malassezia pachydermatis is a basidiomycotic yeast frequently found in the ears of dogs, where a
high load is associated with otitis externa and atopic dermatitis. M. pachydermatis is culturable on
Sabouraud dextrose agar containing Tween 80 and species identification is usually possible using
MALDI-TOF MS. However, we found a number of isolates that could not be identified using the
standard Bruker library.
Eight of these isolates were used to establish MALDI-TOF reference spectra and subsequently
subjected to whole genome sequencing. In addition, minimal inhibitory concentrations of antifungals
were determined using Sensititre™ YeastOne YO10 system with small broth modifications.
Wilcoxon rank sum test was applied to compare MIC values. Phylogenetic analysis of whole
genome sequences revealed three clusters, two for the here described isolates (designated cluster-
15KM2158 and cluster-21KM0316), and one containing the reference strain (NR 126114.1). A
similar clustering was observed based on the MALDI-TOF reference spectra. Isolates of a same
cluster generated MALDI-TOF scores above 2.0 (the threshold for species identification), whereas
the score was below 2.0 when the reference spectra of another cluster were used. MALDI-TOF
identification can therefore be difficult if one of the references is missing in the database.
Interestingly, isolates belonging to cluster-15KM2158 (n=7) had significantly (p=0.002) higher MICs
(8 -64 mg/L) for fluconazole compared to isolates of cluster-21KM0316 (n=7) (2-8 mg/L).Our results
show that there is considerable variation among M. pachydermatis isolates from dogs, which can
complicate diagnostics. Further research is necessary to determine if strains of cluster-15KM2158
consistently show higher fluconazole MICs.
OA2
NOVEL POTENTIAL ANTIFUNGAL COMPOUNDS WITH DUAL MECHANISM OF ACTION

SELECTIVELY ACTING AGAINST MALASSEZIA SPP.
C. Spadini1, C. S. Cabassi1, F. Carta2, A. Angeli2, S. Selleri2, C. T. Supuran2
1Department of Veterinary Science, University of Parma, Parma, Italy
2NEUROFARBA Department, University of Florence, Florence, Italy
Malassezia spp. infections and azole drug resistance phenomena are of great concern in both
human and veterinary medicine. Malassezia spp. cause severe human and animal skin disorders,
with a zoonotic potential for M. pachydermatis, which could include it on WHO fungal pathogens
priority list. Inorganic SeS2 is used as topical treatment, but its mechanism of action on fungal sterol
pathways has not been fully revealed, due to the great variability of lipidome among
Malasseziomycetes. In this work we evaluated antifungal activity by microdilution broth assay of
novel compounds with acyl/selenoureido moieties and primary/secondary sulfonamide groups with
a dual mechanism of action: (i) a selective organic selenium fungal toxicity and (ii) the inhibition of
a new antifungal target metalloenzyme, the Carbonic Anhydrases (CAs, EC 4.2.1.1). Minimal
Inhibitory Concentration (MIC) values of selenium-containing compounds showed very high activity
on M. pachydermatis (0,5-3,33 µg/ml) instead of C. albicans and C. glabrata (3,33-256 µg/ml).
Suppression of antifungal activity was noted when selenium was replaced with either chalcogen
isosteric elements oxygen and sulfur. Compounds library was also tested on M. furfur and
M. globosa showing preferential activity on M. pachydermatis, with only a few candidates more
active on M. furfur. Cytotoxicity properties of selected compounds against MDBK and HaCat cells
were assessed, which showed safety profile at MIC values, better than SeS2. KI values on
Malassezia spp. CAs of compounds bearing primary or secondary sulfonamide moiety was in the
low-medium nanomolar range, demonstrating a multitarget selective activity on Malassezia spp.,
probably depending on lipidome constitution.
OA3
VULTURES’ ORAL MICROBIOME: KEYSTONE YEAST COMMENSALS AND CROSS-

INTERKINGDOM INTERACTIONS
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M. P. Couto1,2, F. Lopes3, M. Liede3, M. Casero4, M. Grilo1,2, E. Cunha1,2, M. Pereira5, R. Dias5,

M. Oliveira1,2
1
CIISA – Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina
Veterinária, Universidade de Lisboa,Portugal
2Laboratório Associado para Ciência Animal e Veterinária (Al4AnimalS),Portugal
3
CERAS - Centro de Estudos e Recuperação de Animais Selvagens, Castelo Branco, Portugal
4
RIAS- Centro de Recuperação e Investigação de Animais Selvagens, Olhão,Portugal
5
BioISI - Biosystems & Integrative Sciences Institute, Faculdade de Ciências, Universidade de
Lisboa,Portugal
Vultures’ oral microbiome may encode valuable data for assessing environmental stress-related
unbalances, host-associated microbiota’s diversity changes, and the efficacy of habitat restorations.
The settings at rehabilitation centers may trigger polymicrobial stress-related reorganizations in
vultures’ microbiome. It is important to characterize, under these specific settings, whether these
oral cross-kingdom interactions are symbiotic or pathogenic for the host. As so, this project aimed
at describing the fungal constituents and the bacterial microbiome of the oral cavity of recovering
vultures (n=23) – namely, Eurasian Griffon Vultures and Eurasian Black Vultures – sampled at
wildlife rehabilitation centers in Portugal. The study was divided in three main topics: 1)
characterization of oral yeast-like species present in vulture’s host-associated microbiota by
performing morphological and biochemical identification (using API20CAUX) of oral yeast-like
colonies isolated from vultures oral samples, and evaluating their biofilm forming capacity (using
Red Congo Agar); 2) evaluation of the bacterial microbiome (16S rRNA profiling by 4th NGS) of
animals with and without gross signs of oral yeast-like infections; and 3) evaluation of the potential
cross-kingdom interactions between yeasts and other microorganisms present in the oral cavity of
the sampled animals.Besides Candida and Trichosporon, other emergent microorganisms exhibited
high relative abundance in vultures’ oral cavity, including Clostridium perfringens and
Paeniclostridium sordelli, whose toxins have been associated with myonecrosis and gangrene in
humans and animals. The detected fungal-bacterial oral interactions linked strictly anaerobic
bacteria growth, in aerobic conditions, to polymicrobial complex biofilms, which may influence the
dynamics and functions of the oral microbiome. Ackowledgements: This work was supported by
CIISA–Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina
Veterinária, Universidade de Lisboa, Project UIDB/00276/2020 (Funded by FCT); and by
Laboratório Associado para Ciência Animal e Veterinária (LA/P/0059/2020 - AL4AnimalS).
keywords: oral commensals, Vultures, Candida sp., Paeniclostridium sordelli., Clostridium
perfringens, polymicrobial biofilm
OA4
DE NOVO SEQUENCING OF PROVIDENCIA ALCALIFACIENS TO STUDY ITS ROLE IN

CANINE ACUTE HEMORRHAGIC DIARRHEA
S. Rodriguez-Campos1, L. Aguilar-Bultet2, A.H. Haaland3, T. L’Abée-Lund4, A.K. Fauske1, E.M.
Soltvedt1, H. Jørgensen5, C. Sekse5, E. Skancke3, S.F. Nørstebø1
1Bacteriology and Mycology Unit, Faculty of Veterinary Medicine, Norwegian University of Life
Sciences, Ås, Norway

2Division of Infectious Diseases and Hospital Epidemiology, University Hospital Basel, Basel,
Switzerland
3University Animal Hospital, Faculty of Veterinary Medicine, Norwegian University of Life Sciences,
Ås, Norway
4Food Safety Unit, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Ås,
Norway
5Norwegian Veterinary Institute, Ås, Norway
In 2019, an outbreak of canine acute hemorrhagic diarrhea (cAHD) was observed in Southeastern
Norway affecting dogs of all ages and breeds, and without known predisposing factors. Recognized
causes including infectious agents and intoxication were ruled out; the only major finding was the
high prevalence of samples that were culture-positive for Providencia alcalifaciens, occasionally in
co-occurrence with Clostridium perfringens. P. alcalifaciens has been associated with diarrhea in
humans and animals and was previously isolated from cAHD cases in Norway in 2005 and 2006.
Twenty-one P. alcalifaciens isolates (dogs from 2019 outbreak, n=16; 2019 healthy control, n=1;
2006, n=2, 2005, n=2) were sequenced by Illumina NovaSeq 6000. Genomes were de novo
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assembled using the Shovill pipeline and an ad hoc core genome Multi Locus Sequence Typing
(cgMLST) was developed using the Ridom SeqSphere+ Software and the FDAARGOS_408 strain
as seed genome. The sequences revealed virulence factors encoding cytolethal distending protein,
flagellar filament structural and motor switch proteins. Thirteen of the 2019 isolates formed a distinct
cluster, and a second related cluster grouped three isolates from 2006 (n=2) and 2005 (n=1). Five
isolates did not cluster. All isolates from cAHD cases with lethal outcome grouped in the two
clusters; no pattern was observed for mild or moderate cases of cAHD. The control strain from the
healthy dog was more distantly related to all other isolates. Our results support a possible role for
P. alcalifaciens as a primary pathogen of cAHD. Unknown virulence factors leading to differences
in pathogenicity remain to be disclosed.
OA5
APPLICATION OF MOLECULAR METHODS AND MASS SPECTROMETRY FOR THE

DETECTION AND IDENTIFICATION OF MASTIDOGENIC MICROORGANISMS IN RAW MILK
PRODUCED IN PUGLIA
L. Del Sambro1, L. Capozzi1, C. Quarato1, E. Catalano1, G. Schino1, D. Ridolfi1, A. Giannico1,
M. Galgano2, V. Manzulli1, L. Pace1, D. Galante1, A. Bianco1.
1Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Foggia, Italia
2Università degli Studi di Bari - Dipartimento di Medicina Veterinaria, Bari, Italia
Mastitis is one of the most common problems on dairy farms.

The aim of the present work was to detect the bacteria present in bovine milk samples, using
multiplex qPCR assay and MALDI-TOF MS after bacteriological growth.In February 2022, we
collected 83 samples of bovine raw milk which showed somatic cells count higher than 300.000
cell/ml. The mutiplex qPCR assay was performed with the VetMAX™ MastiType Multi Kit. In
addition, a loop of milk was streaked on selective growth media; colonies were identified by MALDI-
TOF MS. Overall, in silico analysis showed a partial agreement with in vitro assays. The presence
of at least one of the pathogens that may be involved in the pathogenesis of the mastitis has been
detected in silico in 98,8% (82/83) of the samples tested, but only 49% (n=41) grew on the isolation
media and were identified by mass spectrometry. In silico investigations found 63 % (n=52) positive
samples for Staphylococcus aureus and 43% (n=36) for Streptococcus agalactiae, confirmed
respectively in 8 and 5 samples analyzed phenotypically; these species are recognized worldwide
as the most important etiological agents causing mastitis. Our results highlights the importance of
using molecular methods for species identification for higher sensitivity than phenotypic methods,
which may be affected by the microbial count in the milk sample if less than the sensitivity of the
culture. In addition, the study can support veterinarians and breeders by providing valuable tools
for managing bovine mastitis in terms of prevention and control.
OA6
ISOLATION AND CHARACTERIZATION OF STREPTOCOCCUS AGALACTIAE FROM BULK

TANK MILK IN THE NETHERLANDS
A.E. Heuvelink1, M.M.C. Holstege1, M.J. Swarts1, R. Peerboom1, G.J. Buter1, T.J.G.M. Lam1,2
1Royal GD, Deventer, the Netherlands
2Utrecht University, Utrecht, the Netherlands
Streptococcus agalactiae (SAG) is one of the first pathogens described as having harmful
consequences for udder health in dairy cows. The recent increase in prevalence of SAG published
in literature and of anecdotal cases in the Netherlands prompted us to initiate more research on
this pathogen.The objective of the current study was to evaluate the performance of BrillianceTM
GBS Agar (GBS) (Thermo Scientific), a commercial chromogenic medium selective for SAG, for
isolation of SAG from bulk tank milk (BTM) in comparison with modified Edward’s medium (Oxoid
Ltd) (mEDW).
BTM samples collected during 10 sampling rounds (August 2020-July 2021) were in addition to the
routinely used agars, also inoculated onto GBS. SAG-isolates of a subset of herds were subjected
to strain-level typing by Fourier Transform - Infra-Red spectroscopy and Whole Genome
Sequencing.
The prevalence of SAG per sampling round (herd prevalence) based on mEDW+GBS varied from
2.3% to 2.8% of samples. Adding GBS agar as a second agar, yielded 24.6% up to 72.2% more
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SAG-positive samples per sampling round, indicating GBS has a significant added value in the
culture of BTM for the presence of SAG. Overall, sample prevalence increased from 1.5% by using
mEDW only to 2.4% (P<0.001) and 2.6% (P<0.001) by using GBS only and both mEDW and GBS,
respectively. First typing results showed that isolates from herds found SAG-positive on GBS only
do not belong to clearly distinct types than isolates from herds SAG-positive on mEDW only or the
combination of mEDW and GBS.
OA7
THE CERVICAL MICROBIOME OF SHEEP BREEDS WITH DIFFERENT FERTILITY RATES

S.F. Nørstebø1, S. Rodriguez-Campos1, Ö.C.O. Umu1, L. Abril-Parreño2, M. Dalland3, G.D.
Gilfillan3, S. Fair2, A. Krogenaes4
1Bacteriology and Mycology Unit, Faculty of Veterinary Medicine, Norwegian University of Life
Sciences, Ås, Norway

2 Laboratory of Animal Reproduction, Department of Biological Sciences, School of Natural
Sciences, Biomaterials Research Cluster, Bernal Institute, Faculty of Science and Engineering,
University of Limerick, Limerick, Ireland
3 Department of Medical Genetics, Oslo University Hospital and University of Oslo, Oslo, Norway
4University Animal Hospital, Faculty of Veterinary Medicine, Norwegian University of Life Sciences,
PB 5003, 1432 Ås, Norway
In sheep, the use of cervical artificial insemination (AI) using frozen-thawed semen is hampered by
low pregnancy rates worldwide, except in Norway where high pregnancy rates are achieved.
Previous studies have shown that differences in pregnancy rates following cervical AI are related to
the inability of frozen-thawed sperm to traverse the cervix of some ewe breeds but not others. To
study the possible effect of the cervical microbiome on sperm transport, we compared the
microbiomes of ewe breeds with known differences in pregnancy rates following cervical AI using
frozen-thawed semen. These were Suffolk and Belclare (low and medium fertility, respectively) in
Ireland and Norwegian White Sheep (NWS) and Fur (both high fertility) in Norway. Cervical swabs
from 10 animals of each breed were collected at the follicular phase of both a natural and
synchronised oestrus in compliance with ARRIVE Guidelines. DNA was extracted with the IndiSpin
Pathogen Kit, and 16S rRNA gene sequencing was performed on an Illumina MiSeq. The R
packages DADA2, decontam, phyloseq and ggplot2 were used for downstream analyses. The Irish
breeds showed higher microbial diversity compared to the Norwegian breeds. Likewise, the
samples taken at a synchronized oestrous showed higher microbial diversity. The top features
leading to microbial differences between breeds and between natural and synchronized oestrous
corrected for breed, belonged to the genera: Histophilus, Ruminococcus, Sphingobium and
Treponema. In conclusion, the higher cervical microbial load in low fertility ewe breeds and in the
follicular phase of a synchronized oestrous may contribute to reduced sperm transport.
OA8
NEXT GENERATION SEQUENCING ON THE VAGINAL MICROFLORA OF MARES

A. Mataragka1, A. Symeonidou1, P. Katsarou1, G. Diakoudi2, F. Pellegrini2, V. Vasinioti2, G.
Lanave2, N. Decaro2, D. Vlachakis3,4,5,6, E. Papakonstantinou3, John Ikonomopoulos1.
1Laboratory of Anatomy and Physiology of Farm Animals, Department of Animal Science, School
of Animal Biosciences, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece
2Department of Veterinary Medicine, University of Bari Aldo Moro, Sp Casamassima Km3, 70010
Valenzano, Bari, Italy

3Laboratory of Genetics, Department of Biotechnology, School of Applied Biology and
Biotechnology, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece

4University Research Institute of Maternal and Child Health & Precision Medicine, School of
Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece

5Division of Endocrinology and Metabolism, Center of Clinical, Experimental Surgery and
Translational Research, Biomedical Research Foundation of the Academy of Athens, 11527

Athens, Greece
6Algorithms and Bioinformatics Group, Informatics Department, Faculty of Natural, Mathematical &
Engineering Sciences, Strand Campus,WC2R 2LS, King’s College London,UK
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Objective: this study was focused to the assessment of the constitution of the vaginal microbial
flora of mares. Materials and Methods: Vaginal swabs were collected between May to September
2020, from 30 mares, 4 to 26 years old, maintained in different parts of the Attika Prefecture, in
central Greece. Before sample collection, the test animals were submitted to clinical examination to
determine the absence of pathologic evidence.Samples were processed for PCR amplification of
16srDNA gene and Nanopore sequencing. Results: Successful reads were retrieved from 28 of the
30 samples (93.5%), which resulted to the presumptive identification of 192 bacterial species
belonging to 87 genera, with diverse genus and species distribution among samples.
Staphylococcus and Acinetobacter spp. were detected in 89% (25 of 28) and 54% (15 of 28) of the
test samples, respectively, of which the former referred primarily to Staphylococcus equorum,
Staphylococcus saprophyticus, Staphylococcus succinus, Staphylococcus cohnii and
Staphylococcus hominis, whereas the latter, to Acinetobacter equi, Acinetobacter bouvetii,
Acinetobacter lwoffii and Acinetobacter variabilis. The following genera were detected in at least
25% of the test samples: Klebsiella (43%), Pseudomonas (36%), Psychrobacter (36%), Bacillus
(32%), Enterococcus (25%), Providencia (25%), and Sphingomonas (25%). Results were compared
with those of similar studies, though to the best of our knowledge there is no previous report of an
NGS analysis on vaginal microflora of mares.Within the context of the microbiomics analysis, the
individuals of the tested population were assigned into groups, based on certain characteristics,
including race, reproduction, vaccination. This analysis produced in many cases similarity and
discriminating traits, primarily at the genus level.Discussion/Conclusion: The NGS analysis of the
vaginal microflora of mares provides an abundance of information that can be used to identify
diagnostic or prognostic indicators and improve health monitoring and protection in female horses.
OA9
DEVELOPMENT OF A MYCOPLASMA HYORHINIS CHALLENGE MODEL IN FOUR WEEK OLD

PIGS
D. Földi1, E. Zsófia Nagy1, N. Belecz1, L. Szeredi2, J.Földi3, Z. Kreizinger 1,4, M. Tenk2, A.
Kollár2, M. Gyuranecz 1,4
1Veterinary Medical Research Institute, Budapest, Hungary
2University of Veterinary Medicine, Budapest, Hungary
3Euvet Bt., Gödöllő, Hungary
4MolliScience Kft., Biatorbágy, Hungary
Objective: Mycoplasma hyorhinis causes polyserositis, arthritis, and in some cases conjunctivitis
and otitis media in piglets. Our aim was to develop a challenge model for the triggering of both
polyserositis and arthritis in four-week-old animals.Method: The pigs were challenged two
consecutive days with freshly prepared M. hyorhinis broth cultures either by intravenous route (10
ml) on both days (group A), or by intravenous route (10 ml) on the first day and by intraperitoneal
route (20 ml) the next day (group B). Results: Differences were detected in the weight gain of the
challenged groups and the control group. Average weight gains were 11.7, 7.4 and 5.7 kg for the
control group, group A and group B, respectively. Swollen joints in the two infected groups were
detected. The most affected articulations were stifles, one or both stifles were swollen in 5/6 animals
in both infected groups. Pericarditis and pleuritis were also detected in the challenged animals. In
the histopathological samples lympho-histiocytic inflammation was detected in the joints, while
thickening of the connective tissue was observed in the serosa. After necropsy, M. hyorhinis was
isolated from two samples of two different animals in group A and from four samples of two different
animals in group B. After genotyping the isolates, we were able to confirm that the challenge strain
was re-isolated.Conclusions: A M. hyorhinis challenge model was established in four-week-old
piglets suitable for evaluation of future vaccine candidates.
OA10
PREVALENCE OF PARATUBERCULOSIS IN DAIRY CATTLE HERDS AND APPROCHES TO

CONTROL THE DISEASE IN SLOVENIA
U. Zajc, T. Knific, J. J. Hodnik, D. Kušar, J. Starič, M. Kavalič, T. Pirš, J. Ježek, M. Pate, M.
Ocepek
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
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Paratuberculosis (paraTB), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a

contagious, incurable and chronic disease that primarily affects the small intestine of ruminants but
also a number of other domestic and wild animal species. It is endemic in cattle populations
worldwide and has a significant negative economic impact on the dairy industry. The aim of this
study was to (i) assess the prevalence of paraTB in large dairy cattle herds using quantitative real-
time PCR (qPCR), (ii) evaluate the impact of livestock trade on the spread of paraTB in herds using
static and temporal network analysis, and (iii) assess the risk of consumer exposure to MAP from
milk and dairy products using stochastic modelling. In 2019 and 2020, pooled fecal samples from
207 Holstein-Friesian cattle herds were analysed by qPCR. In total, 4.35% of herds tested positive
for MAP. In the positive herds, faecal and blood samples were collected from all animals older than
two years (n=1015). The average prevalence of MAP within herds was 13.6%. The prevalence of
paraTB at the herd level was relatively low. Data on animal trade allowed identification of high-risk
farms with intensive trade and may therefore help to implement targeted control measures. The risk
of consumer exposure to MAP via pasteurized milk and dairy products was generally low, with the
exception of individuals consuming raw milk and dairy products from paraTB-infected herds.
OA11
PHENOTYPIC SURVEILLANCE FOR ANTIMICROBIAL SUSCEPTIBILITY IN VETERINARY

MASTITIS PATHOGENS FROM IRELAND
A. Naranjo-Lucena 1, G. Madigan 1, A. Johnson 2, C. Sánchez-Miguel 2, S. McGettrick 2, S.
Fagan 2, M. Sheehan 2, M. Casey 2, M. Wilson 1, M. McElroy 1, R. Slowey 1
1Department of Agriculture, Food and the Marine (DAFM) Laboratories Backweston, Ireland
2
DAFM Regional Veterinary Laboratories, Ireland
Surveillance for antimicrobial resistance in veterinary clinical pathogens is not as well developed as
for zoonotic and indicator bacteria. Mastitis-causing pathogens isolated from milk submitted to
DAFM laboratories in Ireland between 2019 and 2021 were screened for antimicrobial susceptibility.
A single isolate per herd of each bacterial species was subject to antimicrobial susceptibility testing
as per CLSI guidelines. Streptococcus uberis was the most common pathogen isolated from
mastitic milk during 2021 (n=151), closely followed by Staphylococccus aureus (n=150), Eschericha
coli (n=128) and finally Streptococcus dysgalactiae (n=28). S. uberis showed 93.38% susceptibility
to β-lactam antimicrobials. S. dysgalactiae, however, were 100% susceptible. S. uberis had lower
levels of susceptibility to erythromycin and pirlimycin (78.7% and 66.2%, respectively) than S.
dysgalactiae (89.3% and 96.4%). No MRSA was detected. Susceptibility of S. aureus to penicillin
was greater in 2021 (56.9%) than in 2019 (46.2%), while susceptibility to erythromycin and pirlimycin
was above 96% all three years. Comparing the results obtained for E.coli isolates in 2021 with 2020,
some trends were evident - susceptibility to amoxicillin-clavulanic was 5.3% lower and susceptibility
to tetracycline was 6.7% lower, whereas susceptibility to enrofloxacin was slightly higher (1.4%) in
2021. There are preliminary indications that the susceptibility of S. aureus to penicillin and
erythromycin and that of E. coli to enrofloxacin may be increasing. The susceptibility of S.
dysgalactiae to erythromycin and of S. uberis to all anitmicrobials declined in 2021. These findings
highlight the importance of establishing a robust surveillance programme to monitor trends and
emerging resistance.
OA12
PREVALENCE AND RISK FACTORS ASSOCIATED WITH FAECAL CARRIAGE OF

EXTENDED-SPECTRUM Β-LACTAMASE-, AMPC- AND CARBAPENEMASE-PRODUCING
ESCHERICHIA COLI IN STRAY CATS
G. Ratti, A. Facchin, A.Stranieri, A. Giordano, S. Paltrinieri, P. Scarpa, D. Maragno, L. Villa,
M. Penati, S. Lauzi
Department of Veterinary Medicine and Animal Sciences, Università degli Studi di Milano, Milan,
Italy
Antimicrobial resistance in pets is gaining attention worldwide. The aim of this study was to
determine the prevalence and risk factors associated with faecal carriage of ESBL-, AmpC-, and
Carbapenemase-producing E. coli in stray cats from Northern Italy. Rectal swabs were collected
between 2020 and 2021 from stray cats admitted to the Veterinary Teaching Hospital of Lodi.
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Samples were microbiologically and molecularly analyzed to investigate the presence of ESBL-,
AmpC-, and Carbapenemase-producing E. coli. Anamnestic data were analyzed to identify risk
factors. In total 20/121 (16.5%) animals were positive for the presence of ESBL-/AmpC-
/Carbapenemase-producing E. coli. Phenotypic test showed 11 (9.1%) ESBL-, 3 (2.4%) ESBL-
/AmpC- and 2 (1.7%) AmpC- producing E. coli. ESBL genetic determinants were blaCTX-M (14,
100%), blaTEM (12, 86.7%) and blaSHV (2, 14.3%) whereas blaCMY-2 gene was the most frequently
detected gene in AmpC-producing E. coli (4, 80%). Moreover, 4 (3.3%) isolates showed
Carbapenemase-producing phenotype supported by the detection of blaNDM gene in all these
isolates. E. coli phylogroups F (6/20, 30%) and B2 (5/20, 25%) were the most frequently detected.
Nineteen (95%) isolates showed multidrug resistance. Risk factors associated with ESBL-/AmpC-/
Carbapenemase-producing E.coli faecal carriage were hospitalization (P <0.0001), previous
antibiotic treatment (P <0.0001) and presence of clinical disease (P <0.0001). Our findings highlight
the need of surveillance programs and antimicrobial stewardship to reduce the emergence and
spread of resistant bacteria. Further studies are needed to define the risk of transmission to humans
and other animals.
OA13
ASSESSMENT OF RELIABILITY AND REPRESENTATIVENESS OF ANTIMICROBIAL

SENSITIVITY TESTING RESULTS FOR DIFFERENT LIVESTOCK SPECIES AND DIFFERENT
BACTERIA IN THE NETHERLANDS FROM 2016 - 2020
J. van Hout1, M. Augustijn1, E. Broens2, T. Derkman1, M. Gonggrijp1, A. Heuvelink1, J. Wiegel1
1Royal GD
2Utrecht University
To further promote prudent use of antimicrobials and to provide solid evidence for policies aimed at
reducing antimicrobial resistance levels in bacteria from animals, it’s extremely important to have
access to validated antimicrobial susceptibility testing (AST) data, representative for specific animal
species/bacterial species and with known bias levels (if present). Therefore this study (financed by
ZonMw) aimed to assess reliability and representativeness of currently at Royal GD for livestock
available AST results of selected antimicrobial/bacterial species combinations. AST results and
additional data were collected over the time period 2016-2020 for: Actinobacillus
pleuropneumoniae/Escherichia coli/Streptococcus suis (pigs), Salmonella Dublin/Salmonella
Typhimurium (veal calves) and Escherichia coli (broilers). Aggregated Minimal Inhibitory
Concentration data were statistically evaluated per animal species/bacterial species
combination.Overall, AST results are fairly representative for the animal and/or farm density per
province (pigs/veal calves) or the number of farms per veterinary practice (poultry). However, the
low number of isolates in the dataset of veal calves led to less representative and less reliable AST
results. Effort should be made to increase the number of isolates from veal calves. In general,
different associations between susceptibility levels and specific factors (i.e., additional information
like farm of origin or age) were demonstrated. These factors should be taken into account when
showing the final susceptibility patterns for use in antimicrobial treatment guidelines and in
veterinary practice. Evaluation of MIC data was hampered by the lack of veterinary clinical
breakpoints, clearly emphasising the need for such breakpoints to further improve prudent use of
antibiotics in the field.
OA15
ROLE OF COMPANION ANIMAL-HUMAN HOUSEHOLDS IN THE DISSEMINATION OF

COLISTIN AND EXTENDED-SPECTRUM BETA-LACTAM RESISTANCE
J. Menezes1,2,3, A. Belas1,2,3, C. Marques1,2,3, J. Moreira da Silva1,2,3, A. Amaral1,2, C. Pomba1,2,3,4
1Centre for Interdisciplinary Research in Animal Health (CIISA), Faculty of Veterinary Medicine,
University of Lisbon, Lisbon, Portugal

