Journal of Infection and Public Health 16 (2023) 194–202

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Journal of Infection and Public Health

journal homepage: www.elsevier.com/locate/jiph

Original Article

Prevalence and characterization of antimicrobial-resistant Escherichia coli

isolated from veterinary staff, pets, and pet owners in Thailand ]]
]]]]]]
]]

Shutipen Buranasinsup a, Anuwat Wiratsudakul b,c, Boonrat Chantong a,

Khuanwalai Maklon b, Sarin Suwanpakdee b,c, Sineenard Jiemtaweeboon b,
⁎
Walasinee Sakcamduang b,
a
Department of Pre-Clinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
b
Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
c
The Monitoring and Surveillance Center for Zoonotic Diseases in Wildlife and Exotic Animals, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom
73170, Thailand

a r t i cl e i nfo a bstr ac t

Article history: Background: Companion animals may act as antimicrobial resistance (AMR) reservoirs. This study in­
Received 10 August 2023 vestigated the prevalence and AMR patterns of Escherichia coli in pets and people in close contact with pets.
Received in revised form 1 November 2023 Methods: A total of 955 samples were collected from veterinary clinics across Thailand by rectal and skin or
Accepted 3 November 2023
ear swabs from dogs and cats and fecal swabs from veterinarians, veterinary assistants, and pet owners. The
minimum inhibitory concentrations (MICs) of the obtained isolates were investigated using Sensititre™ MIC
Keywords:
plates against 21 different antimicrobial drugs.
Antimicrobial resistance
Companion animals Results: Escherichia coli from pets was frequently resistant to ampicillin (100%) and amoxicillin–clavulanic
Escherichia coli acid (100%), whereas E. coli from pet owners, veterinarians, and veterinary assistants was mostly resistant to
Veterinary tetracycline. The multiple antibiotic resistance index revealed that multidrug-resistant E. coli isolates were
frequently found in dogs (34.92%), cats (62.12%), veterinarians (61.11%), veterinarian assistants (36.36%), and
pet owners (47.62%). The most common AMR genes identified in this study were blaCTX-M, blaTEM, tetA, and
tetB, which were associated with the antimicrobial susceptibility results. Additionally, extended-spectrum
beta-lactamase (ESBL)-associated genes (i.e., blaCTX-M, blaTEM, and blaSHV) were found in 21.69%, 71.97%,
27.78%, and 21.43% of E. coli isolated from dogs, cats, veterinarians, and pet owners, respectively.
Conclusions: Our findings demonstrated the presence of AMR genes, particularly ESBL-associated genes, in
E. coli isolated from healthy pets and veterinarians. This implies that these sources of E. coli could potentially
be reservoirs for antibiotic resistance, thereby increasing the risk of harm to both humans and animals.
These findings highlight the importance of implementing effective AMR control measures in veterinary
practices, as bacteria resistant to commonly used antimicrobials are present in humans and animals.
© 2023 The Author(s). Published by Elsevier Ltd on behalf of King Saud Bin Abdulaziz University for Health
Sciences. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/li­
censes/by-nc-nd/4.0/).

Background

Antimicrobial resistance (AMR) is among the most challenging

problems facing the world, and it is a frequent issue across both
human and veterinary medicine. As many antimicrobial drugs used
Abbreviations: AMR, antimicrobial resistance; CLSI, Clinical and Laboratory in human medicine are also utilized in veterinary medicine, anti­
Standards Institute; CPE, carbapenemase-producing Enterobacteriaceae; ECDC, the microbial underuse, overuse, and misuse can result in therapeutic
European Centre for Disease Prevention and Control; E. coli, Escherichia coli; ESBL, failure and increased health consequences in both animals and hu­
extended spectrum beta-lactamase; MAR, multiple antibiotic resistance; MDR, mul­ mans [1–3]. Antimicrobial drugs, when misused, can exert selection
tidrug resistance; MIC, minimum inhibitory concentration; MRS, methicillin-resistant
pressure on both pathogenic bacteria and normal flora in people and
staphylococci; MRSA, methicillin-resistant Staphylococcus aureus; VRE, vancomycin-
resistant enterococci; WHO, World Health Organization animals, and resistant commensal bacteria serve as reservoirs of
⁎
Corresponding author. resistance genes for potentially pathogenic bacteria [4–6].
E-mail address: [email protected] (W. Sakcamduang).