2AL4Animals, Associate Laboratory for Animal and Veterinary Sciences
3ENOVAT—European Network for Optimization of Veterinary Antimicrobial Treatment (COST
ACTION CA18217).
4Molecular Veterinary Diagnostic Laboratory - Genevet, Rua Quinta da Nora Loja3B, 2790-140
Carnaxide, Portugal
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Objectives: To determine the colonization and sharing of colistin resistant, extended-spectrum

beta-lactamases (ESBL)- and carbapenem-producing Enterobacterales (CPE) between healthy
companion animals and its co-habiting humans. Methods: Forty-one Portuguese households (58
humans, 40 dogs, 18 cats) with at least one human–animal pair were studied. Informed consent
was obtained. Fecal samples were inoculated on MacConkey agar plates supplemented with 1.5
µg/mL cefotaxime, 1.0 µg/mL meropenem and SuperPolymyxin medium plates. Microdilution
susceptibility testing was performed according to EUCAST 2021 guidelines. Beta-lactam and
plasmid-mediated colistin resistance (mcr-type) genes were screened by PCR and sequenced.
Species identification was performed by PCR. Animal-owner pair strains genetic relatedness was
estimated by WGS (Illumina NovaSeq), producing paired-end libraries with 150 bp. Results: Three
dogs (7.5%) presented multidrug-resistant Escherichia coli strains harbouring the mcr-1 gene, with
colistin MICs of 4 mg/L. No CPE was found. Twenty-five ESBL/AmpC-producing Enterobacterales
(1 Citrobacter freundi, 2 Klebsiella pneumoniae, 22 E. coli) were detected in 15 households (20.7%
from humans and 13.8% from animals); 10 strains presented a multidrug-resistant profile. WGS
SNPs analysis suggests human–dog pair shared E. coli strains in 2 households: i) ST93 strains
harbouring blaCTX-M-15 and blaTEM-1 genes isolated from a cat and its owner; ii) ST457 strains,
harbouring blaCTX-M-55 and blaCMY-2 genes, and ST410 strains, harboring blaCTX-M-15 and blaOXA-1
genes isolated from a dog and its owner. Conclusions: Our findings point toward the role of CA
in the dissemination of clinically important genes to humans since they live closely together.
Therefore, household hygiene and antimicrobial surveillance is recommended in this community
setting.
OA16
HETERORESISTANCE IN A MULTI-RESISTANT CLINICAL ENTEROBACTER CLOACAE

COMPLEX STRAIN DUE TO AMPLIFICATION OF AN AMPC Β-LACTAMASE
J. Kupke1, J. Brombach1, F. Ghazisaeedi1, D. Hanke1, T. Semmler2, K. Tedin1, N. Nordholt3, F.
Schreiber3, A. Lübke-Becker1, M. Fulde1
1Institute of Microbiology and Epizootics, Centre for Infection Medicine, Department of Veterinary
Medicine, Freie Universität Berlin, Berlin, Germany

2Robert Koch Institute (RKI), MF2-Genome Sequencing and Genomic Epidemiology, Berlin,
Germany
3Federal Institute for Materials Research and Testing (BAM), Department of Materials and the
Environment (Dpt. 4), Berlin, Germany
Introduction: Heteroresistance (HR) describes the phenomenon of a subpopulation that grow in

the presence of inhibitory antibiotic concentrations. We found HR to ceftazidime in an Enterobacter
cloacae complex (ECC) strain (IMT49658) isolated from the wound of a horse. Materials &
Methods: For phenotypic characterisation we conducted population analysis profiles and stability
analysis of resistance. Quantification of β-lactamase genes was done with qRT-PCR. Whole
genome sequencing (WGS) shed light on the genome of resistant and susceptible population
entities. Furthermore, we exposed IMT 49658 to long-term heat stress. Competition assays and
ScanLag assessed growth traits of this strain. Results: PAPs confirmed HR with growth up to 16-
fold the breakpoint concentration. The resistant subpopulation was reversible after subcultivation
on non-selective media and comprised a gene-amplification region with the AmpC β-lactamase
blaDHA-1, detected with WGS. No HR could be detected after heat stress led to gene loss of blaDHA-
1. Competition assays revealed a loss of biological fitness due to the gene-amplification.
Heterogeneous lag times for resistant subpopulations growing on double breakpoint concentration
of CAZ in ScanLag revealed that CFUs appearing early maintained higher copy numbers of blaDHA-
1 than late occurring CFUs, defining the amplifications as the reason for the plasticity of this strain.
Conclusion: In this ECC strain AmpC β-lactamase blaDHA-1 and its amplification lead to a
heteroresistant phenotype. In order to combat HR infections in veterinary medicine, strong effort for
the investigation of HR is necessary.
OA17
THE ENOVAT SURVEY ON CURRENT METHODOLOGIES USED FOR BACTERIAL

IDENTIFICATION AND ANTIMICROBIAL SUSCEPTIBILITY TESTING IN EUROPEAN
VETERINARY DIAGNOSTIC LABORATORIES
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D. Timofte1, I. Cvetkovikj2, F. Zendri1, S. Blum3, S.C. Chaintoutis4, P.A. Kopp5, C. Hare6, Z.

Štritof7, S. Kittl8, J. Goncalves9, I. Zdovc10, E. Paulshus11, T. Koritnik12, A. Laconi13, E.
Broens14,
P. Damborg15
1Institute of Infection, Veterinary and Ecological Sciences, Leahurst Campus, University of
Liverpool, Neston, United Kingdom

3Department of Bacteriology and Mycology, Kimron Veterinary Institute, Bet Dagan, Israel
4Diagnostic Laboratory, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle
University of Thessaloniki, Thessaloniki, Greece

5IDEXX Vet Med Labor GmbH, Kornwestheim, Germany
6Department of Veterinary Medicine, University of Cambridge, Cambridge, UK
7Department of Microbiology and Infectious Diseases with Clinic, Faculty of Veterinary Medicine,
University of Zagreb, Croatia

8Department of Infectious Diseases and Pathobiology, Institute of Veterinary Bacteriology,
University of Bern
9Institute of sustainable processes, University of Valladolid, Spain
10Institute of Microbiology and Parasitology, Veterinary Faculty of Ljubljana, Croatia
11Department of Analysis and Diagnostics, Microbiology, Norwegian Veterinary Institute, Ås,
Norway
12Department for Public Health Microbiology Ljubljana, Centre for Medical Microbiology, National
Laboratory of Health, Environment and Food, Ljubljana, Slovenia

13Department of Comparative Biomedicine and Food Science, University of Padua, Legnaro (PD),
Italy
14Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University,
Utrecht, the Netherlands

15Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences,
University of Copenhagen, Frederiksberg, Denmark.
Background: Veterinary diagnostic laboratories (VDLs) play a key role in the identification of
infectious agents and antimicrobial stewardship. However, there is still a lack of harmonisation of
methodologies and procedures used in European VDLs (1,2). Methods: The European Network for
Optimization of Veterinary Antimicrobial Treatment (ENOVAT) designed a survey, which was
distributed via a online platform to VDLs in 34 European countries. The survey focused on practices
and interpretive criteria used for culture and identification (C&ID), and antimicrobial susceptibility
testing (AST) of veterinary bacterial pathogens. Results: Two hundred and ninety laboratories
responded, representing a mixture of academic (39%), government (33%), and private (28%)
laboratories. Average C&ID turnaround varied from 1-2 days (78%) to 3-5 days (20%), and 6-8 days
(0.5%). For AST, similar time frames were achieved by 63%, 60%, and 0.5% of VDLs, respectively.
Only 57% of laboratories attempted bacterial ID to species level. Biochemical ID systems (e.g., API
kits) were the most used (56%) followed by MALDI-TOF MS (46%). For AST, Kirby-Bauer disc-
diffusion (DD) and MIC determination were conducted by 44% and 33% of laboratories,
respectively. A combination of EUCAST and CLSI clinical breakpoints was most commonly used
for interpretation of both DD (41%) and MIC (47%). Only 48% and 46% of VDLs routinely screened
isolates for methicillin resistance and ESBL production, respectively. Conclusion: A variety of
methodologies were identified for C&ID and AST in European VDLs. Our results emphasize the
need to harmonise diagnostic methodologies to benefit rational antibiotic use and ultimately improve
animal and public health.
References:
1. Timofte D, et al. Driving Laboratory Standardization of Bacterial Culture and Antimicrobial
Susceptibility Testing in Veterinary Clinical Microbiology in Europe and Beyond. Journal of Clinical
Microbiology 2021;59(6):e02572-20.
2. Morency-Potvin P, Schwartz DN, Weinstein RA. Antimicrobial stewardship: how the microbiology
laboratory can right the ship. Clinical Microbiology Reviews 2017; 30:381–407.
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OA18
THE PERSISTENCE OF METHICILLIN-RESISTANT STAPHYLOCOCCUS

PSEUDINTERMEDIUS IN PETS WITH SKIN AND SOFT TISSUE INFECTIONS: WHICH IS THE
ROLE OF THE COHABITANTS AND THEIR LIVING ENVIRONMENT?
E. Spagnolo1, M. Berlanda2, L. Ceglie1, I. Drigo1, K. Trevisiol1, M. Cocchi1, R. Perin1, C.
Losasso1, K. Capello1, L. Decastelli3, D. Pasotto2, M. Corrò1.
1 Istituto Zooprofilattico Sperimentale delle Venezie, Padua, Italy
2Università degli Studi di Padova, Dipartimento di Medicina Animale, Produzioni e Salute, Padua,
Italy
3Laboratorio Nazionale di Riferimento per gli Stafilococchi Coagulasi Positivi, Turin, Italy
Staphylococcus pseudintermedius (SP), usually belonging to the human and animal saprophytic
bacterial flora, could be associated with skin and soft tissue infections (SSTIs) in pets, as well. The
presence and the spread of methicillin-resistant SP (MRSP) strains are a matter of concern due to
the difficulty of handling SSTIs in pets. In order to understand the cross-contamination between
patients and their animal cohabitants, as well as the potential persistence of MRSP in the
environment, the sampling was scheduled from armpit, mouth and genitals in all animals, from
lesions in pets with SSTIs, and from the environment (bowl, kennel and sofa or bed if shared with
the owners) periodically. Three hundred twenty-seven swabs were collected from 24 patients, their
10 cohabitants and the environment. SP and MRSP colonies were identified by phenotypic
methods; methicillin resistance gene (mecA) was confirmed by PCR. Species identification was
performed by MALDI-TOF mass spectrometry. SP was isolated in 221/327 samples (67%) and 124
of these strains were mecA positive (56%).Interestingly, 26/124 (21%) were strains isolated from
the same cohabitants or from the same environment repeatedly.This study reports the persistence
of MRSP strains in healthy cohabitants and in the environment. Such persistence could represent
an important source of recontamination for the animals and therefore its investigation should be
considered in patient management. Moreover, the close sharing of the living environment with
humans may represent a health concern due to the risk of human-animal cross-contamination. Work
financed by the Ministry of Health (IZSVE 16/18 RC).
OA19
RECOMMENDATIONS FOR THE THERAPY OF LOWER URINARY TRACT INFECTIONS IN

CATS AND DOGS
A. Bethe1,2, C. Weingart2,3, A.-K. Schink1,2, J. Brombach1,2, B. Walther4, R. Köck5, R. Merle6, W.
Bäumer7, B. Kohn2,3, A. Lübke-Becker1,2
1Institute of Microbiology and Epizootics, FU Berlin, Berlin, Germany
2Veterinary Centre for Resistance Research (TZR), FU Berlin, Berlin, Germany
3Small Animal Clinic, FU Berlin, Berlin, Germany
4Robert Koch-Institute, Berlin, Germany
5DRK Kliniken Berlin, Berlin, Germany
6Institute for Veterinary Epidemiology and Biostatistics, FU Berlin, Berlin, Germany
7Institute of Pharmacology and Toxicology, FU Berlin, Berlin, Germany
Objective: Lower urinary tract infections (UTIs) are a common reason for antimicrobial therapy in
small animal practice. To minimize and improve the use of antimicrobial agents, we aimed to
develop treatment recommendations based on detection rates and antimicrobial susceptibility
testing (AST) of the respective causative pathogens in clinical samples of cats and
dogs.Material/Methods: Data on 470 urine samples were collected from dogs (n=258) and cats
(n=212), mainly suffering from sporadic (dogs: n=117, cats: n=96) or recurrent cystitis (dogs: n=108,
cats: n=105). Minimal inhibitory concentrations (MIC) for eleven antimicrobial agents licensed to
treat UTIs in Germany were determined for 308 isolates of clinical relevance (dogs: n=191; cats:
n=117) following CLSI guidelines.Results: In total, 214 samples were culture-negative or revealed
nonspecific bacterial growth. Escherichia coli (dog: n=89, cat: n=55) dominated the sample
collection. While additionally, Staphylococcus pseudintermedius (n=31) and Streptococcus canis
(n=16) were frequently associated with canine samples, Enterococcus faecalis (n=14) and
Staphylococcus felis (n=14) were common in feline samples. Evaluation of the AST results of the
relevant pathogens revealed that potentiated aminopenicillins and 1st generation cephalosporins
seem suitable as first line therapeutic choices in sporadic cases of canine and feline lower
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UTI.Discussion: The high proportion of culture-negative urine samples and the cumulative
susceptibility patterns of uropathogens highlight the importance of a sound bacteriological
examination including urine sediment analysis to prevent unnecessary treatment as well as
treatment failure.
OA20
PROFILING OF THE MICROBIOTA FROM APULIAN COW MILK MOZZARELLA PRODUCED

WITH WHEY
D. Simone, C. Trisolini, V. Laera, G. Caldarola, A. Miccolupo, E. Catalano, A. Caffò, A. M.
Derobertis, V. Nardelli, M. Iammarino, A. Parisi
Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Via Manfredonia, 20, 71121
Foggia (Italy)
The Mozzarella di Gioia del Colle PDO is a production excellence of Apulia and Italy and its
production is based on whey. Microbial communities play a pivotal role in the quality and features
of the mozzarella production chain, but to date they are poorly investigated. In this study, we have
profiled microbial communities from mozzarella cheese samples together with the wheys used for
their production with targeted metagenomics (V3V4 16S). Our survey revealed the presence of taxa
commonly associated with mozzarella production (lactobacilli, staphylococci). Additionally, we
detected other taxa commonly related to other dairy production chains (e.g. Thermus). Overall,
within the same production lot, the same non-lactobacilli taxa were more abundant in the mozzarella
compared to the whey. We were also able to detect possible contaminants of the production chain
with very low abundance. We showed that targeted metagenomics is a valuable tool to profile
microbial communities in mozzarella and whey and to identify pathogens that can enter the
production line.
OA21
EVIDENCE OF THE WIDE CIRCULATION OF MULTI-DRUG RESISTANT ENTERIC

STRAINS IN LESSER KESTREL (FALCO NAUMANNI)
R. Samarelli, N. Pugliese, A. Schiavone, E. Circella, I. Siddique, M. Saleh, C. Zizzadoro,
G. Crescenzo, A. Camarda
Università degli Studi di Bari “Aldo Moro”, Bari, Italy
This study was aimed to assess the circulation of antimicrobial-resistant, Gram-Negative

enteric bacteria in wild birds. Specifically, cloacal swabs were collected from 32 lesser
kestrels (Falco naumanni) hosted in the apulian wildlife rescue centre in July 2021. All
samples were collected upon admittance as part of the routine diagnostic procedures. Swabs
were incubated in tryptic soy broth (TSB) with enrofloxacin (enr), selected on McConkey agar
with enr, and identified biochemically or by MALDI-TOF. Susceptibility of isolates to the most
common antibiotic classes was ascertained by the disc diffusion method. Only one out of 32
samples (3,12%) showed no bacteria growth. From the remaining 31 birds, 41 enr-resistant
strains were isolated, specifically 23 Escherichia coli, 10 Proteus mirabilis, 6 Klebsiella
pneumoniae, 1 Citrobacter freundii, and 1 Enterobacter cloacae. Out of them, 39 (95.12%)
were resistant to three or more antibiotics.In detail, 35 strains were resistant to tet, 32 to amp,
and 25 to sxt or streptomycin (str). No cst-resistant strain was isolated except for P. mirabilis,
naturally resistant. No strain was resistant to ipm, but 7 were resistant to the fourth-generation
cephalosporin cefepime (fep). Those data suggest a wide circulation of multi-drug resistant
(MDR) strains, some of them with clinical relevance, in birds that were never treated with
antibiotics. Considering its migratory habits, and its position at the top of the food chain, F.
naumanni should be considered an important indicator of the flow of MDR bacteria between
anthropic and wild environments, providing useful information in a One Health view. "The
antibiotic names have been abbreviated according to the indications of the British Society for
Antimicrobials Chemotherapy, unless otherwise specified"
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ORAL- AULA B
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OB1
PATTERNS AND DETERMINANTS OF PORCINE REPRODUCTIVE AND RESPIRATORY

SYNDROME VIRUS (PRRSV) IN EUROPE: A PHYLODYNAMIC AND PHYLOGEOGRAPHIC
APPROACH
G. Franzo, G. Faustini, M. Legnardi, M. Cecchinato, M. Drigo, CM. Tucciarone
Department of Animal Medicine, Production and Health (MAPS), University of Padua, Legnaro (PD),
Italy
Porcine reproductive and respiratory syndrome (PRRS) is among the most devastating diseases
affecting the pig industry. The remarkable genetic variability of this virus, leading to poor cross-
protection, has limited the vaccine efficacy and other measures must be adopted to effectively
control the viral circulation. Some recent studies have investigated the factors involved in viral
spreading and persistence, at least at the local level. However, despite the topic's relevance, the
variables involved in PRRSV epidemiological success at a broader scale, like the European one,
have never been investigated using a robust statistical approach, linking classical and molecular
epidemiology. In the present study, more than one thousand ORF5 sequences were analyzed
through a phylodynamic and phylogeographic approach to investigate the history, dynamics and
spreading patterns of PRRSV within European borders. Moreover, several potential predictors
representative of swine population features and trade, human population, economy and geographic
characteristics, were evaluated through a specifically designed generalized linear model (GLM) to
quantify their relevance on viral migration rate between countries over time. Although pig stock
density, mean PRRSV strains genetic diversity, investments in agriculture (including a likely role of
vaccination) and farmer education were involved to a certain extent, the major determinant was
proven to be by far the live pig trade. The present study, which provides a robust depiction of PRRSV
European molecular epidemiology patterns and determinants, could contribute to a more rational
allocation of resources based on effective prioritization of control measures.
OB2
CANCELLED
OB3
ENTERIC VIRAL CO-INFECTIONS IN ACUTE GASTROENTERITIS OF CATS

A. Palombieri1, V. Sarchese1, F. Di Profio1, P. Fruci1, V. Martella2, B. Di Martino1
1Faculty of Veterinary Medicine, Università degli Studi di Teramo, Teramo, Italy
2Department of Veterinary Medicine, Università Aldo Moro di Bari, Valenzano, Italy
Acute gastroenteritis (AGE) is a common clinical problem of cats living in densely housed
environments that may recognize a multi-agent aetiology, with more than one virus sequentially or
synergistically being involved. The aim of this study was to perform a molecular survey on 62 cats
with diarrhoea admitted during 2021 to the veterinary hospital of the Faculty of Veterinary Medicine
of Teramo (Italy). Feline enteric samples were tested for selected viral pathogens including feline
panleukopenia parvovirus (FPV), feline coronavirus (FCoV), feline kobuvirus (FeKoV), feline
chaphamaparvovirus (FeChPV), feline picornavirus (FePV), feline calicivirus (FCV) and norovirus
(NoV) using specific or broadly reactive consensus PCR assays at the family, sub-family and/or
genus level. The virome composition of eight specimens were further assessed by combining a
sequence-independent single-primer amplification (SISPA) approach with Oxford Nanopore
Technologies (ONT) sequencing platform. At least one viral pathogen was detected in 67.7%
samples with FPV being the most frequently detected virus (35.4%), followed by FCoV (33.8%).
FeChPV and FCV were found with rates of 12.9% and 6.4%, respectively. FeKoV, FePV and FeNoV
were all detected with a rate of 1.61%. Single infection was identified in 26 samples (41.9%), mainly
related to FPV (84.6%). Co-infections were present in 25.8% samples and mainly accounted for by
FPV and FCoV. By ONT analysis a picornavirus genetically close to a Feline sakobuvirus A (FeSA)
was sequenced. FeSA was first detected in 2012 during a metagenomic study in a healthy cat in
Portugal but, since then, it has not been reported again in cats.
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OB4
GENOMIC AND TRANSCRIPTOMIC OF RANID HERPESVIRUS 3 AND BUFONID

HERPESVIRUS 1 IN NATURALLY INFECTED HOSTS: LINKING MOLECULAR SIGNATURE
AND PATHOLOGY PHENOTYPE
F.C. Origgi
Institute of Animal Pathology (ITPA) Vetsuisse Faculty, University of Bern, Bern, Switzerland
Two novel herpesviruses have been recently discovered in free-ranging amphibian populations.
Ranid herpesvirus 3 (RaHV3) and Bufonid herpesvirus 1 (BfHV1) have been shown to be
associated with a remarkably similar, but distinct proliferative skin diseases in their respective hosts,
the common frog (Rana temporaria) and common toad (Bufo bufo). Interestingly, the disease in
both frogs and toads is associated with an inconspicuous or absent detectable, cellular immune
response. We have completed the full sequencing of the genome of both viruses and we have
analyzed the transcriptomic data obtained from naturally infected frogs and toads. The genome of
RaHV3 is 207,914nt long and encodes for 186 predicted protein. The genome of BfHV1 is 158250nt
long and encodes for 152 predicted proteins. Both genomes encode for predicted proteins with
putative immunomodulatory activity including those known to interfere with macrophage and T-cell
activity, all immune cells of critical relevance for antiviral activity. At the same time, an immune
modulation appears to occur in the infected host as well. Paralleling this, the transcription of the
genes involved in signaling and cell remodeling is the mainly impacted in the infected hosts,
suggesting a relevant role in the disease. Furthermore, the genome of RaHV3 contains a predicted
gene whose putative encoded protein shares similarities with FOXM1, known to be a critical
transcription factor involved in the development of carcinomas in humans. Remarkably, the
observed disease phenotype in the infected hosts appears to be the product of a tight interaction
between the host and pathogen gene armories
OB5
DO DEFECTIVE KOALA RETROVIRUS SEQUENCE OFFER SELECTIVE ADVANTAGE TO

KOALAS?
S. Dudeja, R.E. Tarlinton
School of Veterinary Medicine and Science, University of Nottingham, United Kingdom
Koala Retrovirus (KoRV) is known to be the youngest endogenous retrovirus still settling to its
genomic parasitic lifestyle. KoRV has both infectious and endogenous (integrated into the host
genome) forms. 100 % of the Koalas in northern Australia are positive for the virus. A higher proviral
copy number per cell has been observed in Northern koalas due to endogenized KoRV compared
to that in the south. The Koalas in southern Australia show a variability in the prevalence rate of the
exogenous virus and a lower rate of KoRV induced disease. Southern Australian koalas earlier
considered to be disease free or only having the exogenous counterparts of the virus, demonstrate
a defective variant of KoRV known as RecKoRV. RecKoRV is formed due to the recombination
between Phascolarctid Endogenous Retroelement (PhER) and KoRV. The presence of RecKoRV
variants particularly in the founder population in the French island calls into question the existence
of KoRV free animals. The difference in the KoRV profiles between the northern and the southern
animals, heighten the chances that these replication defective transcripts may be interfering with
full length transcripts of the replication competent KoRV. The research objectives of this project
involve screening samples from southern animals for polymorphism of RecKoRV loci using
integration site specific PCRs to explore whether these are fixed or variable in the population. Follow
on work will then focus on whether there are any links between polymorphic loci and infectious
KoRV prevalence and clinical disease incidence.
OB6
A COLD CASE OF EQUINE INFLUENZA DISENTANGLED WITH NANOPORE SEQUENCING

F. Pellegrini1, G. Lanave1, A. Hassan Omar1, G. Diakoudi1, R. Augelli2, M. Camero1, N. Decaro1,
G. Elia1, C. Buonavoglia1, V. Martella1
1Dipartimento di Medicina Veterinaria, Università degli Studi di Bari Aldo Moro, Italy
2Ministero della Salute, Ufficio UVAC Bari, Italy
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Influenza A viruses (IAVs), family Orthomyxoviridae, are enveloped viruses with a ssRNA(-)
genome of 13.5Kb, made up of 8 segments. IAVs are classified in subtypes based on the surface
proteins hemagglutinin (HA) and neuraminidase (NA), for a total of 18 H and 11 N subtypes. IAVs
can infect various avian and mammalian species, causing major acute febrile respiratory disease
outbreaks. Equine IAVs (EIAVs) are either H7N7 or H8N3, although H7N7 seems to have
disappeared. We determined the whole genome sequence of a H3N8 equine influenza virus (strain
Bari/05) identified from a 2005 outbreak in Apulia, Italy, seemingly linked to a 2003 outbreak in a
horse racetrack in Rome. After reverse transcription and PCR-based enrichment, a library was
generated with the Ligation sequencing kit 1D SQK-LSK110 (ONT) and used for sequencing on
MinION Mk1C. The fastq data were assembled to generate contigs of the 8 genome segments and
the sequences were analyzed with BLASTn, unveiling a tight correlation of strain Bari/05 to strain
A/equine/Pulawy/1/2005(H3N8), identified in Poland in 2005. On interrogation of the national
register for animal trading (NSIS), we observed that in 2005 nearly half of the meat (DPA) horses
imported in Apulia were from Poland. A small flux of non-DPA horses from Poland was also
documented, supporting the hypothesis of introduction of EIAV with horse trading, and ruling out a
correlation with the 2003 outbreak. Generating a large database of EIAV genome sequences,
including archival viruses, will help understand better the molecular evolution and phylodynamic of
EIAV.
OB7
THE EPIDEMIOLOGY OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME

(PRRS) IN AN INTEGRATED PIG COMPANY OF NORTHERN ITALY: A MULTIPLE THREATS
REQUIRING MULTILEVEL INTERVENTIONS
G. Franzo1, G. Barbierato1, P. Pesente2, M. Legnardi1, C. M. Tucciarone1, G. Sandri2, M. Drigo1
1Department of Animal Medicine, Production and Health (MAPS), University of Padua, Legnaro
(PD), Italy
2Laboratorio Tre Valli, 37132 Verona, Italy
Despite the remarkable efforts paid in terms of vaccination administration and biosecurity,
eradication and long‐term control of Porcine reproductive and respiratory syndrome virus (PRRSV)
have often been frustrating. Unfortunately, few studies are currently available that objectively link,
using a formal statistical approach, viral molecular epidemiology to the risk factors determining the
observed scenario. The present study takes advantage of a remarkable dataset of approximately
one‐thousand ORF7 sequences, obtained from strains collected between 2004 and 2021 from the
largest Italian pig company, which implements strict compartmentalization among independent
three‐sites (i.e., sow herds, nurseries and finishing units) pig flows. The history and dynamics of the
viral population and its evolution over time were reconstructed and linked to managerial choices
using a phylodynamic approach. The viral fluxes within and among independent pig flows were
evaluated, and the contribution of other integrated pig companies and rurally risen pigs in mediating
such spreading was investigated. Finally, viral circulation in Northern Italy was reconstructed using
a continuous phylogeographic approach, and the impact of several environmental features on
PRRSV strain persistence and spreading velocity was assessed. PRRSV epidemiology was shaped
by a multitude of factors, including pig herd management (e.g., immunization strategy),
implementation of strict‐independent pig flows, and environmental features (e.g., climate, altitude,
pig density, road density, etc.). Small farms and rurally raised animals also emerged as a potential
threat for larger, integrated companies. Therefore, a multidimensional approach, ranging from
individual herd management to collaboration and information sharing among different companies,
is mandatory for effective infection control.
OB8
EPIDEMIOLOGICAL INVESTIGATION OF DOMESTIC CAT HEPADNAVIRUS IN HONG KONG

P. Capozza1, M. Carrai2, G. Lanave1, V. Martella1, J. A. Beatty2, V. R. Barrs2
1Department of Veterinary Medicine, University of Bari, Valenzano (Ba), Italy.
2Dept Veterinary Clinical Sciences & Centre for Animal Health and Welfare, Jockey Club College of
Veterinary Medicine and Life Sciences (JCC), City University of Hong Kong ,Hong Kong
Domestic cat hepadnavirus (DCH) is an emerging virus related to hepatitis-B virus (HBV) of
humans. On molecular inestigation from Australia, Italy, Malaysia, Thailand, Japan and UK, DCH
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prevalence in feline blood/serum ranges from 0.78 to 12.3%. The aim of this study was to assess
the prevalence of DCH in Hong Kong (HK) and to gather information on sequence diversity of HK
DCH strains. DCH DNA was detected with virus-specific real-time PCR in 81 (15.8%) of 513 feline
blood samples tested, a prevalence higher than reported elsewhere. The mean and median values
of DCH in the feline blood were 2.43x105 DNA copies/10 μl and 1.95x101 DNA copies/10μl (range
2.55x100-1.96x107 DNA copies/10μl), apparently without any significant correlation with the main
risk factors considered, i.e. age, sex, breed, neuter status and alanine amino-transferase (ALT).
The complete genome sequence of 12 DCH strains was obtained, using a primer walking strategy.
On phylogenetic analysis, all the HK DCH strains clustered together with viruses from Australia and
Asia (clade A), separate from viruses from Europe (clade B) and from a divergent virus from Japan
(clade C). Based on our findings, the DCH strains circulating in HK appear as a continuum of the
Asiatic strains.
OB9
CORONAVIRUS DIVERSITY IN UK WILDLIFE

T. Apaa1, R. Tarlinton1, C. Staley1, A. Withers 1, E. Chadwick2, F. Hailer2, L. Mckenzie5,
X. Lambin5, F. Mathews3, S. Harrison4, S. Bremner-Harrision4, M. Loose1,
A. Blanchard1, M. Bennett1
1
University of Nottingham, Nottingham, UK.
2
Cardiff University, Cardiff, UK.
3
University of Sussex, Brighton, UK.
4
Nottingham Trent University, Nottingham, UK.
5
University of Aberdeen, UK.
Horseshoe bats (Rhinolophidae) are known to be the natural reservoir hosts for
Sarbecoviruses, and SARS-CoV-2 is likely to have originated from these bat species.
Computational analysis and animal studies have demonstrated the presence of high
binding affinity of SARS-CoV-2 to ACE-2 receptors in multiple potential animal host
species. Continuous field detection of SARS-CoV-2 in a wide range of animal species is
reported. Large numbers of cross species transmission events have been recorded in
White tailed deer, cats, and mink. The emergence of further reservoir hosts and SAR-CoV-
2 variants with potential spill-back to humans is likely. The United Kingdom is home to
several wild mustelid, bat and cricetid rodent species that are potentially susceptible to
SARS-CoV-2 however, insufficient data on SARS-CoV-2 spill-over/circulation in wild
animals is available. The aim of this project was to determine Coronaviruses (CoVs)
circulating in UK wildlife and SARS-CoV-2 spill-over from humans into wildlife. A total of
960 samples (faecal, oronasal or rectal swabs, head and neck lymph nodes, and lung
samples) collected from UK wildlife (water voles, stoat, otter, pine marten, weasel, mink,
badgers, fox, and four UK bat species) have been screened using published generic CoVs
pol and Sarbecovirus E gene PCR protocols. Meta-transcriptomics sequencing and
bioinformatic analysis shows the presence of ferret and bat coronaviruses.
OB10
CROSS-TRANSMISSION OF FELINE LEUKEMIA VIRUS (FELV) BETWEEN CHILEAN

DOMESTIC AND WILD FELIDS
C. Castillo1, E. Hidalgo2, C. Napolitano3, A. Blanchard1, R. Tarlinton1
1University of Nottingham, Nottingham, UK
2 Fundación Buin Zoo, Santiago, Chile
3Universidad de Los Lagos, Osorno, Chile
Feline Leukaemia Virus (FeLV) is a gammaretrovirus of cats. FeLV is widespread among domestic
cat populations and can infect many other felids and can produce a serious disease threat to several
species of endangered felids. The main consequences of FeLV infection are hematopoietic
disorders and neoplasia. Domestic cats have both endogenous (copies of virus in the cat genome)
and exogenous variants of FeLV, these frequently recombine producing variants with recombinant
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envelope genes and alternate receptor usage. Until recently inter-cat transmission was presumed
to be solely the exogenous (FeLV-A) variant but recent studies in domestic cats and in wild pumas
in North America have demonstrated that the recombinant FeLV-B variants are also transmitted
horizontally. This study applied envelope gene PCR and NGS (Illumina) to determine the envelope
gene diversity and transmission dynamics of FeLV variants in Chilean domestic cats obtaining
sequences from FeLV-A, B and endogenous FeLV. FeLV-A sequences obtained, are clustered in
a Chilean group closer to sequences from the United States and Brazil. The next step will be to
apply the same methodology to Chilean wild felids including: free-range wild felids (Leopardus
guigna), and felids held in zoological parks (Caracal caracal, Leopardus guigna, Panthera leo,
Panthera uncia, Leopardus pardalis, Panthera onca, Puma concolor and Panthera tigris). The
primary aims of the study are to determine if FeLV B (or other envelope variants of FeLV) are being
disseminated between wild and domestic felids.
OB11
EVIDENCE OF CIRCULATION OF NEWLAVIRUS IN FOXES IN SOUTHERN ITALY

G. Lanave1, L.A. Ndiana1, F. Pellegrini1, B. Di Martino2, G. Sgroi1, V. Vasinioti1, M. Camero1,
D. Otranto1, N. Decaro1, C. Buonavoglia1, V. Martella1
1Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy.
2Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy
In 2021, a novel protoparvovirus, provisionally named as Newlavirus (NLV), was reported with a
prevalence as high as 44% in foxes in Canada. The virus exhibited a high degree of genetic diversity
with low sequence identities in the VP1 gene (70.5–87.8%) and variable detection rates in different
fox tissues. Since NLV were detected in other animal species tested, foxes were supposed to be
the natural reservoir of the virus.
We assessed the prevalence and genetic diversity of NLV in a collection of fox samples in Italy.
One hundred lymph node samples were collected from necropsied foxes (Vulpes vulpes) in
Southern Italy over the 2013-2016 period. NLV DNA was detected by real time PCR in 71/100 (71%)
samples. NLV load ranged between 2.3x101 and 4.0x106 DNA genome copies/10μl (mean = 2.2x105
DNA genome copies/10μl, median=4.9x103 DNA genome copies/10μl). The complete genome
sequence of 7 NLV strains and the full-length VP2 sequence of other 14 NLV strains were obtained.
Whilst in the NS1 region the 7 Italian strain clearly segregated apart from the Canadian viruses, in
the partial (663 bp) VP1/VP2, the 21 Italian NLV strains were intermingled with the Canadian strains
throughout three distinct genetic clusters, a pattern consistent with recombination. Overall, the NLV
prevalence in foxes in Italy was higher than expected. The high genetic diversity observed in the
VP1/VP2 gene (70.2-99.7%) of NLVs collides with the high genetic conservation of carnivore
protoparvovirus species 1 (CPV/FPV), suggesting complex evolutionary dynamics of this novel
protoparvovirus in wildlife.
OB12
MOLECULAR PREVALENCE OF EQUINE PARVOVIRUS-HEPATITIS IN ITALIAN HORSES

R. Cardone, G. Diakoudi, G. Lanave, V.I. Vasinioti, V. Martella, C. Buonavoglia,
G. Elia
Department of Veterinary Medicine, University of Bari “Aldo Moro”, Italy
In 2018 a novel equine parvovirus (EqPV-H) was identified in horses with hepatitis and tentatively
associated with Theiler’s disease. Since then, surveillance studies have demonstrated a global
distribution of EqPV-H in horses with subclinical to severe hepatitis. To assess the distribution of
EqPV-H infection in equid population in Italy, a collection of 1090 serum samples was screened for
the presence of viral DNA using a specific real time PCR assay. Viral DNA was extracted using a
commercial Kit (Qiagen S.p.A., Milan, Italy). Pooled serum samples were initially screened and
EqPV-H positive pools were separated and re-screened in order to identify the virus-positive
samples. Overall, 61/1090 (5.6%) sera tested positive for EqPV-H. EqPV-H transmission still
remains unclear, even if iatrogenic transmission via contaminated biological products of equine-
origin has been demonstrated experimentally. Interestingly, a cluster of infections was identified in
the same stable, in Apulia, South Italy, with 7/14 contact horses being positive to EqPV-H. This
finding hints to a contagious nature of EqPV-H and highlights the need for further investigation on
the modalities of virus transmission.
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OB13
STANDARDISATION OF MOLECULAR AND MICROBIOLOGICAL METHODS FOR THE

DETECTION AND IDENTIFICATION OF PATHOGENS RESPONSIBLE FOR BOVINE
RESPIRATORY DISEASE (BRD)
L. Capozzi1, A. Bianco1, D. Simone1, C. Quarato1, C. Bulzachelli1, M. Solito1, G. Schino1, L.
pace1, M. Galgano2, A. Pratelli2, L. Del Sambro1
1Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Foggia , Italy
2Department of Veterinary Medicine, University Aldo Moro of Bari, Valenzano (BA), Italy
Bovine Respiratory Disease (BRD) is one of the most common causes of morbidity and mortality in
cattle. It affects the lower respiratory tract, is characterized by severe clinical conditions and high
contagiousness. BRD is sustained by several pathogens, both viral and bacterial, and mixed
infections are also frequently identified in pneumonia cases.
The aim of the present study was to identify, using microbiological and molecular methods, the
pathogens responsible for pulmonary lesions detected in cattle at the abattoir, related to BRD.
Sampling was carried out during the slaughter phase, exclusively on carcasses presenting
pulmonary lesions referable to BRD, at two slaughterhouses located in the province of Bari. All the
samples taken (n.44) were subjected to culture examinations by sowing on selective media, in order
to search for the micro-organisms responsible for the disease. Isolated colonies were identified by
biochemical and molecular methods (Table 1). Next generation sequencing methods allowed the
investigation of virulence and antibiotic resistance characteristics. Nucleic acids extracted from
tissue fragments taken from lesions were tested by Real-Time PCR for specific targets of the main
viral and bacterial pathogens responsible for BRD (Table 2).
Microbiological and molecular tests revealed in 88.63% of samples tested (39/44) the presence of
at least one of the pathogens that can be involved to the pathogenesis of BRD.
The outcomes obtained from this surveillance study may lay the foundations for promoting
therapeutic and prophylactic strategies, aimed at reducing the spread of BRD as well as assessing
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the economic impact on regional animal husbandry.
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OB14
GENETIC DIVERSITY OF SALMONELLA INFANTIS ISOLATES FROM BROILERS, HUMANS

AND FOOD, 2007–2021
B. Papić1, D. Kušar1, J. Mićunović1, M. Pirš2, M. Biasizzo1, M. Kavalič1, M. Ocepek1,
J. Avberšek1
1University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
2University of Ljubljana, Faculty of Medicine, Ljubljana, Slovenia
Salmonella Infantis is the most common Salmonella serotype in broilers and is among top five
serotypes isolated from human patients. A decade ago, S. Infantis clone harboring a mosaic
megaplasmid named pESI was first reported, associated with increased virulence, biofilm formation
and multidrug resistance. Since then, S. Infantis clones with pESI-like plasmids have been reported
worldwide, replacing pESI-free clones. Here, 212 S. Infantis isolates from Slovenia, 2007–2021,
underwent whole-genome sequencing to compare the population structure of S. Infantis from
different sources: broilers (n = 150), humans (n = 30) and meat (n = 32). Core genome multilocus
sequence typing (cgMLST) revealed that all isolates were highly genetically homogeneous and
belonged to sequence type (ST) 32, except for one human isolate, which was assigned to a novel
ST. Out of 30 human isolates, five were closely related (≤7 allele differences) to broiler and/or food
isolates, indicating possible zoonotic transmission. Several additional clusters of non-human
isolates were observed and were mostly supported by the underlying epidemiological data. On
farms where multiple broiler isolates were analyzed, persistence of a single clone was observed;
however, reintroduction of additional clones was also noted. pESI-positive clones predominated in
all three groups. Because the spread of S. Infantis clones was observed across all three analyzed
sources, the Slovenian broilers and meat are the source of infection for humans in some cases.
Nevertheless, the source of infection for most human cases remains unknown.
OB15
METAGENOMICS INVESTIGATION IN A MULTI-PATHOGEN FOODBORNE OUTBREAK OF

ACUTE GASTROENTERITIS USING NANOPORE TECHNOLOGY.
G. Diakoudi1, G. Lanave1, F. Pellegrini1, F. Beikpour1, E. Suffredini2, C. Catella1, D.
Loconsole3,
M. Chironna3, V. Martella1
1Dipartimento di Medicina Veterinaria, Università degli Studi di Bari Aldo Moro, Italy
2Istituto Superiore di Sanità, Rome, Italy
3Dipartimento di Scienze Biomediche, Università degli Studi di Bari Aldo Moro, Italy
Syndromic diagnostics have unveiled that the aetiology of acute gastroenteritis (AGE), in either
sporadic cases or in outbreaks, can be multifactorial, thus posing a challenge for the current
diagnostic approaches. In April 2018, an AGE outbreak related to consumption of raw seafood in
Bari, Italy, was identified. Up to twelve different viral strains of norovirus, astrovirus and aichivirus
were detected from both symptomatic and asymptomatic persons, suggesting massive
contamination of the food source. Accordingly, these samples were considered an excellent target
for metagenomics investigation using Nanopore sequencing technology. The fecal sample of a
symptomatic patient was used to generate a library with SISPA protocol, producing ~5.8 GB of data
(> four million reads) in MinION Mk1C flow cell. About 12% of the reads were recognized as viral
on Genome Detective virus tool. We generated the complete genome sequence of a human
mamastrovirus (family Astroviridae), the nearly complete genome sequence of a human aichivirus
A (family Picornaviridae) and reads of norovirus genogroup I (family Calicivirdae), all associated
with AGE in humans. Also, we detected reads of picobirnaviruses (family Picobirnaviridae) and of
salivirus A (family Picornaviridae), whose role in AGE is unknown/unclear. In addition, reads of 4
plant viruses and of 138 phages were identified. Overall, the experiment unveiled the complexity of
fecal virome in the patient and confirmed the importance of metagenomics approach in investigation
of foodborne outbreaks.
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OB16
GENOME SEQUENCING OF A RABBIT ROTAVIRUS A STRAIN WITH NANOPORE PLATFORM

A. Hassan Omar, F. Pellegrini, G. Lanave, C. Catella, G. Diakoudi, A. Cannarozzi, G. Casalino,
E. Circella, F. D’Amico, C. Buonavoglia, V. Martella
Dipartimento di Medicina Veterinaria, Università degli Studi di Bari Aldo Moro, Italy
Group A Rotaviruses (RVAs), family Reoviridae genus, are important gastroenteric pathogens
associated with acute gastroenteritis in humans and animals. RVA genome is composed of 11
segments of double-stranded RNA (dsRNA) for a total of 18.5kb. A genome-based classification
was developed in 2008 to compare effectively different RVA strains and explore their genome make-
up. RVA are associated with enteric diseases in rabbit, mainly in post-weaning animals.
We identified the RNA of RVA by a quantitative TaqMan real time RT-PCR (qRT-PCR) in a group
of young rabbits (9 out of 11 stool samples, mean CT= 25.88, range CT=15.33-31.34) with enteritis
complex from Santeramo in Colle (Bari, Italy).
By using a PCR-based enrichment protocol, the complete genome of the rabbit RVA strain was
reconstructed. The PCR product was used for library preparation with a Ligation sequencing kit
SQK-LSK110 (Oxford Nanopore Tech.) and sequenced on a MinION MK1C device. The fastq files
were analysed with Genome Detective. The 96.8% of the genome was assembled, using 62,1268
reads, with a 69,739.5-depth coverage. The virus showed the genotype constellation G3-P[14]-I2-
R2-C2-M3-A9-N2-T6-E5-H3, with the highest nucleotide identity in BLAST to either RVA/Human-
wt/BEL/B4106/2000/G3P[14] or RVA/Human-wt/BEL/BE5028/2012/G3P[14], in all the genome
segments. These human viruses were detected in children in Belgium in 2000 and 2012 and were
regarded as zoonotic viruses of rabbit origin. Gathering sequence data on animal RVA is pivotal to
reconstruct the origin of unusual human RVAs of zoonotic origin.
OB17
CANCELLED
PSA 23 – OB18
A SUSPECTED CASE OF CONTAGIOUS BOVINE PLEUROPNEUMONIA IN THE

NETHERLANDS – A RARE DISEASE REVISITED
H. Graham1, M. van der Most1, R. Dijkman2, L. Harkema2, L. Labout3, P.H.M.M. Jacobs3, M.A.H.
Spierenburg3, M. Heijne1
1Wageningen Bioveterinary Research, Lelystad, The Netherlands
2Royal GD, Deventer, The Netherlands
3 Netherlands Food and Consumer Product Safety Authority (NVWA), Utrecht, The Netherlands
The histologic features of lung tissue of a bull that was euthanized due to severe respiratory
symptoms shortly after import in the Netherlands, were indicative of contagious bovine
pleuropneumonia (CBPP), a notifiable disease caused by Mycoplasma mycoides subspecies
mycoides (Mmm). The Netherlands had been free of this rare disease since 1886. The suspicion
prompted an investigation, which involved the veterinary authorities and the national reference
laboratory (NRL). A real-time PCR was performed on tissue samples of the lung and bronchial tree,
yielding a negative result. Furthermore, a farm visit was made to inspect the health of the herd and
to collect serum samples from the co-housed animals. One out of twenty serum samples tested
positive in an in-house CBPP complement fixation test (CFT), which led to a second farm visit to
repeat the sampling. In addition, tissue samples of the bull and the collected serum samples were
forwarded to an OIE reference laboratory for confirmation. The OIE reference laboratory confirmed
the absence of Mmm DNA in the samples of the bull and sequencing revealed the presence of
Mycoplasma bovis. Because of this and by performing the cELISA for CBPP on the serum samples,
the initially positive CFT result was considered to have been caused by cross-reactivity. This case
may be closed, but is a valuable reminder for laboratories and microbiologists not to forget this
disease and be prepared with the right set of specific diagnostics tests: not only to diagnose CBPP,
but more importantly, to be able to exclude it.
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OB19
NOVEL CORYNEBACTERIUM SPECIES ISOLATED FROM A CORNEAL ULCER AND

URINE IN DOGS
M. Tresch1, C. Watté2, M. Stengard2, S. Feyer1, E. Akdesir1, C. Ritter3, S. Kittl1
1
Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Switzerland
2
Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern,
Switzerland
3
Veterinary Office Villereuse, Geneva, Switzerland
Background: Corynebacteria are Gram-positive club-shaped bacteria, which cause

various diseases in animals. Though corynebacteria can occur in conjunctival sacs of
healthy dogs, species such as C. pseudodiphtheriticum and C. renale have also been
isolated from dogs with corneal ulceration. C. urealyticum is described as a rare cause of
cystitis in dogs. Methods: A swab from a corneal ulcer and a urine sample from two canine
patients were routinely cultured in our laboratory. Coryneform bacteria were isolated which
could not be identified using MALDI-TOF MS. Therefore, 16s rRNA gene sequencing was
performed. Antimicrobial susceptibility was determined by broth microdilution (Sensititre®)
following CLSI guidelines. The isolates were subjected to whole genome sequencing
(Illumina®) followed by digital DNA-DNA hybridization (dDDH) and Genome Blast Distance
Phylogeny analysis (Type Strain Genome Server). Furthermore, average nucleotide
identity (ANI) was determined using OAT v0.93.1. Results: While 16s rRNA sequencing
preliminarily identified the isolates as Corynebacterium oculi, first isolated from ocular
specimens in humans, analysis of the whole genome sequencing data indicated that these
isolates cluster together as a potentially new, closely related species. ANI with the most
closely related species C. oculi was only 87%, confirming this finding. While the ocular
isolate showed low MIC ranges for all tested antibiotics, the isolate from urine showed high
MIC values for fluoroquinolones and a mutation in the gyrA gene was detected.
Conclusion: As a versatile and potentially fluoroquinolone-resistant pathogen in dogs,
this novel species should be further investigated. More isolates are needed for further
characterization.
OB20
ARYL HYDROCARBON RECEPTOR IS ACTIVATED BY INFECTION WITH CANINE

CORONAVIRUS
C. Cerracchio1, M.G. Polverino2, V. Iovane3, A.R. Attili4, M.G. Amoroso2, F. Fiorito1,4
1Department of Veterinary Medicine and Animal Production, University of Naples Federico II, 80137
Naples, Italy
2Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055, Portici, Portici (Naples), Italy
3Department of Agricultural Sciences, University of Naples Federico II, Portici, Naples, Italy
4BAT Center-Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology,
University of Naples Federico II, Portici, Naples,Italy
Objectives: The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that
interacts with endogenous and exogenous substrates including bilirubin, biliverdin, tryptophan
metabolites, environmental pollutants, and microbial metabolites. The activation of AhR by these
substances induces the control of the expression of target genes such as AhR repressor, detoxifying
monooxygenases, and cytokines. Recent advances reveal that AhR signaling regulates aspects of
the intrinsic, innate and adaptive immune response to diverse microorganisms. AhR is involved in
the host response to Coronaviruses (CoVs) (i.e. MCoV, SARS-CoV-2, HCoV 229E) infection.
Particularly, AhR agonists decrease the expression of ACE2 via AhR activation, resulting in
suppression of SARS-CoV-2 infection in mammalian cells. Here, we report that AhR is activated by
infection with genotype II of canine coronavirus (CCoV-II), an alphacoronavirus. Moreover,
pharmacological inhibition of AhR suppresses in vitro replication of CCoV.
Methods used: Infection of CCoV (378/strain) in canine fibrosarcoma (A72) cell line was performed
in the presence of CH223191, an AhR antagonist. Bioscreen, immunofluorescence, and virus yield
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analyses were carried out.

Results: Following CCoV infection, we found a considerable stimulation of AhR, a receptor
expressed in A72 cells. At non-toxic concentration, CH223191 noticeably reduced cell death signs
and increased cell viability. Furthermore, the AhR antagonist induced a meaningful decline in virus
yield, accompanied by the inhibition of the expression of viral nuclear protein.
Conclusions: Taken together, our findings show that infection with CCoV activates AhR.
Furthermore, pharmacologic AhR inhibition reduces CoVs replication in vitro, identifying AhR as a
possible candidate target for antiviral therapy..
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POSTER SESSION A
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PSA01
ANTIBODIES AGAINST TESTUDINID HERPESVIRUSES 1 AND 3 IN PET TORTOISES IN

EUROPE
R. E. Marschang, E. Müller, C. Leineweber
LABOKLIN GmbH & CO KG, 97688 Bad Kissingen, Germany
Herpesviruses can cause serious disease in turtles and tortoises. Four genetically distinct
herpesviruses have been described in tortoises, testudinid herpesvirus (TeHV) 1-4. Types 1 and 3
are by far most commonly described. Both have been isolated in cell culture and virus neutralization
tests (VNT) can be used to detect antibodies against each in plasma or serum from infected
tortoises. In a retrospective study, antibody detection by VNT in samples from 1728 captive tortoises
submitted to our laboratory between 2016 and 2020 were evaluated. Antibodies against at least
one of the viruses were found in 122 animals (7.06%; 95% CI 5.95-8.37%), including 41 (2.37%;
95% CI 1.75-3.20%) with antibodies against TeHV1 only, 62 (3.59%; 95% CI 2.81-4.58%) against
TeHV3 only and 19 (1.1%; 95% CI 0.71-1.71%) against both viruses. Detection rates differed
significantly between different species (P < 0.001) with the highest rates against TeHV1 found in
Russian tortoises (Testudo horsfieldii) (22.92%) and the highest rates against TeHV3 found in
marginated (T. marginata) (22.47%) and spur-thighed tortoises (T. graeca) (12.23%). TeHV3
detection rates also differed significantly depending on the country from which the samples were
submitted (P = 0.0014), while those for TeHV1 did not (P = 0.5567). Compared to results of virus
detection studies in pet tortoises in Europe, antibodies were detected in a significantly (P = 0.0059)
lower percentage of Hermann’s tortoises (T. hermanni) compared to virus detection rates. This
supports the hypothesis of previous authors that immune response to herpesvirus infection is
dependent on host species in tortoises. Key words: Russian tortoise, Hermann’s tortoise, spur-
thighed tortoise, testudinid herpesvirus, Testudinid alphaherpesvirus 3, Testudo, serology
PSA02
DETECTION OF PORCINE CIRCOVIRUS IN AN ITALIAN FOX

F. Alfano1, M. G. Lucibelli1, N. D’Alessio1, C. Auriemma1, A. Gallo1, E. De Carlo1, N. Decaro2,
V. Martella,2 G. Fusco1
1Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici (NA), Italy
2 Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, Valenzano (BA), Italy
The aim of our work was to study the circulation of porcine circovirus (PCV-2) in domestic and wild
carnivores in Italy.Out of 206 carcasses (100 dogs, 77 cats, 14 wolves, 15 foxes) analysed in our
investigation, 5 tested positive for PCV-2 (2 dogs, 1 cat, 1 fox, 1 wolf). A single PCV-2 infection was
found in the brain of a fox (#119493), whereas in the other PCV-2 positive animals, co-infections
with other pathogens were observed. The localization of circoviruses in the brain is not unusual in
animals and has recently been reported in humans with paraplegia and acute central nervous
system infections (Smits et al. 2013; Tan et al. 2013). In 2016 in southern Italy a canine circovirus
was found in the brain of dogs and wolves, but not in foxes and it was associated to neurological
and enteric signs (Zaccaria et al. 2016). According to our knowledge, this is the first report of porcine
circovirus in the brain of a fox.
The results of our study highlight the need to monitor the PCV-2 circulation in carnivores, as well as
the clinical forms associated to this viral agent in animals different from swine.
References
1. Smits SL et al. 2013 Emerg Infect Dis 19:1511-3
2. Tan LV et al. 2013 MBio. 4:e00231–13
3. Zaccaria G, et al. 2016. Virology. 490:69-74.
PS03
EFFECTS OF FARMACOLOGICAL INHIBITION ON THE AUTOPHAGIC MACHINERY DURING

FELINE HERPESVIRUS (FHV-1) INFECTION IN VITRO.
G. Ferrara1, V. Iovane2, F. P. Nocera1, C. Longobardi3, U. Pagnini1, S. Montagnaro1
1Department of Veterinary Medicine and Animal Productions, University of Naples “Federico II”,
Naples, Italy
2Department of Agricultural Sciences, University of Naples “Federico II”, Naples, Italy.
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3Department of Mental, Physical Health and Preventive Medicine, University of Campania “Luigi
Vanvitelli”, Naples, Italy.
Feline herpesvirus type 1 (FeHV-1) is a common virus that causes rhinotracheitis and conjunctivitis
in cats. This virus, like other members of the alphaherpesviridae subfamily, interacts with
autophagic machinery to complete its biological cycle. We investigated the effects of
pharmacological autophagy inhibition on viral progeny production in this study.
Permissive CRFK cells were infected with FeHV-1, in the presence of different autophagy inhibitors
(3-Methyladenine, LY294002, bafilomycin A1, and chloroquine diphosphate). Following viability
evaluation (MTT assay), we investigated autophagic flux using autophagy markers (LC3 and
p62/SQSTM1) by western blot analysis. Furthermore, we used viral titration (TCID50) and real-time
PCR to investigate viral production in the absence and presence of inhibitors. Bafilomycin and
chloroquine treatment reduced the conversion of LC3I in II as well as the degradation of
p62/SQSTM1. Autophagy inhibition was accompanied by lower viral titers and viral gene
expression. Using other inhibitors, no effects were observed.
These results were confirmed when siRNA was used to interfere with the late stages autophagy
related gene ATG5. On the other hand, the use of rapamycin (an autophagy inducer) had the
opposite effect on the viral titers as well as on the viral gene expression. These results suggest the
importance of basal autophagy for the accomplishment of the FeHV-1 cycle, as well as the reduction
of viral spread caused by the use of late-stage autophagy inhibitors. These compounds require
additional experiments to determine their potential therapeutic use, considering the in vivo
cytotoxicity of bafilomycin and the wide application of chloroquine in humans.
PS04
THE IMPORTANCE OF INDIRECT ELISA TESTS IN THE DETECTION OF SMALL

RUMINANTS LENTIVIRUS INFECTION IN PORTUGAL
J. Jacob-Ferreira1, A.C. Coelho1, A. Grau Vila2, O. Mínguez González2, D. Lacasta3, R.
Valentim4, H. Quintas4
1CECAV – Animal and Veterinary Research Centre; Associate Laboratory for Animal and
Veterinary Sciences (AL4AnimalS), Portugal

2Servicio de Sanidad Animal, Dirección General de Producción Agropecuaria e
Infraestructuras Agrarias, Consejería de Agricultura y Ganadería, Junta de Castilla y León,

Valladolid, Spain,
3Animal Pathology Department, Instituto Agroalimentario de Aragón-IA2, Universidad de
Zaragoza-CITA, 50013 Zaragoza, Spain

4Mountain Research Centre (CIMO), School of Agriculture, Polytechnic Institute of Bragança
(IPB), Campus de Santa Apolónia, Bragança, Portugal
OBJECTIVES: Small ruminant lentiviruses (SRLVSs) are the group of viruses responsible
for Maedi-Visna in ovine and for Caprine Arthritis Encephalitis in caprine species. Theses
diseases result of progressive and chronic infections as well as are one of the major causes
of severe economic loss. On current days, in Portugal, there is few information about SRLV
infection. The main aim of this research is to quantify the seroprevalence associated to SRLV
in Portugal. MATERIAL AND METHODS: Seroprevalence of SRLV research was done in
small ruminant herds of the Portugal. Between 14 to 19 blood samples of different ages,
based on the total number of animals in each herd were collected. The infection by SRLV in
each sample was made using a commercial test of indirect ELISA (ID Screen® MVV/CAEV
Indirect). The herd was considered positive if at least one animal was seropositive.
RESULTS: We collected samples in a total of 102 herds, having 75 of ovine, 18 of caprine
and 9 of mixed species. We verified that 91(89.22%) herds were positive to SRLV, of which
66 (88%) of ovine, 16 (88.89%) of caprine and 9 (100%) of mixed species. Of 1774 blood
samples, 745 (42%) were positive, with 519 (39.74%) positive animals in ovine herds, 160
(51.78%) in caprine herds and 66 (41.51%) in mixed species herds. CONCLUSION: There
is a highly seroprevalence of SRLV infection in Portugal. These results highlight the
importance of using laboratorial methods such as ELISA tests for the early detection of
diseases in small ruminant herds.
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PS05
ANTIVIRAL ACTIVITY OF FUNGAL SECONDARY METABOLITE 6-PENTYL-a-PYRONE

AGAINST CANINE CORONAVIRUS INFECTION
C. Cerracchio1, M.G. Polverino2, A. Staropoli3,4, F.P. Nocera1, L. De Martino1, MG Amoroso2,
F. Vinale1,5, F. Fiorito1,5
1Department of Veterinary Medicine and Animal Production, University of Naples Federico II, 80137
Naples, Italy
2Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055, Portici, Portici (Naples), Italy
3Department of Agricultural Sciences, University of Naples Federico II, Portici, Naples, Italy
4Institute for Sustainable Plant Protection, National Research Council, Portici, Naples, Italy
5BAT Center-Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology,
University of Naples Federico II, Portici, Naples, Italy
Objectives: Genotype II of canine coronavirus (CCoV-II), an alphacoronavirus, can cause self-

limiting enteric disease in dogs. The remarkable plasticity of CoVs occurs through mutation and
recombination processes. The current SARS-CoV-2 pandemic as well as the recent detection of a
novel canine-feline recombinant alphacoronavirus isolated from a human patient highlights the
cross-species transmission ability of CoVs. In this scenario, antiviral compounds are considered
necessary to fight CoVs infections. Fungi produce secondary metabolites (SMs), often developed
as antibiotics, fungicides, hormones, and plant growth regulators. Screening performed on benzo-γ-
pyrone 3-O-methylfunicone, a SM produced by Talaromyces pinophilus, showed that it reduces
infectivity of hepatitis C virus and bovine herpesvirus 1. 6-pentyl-α-pyrone (6PP), a SM produced by
Trichoderma genus, displays activity against plant pathogens, and anti-biofilm-producing bacteria.
Up to now, no studies about the potential antiviral activity of 6PP are known. So, here antiviral ability
of 6PP was assessed against CCoV-II infection. Methods used: During CCoV (378/strain) infection
in canine fibrosarcoma (A72) cell line, bioscreen, immunofluorescence staining, cytomorphological
and virus yield analyses were performed. Results: Following CCoV infection, the non-toxic
concentration of 0.1 µg/mL of 6PP markedly increased cell viability and cell proliferation. These
results were accompanied by reduced cell death signs. Moreover, 6PP significantly reduced virus
yield and the expression of the nucleocapsid protein NP. Conclusions: Overall, our findings
suggest that nontoxic concentration of 6PP shows potential activity against CCoV infection.
Noteworthy, in the screening of potential antivirals, in vitro animal model of CoVs avoids the
manipulation of extremely hazardous CoVs (SARS-CoVs, MERS-CoV).
PS06
DETECTION OF SAPOVIRUSES IN ITALIAN CARNIVORES

F. Alfano1, M. G. Lucibelli1, G. Miletti1, N. D’Alessio1, A. Gallo1, C. Auriemma1,
R. Brunetti1, M. P. Valentino1, L. Baldi1, E. De Carlo1, N. Decaro2, V. Martella,2 G. Fusco1
1Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici (NA), Italy
2Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, Valenzano (BA), Italy
The aim of our work was to study the circulation of sapoviruses (SaVs) in carnivores in Italy. Out of
206 carcasses (100 dogs, 77 cats, 14 wolves, 15 foxes) analysed in our investigation, 9 (5 dogs, 3
cats, 1 fox) tested positive for SaV RNA. Single SaV infection was found in only 2 dogs (#3354 and
#23021), whereas in the other SaV-positive animals, co-infections with other pathogens were
observed. Dog #23021 displayed gastroenteric lesions, as commonly observed in SaV infections in
humans (4) and dogs (1). In contrast, in dog #3354 there was no gastroenteric involvement and the
observed lesions were referable to a uremic syndrome (US). Whether the SaV infection in the dog
was associated with US or it was a mere coincidence could not be assessed. Also, since we did not
use an open diagnostic system, we cannot rule out the presence of other undetected pathogens. In
humans, US has been reported in patients with bacterial gastroenteritis (3). However, there is also
a limited number of cases not related to gastroenteritis. These "atypical" forms of US recognize
various causes, including viral infections (2). The results of our study highlight the need to monitor
the SaV circulation in dogs as well as the clinical forms associated to this viral agent.
References
1. Bodnar 2016. Infect Genet Evol, 38:8-12.
2. Noris and Remuzzi 2005. J Am Soc Nephrol Apr;16(4):1035-1050.
3. Sugimoto et al. 2007. Nephrol Dial Transplant 22: 2098.
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4. Usuku et al. 2008. Jpn J Infect Dis, 61:438-41.
PS07
MOLECULAR SURVEILLANCE FOR NOROVIRUS AND ROTAVIRUS INFECTIONS IN

DOMESTIC CARNIVORES IN ITALY
F. Mira1, F. Gucciardi1, G. Schirò1, G. Chiaramonte1, G. Purpari1, D. Vicari1, F. Antoci1, V.
Martella2, A. Guercio1, S. Di Bella1
1Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Palermo, Italy
2Department of Veterinary Medicine, University of Bari, Valenzano, Italy
A One Health approach is strongly suggested for management of infectious diseases in humans
and animals. Companion animals, dogs and cats, have a close interaction with humans, sharing the
same environment. This generates occasional exposure of humans to potential zoonotic enteric
viruses, such as norovirus (NoV) and rotavirus (RoV) and viceversa of domestic carnivores to
human enteric viruses. We investigated the epidemiology of NoVs and RoVs in dogs and cats in
Italy. Intestines, faeces or rectal swab were collected from n=170 dogs and n=47 cats in the period
2020-22. NoV was detected in 4/170 (2.3%) dogs and 3/47 (6.4%) cats, whilst RoV was detected
in 2/170 (1.1%) dogs and 3/47 (6.4%) cats, either alone or in mixed infections with other viral
pathogens. None of the NoV- and RoV-positive animals lived in a household. All but two (2 and 3
years old) were young animals (10 days - 3 months old). The amplicon generated from the
diagnostic region B (ORF1) of NoVs was sequenced. The canine NoVs displayed the highest
nucleotide (nt) identities to 2018/19 GVI.2 strains from China (95.72%), to 2007 GIV.2 strains from
Italy (93.90%) and 2008 NoVs from Greece (93.89-94.86%). Feline NoVs displayed the highest
identities to 2013 GVI.2 strain (91.95%), to 2010 GIV.2 strains from USA (90.10-94.79%), Italy 2006
(91.75-92.33%), and Japan 2017 (91-91.95%). Gathering sequence data on enteric viruses of
domestic carnivores is useful to define better the plasticity of the enteric virome at the human/animal
interface.
PS08
HEPATITIS E VIRUS IN TUSCANY: EVIDENCE OF PRESENCE IN CO-HABITING WILD AND

DOMESTIC ANIMALS
M.I. Pacini, M. Forzan, N. Fonti, M. Mazzei
Department of Veterinary Sciences, University of Pisa, Italy
Hepatitis E virus (HEV) is a non-enveloped ssRNA+ virus belonging to the Hepeviridae family
responsible for sporadic and epidemic outbreaks in industrialized countries. Although suids are
recognized as the main reservoirs of HEV, other species may act as HEV hosts, including domestic
and wild animals. In central Italy, Tuscany represent a model for studying HEV epidemiology in
complex ecosystems since it is one of the Italian regions with the largest number of wild animals
and hunting activity is largely diffused.The aim of our research is to investigate and analyse the
circulation of HEV in several wild and domestic animals sharing the same ecosystem in Tuscany.
From 2014 to 2021, serological and molecular investigations were carried out in wild species mostly
diffused and hunted in Tuscany (wild boar, deer, brown hares, and wild rabbits), and in domestic
animals living in close contact with wildlife (hunting dogs and semi free-range cattle). Serological
analysis recorded variable seroprevalences with values ranging from 50% (wild boar, 186/374) to
5% (rod deer, 2/43); no serological positivity was found in cattle (0/10), fallow deer (0/13) and brown
hares (0/103). Moreover, HEV RNA was detected in liver samples collected from wild boar, wild
rabbit, roe deer and fallow deer and in swabs sampled from wild boar carcasses; no molecular
positivity was found from hunting dogs’ rectal swab. In conclusion our data indicate a diffuse
circulation of HEV in a wide range of animals living in an area characterised by a wide interface
between wildlife and domestic animals.
PS09
DETECTION OF INFLUENZA A VIRUS-SPECIFIC ANTIBODIES IN HOUSE CATS AND STRAY

CATS IN THE NETHERLANDS, 2020-2022
M.B.H.M. Duijvestijn1*, N. Schuurman1, T.F. Verweij1, J.M.A. van den Brand1, J.A. Wagenaar1,2,
F.J.M. van Kuppeveld1, C.A.M.de Haan1, H.F. Egberink1 , J.H.Verhagen1.
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1Utrecht University, Faculty of Veterinary Medicine, Department of Biomolecular Health Sciences,

Utrecht, The Netherlands.
2Wageningen Bioveterinary Research (WBVR), Lelystad, The Netherlands.
Objective: Cats have been shown to be susceptible to infection and disease with highly pathogenic
avian influenza (HPAI) H5 viruses, yet limited knowledge is available about cats’ susceptibility to
different influenza A virus (IAV) subtypes. Haemagglutinin of IAV is a major target for protective
antibodies. Here we used sera from house cats and stray cats sampled in the Netherlands (2020-
2022) to investigate pre-exposure to H1, H3, H5, H7 and H9 IAVs. Materials and Methods: A total
of 296 house cats (95 municipalities) and 476 stray cats (52 locations) were tested for presence of
antibodies against human origin (H1N1pdm2009, H1N12007, H3N2), canine origin (H3N8), and avian
origin (H5N8, H7N9, H9N2) IAV using an in house developed ELISA based on recombinant
haemagglutinin proteins. Results: The seroprevalence of antibodies against H1pdm2009 in house cats
was 7.1% (21/296) and 3.2% (15/476) in stray cats. The H5 seroprevalence was 0.7% (2/296) in
house cats and 6.9% (33/476) in stray cats. Seroprevalence for H12007 and H3 in both groups was
<1%. Less than 1% (2/296, 1/296) of house cats had H7- or H9-specific antibodies, while stray cats
had 1,5% (7/476) H7- and 1,7% (8/476) H9-specific antibodies. The H5 seroprevalence in stray cats
varied by location, ranging from 0% to 100% (8/8). Three cats <1 year had H5N8-specific antibodies,
reflecting recent exposure.Conclusion: House cats showed higher seroprevalence to human origin
H1pdm2009 IAV and stray cats showed higher seroprevalence to avian origin H5N8 IAV. A role of cats
as mixing vessel for IAVs needs further research.
PSA10
VAGINAL ADMINISTRATION OF A BOHV-4-BASED VECTOR EXPRESSING CpHV-1 gD

PROTECTS GOATS AGAINST CpHV-1 INDUCED PATHOLOGY
A.E. Odigie1, M.S. Lucente1, C. Catella1, D. Mrenoshki1, I.C. Ugochukwu1, F. Pellegrini1, P.
Capozza1,A.A.K Zarea1, V. Franceschi2, G. Greco1, A. Pratelli1, G. Donofrio2, M. Tempesta1,
1Department of Veterinary Medicine, University of Bari “Aldo Moro”, SP 62 Casamassima km 3,
Valenzano, Italy
2Department of Veterinary Medical Sciences, University of Parma, Via del Taglio 10, Parma, Italy
Objectives: Caprine herpesvirus 1 (CpHV-1), an Alphaherpesvirus of the Herpesviridae family and

Varicellovirus genus, is a globally occurring non-vaccine preventable genital infection linked with
major economic losses due to returns in service, abortions and stillbirths. We evaluated the efficacy
of a recombinant BoHV-4 virus vector expressing CpHV1 gD (BoHV-4-A-gD(cp)gD(106)ΔTK),
already validated when inoculated parenterally, vaginally administered as a potential vaccine
candidate against CpHV-1 infection in goats. Methods: Four CpHV1-seronegative goats received
via vaginal mucosa three dose of the BoHV-4-A-gD(cp)gD(106)ΔTK at intervals of three weeks
while three other goats remained unvaccinated as controls. Two weeks after the third vaccination,
all goats were intravaginally challenged with a highly pathogenic dose of BA.1 strain of CpHV-1 and
underwent a 20-day observation for clinical signs. Vaginal swabs, vaginal washes and sera were
collected for serology and virological tests. Results: All vaccinated goats were clinically protected
against CpHV-1 induced genital disease with short duration of mild hyperemia, minimal viral
shedding, and very low to absent humoral response. In contrast, the unvaccinated controls
developed severely graded hyperemic and oedematous vaginal lesions with prolonged high titer
viral shedding. Conclusion: This study validated BoHV-4-based vector administered mucosally as
a safe and effective vaccine candidate against CpHV-1 induced genital disease in goats with
potentially diverse applications.
PSA11
SARS-CoV-2 INFECTION IN DOGS AND CATS IS ASSOCIATED WITH CONTACT TO COVID-

19 POSITIVE HOUSEHOLD MEMBERS
M.M. Kannekens-Jager1, M.M.T. de Rooij2, H.S. Kooistra3, H.F. Egberink1, J.A. Wagenaar1,
E.M. Broens1
Utrecht, The Netherlands

2Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht University,
Utrecht, The Netherlands
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3Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The
Netherlands
The aim of this research was to assess the SARS-CoV-2 prevalence amongst cats and dogs (PCR-
and/or antibody positive) and potential risk factors in two different populations in the Netherlands
during the period July 2020 – April 2021. Samples were collected from dogs and cats living in a
household with at least one confirmed COVID-19 positive person (household study; 156 dogs and
152 cats) and dogs and cats visiting a veterinary clinic (veterinary clinic study; 183 dogs and 140
cats). Collected oropharyngeal and rectal swabs were tested for presence of virus (PCR) and blood
samples were tested for antibodies (ELISA). PCR-positive animals were followed over time and
retested. In the household study, 18.8% (27 dogs, 31 cats) tested SARS-CoV-2 positive, whereas
in the veterinary clinic study SARS-CoV-2 prevalence was 4.6% (6 dogs, 9 cats). SARS-CoV-2
prevalence amongst dogs and cats was significantly higher in multi-person households with two or
more COVID-19 positive persons compared to multi-person households with only one COVID-19
positive person. In both study populations, no associations could be identified between SARS-CoV-
2 status of the animal and health status, age or sex of the animal. During follow-up of PCR-positive
animals, no other pets in the household were tested positive despite long-lasting SARS-CoV-2 RNA
positivity in cats (up to 35 days). In conclusion, SARS-CoV-2 infection in dogs and cats suggests
no severe clinical signs and SARS-CoV-2 infection in dogs and cats appeared to be clearly
associated with confirmed COVID-19 positive status of the household.
PSA12
VIRULENCE PROFILE OF ORAL ENTEROCOCCI OBTAINED FROM DOGS

E. Cunha1,2, L.M. Carreira1,2, M. Videira3, A.F. Ferreira1,2, R. Abreu1,2, l. Chambel4, L.
Tavares1,2, M. Oliveira1,2
1CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina
Veterinária, Universidade de Lisboa, Lisboa, Portugal
2Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Portugal
3Casa dos Animais de Lisboa, Câmara Municipal de Lisboa, Lisboa, Portugal 4BioISI – Biosystems
and Integrative Sciences Institute, Faculdade de Ciências da Universidade de Lisboa, Lisbon,

Portugal
Background: Enterococci are ubiquitous bacteria with high genome plasticity. As opportunistic
bacteria and according to their virulence signatures, enterococci are a leading cause of difficult
to treat healthcare infections in both humans and animals. This work aimed to isolate, identify
and characterize the virulence profile of enterococci obtained from the oral cavity of healthy
dogs. Materials/methods: Twenty dogs were selected from an official animal’s rescue institution.
All procedures were approved by the Ethical Committee of the Faculty of Veterinary Medicine,
University of Lisbon. Oral swabs were obtained from each animal and conventional bacteriological
techniques were used for enterococci isolation, followed by PCR identification as described by
Jackson et al 2004. The phenotypic virulence profile of each isolate was assessed by evaluating
lipase, gelatinase, lecithinase, protease and DNase activity, haemolysin’s’ production and biofilm-
forming ability. Results: A total of 68 enterococci were recovered from 17 animals. PCR
identification revealed that 35.29% of the isolates were identified as E. faecalis, 13.24% as E.
faecium, 5.88% as E. hirae, and 45.59% as Enterococcus spp. Virulence evaluation revealed that
55.88% of the isolates were able to produce lipase, 27.94% gelatinase, 27.94% lecithinase,
26.47% protease, 0% DNase, 51.47% haemolysins, and 92.65% biofilm. Isolate´s virulence
index ranged from 0.14 to 0.86. Conclusions: Enterococci with high virulence index are present in
the oral cavity of most dogs, having potential to became opportunistic pathogens. Our results
reinforce the establishment of enterococci as an important reservoir of virulence determinants,
being a relevant bacterial model in veterinary bacteriology. Ackowledgements: This work was
supported by CIISA–Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de
Medicina Veterinária, Universidade de Lisboa, Project UIDB/00276/2020 (Funded by FCT); and by
the Associate Laboratory for Animal and Veterinary Sciences (LA/P/0059/2020 - AL4AnimalS).
PSA13
GENOMIC ANALYSIS OF ESCHERICHIA COLI ISOLATES FROM A CHICKEN HATCHERY

A. Papoušková1,2, M. Masaříková1.2, J. Palkovičová2, A. Čížek1,2
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1Department of Infectious Diseases and Microbiology, Faculty of Veterinary Medicine, Veterinary

University Brno, Brno, Czech Republic
2Central European Institute of Technology, Veterinary University Brno, Brno, Czech Republic
E. coli is one of the most important bacterial pathogens in commercial poultry. Recent research
shows that pathogenic poultry strains (APEC) form a dynamic variable population in which "risk"
genotypes and clonal lines play an important role. These can also spread vertically from parent
farms to offspring, potentially from poultry products to consumers. Hatcheries appear to be a key
site of cross-contamination and primary colonization of newly hatched chickens. Samples of shells
and dust were taken from hatchery boxes in a commercial hatchery in the years 2020-21. Based on
preliminary characterization, 74 E. coli isolates were selected for whole genome sequencing by
Illumina. The acquired data were analyzed by available tools to obtain a complex genetic
characterization (VirulenceFinder, ResFinder, SeroTypeFinder, PlasmidFinder) and phylogenetic
analysis (MLST, pMLST). In contrast to previously analyzed clinical isolates, the B1 phylogenetic
group dominated among the hatchery population, with a high prevalence of ST155 and ST162.
Typical APEC genotypes, especially ST23 and ST117, were detected sporadically, which proves
their occurrence in the hatchery environment, where they readily colonize day-old chicks. Repeated
detection could indicate their persistence in parent flocks or their spread between farms,
presumably through hatcheries. Whole genome sequencing makes a fundamental tool for
monitoring the occurrence of risk lines in the production chain of commercial poultry. Only long-term
sampling at different levels of the pyramid with detailed genetic analysis will provide sufficient data
to assess the significance of E. coli risk clonal lines in poultry.
PSA14
ISOLATION AND CHARACTERIZATION OF FOUR NEW BACTERIOPHAGES AGAINST

AEROMONAS SALMONICIDA, THE CAUSATIVE AGENT OF FURUNCULOSIS
S. Desmecht1, C. Antoine1, F. Laforêt1, J.-N. Duprez1, A. Schonbrodt2, F. Lieffrig3, D. Thiry1
1Bacteriology, Departement of parasitic and infectious diseases, FARAH, ULiége, Liége, Belgium.
2Independent fish farm veterinarian, Hargimont, Belgium.
3Fish Pathology Laboratory, CER groupe, Marche-en-Famenne, Belgium.
Aeromonas (A.) salmonicida, a Gram-negative bacteria belonging to the Aeromonadaceae family,

is a primary fish pathogen that causes furunculosis in salmonids, carp and perch, as well as
septicemia in a variety of fish. This species is considered as one of the main bacterial pathogens
responsible for important economic losses in aquaculture industry. Large amounts of antibiotics
such as oxytetracycline, quinolones and sulfonamides are used to treat this infection, which highly
contributes to the emergence of antibiotic-resistant strains. The application of bacteriophages
(phages) in aquaculture seems to be a promising solution to control pathogenic bacteria in this field
because these organisms are well adapted to aquatic environments. The aim of this work was to
isolate and characterize new lytic phages against A. salmonicida. The enrichment method used to
isolate phages consists in mixing a centrifuged and filtered water sample with a bacterial culture in
exponential phase. When clarification of the medium was observed, the supernatant of this mixture
was spread on the surface of LB agar and covered with a bacterial overlay in exponential phase.
Phages present in distinct clear lysis plaques were then purified three times by subculturing. For
this purpose, a sampling campaign of water from fish farming ponds and natural aquatic
environments in the south of Belgium was carried out in early 2022. Out of 48 water samples, four
new lytic phages were isolated. A preliminary host spectrum test showed that three of these four
phages were active against other wild A. salmonicida strains while the fourth one showed a narrower
host spectrum. These four phages were not active against any of the A. hydrophila strains tested.
After having determined temperature and pH stabilities, adsorption times and kinetics of these four
new phages, further studies are needed to analyse their genomes and to assess the in vivo safety
and efficacy of these phages.
PSA15
DISSEMINATED NOCARDIOSIS CAUSED BY NOCARDIA FARCINICA IN TWO PUPPY

SIBLINGS
F. Zendri, P. Richards-Rios, I. E. Maciuca, E. Ricci, D. Timofte
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Department of Veterinary Anatomy, Physiology and Pathology, Institute of Infection, Veterinary and
Ecological Sciences, Leahurst Campus, University of Liverpool, Neston, United Kingdom.
Background: Systemic nocardiosis due to Nocardia farcinica has not been reported in canine
outbreaks. Case presentation: Two 14-week-old female Dogue de Bordeaux siblings presented
with fever and severe, acute onset limb lameness; traumatic lesions with evidence of infection were
identified over the lame limbs of both dogs. The patients had to be euthanised due to poor response
to therapy and rapid escalation to systemic infection with severe central nervous system
manifestations. The post-mortem changes consisted of multiple disseminated abscesses, mainly
affecting the skin and subcutis of the wounded skin, lung, kidney and brain, with regional
suppurative lymphadenitis. Bacterial culture isolated Nocardia farcinica from several of these sites
in both dogs, whose identification was achieved via MALDI-TOF MS and confirmed by 16sDNA
sequencing. Clinical significance of the isolate was supported by cytology of the post-mortem
organs’ impression smears showing numerous branching filamentous bacteria associated with
purulent inflammation. The organism displayed marked multidrug-resistance including to
doxycycline and imipenem. No history of immunosuppression was available neither
histopathological signs of lymphopenia were detected, although further testing would be required to
rule out viral pathogens as canine distemper and parvovirus. Conclusion: N. farcinica should be
considered as a potential differential cause of sudden lameness and systemic infection in dogs with
traumatic skin lesions. This is the first reported small-scale outbreak of systemic nocardiosis in dogs
due to N. farcinica.
PSA16
CHARACTERIZATION OF FLAVOBACTERIUM PSYCHROPHILUM ISOLATES FROM CZECH

AQUACULTURE
V. Nováková, M. Palíková, A. Čížek
University of Veterinary Sciences Brno, Brno, Czech Republic
Flavobacterium psychrophilum is an important fish pathogen causing economical losses mainly in

salmonids all over the world, including the Czech Republic. Several studies have characterized F.
psychrophilum isolates from various countries, but none from the Czech Republic. Aim of this study
was to characterize isolates gained in 2012-2019 from aquaculture in the Czech Republic. Seventy
isolates from 11 farms were included in this study. Analysis of antimicrobial susceptibility to most
frequently used antibiotics in aquaculture using broth microdilution method, presence of selected
virulence factors using culture methods and serotyping using mPCR protocol was performed.
Antimicrobial susceptibility testing showed high MIC for oxolinic acid, oxytetracycline, flumequine,
enrofloxacin and increasing resistance to florfenicol, which is currently drug of choice for
flavobacteriosis treatment in many countries. By mPCR-based serotyping type 1, corresponding to
serotype Fd, was most frequently identified, less often type 2, corresponding to serotype Th, and
only sporadically types 3 and 4, corresponding to serotype FpT. No type 0 was detected. All isolates
were able to create visible biofilm, high proportion created sediment and hydrolyzed gelatin, but
only one third of isolates was positive for gliding motility. Characterization of isolates is essential for
revealing ways of introduction of the pathogen in the farms and enables to create effective protective
measures in the aquaculture. This study showed increasing resistance to antibiotics similarly as in
other countries, meaning that alternative therapy methods are necessary to be studied.
PSA17
ANTIMICROBIAL SUSCEPTIBILITY AND MULTILOCUS SEQUENCE TYPING OF

CAMPYLOBACTER JEJUNI ISOLATES FROM DOGS WITH DIARRHEA
M. Cvetnić, V. Mojčec Perko, S. Pintarić, S. Hađina, J. Habuš, I. Benvin, Z. Štritof
Department of Microbiology and Infectious Diseases with Clinic, Faculty of Veterinary Medicine,
University of Zagreb, Croatia
Campylobacteriosis is one of the most common zoonoses worldwide, with dogs being one of the
possible sources of infection. The aim of this study was to investigate the antimicrobial susceptibility
and sequence types of Campylobacter jejuni isolated from the feces of dogs with diarrhea.
Campylobacter spp. were isolated on modified charcoal-cefoperazone-deoxycholate agar
(mCCDA). Species was identified by MALDI TOF MS. Antimicrobial susceptibility of isolates was
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tested using the disk diffusion method according to the EUCAST. Multi-locus sequence typing
(MLST) was performed according to Dingle et al (2001).
From 248 dogs, 46 (18.5%) Campylobacter spp. were isolated. Of these, 26 (56.5%) were
C. jejuni. Resistance to ciprofloxacin was 57.7%, to tetracycline 23.1%, and to erythromycin 3.8%.
Five (19.2%) isolates were resistant to two antimicrobials; four to ciprofloxacin and tetracycline and
one to ciprofloxacin and erythromycin. Fifteen isolates were randomly selected for MLST and
classified into six clonal complexes (CC-21, CC-206, CC-403, CC-45, CC-443, and CC-48) and 10
sequence types; three each from ST-1943 and ST-10039, two from ST-538, and one each from ST-
3156, ST-19, ST-2086, ST-572, ST-3335, ST-475, and ST-51. Six of these STs have not been
previously detected in dogs. Six STs have not been confirmed in Croatia to date. High genetic
diversity was found among the typed isolates. Given the increasing contact of dogs and humans,
continuous epidemiological studies of campylobacteriosis and antimicrobial susceptibility of isolates
from all sources are needed.
Acknowledgement
The participation in the “4th ICECVM Conference 2022” is supported by COST 18217 European
Network for Optimization of Veterinary Antimicrobial Treatment.
PSA18
SEROPOSITIVITY OF COXIELLA BURNETII IN WILD BOAR (SUS SCROFA) AND RED DEER
(CERVUS ELAPHUS) IN PORTUGAL
H. Pires1, L. Cardoso2,3, A.P. Lopes2,3, M.C. Fontes2,3, M. Matos4, C. Pintado1, L. Figueira5,6,
J.R. Mesquita7,8,9, A.C. Matos5,6, A.C. Coelho2,3
1Polytechnic Institute of Castelo Branco, Castelo Branco, Portugal
2Animal and Veterinary Research Centre, Vila Real, Portugal, Department of Veterinary Sciences,
University of Trás-os-Montes e Alto Douro (UTAD), Portugal

4Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB),
UTAD, Portugal
5Centre Research for Natural Resources, Environment and Society, Polytechnic Institute of Castelo
Branco, Castelo Branco, Portugal,

6Researcher at Q-RURAL – Quality of Life in the Rural World, Polytechnic Institute of Castelo
Branco, Castelo Branco, Portugal

7ICBAS – School of Medicine and Biomedical Sciences, Porto University, Porto, Portugal
8Epidemiology Research Unit (EPIUnit), Instituto de Saúde Pública da Universidade do Porto, Porto,
Portugal
9Laboratório para a Investigação Integrativa e Translacional em Saúde Populacional (ITR), Porto,
Portugal
OBJECTIVES: Coxiella burnetii is a zoonotic microorganism that infects a wide range of wild and
domestic species, causing the disease Q fever, frequently involving ticks as vectors. To better
understand the occurrence of C. burnetii infection in wild boar (Sus scrofa) and red deer (Cervus
elaphus), an epidemiological study was conducted in the Centre region of Portugal. MATERIAL
AND METHODS: A serological survey was performed on samples from 377 wild boar and 240 red
deer, totalizing 617 animals from the Centre of Portugal. Only adult animals were sampled in this
study. Antibodies specific to C. burnetii were detected with a commercial enzyme-linked
immunosorbent assay (ELISA; IDVet®, Montpellier, France) according to the manufacturer
instructions.RESULTS: A total of 9/617 samples (1.5%, 95% confidence interval [CI]: 1.1-1.9%))
were reactive to C. burnetti and regarded as positive. The seropositivity for wild boar was found to
be 1.1% (4/377, 95% CI: 0.6-1.6%), and for red deer 2.1% (5/240, 95% CI: 1.5-2.7%).
CONCLUSION: Results indicate that wild boar and red deer from the Centre of Portugal are
exposed to C. burnetii. This study demonstrates that wild boar and red deer can be reservoirs of
infection for both livestock and humans.
PSA19
USEFULNESS OF MALDI-TOF MASS SPECTROMETRY IN THE DIAGNOSIS OF BOVINE

MASTITIS
A. Bellato, P. Robino, M. Furlan, F. Traverso, C. Pegorin, A. Mannelli and P. Nebbia
Department of Veterinary Sciences, University of Turin, Italy
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In udder microbiology, MALDI-TOF MS method could become the reference method due to its ability
to identify bacteria at the species level faster and at a lower cost than classical methods. Accurate
and fast diagnosis can result in a timely and better treatment plan. Information about the strains
involved is of crucial importance due to the difficulty in treating the disease, whether clinical or
subclinical. The aim of this work was to explore the use of the MALDI-TOF for the diagnosis of
subclinical mastitis and providing more information regarding the differences among the bacterial
strains circulating in the herd. The study involved 256 clinically healthy cows; milk samples were
cultured on conventional media and a colony representing each population was analysed, obtaining
a high-quality mass spectrum. Peak-intensity matrices were produced from mass spectra analysis,
thus dendrograms were obtained using R coding language to measure the dissimilarities among
strains of the same species. Our findings confirmed that the MALDI-TOF MS method easily
identifies the pathogens usually involved in subclinical mastitis. The cluster analysis, despite having
limitations in generalizability due to the small sample size and the lack of standardisation, can be
useful in differentiating bacterial strains between different herds. Differences in the reliability of
cluster analysis are likely linked to the bacterial species. Future developments concerning the
diagnosis of mastitis with the MALDI-TOF method should be the implementation of the mass-
spectra database in order to improve the accuracy of the method, and the possible identification of
additional species. Key words: subclinical mastitis, MALDI-TOF, cluster analysis
PSA20
LOAD OF CAMPYLOBACTER SPP. IN THE ENVIRONMENT OF POULTRY FARMS IN

GERMANY
B. Reichelt, V. Szott, U. Roesler, A. Friese
Institute for Animal Hygiene and Environmental Health, Freie Universität Berlin, Berlin, Germany
Introduction:Campylobacter (C.) jejuni is the most common cause of campylobacteriosis in humans,

and broiler meat is considered one of the major sources. The poultry house environment may be a
reservoir for Campylobacter spp. to some extent. However, to date, there are limited data on the
spillover of Campylobacter from houses with Campylobacter-positive flocks and the buildup of
environmental reservoirs. In addition, Campylobacter spp. are capable to transit into a viable but
non-culturable (VBNC) state as a result of various extrinsic stress factors. In order to identify
possible reservoirs of persistent and VBNC-Campylobacter and thus uncover relevant transmission
pathways, a longitudinal study was conducted. Material/Methods: Three Broiler farms and their
environment close to the barn were intensively investigated at the end of two consecutive fattening
cycles in summer and winter over three years. The selected farms were also examined after
cleaning and disinfection. All samples were processed according to the semi-quantitative method
for the detection and enumeration of Campylobacter spp. (ISO/TS 10272-3). A systematic selection
of isolates from all sampling collections was examined by whole genome analyses. Moreover,
environmental and selected broiler house samples were treated with propidium monoazide (PMA)
and analyzed by live/dead discrimination using real-time PCR (qPCR). Results: In two out of three
farms, Campylobacter was frequently detected in high amounts in the chicken barns, especially in
summer. However, the pathogen was only occasionally detectable in the environment, particularly
in water-associated matrices, especially in winter. However, Campylobacter could not be isolated
in broiler houses after cleaning and disinfection. The emission source of culturable Campylobacter
was found to be primarily contaminated chicken manure. C. jejuni proved to be the dominant species
of the isolates examined. PMA-qPCR revealed no detection of VBNC-Campylobacter in selected
barn and environmental samples. In contrast, Campylobacter DNA was more frequent detected in
environmental samples. Discussion: The present study provides insight into the significance of
Campylobacter in the environment in relation to prevalence in the broiler farms investigated in
Germany. The results established indicate sporadic environmental findings in the immediate vicinity,
suggesting spread, persistence and possible reintroduction. C. jejuni was found in nearby water
bodies, indicating that the pathogen is ubiquitous by spread and circulation. Although the findings
were sporadic and no significant source of transmission has yet been identified, it should be kept in
mind that even very low levels of Campylobacter may initiate the colonization of whole poultry flocks.
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PSA21
VIRULENCE CHARACTERIZATION OF PSEUDOMONAS AERUGINOSA FROM CANINE

OTITIS
R. Abreu1,2, F. Monteiro, E. Cunha1,2, M. Lourenço, L. Tavares1,2, M. Oliveira1,2

2Laboratório Associado para Ciência Animal e Veterinária (AL4AnimalS), Portugal
Background: Otitis externa is a recurrent problem in small animal general practice. Despite of
being a multifactorial disorder, secondary infections are common. Pseudomonas aeruginosa is one
of the most frequent gram-negative pathogens involved in infectious otitis. These ubiquitous
bacteria are found in water and decaying matter, but when present in the ear canal they can cause
disease. It presents multiple intrinsic resistances and produces multiple virulence factors; thus, its
treatment can be challenging. This work aimed to identify and characterize the virulence profile of
P. aeruginosa obtained from the ear canal of dogs with otitis externa. Materials/methods: A total
of 48 P. aeruginosa isolates from the collection of clinical isolates from the Laboratory of
Bacteriology, Faculty of Veterinary Medicine, University of Lisbon, Portugal (2016-2021) were
analyzed, by evaluating various phenotypic virulence traits (biolfim and haemolysins production,
lipase, lecithinase, DNAse protease and gelatinase activity) using specific media. The virulence
index (V. Index) values were determined for all isolates. Results: All isolates produced lipase and
haemolysins (100%), 20 isolates (41.67%) produced gelatinase, 26 isolates produced lecithinase
(54.17%), 46 isolates produced protease (95.83%), none of the isolates produced DNAse (0%) and
6 were able to produce biofilm (7.69%). P. aeruginosa V. Index ranged between 0.28 to 0.86, with
a 0.58 media and a 0.57 median. Conclusions: P. aeruginosa isolates tested presented a high
virulent index, demonstrating the importance of adequate treatment for related infections, avoiding
their recurrence and reducing the harmful effects on the health and welfare of the patient.
Ackowledgements: This work was supported by CIISA–Centro de Investigação Interdisciplinar em
Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Project
UIDB/00276/2020 (Funded by FCT) and by Laboratório Associado para Ciência Animal e
Veterinária (LA/P/0059/2020 - AL4AnimalS).
PSA22
DIETZIA SP. B32 INFECTION IN A HORSE: FIRST GENETIC CHARACTERIZATION

A.-R. Attili1, R. Gonçalves dos Santos2, R. Hurtado Castillo2, D.L. Neres Rodrigues2,
A. Lima2, W. Ferreira dos Anjos3, C. Rifici4, S. Tiwari2, A. Kumar Jaiswal2, T.L. de Paula
Castro2,5, N. Seyffert2,6, A. Santos3, A. Góes-Neto7, T. de Jesus Sousa2, V. Azevedo2
1School of Biosciences and Veterinary Medicine, University of Camerino, Matelica Italy
2Cellular and Molecular Genetics Laboratory, Institute of Biological Sciences, Federal
University of Minas Gerais, Belo Horizonte, MG, Brazil

3Department of Computer Science, Federal University of Uberlandia, Uberlandia, Brazil
4Department of Veterinary Science, University of Messina, Messina, Italy
5Institute of Health Sciences, Federal University of Bahia, Salvador, BA, Brazil
6Institute of Biology, Federal University of Bahia, Salvador, BA, Brazil
7Molecular and Computational Biology of Fungi Laboratory Department of Microbiology, Institute of
Biological Sciences, Federal University of Minas Gerais, Brazil
Environmental and opportunistic Dietzia species belong to the Dietziaceae family, order
Mycobacteriales, class Actinobacteria. In the absence of accurate techniques, Dietzia spp. could
be misidentified as Rhodococcus hoagii, formerly Rhodococcus equi. Objectives: In order to study
the complete genome sequence of Dietzia strain isolated from diffuse subcutaneous nodules of a
mare in Reggio Calabria (Italy), comparative and phylogenetic analyses, searches for antibiotic
resistance genes, virulence factors, and genomic islands were performed. Methods: Analyses were
performed for 49 genomes of Dietzia species (NCBI), and one genome was sequenced using Hiseq
(Illumina), Dietzia sp. isolate. All genomes were homogenized by Prokka software. Phylogenomic
trees, presence of antibiotic resistance and virulence genes were predicted by MEGAX software,
CARD and VFDB databases, respectively. Images were created using iTOL. Islands of
pathogenicity (PAIs), resistance (RIs), metabolic (MIs) and symbiotic islands (SIs) were predicted
using GIPSy software.
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Results: Dietzia sp. B32 resulted as a new species not yet described because of similarity values
below 90.1% and 41 unique genes involved in “cellular component” and “molecular function. Four
genes (rpoB, mtrA, rbpA, and APH(3’)-Iia) related to the antibiotic resistance, and six genes
associated with virulence factors (phoP, phoR, icl, tufA, relA, and cap8E) were described. The
mutation in the rpoB gene confers resistance to rifampicin. Five pathogenicity (PAIs), five resistance
(RIs), two metabolic (MIs), and one symbiotic island (SI), were found. Conclusions: As the true
pathogenic potential of Dietzia species is not known, the comparative analysis of Dietzia B32
contributes to better understand this increasingly emerging genus.
PSA24
RESULTS OF URINARY BACTERIAL CULTURE AND ANTIMICROBIAL SUSCEPTIBILITY

TESTING OF DOGS AND CATS FROM A VETERINARY LABORATORY IN PORTUGAL IN 2021
V. Campos da Silva, H. Carvalho, N. Carvalho
Cedivet – Centro Diagnóstico Veterinário, Oporto, Portugal
Objectives: The aim of this study was to investigate the aetiology and antimicrobial resistance of
pathogens isolated from urinary samples of dogs and cats submitted to analysis in a private
veterinary clinical laboratory in Portugal. Materials and methods: Data from urinary samples,
submitted as part of a urinary profile investigation, between January and December of 2021 were
used (n=4001). Results: A total of 601 samples showed significant bacteraemia (>100,000 UFC/mL)
and 61,1% of urine samples from dogs and 38,9% from cats had significant cultural growth. E. coli
was the most frequently isolated microorganism in both dogs (49,7%) and cats (49,3%), followed
by Proteus mirabilis in dogs (23,3%) and coagulase-negative staphylococci in cats (11,3%).
Amoxicilin-clavulanic acid was the drug with the lowest percentage of resistance (11,4%),
fluoroquinolones showed levels of resistance above 10% (enrofloxacin 16,3%, marbofloxacin
16,1%, pradofloxacin 15,0%) and trimethoprim-sulfamethoxazole showed a total level of resistance
of 20,8%. Conclusions: Our work confirms E. coli as the most common cause of bacterial urinary
tract infections in both dogs and cats, provides evidence of high levels of resistance to
fluoroquinoles and enhances the need of surveillance of antimicrobial resistance in the
establishment of guidelines at a national level.
PSA25
PRELIMINARY EVALUATION OF THE EFFICACY OF A NEW INACTIVATED ORAL VACCINE

AGAINST ENTEROTOXIGENIC E. COLI INFECTIONS IN POST-WEANING PIGLETS
A. Inglesi1, G.A. Clemente2, S. Draghi1, J. Filipe1, A. Maisano3, F. Riva1, M. Amadori2, F.
Guarneri2, V. Lorenzi2, C. Montagnin2, L. Bertocchi2, F. Fusi2
1Department of Veterinary Medicine and Animal Sciences, University of Milan, Lodi, Italy
2Reparto produzione e controllo materiale biologico, Istituto Zooprofilattico Sperimentale della
Lombardia e dell’Emilia Romagna "Bruno Ubertini", Brescia, Italy

3Animal Health and Welfare, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia
Romagna 'Bruno Ubertini', Lodi, Italy
Weaning is a highly stressful period for piglets, which become more susceptible to diverse diseases.
At the intestinal level, weaned piglets show severe inflammatory responses that impair the gut
epithelial barrier and predispose them to pathogen colonization and invasion. In post-weaning
piglets several E. coli strains cause diarrhea leading to the reduction of production performances
and the increased use of drugs. In order to limit and mitigate the impact of E. coli infections in post-
weaning piglets, we developed a new oral vaccine to be easily administrated to the animals via
drinking water. The vaccine is based on heat-inactivated enterotoxigenic E. coli strains expressing
F4 and F18 fimbriae for 26 days. The efficacy of our vaccine has been preliminarily evaluated in
terms of mucosal anti-F4 and anti-F18 IgA production in the saliva (IgAF4sal and IgAF18sal) by
ELISA analysis and the number of IgA-secreting cells in mesenteric lymph nodes (IgAcell) by
ELISPOT assay. The level of IgAF4sal was significantly higher in vaccinated piglets compared to
the unvaccinated control ones, whereas no differences were observed for IgAF18sal. Based on the
increment of IgAF4sal production the vaccinated animals were classified as low, medium and high
responders. Interestingly, only high responders showed a significant increase in the number of
IgAcells. These encouraging preliminary results suggest a possible efficacy of our new vaccine and
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highlight diverging immune responses to different E. coli fimbrial antigens. This work was supported
by the Italian Ministry of Health: Grant n. PRC2019001.
PSA26
VIABLE MICROBIAL COMMUNITY COMPOSITION OF HEALTHY AND DAMAGED SKIN IN

STRAY DOGS AND CATS.
F. Paola Nocera1, C. Di Palma1, S. Cappiello2, A. Masullo1, M. Pompameo2, B. Lamagna1, Luisa
De Martino1,3
1Department of Veterinary Medicine and Animal Production, University of Naples “Federico II”, Via
F. Delpino 1, 80137 Naples, Italy

2Veterinary Hospital, ASL Napoli 1 Centro, via M. Rocco di Torrepadula 13, 80145 Naples, Italy
3Task Force on Microbiome Studies, University of Naples “Federico II”, Naples, Italy
Objectives: The purpose of this report was to describe the microbial community composition
regarding three sites of healthy skin and the site of traumatic wounds in stray dogs and cats, and to
verify the correlation between the bacteria resident on healthy skin and those of the damage skin
of the same animal Materials and methods: Skin swabs were collected from stray dogs and cats
presenting traumatic wounds. Precisely, swabs were performed on injured skin; furthermore, a
collection of nasal and ear swabs as samples of skin cavities, and nasal spine swab as sample of
external skin, representing samples of healthy skin sites, were performed for each animal. Samples
were plated onto different types of solid culture agar media and incubated aerobically at 37°C for
24-48 h. Once bacterial growth was detected, the identification was performed by MALDI-TOF-MS
Results: Stray cats presented a larger bacterial diversity in skin samples than stray dogs.
Staphylococcus pseudintermedius was the most frequently identified bacterium in stray dogs both
from damaged and healthy skin samples, whereas in stray cats S. pseudintermedius represented
the third most frequently isolated bacterial species. However, in both animals S. pseudintermedius
isolation was significantly higher in damaged skin samples than healthy ones (P < 0.05). In stray
cats the Gram-negative E. coli resulted to be the most common identified bacterium, and its
presence was significantly higher in damaged skin samples than healthy ones (P < 0.05)
Conclusions: According to this study, traumatic wounds provide an opportunity for microorganisms
from the skin microbiota, to enter the underlying tissues and find the optimal conditions for their
overgrowth
PSA27
DITHIOTHREITOL PROTECTION ON MICROBICIDAL ACTIVITY OF INNOVATIVE LIGHTS: AN

IN VITRO STUDY
M. Ambrosio1,2, F. P. Nocera1, C. Cerracchio1, F. Fiorito1, M. Hohenegger2, L. De Martino1,3
1Department of Veterinary Medicine and Animal Production, Infectious Diseases Unit, University of
Naples “Federico II”, Naples, Italy

2Institute of Pharmacology, University of Medicine, Vienna, Austria
Objectives: Innovative mechanisms using the microbicidal photodynamic effect of light energy are
used in bacterial eradication strategies. The light is emitted in the visible light spectrum (VIS) and
is safe for mammalian cells. When microorganisms are exposed to light, excitation of porphyrins,
photosensitizing chromophores endogenous to cells and sensitive to VIS light, is created. The
resulting excitation leads to the production of reactive oxygen species (ROS), which completely
damage the cell. We tested the Biovitae ® light, which emits white light from a light-emitting diode
(LED) and investigated the effect of a sulfhydryl compound, dithiothreitol (DTT), a protective reagent
against ROS, on lytic activity of the lights. Materials and Methods: The lights (by company
Nextense) have a special combination of frequencies and intensities: 400-420 nm, 400-450 nm,
400-700 nm. 96-well plates containing Escherichia coli (E. coli) and Staphylococcus aureus (S.
aureus) were exposed for six hours, and bacterial growth was monitored over 24 hours by optical
density (OD) measurement. Bacterial strains were also exposed to light in combination with DTT.
Results: During the six hours light exposure, a slight increase in growth was observed for both
bacteria strains. Bacterial growth was significantly (P ≤0.005) decreased 18 hours after the end of
exposure. DTT significantly protected (P ≤0.05) both E. coli and S. aureus, when exposed to light.
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Discussion: These results confirmed the microbicidal photodynamic effect of Biovitae ® light
against bacteria. Since DTT protected bacterial strains from antimicrobial light effects, these results
are indicative for a redox-sensitive mechanism of action.
PSA28
LEPTOSPIRA INTERROGANS SEROGROUP SEJROE SEROVAR HARDJO IN A DAIRY

CATTLE FARM IN SOUTH OF ITALY (SICILY)
G. Macaluso1, V. Blanda1, R. D’Agostino1, C. Sciacca1, F. Arcuri1, I. Giacchino1, A.
Stancanelli2, A. Torina1, F. Grippi1
1Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Palermo, Italy
2Istituto Zooprofilattico Sperimentale della Sicilia, Caltanissetta, Italy
Bovine leptospirosis is a zoonoses causing economic losses in livestock. This work reports an
outbreak of Leptospira interrogans serogroup Sejroe serovar Hardjo in a officially free from
Brucellosis, Tuberculosis and Enzootic Bovine Leucosis cattle farm in province of Enna (Sicily). In
February 2022, 2 cows showing full-term abortion from three months without placental retention,
were sampled at three times: on day 0 (T0), eight days after the first abortion signs, on day 30 (T1)
and on day 60 (T2), after antibiotic treatment. At T1 and T2 42 other animals were sampled. Blood,
milk and stool samples were also collected at T2. Research of antibodies against pathogenic
Leptospira species was carried out by Micro Agglutination Test (MAT); ELISA against the main
abortion agents (Chlamydia abortus, Coxiella burnetii, Neospora caninum were also performed.
Blood, milk and drinking water samples were subjected to Real Time-PCR to detect Leptospira
pathogenic species and the main abortion agents (Chlamydia abortus, Coxiella burnetii, Neospora
caninum, Coronavirus, Cryptosporidium spp., Escherichia coli K99, Rotavirus). No animals resulted
positive at differential diagnosis except for two ones seropositive for N. canimun and C. burnetii,
without showing signs of abortion. At T0 one of the two sera resulted positive at MAT. Thirty and
twenty-five sera resulted MAT positive at T1 and T2 respectively. This study describes clinical
manifestations, diagnostic implications and epidemiological characteristics of an outbreak in cattle
due to L. interrogans serogroup Sejroe serovar Hardjo, confirming that L. interrogans plays a role
in determining leptospirosis infection in cattle reared in Sicily
PSA29
BOVINE TUBERCULOSIS (MYCOBACTERIUM BOVIS/MYCOBACTERIUM CAPRAE) IN

SLOVENIA: A REVIEW 2015-2021
T. Pirš, U. Zajc, M. Ocepek
University of Ljubljana, Veterinary Faculty, National Veterinary Institute, Gerbičeva 60, SI-1115
Ljubljana, Slovenia
Slovenia has been officially free of bovine tuberculosis (bTB) since 2009. However, the country
achieved less than 0.1% infected cattle herds for six consecutive years as early as the mid-1970s.
Since 2009, routine tuberculin testing in line with the relevant EU legislation for maintaining officially
free status was conducted. In the last 20 years, only four outbreaks of bTB in cattle have been
confirmed in Slovenia. Most of the animals were traded from other EU member states, only in one
case we couldn’t determine the origin of the animal involved. From 2015 to 2021, a total of 1098
samples from cattle were submitted for bacteriological testing. In 2015, 14 cases caused by M.
caprae were detected in a single herd of fattening bulls. M. caprae was isolated from suspicious
lesions of the lungs and adjacent lymph nodes, detected at regular slaughter. In 2018, another case
was confirmed following the postmortem examination of an intradermal tuberculin test reactor. In
addition to regular surveillance of cattle, comprehensive disease control also requires surveillance
of wildlife species. From 2015 to 2021, 1860 samples of wildlife were analysed as part of annual
state programmes, most of which were from wild boar (1300 samples from 625 animals) and deer
(411 samples from 211 animals). The presence of M. caprae was comfirmed in one red deer
(Cervus elaphus) in 2018 and the presence of M. bovis was confirmed in one wild boar (Sus scrofa)
in 2019. Data were obtained from regular disease surveillance programme financed by the
Administration of the Republic of Slovenia for Food Safety, Veterinary Sector and Plant Protection.
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PSA30
SUBCUTANEOUS NOCARDIOSIS IN A 3-YEAR-OLD FEMALE CAT IN ITALY: A CASE

REPORT
M. Medardo¹, F. Clemente², M. Marino¹, C. Lisi¹, G. Cocciolo¹, P. A. Martino3, N. Decaro4
¹Laboratorio di Analisi Veterinarie MYLAV, Passirana di Rho, Italy
²Ambulatorio Veterinario San Luca, Bologna, Italy.
³One Health Unit - Department of Biomedical, Surgical and Dental Sciences, University of Milan,
Milan, Italy.
4Department of Veterinary Medicine, University of Bari, Valenzano, Italy.
Introduction: Nocardia spp. includes worldwide, ubiquitous zoonotic bacteria that can cause
suppurative to pyogranulomatous infections, with localized or disseminated lesions. Nocardia spp.
is considered an opportunistic pathogen, with the majority of clinical cases occurring in
immunocompromised hosts. Case report information: A 3-year-old European cat presented with
severe pododermatitis with fistulas in the left limb. Intervention/Treatment: An immunosuppressive
therapy was administered after two tentative histopathological diagnoses of pemphigus foliaceus.
After initial regression, lesions became more severe in the following 15 days, so that surgical
curettage and other biopsies for histology and culture were carried out. Results: Histopathology
was suggestive of severe pyogranulomatous dermatitis; microbiological culture showed positive
results with presumptive diagnosis of Nocardia spp. infection, which was confirmed by modified
Ziehl-Neelsen staining and Nocardia genus-specific PCR. Drug sensitivity was determined by the
Kirby-Bauer disk-diffusion method and the isolate was susceptible to amikacin, doxycycline,
trimethoprim-sulfamethoxazole and chinolones. Immunosuppressive therapy was discontinued and
antibiotic therapy with doxycycline and enrofloxacin was administered. Follow up in July 2022
showed a remarkable improvement of the clinical conditions.Discussion and conclusion: This
case report aims to highlight the importance of a multi-level steps for correct diagnosis and
tempestive treatment of feline nocardiosis, which should be regarded as a potential zoonosis.
PSA31
PHAGE-MEDIATED SHIGA-TOXIN (STX2D) GENE TRANSDUCTION FROM O80:H2 SHIGA

TOXIGENIC ESCHERICHIA COLI (STEC) TO NON-STEC STRAINS AND IN VIVO VIRULENCE
ASSESSMENT
A. Habets1, C. Antoine1, J. Wagemans2, M. Vermeersch3, F. Laforêt1, R. Lavigne2, J. Mainil1,
D. Thiry1
1Bacteriology, Department of Parasitic and Infectious Diseases, FARAH, ULiège, Liège, 4000,
Belgium
2Department of Biosystems, Laboratory of Gene Technology, KU Leuven, Leuven, 3000, Belgium
3Center for Microscopy and Molecular Imaging, Electron microscopy laboratory, Gosselies, ULB,
6041, Belgium
Shiga toxin-producing Escherichia coli (STEC) are major foodborne pathogens that cause human
diseases ranging from diarrhea to life-threatening complications including hemolytic–uremic
syndrome (HUS). Virulence of STEC strains and their ability to cause severe diseases are
associated with the activity of prophage-encoded Shiga toxins (Stxs). The first objective of this work
was to isolate and characterize the Stx2d phage from STEC O80:H2 and study the transfer of this
phage in non-STEC strains. The second objective was to assess the survival of Galleria mellonella
larvae inoculated with these transduced trains. One bacteriophage isolated from a STEC O80:H2
strain was used to infect five non-STEC strains. Three strains (K12-MG1655, K12-DH5α and
O80:H26) were successfully converted. The genome of the phage was analyzed. A stability study
was performed and showed that this phage was stable in the new STEC strains after three
successive subculturing steps. This phage (vB_EcoS_ULI-O80_Stx2d) presents resistance to high
temperature (60°C) and low pH (2-8). The phage belongs to the Caudoviricetes class (unclassified
genus and family) and to the siphovirus morphology. It encodes several proteins involved in the
lysogenic cycle. Galleria mellonella experiments showed that mutant strains caused significantly
higher mortality rates than the corresponding non-STEC strains. In conclusion, this study showed
that stx2d gene from O80:H2 E. coli can be transferred to non-STEC strains and contributes to their
virulence. This research was funded by the University of Liège (project HYBRID_COLI_O80)
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PSA32
IN VITRO CHARACTERIZATION AND ASSESSMENT IN GALLERIA MELLONELLA OF NEWLY

ISOLATED PHAGES AGAINST ESCHERICHIA COLI K1
C. Antoine1,2, F. Laforêt1,2, B. Blasdel3, A. Fall4, J.N. Duprez1, J. Mainil1, V. Delcenserie2,
D. Thiry1
1Department of Infectious and Parasitic Diseases, Bacteriology and Pathology of Bacterial
Diseases, FARAH, ULiège.Belgium

2Department of Food Science, Quality management in the food chain, FARAH, ULiège.Belgium
3Vésale Bioscience, 5310 Noville sur Mehaigne, Belgium
4Genalyse Partner SA, En Hayeneux 62, 4040 Herstal, Belgium
Extra-intestinal Escherichia coli express several virulence factors that increase their ability to
colonize and survive in different localizations. The K1 capsular type is involved in several infections,
including meningitis, urinary tract, and bloodstream infections. The aims of this work were to isolate,
characterize, and assess the in vivo efficacy of phages targeting avian pathogenic E. coli (APEC)
O18:K1, which shares many similarities with the human strains responsible for neonatal meningitis.
Eleven phages were isolated against APEC O18:K1, and four of them presenting a narrow spectrum
targeting E. coli K1 strains were further studied. The newly isolated phages vB_EcoS_K1-ULINTec2
were similar to the Guernseyvirinae subfamily, and vB_EcoP_K1-ULINTec4, vB_EcoP_K1-
ULINTec6, and vB_EcoP_K1-ULINTec7 to the Autographiviridae family. They are capsular type
(K1) dependent and present several advantages characteristic of lytic phages, such as a short
adsorption time and latent period. vB_EcoP_K1-ULINTec7 is able to target both K1 and K5 strains.
This study shows that these phages replicate efficiently, both in vitro and in vivo in the Galleria
mellonella model. Phage treatment increases the larvae survival rates, even though none of the
phages were able to eliminate the bacterial load.This research was funded by the Walloon Public
Service, BIOWIN project: Inteliphages
PSA33
THE EFFECTIVENESS OF THYME VULGARIS ESSENTIAL OIL ON ANTIBIOTIC RESISTANCE

BACTERIA ISOLATED FROM POULTRY LITTER
M. Galgano1, F. Pellegrini1, G. Fracchiolla2, D. Mrenoshki1, A. Attia Koraney Zarea1,3, A.
Bianco 4, L. Del Sambro4, L. Pozzi4, A. Schiavone1, A. Assan Omar1, A. Camarda1, F. Cirone1,
M. Tempesta1, D. Buonavoglia1, A. Pratelli1
1Department of Veterinary Medicine, University Aldo Moro of Bari, Sp Casamassima Km 3, 70010
Valenzano (Ba), Italy

2Department of Pharmacy-Drug Sciences, University Aldo Moro of Bari, Via Orabona 4, 70125, Bari,
Italy.
3Department of Microbiology and Immunology, Veterinary Research Institute, National Research
Centre (NRC), Egypt

4Istituto Zooprofilattico della Puglia e della Basilicata, Contrada San Pietro Piturno, 70017,
Putignano (Ba), Italy
The development and spread of antibiotic-resistanceamong bacteria is a major cause for concern
worldwide. The indiscriminate use of antimicrobial agents in poultry farms is linked to the increase
of multi-resistant bacteria and their spread in the environment. Furthermore, the presence of
antimicrobial residues in poultry products is also documented. Accordingly, this study aims to
identify bacteria in chicken litter, test their resistance to antibiotics and evaluate the antimicrobial
efficacy of Thyme Essential Oil (TEO). Litter sample bulk was collected from a broiler farm and was
subjected to qualitative and quantitative investigations of microorganisms. The litter was treated
with aqueous solutions of TEO at different concentrations (5% to 1.25%). The pathogens detected,
Lactose positive Enterobacteriaceae and Mannitol salt-positive Staphylococcaceae, was identified
as Escherichia Coli and Mammaliicoccus lentus, respectively. These pathogens showed phenotypic
and genotipic resistance to different class of antibiotics (b-lactams, macrolids, glycopeptides,
lincosamids, polimixine).The results showed that TEO is able to reduce the total number of bacteria,
expressed as % reduction of log10CFU/g of litter, from 76.63% to 54.85% based on the concentration
of use. In particular, the strongest antibacterial action was observed against Enterobacteriaceae
with a reduction, at the lowest concentration, of 73.3% compared to the positive control, while
Staphylococcaceae showed a reduction of 50%. In the One Health perspective, the antibacterial
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action of TEO against multidrug-resistant bacteria and the absence of any oil residues, unlike what
happens with antibiotics and disinfectants, represent fundamental advantages for encouraging and
supporting the use of oils as antibacterialin animal husbandry.
PSA34
ANTIMICROBIAL RESISTANCE ANALYSIS OF ACINETOBACTER SPP. STRAINS ISOLATED

FROM A DOMESTIC CAT DURING ROUTENELY VETERINARY ACTIVITY ON ANIMAL
CARCASSES
D. Vicari, A. Carrozzo, M. La Giglia, F. Mira, V. Randazzo, G. Schirò, F. Antoci, M. Vitale
Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Palermo, Italy
During animal carcasses inspection to establish the cause of death, microbiology analyses are
routinely performed from different organs in search of pathogenic bacteria. During this activity,
strains of Acinetobacter spp. were isolated from different organs of the carcass of a domestic female
Bengala cat. Before death, the cat had been treated with antibiotic and prostaglandine following an
abortion. After the treatment however, a symptomatology of restlessness and convulsions began
which within 6 days determine the death. Necroscopic examination revealed the presence of
necrotized material inside the uterine horns and pulmonary oedema. The microbiological analysis
showed the isolation of Acinetobacter spp. from the lung, liver, brain, intestine, heart. The
identification of strains was performed through Api 20 Gallery (Biomerieux) and antibiogram by Kirby
Bauer method was performed to establish antimicrobial resistance and multidrug resistance was
detected toward ampicillin, cephuroxime, aztreonam, cephixime and trimethoprim. The presence of
a multidrug resistant strain of Acinetobacter spp. in a pet is a serious concern for human health also
and highlight the importance of one health approach to manage antimicrobial resistance (AMR)
avoiding its spreading. The cat was hospitalized in a veterinary clinic, with the risk of MDR strains
diffusion for health operator or other animals. The strain was resistant also to third generation
cephalosporin. The bacteria has been identified as Acinetobacter baumanni but since API gallery is
not valid for the correct identification at specie level, further molecular analysis will be performed for
the fully characterization.
PSA35
COMBATING CAMPYLOBACTER IN THE DUTCH BROILER MEAT PRODUCTION – A JOINT

VENTURE
M. van der Most1, E. Pacholewicz1, M.A. Dame-Korevaar1, J. Schreuder2, M.L. den Hartog3, J.
Visser4, M.G.J. Koene1
1Wageningen Bioveterinary Research, Lelystad, The Netherlands
2Wageningen Livestock Research, Wageningen, The Netherlands
3NEPLUVI, Veenendaal, The Netherlands
4AVINED, Nieuwegein, The Netherlands
Campylobacter is the most reported foodborne pathogen in the European Union and chickens are
the main reservoir for human campylobacteriosis1. This emphasizes the need for enhanced
Campylobacter control at various stages of poultry meat production. In the Netherlands, shared
responsibility of managing food safety led to a public-private partnership (PPP) between
government, private stakeholders and research institutes to reduce Campylobacter prevalence in
broiler meat production. In a PPP all parties show commitment to an applied research project, either
financially or by an in-kind contribution. Since 2015, the Ministry of Agriculture, Nature and Food
Quality, the Ministry of Health, Welfare and Sport, the Dutch Association of Poultry Processing
Industries and representatives of the primary broiler production sector are working together with
research institutes towards reducing Campylobacter at farm level and in slaughterhouses. After
almost 8 years filled with a wide variety of studies, it’s time to make up the balance. Valuable
achievements of the PPP are research projects on intervention strategies like vaccination and
biosecurity measures, maternal immunity and early detection methods. Leads for further research,
for example towards differences between conventionally and alternative production systems, were
provided by a multi-year monitoring study into Campylobacter presence on Dutch broiler farms and
associated risk factors. Whether it’s attributable to the PPP or not, with 43,1% in 2015 and 27,9%
in 2021, monitoring on Dutch slaughterhouses shows a decreasing trend in the percentage of
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Campylobacter positive flocks2. Although consistent effort is needed, slow and steady might win the
race!
References
1EFSA, 2020. Update and review of control options for Campylobacter in broilers at primary
production. EFSA Journal 18, e06090.

2 NEPLUVI, Monitoring report 2021,
https://www.nepluvi.nl/dynamic/media/1/documents/Campylobacter/2022-
037_eindrapportage_campylobactermonitoring_2021_NL_vleeskuikenslachterijen.pdf
PSA36
VIRULENCE PROFILES OF KLEBSIELLA SPP. FROM DIFFERENT SMALL ANIMAL

INFECTION SITES
L. Fernandes1,2,3, J. M. da Silva1,2, J. Menezes1,2, C. Marques1,2,5, C. Caneiras6,7 and C.
Pomba1,2,3
1CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine,

2AL4AnimalS - Associate Laboratory for Animal and Veterinary Sciences, Portugal
3Molecular Veterinary Diagnostic Laboratory - Genevet, Rua Quinta da Nora Loja 3B, Carnaxide,
Portugal
5Faculty of Veterinary Medicine, Lusófona University, Lisbon, Portugal
6Laboratory of Microbiology Research in Environmental Health (EnviHealthMicro Lab), Institute of
Environmental Health (ISAMB), Faculty of Medicine, University of Lisbon, Lisbon, Portugal

7Institute of Preventive Medicine and Public Health, Faculty of Medicine, University of Lisbon,
Lisbon, Portugal
Objectives: To characterize antimicrobial-resistance and virulence of Klebsiella spp. isolated from

different small animal infection sites. Materials and Methods: During 2020, sixty-two Klebsiella
spp. isolates were obtained from clinical samples from small animals and categorized according to
infection site. Antimicrobial susceptibility testing followed the EUCAST guidelines. Beta-lactamase
encoding genes were characterized by PCR and sequencing. Virulence phenotyping included the
string test and microplate crystal violet biofilm quantitation. Virulence genotyping screening by PCR
included 15 virulence encoding genes. Results: Sixty-one isolates were multidrug-resistant, and
most strains were from urinary tract infection (UTI)(n=31/62). Forty-five isolates carried the blaCTX-
M-15 gene (n=45/62), three the blaKPC-3 and one the blaOXA-181. Two Klebsiella pneumoniae blaCTX-M-
15 carriers were positive on the string test but rmpA negative. The more frequent virulence genes
were fimH-1, entB and mrkD (85.5%, 88.7% and 79.0%, respectively), followed by kpn (53.2%),
ycfM (41.9%), kfu (40.3%) and traT (38.7%). allS was absent. The Yersiniabactin High-
Pathogenicity Island (YHPI) was present in 22.6% of the isolates. No apparent relationship between
infection site and biofilm production was found. Nevertheless, 64.5% of the isolates were at least
moderate biofilm producers, of which a slightly higher proportion of non-UTI isolates were moderate-
to-strong producers compared to UTI isolates (35.5% versus 29%). Conclusions: Klebsiella spp.
from different types of small animal infections can harbor virulence genes associated with increased
invasiveness, such as YHPI, and be multidrug-resistant, constituting an emerging problem in small
animal practice. The two K. pneumoniae showing a hypervirulent suggestive phenotype (positive
string test) require further studies, highlighting the need for continuous surveilling of this serious
phenotype.
PSA37
PREDATORY ATTEMPTS ON BATS: INJURY FREQUENCIES, BACTERIOLOGICAL

PATTERNS OF WOUNDS AND ESCAPE ABILITY OF BATS
Ciocanau M.A., Danes D.
University of Agronomic Sciences and Veterinary Medicine, Bucharest Romania
The interactions between bats and their predators are difficult to observe, one of the factors that are
restricting bats to their nocturnal activity being exactly the predation risk exposure. However,
predator-inflicted wounds can offer insights into the predation risk faced by a range of bat taxa.
During 4 years of observation, we investigated the predatory attempts on active bats and hibernating
bats. Furthermore, we investigated the ecological connectivity and habitat use of adults and juvenile
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bats, to estimate their escape ability. Bite-related wounds during the hibernation period were more
frequent on bats nesting in wild areas, in caves with large entrances and less frequent in those
inhabiting urban areas. In contrast, during the active period, predatory bite-injuries were more
frequently observed in bats inhabiting in urban areas rather than those from wild areas. The escape
ability is a factor related with ecological connectivity, with the habitat and roosting activity during the
hibernation and during the active period. The bite injuries were contaminated with specific oral
microbiota members of the predator or with unusual polymicrobial association of oral flora, skin/fur
microbiom and environmental microorganisms. The bacteria present were quite diverse, counting
from 4 to 13 genra isolated. The most common cultural isolates reveal: Staphylococcus sp.,
Pasteurella sp., Escherichia sp., Streptococcus sp., inconsistant isolates were Enterococcus avium,
Bacillus subtilis, Enterococcus faecalis, Proteus vulgaris, Pseudomonas sp., Lactobacillus sp.,
Klebsiella sp., Campylobacter sp. In urban areas the species most frequently responsable for
predation attempts on bats were represented by Felidae during the active period of bats: the wild
bats face a higher risk of predation from Accipitriformes, Falconiformes and Mustelidae more
frequently during the hibernation period. Our results show that the bacterial species identified in
the bite inflicted injuries on bats might give an insight upon the predators. The pathogenic
significance of these isolates request further investigations.
PSA38
OCCURRENCE OF ALTERNARIA ALTERNATA IN THE FUR OF DOGS AND CATS – AN
IMPORTANT FUNGAL ALLERGEN IN A ONE HEALTH APPROACH
P. Afonso1,2,3, H. Quintas3,4, A.F. Vieira5, E. Pinto5, M. Matos6,7, A.S. Soares1,2, L. Cardoso1,2,5,
A.C. Coelho1,2,5
1CECAV – Animal and Veterinary Research Centre, Vila Real, Portugal
3Agrarian School, Instituto Politécnico de Bragança, Bragança, Portugal
4Mountain Research Center (CIMO), Instituto Politécnico de Bragança, Bragança, Portugal
5Department of Veterinary Sciences, University of Trás-os-Montes e Alto Douro (UTAD), Vila Real,
Portugal
University of Trás-os-Montes and Alto Douro, Vila Real, Portugal 7Department of Genetics and
Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
OBJECTIVES: Alternaria alternata is an important human allergen that can be found in the fur of
animals. Inhalation of A. alternata spores is associated with respiratory hypersensitivity, asthma,
and allergic fungal rhinosinusitis. The aim of this work was to study the occurrence of A. alternata
in the fur of dogs and cats from 13 clinic and three shelters in northeast Portugal, to understand
their significance for public health under a One Health approach. MATERIAL AND METHODS: Fur
samples were collected using the toothbrush technique. A convenience sample of 412 animals
belonging to a shelter was examined for the presence of A. alternata in the fur. Samples were
inoculated in Potato Dextrose Agar medium and Sabouraud Dextrose Agar medium and incubated
at 25ºC and 37ºC, for 3-7 days. Identification of A. alternata was based on the morphology of their
colonies and micromorphology of spore and hyphae. RESULTS: Alternaria alternata was identified
in culture in 34 out of the 412 sampled animals (8.3%; 95% confidence interval [CI]: 7.8–8.7%).
Alternaria alternata was identified in 28 animals from the shelters out of 156 ones (17.8%; 95% CI:
17.0-18.6%) and in six animals from clinics out of 256 (2.3%; 95% CI: 5.4–6.6%). CONCLUSION:
Occurrence of A. alternata was high in this study. Since this agent is an important allergen, more
studies are required to better understanding the relevance of the isolation of this fungus in the fur
and its significance for public health.
PSA39
MICROSPORUM CANIS CARRIAGE IN STRAY CATS AND DOGS FROM SHELTERS AND
CLINICS IN THE NORTH OF PORTUGAL
P. Afonso1,2,3, H. Quintas3,4, A.F. Vieira5, E. Pinto5, M. Matos6,7, A.S. Soares1,2, L. Cardoso1,2,5,
A.C. Coelho1,2,5
1CECAV – Animal and Veterinary Research Centre, Vila Real, Portugal
3Agrarian School, Polytechnic Institute of Bragança, Bragança, Portugal
4Mountain Research Center (CIMO), Polytechnic Institute of Bragança, Bragança, Portugal
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5Department of Veterinary Sciences, University of Trás-os-Montes e Alto Douro (UTAD), Vila Real,
Portugal
University of Trás-os-Montes and Alto Douro, Vila Real, Portugal 7Department of Genetics and
Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
OBJECTIVES: Dermatophyte carriage data are crucial for assessing epidemiology and designing
potential control strategies. To study the occurrence of dermatophytosis, a survey was carried out
in dogs and cats, from October 2021 to June 2022 in shelters and clinics of northeast Portugal.
MATERIAL AND METHODS: Fur samples (n = 412) were collected from three shelters and 13 pet
clinics using the toothbrush technique. Dermatophyte culture was performed using Dermatophyte
test medium®. Petri dishes were handled under sterile conditions and incubated at 28°C for up to
21 days. RESULTS: Microsporum canis was identified in seven animals (5 dogs and 2 cats) out of
the 412 animals (1.7%; 95% confidence interval [CI]: 1.2–2.2%). Dermatophytes were identified in
two animals from the shelters (1 dog and 1 cat) out of the 156 animals (1.3%; 95% CI: 0.5-2.1) and
in five animals from clinics (4 dogs and 1 cat) out of 256 (1.9%; 95% CI: 1.3-2.5%). CONCLUSION:
These results suggest that dermatophyte shedding is rare in animals admitted to the shelters and
clinics under study. Considering the scarcity of epidemiological reports in Portuguese shelters and
clinics, these results could be a useful contribution towards diagnosis and prevention.
PSA40
CUTANEOUS MYCOBIOTA OF PET RABBIT (ORYCTOLAGUS CUNICULUS)

AND GUINEA PIG (CAVIA PORCELLUS)
R. Abreu1,2, A. Ramos1, A. T. Reisinho1, E. Cunha1,2, L. Tavares1,2, M. Oliveira1,2

2Laboratório Associado para Ciência Animal e Veterinária (AL4AnimalS),Portugal
Background: Rabbits and guinea pigs are frequently adopted as companion animals. We aimed
to characterize the cutaneous mycobiota of these species and to evaluate their potential role as
dermatophyte carriers. Materials/methods: Samples of hair and scales (n=102) were obtained
from 32 rabbits and 19 guinea pigs using two different methods: by pulling hairs surrounding lesions
and collecting scales (in case of dermatological localized lesions) or along the body of the animal
(in case of absence of dermatological signs) (n=51); and using the Mackenzie’s method (brushing
the animal with a toothbrush) (n=51). Samples were inoculated in Sabouraud Chloramphenicol Agar
and Dermatophyte Test Media and observed daily during the incubation period. Finally, isolated
fungal species were identified through their microscopic and macroscopic morphology. Results: It
was possible to obtain 168 isolates, the majority being saprophytic moulds (83.96%), belonging to
the genera Aspergillus (26.8%), Scopulariopsis (15.5%), Penicillium (14.9%), Mucor (7.1%),
Rhizopus (7.1%), Cladosporium (4.8%), Alternaria (4.2%), Phoma (1.2%) and Chaetomium (0.6%).
Yeast growth was observed in some samples (5.89%), corresponding to Candida spp. (5.4%) and
Rhodotorula spp. (1.2%). It was not possible to identify 19 fungal colonies (11.3%), or isolate any
dermatophytes. Conclusions: The three fungal genera Chaetomium, Phoma and Rhodotorula
were identified for the first time in pet rabbits and guinea pigs. The majority have already been
reported in the hair and skin of rabbits, guinea pigs and other domestic animals, such dogs and
cats. Some species have already been reported to cause infection in human and animals, especially
in immunocompromised individuals. Ackowledgements: This work was supported by CIISA–
Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária,
Universidade de Lisboa, Project UIDB/00276/2020 (Funded by FCT) and by Laboratório Associado
para Ciência Animal e Veterinária (LA/P/0059/2020 - AL4AnimalS)
PSA41
HYPERBETAGLOBULINEMIA IN 3 DOGS WITH SYSTEMIC MYCOSIS

A.Grassi1, S. Rossi2, M. Pantoli1, R. Ferriani2, M. Gambini1, C. Cafarchia3
1I-Vet laboratorio analisi, Flero- Brescia- Italy
2Ospedale Veterinario San Francesco, Milan- Italy
3Dipartimento di Medicina Veterinaria, Università degli Studi Aldo Moro, Bari- Italy
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Introduction: Serum protein electrophoresis (SPE) is a technique used to determine circulating

proteins composition, to identify typical pathological patterns and, and to evaluate the patient’s
health status when combined with haematological and biochemical profiles There are no published
data about SPE changes in dogs with systemic mycosis (SM), which are difficult to diagnose and
require specific laboratory tests. The aim of this study is to analyse SPE of 3 dogs with MS and their
follow-up after antifungal therapy. Material and Methods: The study included a Giant Schnauzer
(P1), Russian Terrier (P2) and Corso dog (P3) ranging from 3 to 4 years-old, suffering from SM by
Scytalidium spp, Aspergillus deflectus and Cladosporium spp. respectively. Diagnosis was made
by means of imaging, cytological examination, culture examination and fungal typing. Patients
underwent antifungal therapy and complete haematological and biochemical analysis at the time of
diagnosis and after clinical improvement as follow-up. Results: All dogs initially showed
hyperproteinaemia with hyperbetaglobulinemia (1- 10.2 g/dl, 46%; 2- 8.8 g/dl, 44%; 3- 8.2 g/dl,
33%); after antifungal therapy and improvement of clinical conditions SPE values decreased (1- 7.2
g/dl, 24%; 2- 7.10 g/dl, 33%; 3- 7.3, 30%). Subsequently dog1 died due to owner’s, dog2 following
heart complications while dog3 survived and was healthy. Conclusion: SPE is already useful
technique in veterinary medicine to support clinical suspicion, diagnosis and monitoring of some
canine diseases caused by etiologic agents (e.g. Leishmaniosis, Ehrlichiosis, Angiostrongylosis).
Hence more studies are needed to correlate SPE changes with SM and use it as a diagnostic and
monitoring tool.
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POSTER SESSION B
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PSB01
EVALUATION OF ANTIBODIES AGAINST PANLEUKOPENIA VIRUS (FPV) IN CATS:

COMPARISON OF TWO METHODS
A. Cavalli, C. Desario, G. Lanave, A. Trotta, M. Corrente, V. Martella
Department of Veterinary Medicine, University of Bari, Valenzano, Italy
Feline panleukopenia virus (FPV) causes acute lethal infection with signs of enteritis and bone
marrow suppression in domestic cats and wild felids. Due to the severity of the disease in cats,
vaccination is strictly recommended in all age groups.
Evaluating the levels of antibodies in the serum of cats to establish the optimal time for vaccination
is important and virus neutralization (VN) and inhibition of hemagglutination (HI) may be used.
Although VN is regarded as the gold standard in FPV serology for its ability to assess the actual
protection status against infection, viral replication in cells must be detected by indirect
immunofluorescence (IFI). HI is universally considered a good proxy for VN but the presence of
non-specific agglutinins in feline sera and artefactual precipitation of red blood cells (RBC) can
cause false negative and false positive results, respectively. In this study, we optimized the HI assay
to quantify precisely FPV-specific antibody titres in 30 serum samples of cats of various age groups
and vaccine status. Overall, removal of nonspecific agglutinins from the sera by adsorption with
RBCs and decreasing the concentration of RBCs from 0.8% to 0.1% improved the performance of
the HI assay. Using the VN-IFI test as a standard, the modified HI test showed 100% sensitivity and
83.3% specificity. The negative and positive predictive values were 90% and 100%, respectively.
When comparing the VN and the modified HI tests, the statistically significant (p <0.001) AUC was
0.917 (CI95%; 0.81; 1.03), highlighting the reliability of the optimized protocol
PSB02
ASSESSMENT OF THE PREVALANCE OF THE MAJOR MICROBIAL AND PARASITIC

PATHOGENS OF BEES IN A CLINICALLY HEALTHY POPULATION MAINTAINED UNDER
CLOSE SERVEILLANCE
A. Mataragka1, P. Katsarou1, A. Symeonidou1, F. Hatzina2, L. Haristos2, T. Papadopoulos3, J.
Ikonomopoulos1
1Agricultural University of Athens, Athens, Greece
2Hellenic Agriculture Org. "DEMETER", Nea Moudania, Greece
3Aristotle University of Thessaloniki, Thessaloniki, Greece
Objective: This study was focused to the application of the polymerase chain reaction (PCR) for
the assessment of the prevalence of major microbial and parasitic pathogens in a clinically healthy
population of bees and investigate for potential diagnostic indicators based on test-positivity
associations.
Materials and Methods: The study was conducted on a swarm of 250 stationary colonies located
in Northern Greece, in which varroosis is considered enzootic. The study population had not shown
clinical evidence of disease and had not received any antimicrobial or antiparasitic agents for a
period of at least six months before sampling. A total of 820 bees were collected from 22 colonies
between July and August 2020, and processed for PCR, qPCR, and RT-qPCR for the detection of
Acute Bee Paralysis Virus (ABPV), Black Queen Cell Virus (BQCV), Chronic Bee Paralysis Virus
(CBPV), Deformed Wing Virus (DWV), Sacbrood Virus (SBV), Varroa Destructor Virus-1 (VDV-1),
European Foulbrood (EFB), Nosema spp., Small Hive Beetle (SHB), Tropilaelaps spp. and Varroa
destructor.
Results: All samples (100%) reacted positively to the PCR assays conducted for the detection of
ABPV and BQCV, whereas 22.7% were also positive to a third pathogen i.e., VDV-1 (18.2%) and
Nosema spp. (4.5%). The prevalence per pathogen was 100% for ABPV and BQCV, 18.2% for
VDV-1, and 4.5% for Nosema spp. Positive test results were not recorded for CBPV, DWV, SBV,
Tropilaelaps spp., Varroa destructor, EFB, and SHB.
The investigation for diagnostic indicators based on test-positivity association did not produce
statistically significant results.
Discussion/Conclusion: In spite close health monitoring and having remained at the same
location for many years, positivity of the target population to ABPV and BQCV was 100%. More
than 1/5th of the study population was co-infected by three pathogens. This study indicates the need
to apply urgently test and removal measures for the control primarily of ABPV and BQCV.
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PSB03
PRELIMINARY SEROPREVALENCE STUDY OF PARATUBERCULOSIS IN DAIRY

HERDS IN APULIA (SOUTHERN ITALY)
M.G. Galgano1, A. Pratelli1, M. Tempesta1, G. Greco1, D. Mrenoshki1, A.A.K. Zarea1,4, A.
Sposato3, M. Beverelli3, C. Trisolini2, V. Laera2, G. Caldarola2, D. Buonavoglia1
1
Department of Veterinary Medicine, University of Bari Aldo Moro, Valenzano (Ba), Italy
2
Istituto Zooprofilattico della Puglia e della Basilicata, Putignano (BA), Italy
3
Istituto Zooprofilattico della Puglia e della Basilicata, Torre Santa Susanna (BR),Italy
4
Department of Microbiology and Immunology, Veterinary Research Institute, National
Research Centre (NRC), Egypt
John’s disease (JD) is a chronic granulomatous disease caused by Mycobacterium avium

subsp. paratuberculosis (MAP) which mainly affects ruminants and whose zoonotic
potential has been debated for over a century. This disease results in huge economic
losses in the cattle livestock.
In Italy, voluntary programs for the control of MAP in infected dairy cattle were implemented
in the Northern and Central part of the country, where many studies have been conducted.
The development of an adequate control plan requires knowledge of the epidemiological
situation in the other non-investigated areas. There are several diagnostic tests to identify
infected animals and/or herds. Currently, ELISA is the most commonly used test for the
detection of antibodies against MAP.
A total of 6.056 serum samples were collected from 341 different farms in the Apulia region
(Italy) during the screening plans for brucellosis, according to the protocol approved by the
Ethics Committee for Animal Experimentation (approved n° 19/21) of the Veterinary
Medicine Department. Sera were tested using a commercial ELISA test. Based on the
performance of the ELISA test (sensitivity 43%, specificity 99.3%), the prevalence rate of
JD detected in Apulia was 1.07% (95%, CI: 0.5 to 1.7%) at animal level and 22.25% (95%,
CI: 15.8 to 30.6%) at herd level.
Our results are in agreement with the data observed in literature and highlight the need to
design and implement effective JD control program at national level to prevent the spread
of the disease.
Table 1: Apparent and true prevalence, with 95% confidence intervals (CI), of
Mycobacterium avium subsp. paratuberculosis in dairy herds of Apulia region.
Number Number
Apparent True
of of
Prevalence Prevalence
positive negative
1.17% 1.07%
Animal level 71 5985
(95%CI: 0.9% to 1.4%) (95%CI: 0.5% to 1.7%)
10.56% 22.25%
Herds level 36 305
(95%CI: 7.7% to 14.3%) (95%CI: 15.8% to 30.6%)
Figure1: Map showing the study areas with the positive and negative cases of M. avium subsp.
paratubercolisis from dairy cattle serum samples.
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PSB04
A POSSIBLE CO-INFECTION RELATIONSHIP BETWEEN CACV AND CPV-2 IN IRANIAN

DOGS
R. Faraji1, M. Sadeghi1, S.-R. Miraei-Ashtiani1, V. Vasinioti2, L. A. Ndiana2, C. Desario2, F.
Beikpour2, N. Decaro2
1Department of Animal Science, College of agriculture & natural resource, University of Tehran,
Karaj, Iran.
2Department of Veterinary Medicine, University of Bari Aldo Moro, Italy.
Since the first detection of canine circovirus (CanineCV), several reports have been published over
the last decade about the worldwide distribution of this emerging virus of dogs. Canine circovirus
(CanineCV) is a small, icosahedral, non-enveloped, and spherical virus, with a circular single-
stranded DNA genome of nearly 2 kb in size (Kapoor, Dubovi et al. 2012) that infects domestic dogs
and wild carnivores (Zaccaria, Malatesta et al. 2016). So far, CanineCV and genetically related
circoviruses have only been reported in a few countries, including the USA (Kapoor, Dubovi et al.
2012, Li, McGraw et al. 2013), Argentina (Kotsias, Bucafusco et al. 2019), Brazil (Weber, Cibulski
et al. 2018), Italy (Decaro, Martella et al. 2014), Germany (Hsu, Lin et al. 2016), China (Sun, Zhang
et al. 2019), Thailand (Piewbang, Jo et al. 2018), Colombia (Giraldo-Ramirez, Rendon-Marin et al.
2020), Norway (Urbani, Tryland et al. 2021), the UK (Bexton, Wiersma et al. 2015) and more
recently in Iran (Beikpour, Ndiana et al. 2022). This study aimed to survey the circulation of
CanineCV in Iranian dogs and also to characterize the detected strains at the molecular level. In
order to investigate the prevalence and genomic features of CanineCV in Iranian dogs, a total of
203 dog faecal samples was collected between February and November 2018 from five different
geographical regions and screened by real-time PCR (qPCR). Thirteen samples (6.4%) from a total
of 203 screened samples were positive for CanineCV by qPCR, while 49 samples (24.13%) were
found to contain the CPV DNA (Table 4). The MGB probe assays showed that all CPV positive
samples contained CPV2a strains. All the CanineCV positive samples were also positive for CPV.
Three partial replicase nucleotide sequences of the detected CanineCV strains were obtained and
compared with the reference sequences deposited in the GenBank depository database. By
sequence analysis, the Iranian CanineCV strains displayed a nt identity of 96.4-98.2 % each to
other and of 88.3-98.2 to CanineCV reference strains. Through analysis of the partial Rep nt
sequences, the Iranian CanineCV strains were more closely related to strains detected in Turkey
(GenBank accession no. MK783223), allowing to hypothesize a possible introduction of the virus
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from a neighbor country. Due to the limited number of analyzed sequences, however, other studies
are needed to better understand the evolutionary pattern of Iranian CanineCVs. Continuous
molecular surveillance for CanineCV should be also carried out to better understand the pathogenic
potential of this emerging virus of the canine enteric tract. The present study provides new insights
into the CanineCV molecular epidemiology and its possible role as a co-infectious pathogen.
Keywords: Dog; Canine circovirus; Genetic analysis; Canine Parvovirus; Iran.
PSB05
DIAGNOSTIC INVESTIGATION FOR THE DETECTION OF MYCOBACTERIA IN SAMPLES OF

FISH FEEDS AND TISSUE FROM SEA BREAM AND SEA BASS WITH SEVERE TUBERCULAR
LESIONS
A. Mataragka1, N. Tzimotoudis2, M. Kolygas3, E. Karavanis4, J. Ikonomopoulos1
1School of Animal Biosciences, Department of Animal Science, Agricultural University of Athens,
Athens, Greece
2Hellenic Army Biological Research Center / Laboratory of Microbiology, Athens, Greece
3Veterinary School, University of Thessaly, Karditsa, Greece
4Histology-Histopathology Laboratory, C’ Army Veterinary Hospital, Camp Makri, Thessaloniki,
Greece
Objective: We report on a case of a sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata)
aquaculture farm in Greece, with many incidents of severe dermal and visceral tubercular lesions
detected during routine inspection and post-mortem examination.
Materials and Methods: The investigation was conducted on samples of fish feeds (n=13) and fish
feed ingredients (n=15) available commercially, and tissue samples (n=51) collected from the
affected population, and consisted of molecular analysis and histopathology. The former was
performed using five PCR assays targeting the main mycobacterial pathogens. Histopathology
examination relied on staining of tissue sections of various visceral organs with hematoxylin-eosin
and Ziehl-Neelsen.
Results: Presence of mycobacteria was demonstrated in the fish feeds (100% and 46.2% positive
for Mycobacterium spp. and Mycobacterium avium, respectively), in the fish feed ingredients (73.3%
and 33.3% positive for Mycobacterium spp. and Mycobacterium avium, respectively) and the tissue
sections (76.5% and 3.9% positive for Mycobacterium marinum and Mycobacterium avium,
respectively) that were examined. Sequence analysis of the PCR products was confirmatory of the
specificity of the amplification process, at a 98% minimum level of identification. Histopathology
revealed in all cases evidence consistent with mycobacterial infection.
Conclusion: The result of the analysis indicates that in addition to contaminated feeds, fish are
exposed to mycobacteria through another source, which is mainly associated with Mycobacterium
marinum. This probably accumulates in large numbers over the years in the organic sediment
collected on the seabed of the aquaculture, causing severe infections.
PSB06
POTENTIAL EFFICACY OF A COMBINATION OF DIFFERENT NON-BIOSECURITY BASED

INTERVENTION MEASURES TO REDUCE C. JEJUNI COLONIZATION OF BROILER
CHICKENS
V. Szott1, E. Peh2, B. Reichelt1, A. Friese1, S. Kittler2 , U. Roesler1
1Institute for Animal Hygiene and Environmental Health, Freie Universität Berlin, Germany
2Institute for Food Quality and Food Safety, University of Veterinary Medicine, Hannover, Germany
Objective: The objective of this study was to evaluate the efficacy of combined non-biosafety
measures to reduce C. jejuni colonization. We combined a CE culture with bacteriophages and an
essential oil (carvacrol) with organic acids. Methods: To examine the effect of carvacrol and organic
acids, broilers were fed daily with 120 mg/kg feed of carvacrol and a mixture of four acids in their
drinking water. To evaluate the efficacy of a CE-culture with bacteriophages, broiler chickens were
treated with the CE-culture twice and received a phage combination of two phages continuously via
drinking water prior to necropsy. Results: Cecal count enumeration demonstrated that the C. jejuni
load was significantly reduced for the group receiving a combination of the CE-culture and
bacteriophages log reduction of 1.0 log10 MPN/g. Likewise, colon counts were significantly
decreased for the group receiving a combination of bacteriophages and the CE-culture (log
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reduction of 1.0 log10 MPN/g). In contrast, although we observed a log reduction of 1.0 log10 MPN/g
in C. jejuni cecal counts in the group receiving a combination of carvacrol and organic acids, this
reduction was nonsignificant. Likewise, there was no significant difference in C. jejuni counts in the
colon. Conclusion: We conclude that a combination of this particular CE-culture and
bacteriophages may be a promising practical approach in broiler production to reduce C. jejuni
colonization in broiler chickens at slaughter age. However, why the combination of carvacrol and
organic acids failed to reduce C. jejuni intestinal colonization is unclear and remains to be
investigated.
PSB07
ZOONOTIC POTENTIAL OF HEPATITIS E VIRUS: THE ROLE OF PIGS IN THE TRASMISSION

OF INFECTION TO HUMANS
F. Gucciardi1, G. Macaluso1, S. Di Bella1, F. Mira1, A. Princiotta1, G. Chiaramonte1, C.
Castronovo1, A. Lastra1, S. Vullo1, G. La Rosa2,A. Torina1, A. Guercio1, G. Purpari1
1Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri” – Palermo – Italy
2Department of Environment and Healt, Istituto Superiore di Sanità – Rome - Italy
Hepatitis E virus (HEV) is responsible for acute entero-transmissible hepatitis in humans. In Italy,
an increase in autochthonous cases of HEV has recently been documented, and several cases
have been epidemiologically linked to the consumption of raw or undercooked wild boar and pork
liver or meat. Strains circulating in pigs and humans have a high sequence similarity, suggesting
that transmission between pigs and humans is common. This study investigated the occurrence of
HEV-RNA in different sample types of pigs intended for human consumption. Faeces,
diaphragmatic muscle, liver, and blood samples were collected from 325 pigs in 46 farms in Sicily,
Italy. The organs were prepared by the Trizol/chloroform homogenization/purification method. A
10% stool suspension in MEM medium was obtained. HEV RNA was detected via Real Time RT-
PCR targenting ORF3, and by nested RT-PCR targeting ORF1 and ORF2 , followed by sequencing
and phylogenetic analysis. Overall, 76/325 (23.38%) animals belonging to 35/46 herds (76.1%),
tested positive for HEV RNA, detected in faeces (16%), livers (3.9%), muscles (9%), and plasma
(1%). Phylogenetic analyses performed on the sequence of ORF1 and ORF2 fragments showed
the circulating of genotype 3 and subtypes 3C and 3F. The data obtained confirm the circulation of
the virus in the suidae in the Sicilian territory. The presence of the virus in the plasma of animals is
important and can contribute to the contamination of meat. As the survival time of the virus in pig
products is not known, precaution are important to prevent HEV infection in animal facilities.
PSB08
EARLY-MYCO: AFLATOXIN B1 EFFECTS ON OFFSPRING GUT IMMUNITY AND

MICROBIOTA
P. Bastos-Amador3, E. L. Duarte1,2*, I. Silva1, R. Assunção5 , P. Alvito, A. T.Caldeira 1,6,7, M.
Ferreira3,4
1School of Science and Technology, Universidade de Évora, Évora, Portugal
2MED-Mediterranean Institute for Agriculture, Environment and Development, Évora, Portugal
3Champalimaud Centre for the Unknown, Lisboa, Portugal
4 Centre for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Biocant Park,
Cantanhede, Portugal
5 CESAM - Centre for Environmental and Marine Studies, Department of Biology, University of
Aveiro, Aveiro, Portugal & Food and Nutrition Department, National Institute of Health Dr. Ricardo
Jorge, Lisbon, Portugal.
6 Hercules Laboratory, Universidade de Évora, Portugal
7City U Macau Chair in Sustainable Heritage, Universidade de Évora, Portugal
Aflatoxin B1 (AFB1) produces acute or chronic deleterious health effects in humans and animals.
Still, long-term effects derived from initial exposure in early life, a critical period for colonization and
development of gut microbiota, has not been fully evaluated. Particularly, aflatoxins could impair gut
microbiota and immunity settlement, as they have been proven to cross the placental barrier and
can be found in breast milk. We investigated the impact of maternal exposure to AFB1 on early-life
microbiota in a mouse model. Females were fed jelly pellets containing 400 µg/kg AFB1 diluted in
DMSO (treated animals n=6) or DMSO vehicle alone (control group n=6) during pregnancy and
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lactation. Faeces from the offspring of both treated and control females were collected immediately
after weaning and faecal DNA was extracted and purified. Bacterial taxa diversity and relative
abundance were assessed by High-Throughput Sequencing performed in an Illumina Miseq®
sequencer, targeting the V3 and V4 regions of the 16S rRNA gene. Operational taxonomic units
(OTUs) were determined by clustering reads to 16S reference databases. A hundred and twenty-
four (N=124) bacterial genera were found in both groups, 5 were only present in AFB1 treated group
and 27 exclusively in control groups. A hundred and fifty-one (N=151) bacterial species were
common to both groups, 15 species exclusively found in AFB1 litters and 34 species were
exclusively found in control litters. To assess abundance and characterize species diversity and
evenness, Shannon diversity index was calculated but no significant differences were found
between groups. Although present in both groups, Akkermansia muciniphila and Bacteroides
acidifaciens were significantly higher in controls. A. muciniphila colonizes the intestinal tract in
childhood and regulates mucus thickness, intestinal barrier integrity and is involved in immune
modulation. B. acidifaciens participates in the metabolism of lipids and sugars and activates some
cytokines and immune cell receptors. Sulfidogenic bacteria recently related to inflammatory bowel
disease such as Desulfovibrio piger and Bilophila wadsworthia were exclusivly found in the treated
litters. Early-life gut microbiome is paramount to trigger the gut immune defences, but is far less
stable than the adult microbiome. Moreover, previous work identified aflatoxins intake as a potential
health hazard in Portuguese children. These preliminary results open an extensive field to further
investigate the association between mycotoxins and microbiome, as the latest is increasingly
recognized as a major player in a wide spectrum of diseases. This work was funded by FCT/MCTES
through national funds, to earlyMYCO (PTDC/MED-TOX/28762/2017), and CESAM
(UIDP/50017/2020+UIDB/50017/2020).
PSB09
RICKETTSIA MASSILIAE CIRCULATION IN SHEEP AND ATTACHED RHIPICEPHALUS

SANGUINEUS IN CENTRAL PORTUGAL
J. R. Mesquita1,2, S. Santos-Silva1, A. de Sousa Moreira1, M. B. Baptista3, R. Cruz3,4, F.
Esteves3,4, H. Vala3,5, P. F. Barradas1,2,6,7
1Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal
2Epidemiology Research Unit (EPIUnit), Instituto de Saúde Pública da Universidade do Porto,
Portugal
3Agrarian School of Viseu, Polytechnic Institute of Viseu (ESAV), Portugal
4Centre for Studies in Education and Health Technologies (CI&DETS), Viseu, Portugal
5Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB)
and Institute for Innovation, Capacity Building and Sustainability of Agri-food Production
(Inov4Agro), UTAD
6Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde (CESPU),
Gandra, Portugal
7Agrarian School, Polytechnic Institute of Coimbra (IPC), Portugal Country,
Rickettsiosis poses a serious public health threat among emerging and re-emerging vector-borne
diseases. Ticks are reservoirs and vectors of Rickettsia having a significant role in the transmission
of rickettsiae. In Portugal, little is known about tick-borne Rickettsia species in sheep. Therefore,
this study aimed to investigate rickettsiae infection in ticks and their sheep host from 27 farms of
four districts of central Portugal, to clarify the role of the sheep host in the circulation of this zoonotic
agent. Between March and May 2021, EDTA blood samples (n=100) of healthy grazing sheep and
their ticks (n=100, one tick per animal) were collected during a herd health program in Central
Portugal. Obtained ticks were all identified as Rhipicephalus sanguineus sensu lato by PCR
targeting a partial sequence of 16S rDNA gene followed by sequence analysis. Rhipicephalus
sanguineus s.l. and host sheep blood were tested for the presence of Rickettsia spp. by
conventional PCR targeting a partial sequence of ompB and ompA genes. From a total of 100 paired
Rhipicephalus sanguineus s.l. ticks and host sheep, Rickettsia massiliae was detected in 62
Rhipicephalus sanguineus ticks and 35 grazing sheep blood samples, collected in central Portugal,
2021. These findings suggest that sheep may develop rickettsiemia and probably, could be capable
of transmitting and amplifying the infection to uninfected ticks maintaining rickettsiae in circulation
in the domestic cycle.
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PSB10
STATUS OF THE CHARACTERIZATION OF PSEUDOMONAS AERUGINOSA ISOLATES AND

ANTIMICROBIAL RESISTANCE FROM HUMAN INFECTIONS
T. de Sousa1,2,3,4, E. Costa5, O. Alves5, M. Hébraud6,7, G. Igrejas1,2,3, P. Poeta3,4,8,9
1Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD),
Vila Real, Portugal;

2Functional Genomics and Proteomics Unit, University of Trás-os-Montes and Alto Douro (UTAD),

3LAQV-REQUIMTE, Faculty of Science and Technology, University Nova of Lisbon, Lisbon,
Caparica, Portugal;
4Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences,
University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal;
5Medical Centre of Trás-os-Montes and Alto Douro, Clinical Pathology Department, Vila Real,
Portugal;
6Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE),
Metabolomic and Proteomic Exploration Facility (PFEM), 60122 Saint-Genès-Champanelle,

France;
7Université Clermont Auvergne, INRAE, UMR Microbiologie Environnement Digestif Santé
(MEDiS), 60122 Saint-Genès-Champanelle, France;

8CECAV – Veterinary and Animal Research Centre, University of Trás-os-Montes and Alto Douro,

9Veterinary and Animal Research Centre, Associate Laboratory for Animal and Veterinary Science
(AL4AnimalS).
P. aeruginosa is an opportunistic pathogen being one of the major opportunistic pathogens. The
general excessive use of antibiotics is increasing their resistance to these substances. The
emergence of multi-drug resistant P. aeruginosa is a serious threat due to the lack of therapeutical
options. This work aims to phenotypically and genotypically determine resistance to different
classes of antibiotics. Thirty-tree P. aeruginosa strains recovered between September 2021 and
February 2022 from human patients infected with this pathogen were used in this study. All strains
were isolated using VITEK 2® COMPACT (bioMérieux) in Medical Centre of Trás-os-Montes and
Alto Douro. The antibiotic susceptibility tests were performed using the Kirby Bauer diffusion test
and the minimum inhibitory concentration (MIC). For the antibiotic colistin, the minimum bactericidal
concentration (MBC) method was also performed. All methods were in accordance with EUCAST
standards. PCR tests performed with the specific primers sets. Of the thirty-tree clinical isolates of
P. aeruginosa, 33% presented resistance to several antibiotic classes. Resistance to imipenem
(n=28, 85%), meropenem (n=11, 33%) and piperacillin-tazobactam (n=10, 30%) were the most
represented. In contrast, strains were very sensitive to the amikacin (n=33, 100%) and tobramycin
(n= 32, 97%). The MBCs was 8 µg/mL (56,25%), 16 µg/mL (37,50%) and 32 µg/mL (6,25%) of the
32 samples (range between 0.25 to 32 µg/mL). Our results confirm that multidrug-resistant P.
aeruginosa is emerging among clinical isolates and, although, colistin remains an effective option,
12% of the isolates are resistant to colistin, which pose a major public health problem.
PSB11
PSEUDOMONAS AERUGINOSA PHENOTYPIC CHARACTERIZATION IN CANIS FAMILIARIS

T. de Sousa1,2,3,4, A. Fortunado3,4,5, M. Hébraud6,7, A. Garcês8,9,10, M. Ribeiro1,2, C. Sabença1,2,3,4,
G. Igrejas1,2,3 P. Poeta3,4,11
1Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD),


3LAQV-REQUIMTE, Faculty of Science and Technology, University Nova of Lisbon, Lisbon,
Caparica, Portugal;
University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal;
5First degree biology student, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real,
Portugal;
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6InstitutNational de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE),

Metabolomic and Proteomic Exploration Facility (PFEM), 60122 Saint-Genès-Champanelle,
France;
7Université Clermont Auvergne, INRAE, UMR Microbiologie Environnement Digestif Santé
(MEDiS), 60122 Saint-Genès-Champanelle, France;

8INNO - Veterinary Laboratory, R. Cândido de Sousa 15, 4710-503 Braga, Portugal;
9Cooperativa de Ensino Superior Politécnico e Universitário, CRL -CESPU, R. Central Dada
Gandra, 1317, 4585-116 Gandra, Portugal;

10CITAB - University of Trás-os-Montes and Alto Douro, Quinta de Prados 5000-801, Vila Real,
Portugal.
(AL4AnimalS).
Pseudomonas aeruginosa is a pathogenic bacterium that has a high natural resistance to several
antibiotics. Companion animals are potential transmitters of this bacterium to humans, and the dog
(Canis familiaris) has a special prominence among this group. Therefore, from a One Health
perspective, it is important to evaluate and to characterize the antimicrobial resistance of P.
aeruginosa from dogs. This work aims to determine phenotypically resistance to different classes
of antibiotics. Twenty-seven P. aeruginosa strains were recovered between September and
December 2021 from dogs infected with this pathogen. All dog samples were collected from the
INNO-Veterinary Laboratory and Veterinary Hospital of Trás-os-Montes and Alto Douro and isolated
using VITEK 2® COMPACT (bioMérieux) to be used in this study. Their identification was confirmed
using the selective medium Pseudomonas CN selective agar. The susceptibility tests were
performed to microbial agents using the Kirby Bauer diffusion test and the minimum inhibitory
concentration (MIC) (EUCAST standards). Eleven antibiotics disk were tested: Ceftazidime,
Cefepime, Amikacin, Gentamicin, Tobramycin, Doripenem, Imipenem, Meropenem, Aztreonam,
Ciprofloxacin and Ticarcillin-clavulanic acid. The antibiotics Enrofloxacin, Ceftiofur, Marbofloxacin
and Cefovecin were performed by MIC. The phenotypic profile of the twenty-seven isolates revealed
a resistance to cefovecin (74%) and ceftiofur (59%), demonstrating a high resistance to the
cephalosporins class of antibiotics. The isolates were susceptible to amikacin (100%) and
tobramycin (100%). In this preliminary study, most of the strains showed intermediate resistance to
the various antibiotics tested, but we confirm that the samples from dogs do not yet show high
resistance to antibiotics. However, their use has to be controlled to minimize the acquisition of
resistances.
PSB12
DETECTION OF ESBL-PRODUCING KLEBSIELLA PNEUMONIAE ISOLATED FROM HUMAN

URINARY TRACT INFECTIONS IN THE NORTH OF PORTUGAL
C. Sabença1-4, E. Costa5, S. Sousa5, T. de Sousa1-4, G. Igrejas2-4, C. Torres6, P. Poeta1,4,7,8
1MicroART-Antibiotic Resistance Team, Department of Veterinary Sciences, University of Trás-os-
Montes and Alto Douro, Vila Real, Portugal

2Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila
Real, Portugal
3Functional Genomics and Proteomics Unit, University of Trás-os-Montes and Alto Douro, Vila Real,
Portugal
4Associated Laboratory for Green Chemistry, University NOVA of Lisbon, Caparica, Portugal
5Hospital Centre of Trás-os-Montes and Alto Douro, Clinical Pathology Department, Vila Real,
Portugal
6Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain
Vila Real, Portugal

(AL4AnimalS) - Vila Real (Portugal)
The most frequent human infections in communities and hospitals are urinary tract infections (UTIs).
Extended-spectrum beta-lactamases (ESBL) producing bacteria is a growing concern because this
resistance mechanism means that fewer antibiotic options are available to treat common infections.
Klebsiella pneumoniae samples were isolated from urinary tract infections in the Medical Centre of
Trás-os-Montes and Alto Douro between the December 7th of 2021 and the February 3rd of 2022.
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K. pneumoniae strains were tested for antimicrobial resistance by the disc diffusion method, and
ESBL production was tested according to the Etest ESBL PM/PML. Finally, resistance genes were
detected by conventional Polymerase Chain Reaction procedure. A total of 85 K. pneumoniae were
isolated from the urinary tract infections. From the ESBL production test, 25 ESBL-producing K.
pneumoniae (29.4%), which were then submitted to the antimicrobial susceptibility test (AST), were
identified. AST has shown that all strains were resistant to ampicillin, cefotaxime, cefepime, and
aztreonam. A high percentage of resistance to ciprofloxacin (96%), ceftazidime (96%), and
trimethoprim-sulfamethoxazole (92%) was verified. Nevertheless, a low percentage of resistance to
imipenem (4%), meropenem (12%), ertapenem (24%), cefoxitin (24%), amikacin (28%), and
tetracycline (24%) was observed. From the 20 resistance genes tested, we detected the blaCTX-M
(92%), and blaTEM (84%) β-lactamase genes. Concerning the other resistance genes, we detected
most frequently the sul2 (88%), aac(3)-II (76%) and aadA1 (64%) genes. In opposition, tetB, and
aac(3)-IV genes were not observed. Results obtained highlighted a worrying percentage of ESBL-
producing K. pneumoniae strains with multidrug resistance isolated from urinary tract infections.
However, the low resistance to some antibiotics gives us hope in fighting this global public health
problem. Close contact with animals and their environments allows zoonotic pathogens, such as K.
pneumoniae, to pass from animals to humans and vice versa. So it is still essential to study these
isolates from human infections since, according to the One Health concept, people's health is
closely linked to the health of animals and our shared environment.
PSB13
SCREENING OF ENVIRONMENTAL SWABS AT A VETERINARY TEACHING HOSPITAL

AFTER ISOLATION OF KLEBSIELLA PNEUMONIAE IN A CAT WITH URINARY TRACT
INFECTION.
N. Mezzasalma, C. Spadini, M. Iannarelli, S. L. Montanaro, F. Fidanzio, A. Corsini,
E. Cataldo1, C. Quintavalla1, C. S. Cabassi
Department of Veterinary Science, University of Parma, Parma, Italy.
Nosocomial infections (NIs) represent an important public health issue of the 21th century, in humans
and animals. On May 2022 a case of NI has been documented in a cat presented at the Veterinary
Teaching Hospital of University of Parma for lower urinary tract disease (FLUTD). The first urine
sample was cultured onto suitable culture solid media and BHI broth, resulting sterile. During the
hospitalization the cat underwent catheterization due to urethral obstruction and developed clinical
signs suggestive for sepsis. Urine and blood samples were cultured, isolating Klebsiella
pneumoniae (KP). Antibiogram (AB) was performed by Kirby-Bauer, testing 20 antibiotics selected
for the treatment of systemic diseases of cats, resulting not effective in vitro. A NI was hypothesized
and environmental screening was performed. Seventy swab samples were collected in intensive
care unit, examination rooms and ultrasound rooms. KP was confirmed in the cage where the
patient was hospitalized and on the keyboard in the ultrasound room, together with other bacteria
agents of nosocomial infections: E.coli, Staphylococcus aureus, Enterobacter cloacae,
Pseudomonas fluorescens, Burkholderia cepacia, Stenotrophomonas maltophilia, Acinetobacter
Iwoffii. Each bacterial strain was evaluated by AB, then 13 strains including KP, resistant to at least
14 antimicrobials, were evaluated through MIC assay. According to Magiorakos et al (2012),
Sweeney et al (2018), referring to antimicrobials used in companion animals, 11 strains were
multidrug-resistant (MDR). These findings highlight the need for active surveillance and improved
hygiene standards to decrease the risk of NI transmission in the setting of a veterinary teaching
hospital.
PSB14
DETERMINING THE PREVALENCE OF ANTIMICROBIAL DRUG RESISTANCE IN WILD

BADGERS IN THE REPUBLIC OF IRELAND
R. Dunne1, G. Madigan1, S. Murphy1, R. Maguire1, E. Bracken1, U. Fogarty2
A. Naranjo Lucena1, R. Slowey1 and M. McElroy1
1Central Veterinary Research Laboratory, Bacteriology and Parasitology division,Co. Kildare,
Ireland.
2Irish Equine Centre, Co. Kildare, Ireland.
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Antimicrobial resistance (AMR) is recognized as a global threat to human and animal health. Under
the WHO - One health approach, regular surveillance and reporting of the incidence of AMR in
humans and livestock enables trends to be identified and monitored. Wildlife is recognized as a
potential vector of AMR to humans and livestock, yet there is little data available in the Republic of
Ireland. This study examined the prevalence of AMR from fecal samples (n=95), collected from
European badgers (Meles meles) throughout the Republic of Ireland. The samples were screened
for commensal Escherichia coli (E. coli), as well as extended spectrum beta lactamases (ESBLs)
using selective agar plates and MALDI-ToF. Commensal E. coli was isolated and identified in 85
samples (80.8%). Anti-microbial susceptibility testing (AST) on the 85 E. coli isolates was then
carried out, identifying 5 samples (4.8%) as ESBLs, including one AMP C producing Beta
Lactamase. A further 5 samples (4.8%) were identified as having resistance to various antimicrobial
agents including ampicillin, tetracycline, sulfamethoxazole, trimethoprim and chloramphenicol. 4 of
these samples (3.8%) were multi-drug resistant. This study provides a snapshot of AMR prevalence
in wildlife in the Republic of Ireland. With the potential of wildlife to act as a reservoir for zoonotic
AMR pathogens, and with the limited amount of data available, further studies are needed to monitor
levels of AMR in wildlife. This will help to establish its significance to humans and livestock as well
as identify potential drivers of AMR.
PSB15
QUANTIFYING TOPICAL ANTIMICROBIAL USE AND EXPLORING THE EFFECT OF

PARTICIPATION IN AN ANTIMICROBIAL STEWARDSHIP PROGRAMME IN DUTCH
COMPANION ANIMAL CLINICS
N.E.M. Hopman1, N. Kardomatea1, L. Portengen2, J.A. Wagenaar1, D.J.J. Heederik2, I.M. van
Geijlswijk2, E.M. Broens1

2Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht University,
Objective: Emergence of antimicrobial resistance in both human and veterinary medicine has
raised concerns, which applies to systemic and topical antimicrobial use (AMU). However, little
information is available on topical AMU. The objective of this study is to quantify topical AMU in 44
Dutch companion animal clinics and to explore the effect of participation in an antimicrobial
stewardship programme (ASP). Methods: Prescription data were obtained (from 2012-2018)
during previous project ‘Antimicrobial Stewardship and Pets’. The Defined Daily Dose for Animals
(DDDA)-method was used, and a mixed effect times series model was applied to monthly topical
AMU data. Clinic characteristics were assessed using a multivariable regression model and the
intervention effect was modelled using a step function with a change in (linear) time trend. Results:
A seasonal topical AMU pattern was observed, with highest AMU in July-August and lowest AMU
in February-March. Topical AMU significantly decreased over time and the proportion of dogs was
positively associated with topical AMU. No strong effects were seen after the introduction of an
ASP, except for second line antimicrobials and for skin products, which both showed a decrease.
Conclusion: This study demonstrated a seasonal pattern and a decreasing time trend in topical
AMU. The effect of an ASP was less clear than for systemic AMU. This might be an effect of the
focus on reducing systemic AMU, resulting in a shift towards topical AMU. As topical AMU appeared
to be a substantial part of total AMU in these clinics, topical AMU should be taken into account when
optimizing AMU.
PSB16
MULTIDRUG-RESISTANT STAPHYLOCOCCUS AUREUS IN CHICKEN STERNAL BURSITIS

V. Silva1-4*, A. Igrejas1, J. Ribeiro1,2, A. Silva1-4, P. Pinto5, M. Vieira-Pinto5-6, G. Igrejas2-4, P.
Poeta1,2,6,7
University of Trás-os-Montes and Alto Douro (UTAD), Portugal.

2Associated Laboratory for Green Chemistry (LAQV-REQUIMTE), University NOVA of Lisbon,
Portugal
3Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Portugal
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Portugal
5Department of Veterinary Science, University of Trás-os-Montes and Alto Douro (UTAD), Portugal
6Veterinary and Animal Research Centre (CECAV), University of Trás-os-Montes and Alto Douro
(UTAD), Portugal
7Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), Portugal
Infectious bursal disease (IBD) is an infectious disease, very contagious in young birds, that affects
the tissues of the animal's immune system, such as Fabricio's bursa. The disease alters the immune
response and leaves infected animals immunosuppressed which may lead to the development of
secondary infections. Furthermore, it is known that chickens with IBD are more susceptible to
develop infections caused Mycoplasma synoviae, Staphylococcus aureus, and Pasteurella spp.
Therefore, this study aimed to isolate S. aureus from chickens with IBD. Swab samples were
collected from 44 chickens with IBD. The swabs were incubated at 37°C for 24 h in BHI broth
supplemented with 6.5% of NaCl. Then, the inoculum was seeded on Baird-Parker agar plates and
incubated at 37°C for 24 h. S. aureus species were identified by MALDI-TOF MS. Antimicrobial
susceptibility testing was performed by the Kirby-Bauer disk diffusion method. From the 44 samples,
24 (54.5%) were positive for S. aureus. Six (25%) isolates showed resistance to at least three
classes of antimicrobials and were classified as multidrug-resistant. All isolates were resistant to
aminoglycosides and resistance to penicillin (n=2), macrolides and lincosamides (n=11),
tetracycline (n=2), trimethoprim-sulfamethoxazole (n=2), fusidic acid (n=5), chloramphenicol (n=2)
and ciprofloxacin (n=2) was also detected. This study demonstrated that S. aureus carrying
resistance to several different antimicrobials is frequently present chicken with IBD which may can
raise a problem since secondary infections can cause significant economic losses. Funding This
work was supported by the Associate Laboratory for Green Chemistry-LAQV, which is financed by
national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020) and by the projects
UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the Portuguese Foundation for Science and
Technology (FCT). Vanessa Silva and Adriana Silva are grateful to FCT (Fundacão para a Ciência
e a Tecnologia) for financial support through the PhD grants SFRH/BD/137947/2018 and
SFRH/BD/04576/2020, respectively.
PSB17
ANTIBIOTIC-RESISTANT ENTEROCOCCI ISOLATED FROM POULTRY’S STERNAL

BURSA LESIONS
J. Ribeiro1,2*, V. Silva1-4, P. Pinto5, M. Vieira-Pinto5-6, G. Igrejas2-4, P. Poeta1,2,6,7
1Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary
Sciences, University of Trás-os-Montes and Alto Douro (UTAD), Portugal

2Associated Laboratory for Green Chemistry (LAQV-REQUIMTE), University NOVA of
Lisbon, Portugal. 3Department of Genetics and Biotechnology, University of Trás-os-Montes

and Alto Douro, Portugal
4Functional Genomics and Proteomics Unit, University of Trás-os-Montes and Alto Douro
(UTAD), Portugal. 5Department of Veterinary Science, University of Trás-os-Montes and

Alto Douro (UTAD), Portugal
6Veterinary and Animal Research Centre (CECAV), University of Trás-os-Montes and Alto
Douro (UTAD), Portugal

Hundreds of thousands of annual deaths worldwide are caused by antibiotic-resistant bacterial

infections, making antibiotic resistance a serious threat to public health. One main contributor to
the spread of antimicrobial-resistant bacteria is the poultry industry since it allows contact between
humans, live animals, and the environment. Poultry can be affected by infection or traumatic injuries
in the sternal bursa that can be responsible for economic losses at the abattoir level due to total or
partial condemnations. Therefore, to determine whether the sternal bursa lesions could represent
a source of antibiotic-resistant bacteria, forty-eight swabs were collected from the bursal lesions of
slaughterhouse chickens. Each sample was seeded onto Slanetz-Bartley agar and Slanetz-Bartley
agar supplemented with vancomycin (4 mg/L) for the isolation of Enterococcus spp. and
vancomycin-resistant enterococci (VRE), respectively. Colonies with typical morphology were
identified by the bile-aesculin reaction. The antimicrobial susceptibility of the isolates was
determined according to the Kirby-Bauer disc diffusion method against ampicillin, vancomycin,
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teicoplanin, erythromycin, tetracycline, ciprofloxacin, chloramphenicol, quinupristin-dalfopristin,

linezolid, and gentamycin. Forty-one (85%) samples presented Enterococcus spp. and their
phenotypic profile revealed that 33% (n=16) were resistant to erythromycin, and 52% (n=25) were
resistant to tetracycline. Furthermore, one (2%) sample presented VRE, and its phenotypic profile
revealed that it was also resistant to erythromycin. These results demonstrate that sternal bursa
lesions in poultry represent a source of antibiotic-resistant bacteria. To prevent the further spread
of these bacteria, precautions should be taken when handling carcasses affected by this lesion.
Funding: This work was supported by the Associate Laboratory for Green Chemistry-LAQV, which
is financed by national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020) and by
the projects UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the Portuguese Foundation for
Science and Technology (FCT). Vanessa Silva is grateful to FCT (Fundacão para a Ciência e a
Tecnologia) for financial support through the PhD grant SFRH/BD/137947/2018.
PSB18
GENOTYPIC AND PHENOTYPIC CHARACTERIZATION OF ESCHERICHIA COLI ISOLATES

RECOVERED FROM THE UTERUS OF MARES WITH FERTILITY PROBLEMS
F. Paola Nocera1, C. Zagaglia2, L. Maurizi2, A. Lucia Conte2, C. Longhi2, L. De Martino1,3
1Department of Veterinary Medicine and Animal Production, University of Naples “Federico II”,
Naples, Italy
2Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome,
Italy
Introduction: Escherichia coli (E. coli) is reported worldwide as one of the main causative agents
of equine endometritis, and its ability to form biofilm within the uterus is a potential cause of
persistent endometrial infections. This study aimed to define the phenotypic antimicrobial resistance
profile, the biofilm formation capacity, and the detection of some virulence genes of equine E. coli
strains isolated from mares suffering from endometritis. Materials and Methods: E. coli strains
were isolated from uterine swabs collected from mares diagnosed with reproductive disorders.
Antimicrobial susceptibility testing was performed by disk diffusion method. Biofilm formation ability
was established by crystal violet staining. The presence of some virulence genes was assessed by
multiplex PCR assay. Results: A total of 24 E. coli strains were collected. Hemolytic phenotypes
were detected in two out of 24 studied strains (8.3%). The antimicrobial resistance profiles showed
high resistance to penicillin (95.8%), ampicillin (95.5%), imipenem (91.7%), amoxycillin/clavulanic
acid and tetracycline (41.7%), whereas the most efficient antimicrobial was amikacin (100% of
cultures), followed by enrofloxacin (95.8%), ceftiofur (86.9%) and gentamicin (75.0%). 54% (13/24)
of the isolated E. coli resulted to be multidrug-resistant strains. Each tested strain exhibited biofilm-
forming capacity. However, most of them were moderate biofilm producers (54.2%), 20.8% strong
biofilm producers and 25.0% weak biofilm producers. A wide genetic diversity about tested virulence
genes was observed. Discussion and Conclusions: In the present study, the worrying prevalence
of multidrug-resistant E. coli biofilm producing strains represents a serious challenge in the
treatment of mares with fertility problems.
PSB19
EPIDEMIOLOGICAL LINK BETWEEN CLINICAL DHA-1-PRODUCING MDR KLEBSIELLA

PNEUMONIAE ST11 OF FELINE AND HUMAN ORIGIN
C. Marques1,2,3 J. Perdigão4, A Spadar5, J Phelan5, S Campino5, T.G. Clark5,
L. Pinheiro1,2,6, P. Cavaco-Silva7, A. Duarte7,8, C. Pomba1,2,6
1CIISA - Centre of Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine,
Universidade de Lisboa, Lisbon, Portugal

2Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal
3 Faculty of Veterinary Medicine, Lusófona University, Lisbon, Portugal
4iMed.Ulisboa - Research Institute for Medicines, Faculdade de Farmácia, Universidade de Lisboa,
Lisboa, Portugal
5Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine,
London, United Kingdom

6Molecular Veterinary Diagnostic Laboratory - Genevet, Rua Quinta da Nora Loja 3B, 2790-140
Carnaxide, Portugal
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7Centro de Investigação Interdisciplinar Egas Moniz (CiiEM), Instituto Superior de Ciências da

Saúde Egas Moniz, Caparica, Portugal
8Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal
Objectives: This study aimed to characterize two clinical multidrug-resistant (MDR) Klebsiella
pneumoniae (KP) ST11 strains obtained from a cat and an unrelated hospitalized human with
urinary tract infection using whole genome sequencing (WGS). Methods: Disk diffusion
antimicrobial susceptibility against 28 antimicrobials was conducted according to CLSI. Both strains
were subjected to WGS (Illumina HiSeq 4K,2x151bp). De novo assembly was carried out from raw
sequence reads and the genome assemblies characterized for its resistome and virulence gene
content. Phylogenetic reconstruction was performed from a set of genome-wide high-quality SNPs.
Results: Both KP ST11 strains were, wzi75 (KL105), DHA-1 producers and had similar MDR
profiles. Other antimicrobial resistance genes detected in both strains included: blaOXA-1, blaSHV-
11, qnrB4, aac(6')-Ib-cr5, aadA2, sul1, arr-3, catB3, fosA, mph(A) and qacE. The replicon detected
in the cat’s strain (IncR1) was also found in the human´s KP strain. According to the phylogenetic
analysis, these strains differed in only 13 SNP, suggesting an epidemiologically link between them
following the 23 SNP cut-off proposed by Sherry et al. (2019). Noteworthy, the unrelated cat and
human were unrelated, but both lived in Lisbon, therefore pointing to a possible scenario of local
hospital-to-community dissemination. Conclusion: By using NGS, this study brings strong
evidence that cats and unrelated hospitalized humans may become infected with closely related
MDR K. pneumoniae ST11 strains. Thus, dissemination of MDR strains seems to be possible even
in the absence of direct contact between humans and pets, highlighting the need for an OneHealth
approach. Financing: CIISA/FCT project-UIDB/00276/2020; AL4AnimalS-LA/P/0059/2020.
PSB20
ENVIRONMENTAL CONTAMINATION WITH MULTIDRUG RESISTANT BACTERIA AND

CARBAPENEMASE-PRODUCING ACINETOBACTER SPP. IN PORTUGUESE SMALL ANIMAL
VETERINARY PRACTICES
J. M. da Silva1,2, J. Menezes1,2, C. Santos3, L. Fernandes1,2,3, A. J. Amaral1,2,
C. Pomba1,2,3
1
CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine,
2AL4AnimalS - Associate Laboratory for Animal and Veterinary Sciences , Portugal
3Molecular Veterinary Diagnostic Laboratory - Genevet, Rua Quinta da Nora Loja3B, 2790-140
Carnaxide, Portugal
Objectives: Little is known about the prevalence and transmission of multidrug resistant (MDR)
bacteria in Veterinary Medicine, particularly in small animal veterinary practices (SAVPs). The aim
of this study is to determine the prevalence of multidrug resistant bacteria in SAVPs. Materials and
Methods: Six SAVPs were studied. Environmental samples from critical surfaces were collected.
Nasal swabs were voluntarily obtained from Veterinarians and staff. All swabs were plated on
specific media selective for resistant bacteria: ESBL- and carbapenemase-producing
Enterobacterales; Methicillin-Resistant Staphylococcus (MRS) and MDR Acinetobacter. Gram
negative isolates were screened by PCR for the presence of major families of beta-lactamases and
carbapenemase genes. Staphylococci isolates were screened by PCR for the presence of mecA
gene. Results: At least one resistant isolate was found in 20.8% (n=26/125) of the analysed
surfaces: i) 58% (n=15/26) were positive for MDR Acinetobacter spp.; ii) 19% (n=5/26) were positive
for MRS Staphylococcus pseudintermedius; iii) 8% of the surfaces (n=2/26) were positive for MRS
Staphylococcus epidermidis. In one SAVP, 18.2% of surfaces analysed (n=4/22) tested positive for
OXA-23-producing Acinetobacter spp. Forty-three percent of human nasal isolates carried the
mecA gene, of which 20% (n=6/30) were MRS Staphylococcus aureus. In one SAVP, one
Veterinarian and one nurse were nasally colonized by a CTX_M-15-producing Klebsiella
pneumoniae. Conclusions: This study demonstrates a concerning level of environmental
contamination, which highlights the need for harmonization of infection, control and prevention
guidelines SAVPs to prevent the dissemination of multidrug resistant bacteria onto the community.
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PSB21
ASSOCIATIONS BETWEEN ENTEROCOCCI SPECIES ISOLATED FROM ORNAMENTAL

ANIMAL FEED AND GENES OF ANTIBIOTIC RESISTANCE
C. Miranda1-4,†, R. Soares1,†, A. Martins5,6, S. Cunha1, L. Ferreira1, G. Igrejas4,7,8, P. Poeta1,4,6,9,
University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal

2Department of Sciences, University Institute of Health Sciences (IUCS), CESPU, CRL, Gandra,
Portugal
3Toxicology Research Unit (TOXRUN), IUCS, CESPU, CRL, Gandra, Portugal
Caparica, Portugal
5Department of Zootechnics, UTAD, Vila Real, Portugal
6Animal and Veterinary Research Center (CECAV), UTAD, Vila Real, Portugal
7Department of Genetics and Biotechnology, UTAD, Vila Real, Portugal
8Functional Genomics and Proteomics Unit, UTAD, Vila Real, Portugal
†
These authors have contributed equally to this work.
Ornamental animals can represent an important reservoir of Enterococcus carrying resistances to

various antibiotics, allowing their transmission to humans by direct or indirect contact through the
environment, fomites and contaminated feces. In this line, the aim of this study was to investigate
the presence of statistical associations between three obtained enterococci species from
ornamental animal feed between February and December 2020 and their antibiotic resistance. Fifty-
six enterococci isolates, E. faecalis (n=22), E. faecium (n=19) and E. gallinarum (n=15) were tested
by PCR assay for the presence of the resistance genes for 13 antibiotics like vancomycin (vanA
and vanB), erythromicyn (ermA, ermB and ermC), gentamicin (aac(6’)-aph(2’’)-Ia), tetracycline
(tetL, tetM and tetK), quinupristin/dalfopristin (vatD and vatE), chloramphenicol (catA) and
streptomycin (ant(6)-Ia). The data was analyzed by the chi-square independence test, using the
SPSS 15® software. A probability level (p) <0.05 was considered statistically significant in the
association of variables. The genotypic characterization showed that the most prevalent resistance
genes detected in enterococci species were ermB (54%), tetL (48%), vanA (37%), tetM (32%), tetK
(30%), and ant(6)-Ia (30%). The statistical analysis showed significant associations for the ermB
(p=0.0001), tetM (p=0.0002), tetL (p=0.0014), vanA (p=0.0205) and ant(6)-Ia (p=0.0015) resistance
genes. Thus, E. faecalis was more likely to be positive for tetM, E. faecium for ermB and tetL, and
E. gallinarum for ermB, tetL, vanA and ant6-Ia. In conclusion, E. gallinarium was the enterococci
species that demonstrated a higher probability of carrying a variety of antibiotic resistance genes.
Acknowledgements: This work was supported by the Associate Laboratory for Green Chemistry -
LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and
UIDP/50006/2020) and by the projects UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the
Portuguese Foundation for Science and Technology (FCT).
PSB22
PREVALENCE STUDY OF ANTIBIOTIC RESISTANCE GENES IN E. faecalis AND E. faecium

ISOLATED FROM BOVINE COLOSTRUM
S. Cunha1, C. Miranda1-4, A. Martins5,6, R. Soares1, M. Maia1, F. Silva6,7, G. Igrejas4,8,9,
P. Poeta1,4,6,7,10,
1Microbiology and Antibiotic Resistance Team (MicroART), University of Trás-os-Montes and Alto
Douro (UTAD), Vila Real, Portugal

2Department of Sciences, University Institute of Health Sciences (IUCS), CESPU, CRL, Gandra,
Portugal
3Toxicology Research Unit (TOXRUN), IUCS, CESPU, CRL, Gandra, Portugal
Caparica, Portugal
5Department of Zootechnics, UTAD, Vila Real, Portugal
6Veterinary and Animal Research Centre (CECAV), UTAD, Vila Real, Portugal
7Department of Veterinary Sciences, UTAD, Vila Real, Portugal
8Department of Genetics and Biotechnology, UTAD, Vila Real, Portugal
9Functional Genomics and Proteomics Unit, UTAD, Vila Real, Portugal
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Enterococcus faecalis and E. faecium, two of the most prevalent nosocomial agents worldwide, are
frequently isolated from animals and animal foods, including bovine colostrum. These bacteria are,
in most cases, resistant to a large number of antibiotics, which they are capable of transmitting to
others. The aim of this study was to assess whether there is a statistical association between the
presence of an antibiotic resistance gene or virulence gene in the enterococci isolate, obtained from
bovine colostrum, and its identified species. Forty-four E. faecalis isolates and eleven E. faecium
isolates were tested, by PCR assay, for the presence of antibiotic resistance genes and virulence
genes. For the statistical analysis, the chi-square (χ²) independence test and Fisher's exact test
were performed, using the SPSS 15® program. A probability level (p) <0.05 was considered
statistically significant in the association of variables. Statistically significant relationships were
obtained for the erm(C) and tet(M) resistance genes (p=0.003 and p=0.048, respectively), in which
E. faecium was more likely to be positive for both. For virulence genes, significant relationships
were observed for ace and gel(E) genes (p=0.007 and p=0.024, respectively), showing that both
are more prevalent in E. faecium. According to this study, E. faecium isolates from calf colostrum
are more likely to act as a reservoir and transmission vehicle for erythromycin and tetracycline
resistance genes, and collagen-binding protein and gelatinase genes, than E. faecalis isolated from
colostrum. Acknowledgements: This work was supported by the Associate Laboratory for Green
Chemistry - LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and
UIDP/50006/2020) and by the projects UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the
Portuguese Foundation for Science and Technology (FCT).
PSB23
EVALUATION OF THE LOWEST CONCENTRATION/TIME RATIO OF THE OXYGEN/OZONE

GAS MIXTURE AGAINST BACTERIA RESPONSIBLE OF METRITIS
A. Trotta, E. Lillo, A. Rizzo, A. Carbonari, M. Cordisco, M. Corrente
Department of Veterinary Medicine, University of Bari “Aldo Moro”, S.P. per Casamassima km. 3
70010 Valenzano (BA),
Ozone is a gas with oxidizing power and showing a strong microbicidal activity on bacteria, fungi,
viruses and protozoa. The aim of this study was to in vitro determine the lowest dose of gaseous
oxygen/ozone (O3/O2) mixture, and the minimum exposition time, showing the optimal antibacterial
effect, on different bacterial species. A pilot study was carried out on reference strains (E. coli
ATCC 25922, Staphylococcus aureus ATCC 25923) starting from 0.5 McFarland suspension,
diluted 1:1000 in 5 ml sterile saline and then plated on Petri dishes, which was exposed to a gaseous
constant flow of an O3/O2 mixture respectively at 50, 35 and 20 μg Ozone/mL (O3/mL) for 5, 3 and
1 minutes. Based on preliminary results, a collection of 17 field isolates from cases of cow metritis
(belonging to Klebsiella pneumoniae, Enterobacter agglomerans, E. coli, S. aureus and Proteus
mirabilis), was subjected to the minimum concentration of O3/mL for the lowest exposition time, and
bacterial growth was evaluated after 24/48 hrs by counting the UFC/ml in Petri dishes. The
experiments highlights the bactericidal activity of the 20 μg O3/mL for 1 minute against all the field
isolates. Ozone appears to have an optimal antibacterial activity on different bacteria and the
promising in vitro low dose/exposition time, encourages the possible in vivo application, as part of
the strategy to limit the use of antimicrobials in bacterial diseases such as metritis in cows.
OA14
ANTIMICROBIAL RESISTANCE, VIRULENCE CHARACTERISTICS AND PHYLOGENETIC

DIVERSITY OF ESCHERICHIA COLI ISOLATED FROM FOOD-PRODUCING ANIMALS
A. Silva.1-6, V. Silva 1-6, R. Cordeiro7, G. Igrejas2,5,6, V. Falco8, P. Valentão 9 and P.Poeta1-4
University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.

2LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology,
Universidade Nova de Lisboa, Caparica, Portugal.

Vila Real, Portugal

4 Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Portugal
78
Italy
5 Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD),
Vila Real, Portugal.
6 Functional Genomics and Proteomics Unit, University of Trás-os-Montes and Alto Douro (UTAD),
Vila Real, Portugal.

7 Intergados, SA, Av. de Olivença, S/N 2870-108 Montijo, Portugal
8 Chemistry Research Centre (CQ-VR), University of Trás-os-Montes and Alto Douro (UTAD), Vila
Real, Portugal.
9REQUIMTE/LAQV, Laboratório de Farmacognosia, Departamento de Química, Faculdade de
Farmácia, Universidade do Porto.
Antimicrobial resistance has emerged in food-producing animals and the antibiotic therapy induces
the selection of resistance bacteria. Not only limited the therapy options but also generate the
spread of resistance bacteria from livestock to humans and cause a major public health problem.
The main objective of this work was to determine the antibiotic resistance profiles of Escherichia
coli isolates from pigs’ farms in Portugal and undertake the presence of virulence and antibiotic
resistance genes, and to carry out a phylogenetic classification of the isolates. Of the 45 E. coli
strains obtained, 8.8% of them belong to the D phylogenetic group, which is implicated in
extraintestinal infection and 57.4% of them belong to the A phylogenetic group and 31.1% belong
to the B1 phylogenetic group, which is indicated that are commensal strains. All the isolates
presented a multi-resistance profile, with high levels of resistance to β-lactams, aminoglycosides,
sulfonamides and tetracycline. Twenty-four resistance genes were detected, the most frequent of
which were ampC and blaTEM conferring resistance to β-lactams and sul3 conferring resistance to
sulfonamides.The presence of genes involved in the expression of aerolysin (aer), necrotizing
factor-type 1 (cnf 1), type1-fimbriae (fimA), haemolysin (hly), typeC-fimbriae (papC) and adhesin
PapG class III (papGIII) was also investigated. In total, 97.7% of the strains carried virulence factors.
This study has revealed a high incidence of resistance to commonly used antimicrobials in animal
production and human medicine. Food-producing animal represent an important reservoir of E. coli
isolates with genes with the potential risk to humans. This work was supported by the projects
UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the Portuguese Foundation for Science and
Technology (FCT). This work was supported by the Associate Laboratory for Green Chemistry -
LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and
UIDP/50006/2020). Vanessa Silva and Adriana Silva are grateful to FCT (Fundação para a Ciência
e a Tecnologia) for financial support through the PhD grant SFRH/BD/137947/2018 and
SFRH/BD/04576/2020, respectively.
OB18
DETECTION OF SALMONELLA ENTERICA SUBSP. ENTERICA SEROVAR KENTUCKY IN

FALCO TINNUNCULUS
A. Bianco1, L. Capozzi1, L. Del Sambro1, D. Simone1, M. Galgano2, R. Fraccalvieri1, L.M.
Difato1, L. Palazzo1, I. Pietragalla1, M. Toce1, A.C. Romano1, A. Parisi1
1Experimental Zooprophylactic Institute of Apulia and Basilicata, Via Manfredonia, 20 - 71121
Foggia,Italy
2Dipartimento di Medicina Veterinaria (DIMEV) dell'Università degli Studi di Bari Aldo Moro,Italy
Wild birds are considered potential sources of zoonoses because they can host enteropathogens
and can became antimicrobial resistance (AMR) reservoirs. In this study we describe the isolation
and the molecular typing of Salmonella spp. from Falco tinnunculus. The Whole Genome
Sequencing (WGS) was performed using MiSeq platform (Illumina) to the aim to characterize the
isolate. Draft genome was assembled using SPAdes v3.15.4 and annotated using Prokka (Version
1.14.6). The serovar and Sequence type (ST) were predicted by Salmonella In Silico Typing
Resource (SISTR) and Multi Locus Sequence type (MLST) analysis, respectively. AMR genes were
detected using biotool ABRIcate (Version 1.0.1). The isolate was typed as Salmonella enterica
subsp. enterica serovar Kentucky (S. Kentucky) belonging to ST152. The S. Kentucky ST152 is
frequently isolated from asymptomatic cattle and poultry in United States, which represent probable
reservoirs, but rarely from wild animals. Generally, in recent years, S. Kentucky has been described
as cause of infection in food animals in several countries. Thus, to safeguard the welfare of domestic
animals and human health, it is therefore important to know the antibiotic resistance of strains
isolated in wild animals. The in silico prediction identified AMR genes correlated with resistance to
beta-lactams, aminoglycosides and fluoroquinolones. The antimicrobial resistance was confirmed
79
Italy
by phenotypic test. Among these, fluoroquinolone is listed as “Critically important antimicrobials” by

the World Health Organization. Our results stressed the importance of a continuous bacteria
surveillance in wild animals.
80

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4th International Conference of the European College of Veterinary Microbiology, 15-17 September 2022, Bari,

DR. LIBBY GRAHAM

PROF. CANIO BUONAVOGLIA

PROF. DR. PATRICK BUTAYE

DR. ANTONIO MARTÍNEZ-MURCIA

ORIGIN AND STUDIES:

In charge of the courses of Veterinary Mycology at Department of Veterinary Medicine and

Hepatitis E virus (HEV) is a member of the Hepeviridae family, Orthohepevirus genus,

MALDI-TOF MS has revolutionized bacterial identification in both human and veterinary

The European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT)

Objectives: Bacterial resistance to antimicrobials and biocides is an increasing threat.

DIFFERENT GENOTYPES OF MALASSEZIA PACHYDERMATIS ISOLATED FROM DOGS

NOVEL POTENTIAL ANTIFUNGAL COMPOUNDS WITH DUAL MECHANISM OF ACTION

VULTURES’ ORAL MICROBIOME: KEYSTONE YEAST COMMENSALS AND CROSS-

M. P. Couto1,2, F. Lopes3, M. Liede3, M. Casero4, M. Grilo1,2, E. Cunha1,2, M. Pereira5, R. Dias5,

DE NOVO SEQUENCING OF PROVIDENCIA ALCALIFACIENS TO STUDY ITS ROLE IN

Sciences, Ås, Norway

APPLICATION OF MOLECULAR METHODS AND MASS SPECTROMETRY FOR THE

Mastitis is one of the most common problems on dairy farms.

ISOLATION AND CHARACTERIZATION OF STREPTOCOCCUS AGALACTIAE FROM BULK

THE CERVICAL MICROBIOME OF SHEEP BREEDS WITH DIFFERENT FERTILITY RATES

Sciences, Ås, Norway

PB 5003, 1432 Ås, Norway

NEXT GENERATION SEQUENCING ON THE VAGINAL MICROFLORA OF MARES

Valenzano, Bari, Italy

Biotechnology, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece

Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece

Translational Research, Biomedical Research Foundation of the Academy of Athens, 11527

Engineering Sciences, Strand Campus,WC2R 2LS, King’s College London,UK

DEVELOPMENT OF A MYCOPLASMA HYORHINIS CHALLENGE MODEL IN FOUR WEEK OLD

PREVALENCE OF PARATUBERCULOSIS IN DAIRY CATTLE HERDS AND APPROCHES TO

Paratuberculosis (paraTB), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a

PHENOTYPIC SURVEILLANCE FOR ANTIMICROBIAL SUSCEPTIBILITY IN VETERINARY

PREVALENCE AND RISK FACTORS ASSOCIATED WITH FAECAL CARRIAGE OF

ASSESSMENT OF RELIABILITY AND REPRESENTATIVENESS OF ANTIMICROBIAL

ROLE OF COMPANION ANIMAL-HUMAN HOUSEHOLDS IN THE DISSEMINATION OF

University of Lisbon, Lisbon, Portugal

Objectives: To determine the colonization and sharing of colistin resistant, extended-spectrum

HETERORESISTANCE IN A MULTI-RESISTANT CLINICAL ENTEROBACTER CLOACAE

Medicine, Freie Universität Berlin, Berlin, Germany

Environment (Dpt. 4), Berlin, Germany

Introduction: Heteroresistance (HR) describes the phenomenon of a subpopulation that grow in

THE ENOVAT SURVEY ON CURRENT METHODOLOGIES USED FOR BACTERIAL

D. Timofte1, I. Cvetkovikj2, F. Zendri1, S. Blum3, S.C. Chaintoutis4, P.A. Kopp5, C. Hare6, Z.

Liverpool, Neston, United Kingdom

University of Thessaloniki, Thessaloniki, Greece

University of Zagreb, Croatia

Laboratory of Health, Environment and Food, Ljubljana, Slovenia

Utrecht, the Netherlands

University of Copenhagen, Frederiksberg, Denmark.

THE PERSISTENCE OF METHICILLIN-RESISTANT STAPHYLOCOCCUS

RECOMMENDATIONS FOR THE THERAPY OF LOWER URINARY TRACT INFECTIONS IN

PROFILING OF THE MICROBIOTA FROM APULIAN COW MILK MOZZARELLA PRODUCED

EVIDENCE OF THE WIDE CIRCULATION OF MULTI-DRUG RESISTANT ENTERIC

This study was aimed to assess the circulation of antimicrobial-resistant, Gram-Negative

PATTERNS AND DETERMINANTS OF PORCINE REPRODUCTIVE AND RESPIRATORY

ENTERIC VIRAL CO-INFECTIONS IN ACUTE GASTROENTERITIS OF CATS