https://doi.org/10.1016/j.jiph.2023.11.006
1876-0341/© 2023 The Author(s). Published by Elsevier Ltd on behalf of King Saud Bin Abdulaziz University for Health Sciences. This is an open access article under the CC BY-NC-
ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
S. Buranasinsup, A. Wiratsudakul, B. Chantong et al. Journal of Infection and Public Health 16 (2023) 194–202

Companion animals such as dogs and cats are increasingly suggested

as potential sources of antimicrobial-resistant bacteria [3,7–9]. An­
timicrobial resistance in companion animals may be caused by
methicillin-resistant Staphylococcus aureus (MRSA), methicillin-re­
sistant staphylococci (MRS), vancomycin-resistant enterococci (VRE),
carbapenemase-producing Enterobacteriaceae (CPE), and extended-
spectrum beta-lactamase (ESBL)-producing bacteria, which present
direct and indirect risks to human health [10]. In addition, Escher­
ichia coli has been identified as a possible source of resistance
transmission [11,12]. The World Health Organization classified E. coli
as one of the 12 species of bacteria that cause human health crises
with continuously increasing antimicrobial resistance [13]. Escher­
ichia coli is a gram-negative bacterium and a member of the En­
terobacteriaceae family present in the intestinal microbiota of
mammals. Most E. coli strains are commensals; however, some
strains are capable of causing life-threatening intestinal and extra-
intestinal infections in humans and animals. Escherichia coli can be
divided into eight phylogroups: A, B1, B2, C, D, E, F, and Clade I, and
phylogroups A, B1, B2, and D are most commonly found [14]. As a
result of frequent exposure to systemic antimicrobial treatment and
high genomic plasticity, the European Centre for Disease Prevention
and Control (ECDC) recommends E. coli as an excellent indicator for
antimicrobial resistance monitoring. Several studies have empha­
sized the importance of monitoring and understanding AMR E. coli
isolated from pets, as they can serve as reservoirs of AMR that may
transmit to humans due to their close contact [7,11,15,16]. In addi­
tion, the prevalence of acquired resistance in commensal E. coli also
indirectly indicates significant selective pressure from the use of
antimicrobials in animal and human populations [4,11,17,18].
Antimicrobial resistance issues in human and animal health can
be sustainably addressed and resolved via a multisectoral approach,
known as One Health, in which multiple health professionals colla­
borate to prevent the development and spread of antimicrobial re­
sistance [19,20]. Companion animals are also important reservoirs of
antimicrobial-resistant bacteria and can act as reservoirs and sen­
tinels of AMR because of their direct interactions with humans, other
animals, and their surroundings [21–24]. In Thailand, the One Health
concept is also used to measure occupational exposure to AMR in pig
Fig. 1. Provinces of Thailand in which the study was conducted.
and poultry farms because animals and humans are colonized by the
same bacterial species and treated with the same antimicrobial
classes [25]. However, more research has yet to be conducted in the their biological samples were collected between June 16th 2020 and
country to determine the presence of E. coli and AMR patterns in May 30th, 2021. Rectal samples from healthy dogs and cats as well as
companion animals and people interacting in the same en­ lesions from skin and ear infections were collected. Fecal swabs from
vironment. healthy humans were also obtained. We have explicitly stated that
This study aimed to explore the prevalence of E. coli in dogs and we collected anonymized data that is not linked to any personally
cats, as well as their antimicrobial susceptibility together with an­ identifiable information to ensure the confidentiality of participants
timicrobial resistance gene pattern, in conjunction with veterinar­ during and after data collection. This research was performed in
ians, veterinary assistants, and the owners of those dogs and cats accordance with ethical guidelines and institutional policies to
from private veterinary clinics/hospitals in Thailand. protect the privacy and confidentiality of the study participants.
The specimens were kept at 4 °C and transported to the labora­
Methods tory within 72 h of collection. Escherichia coli isolates were grown on
the selective medium, MacConkey agar (Oxoid, UK), and identified
Sample collection based on their morphological (Gram stain), and biochemical char­
acteristics following standard microbiological techniques [26]. We
In this study, samples were collected from both humans (veter­ then cultured E. coli for further identification. Subsequently, anti­
inarians, veterinary assistants, and pet owners) and animals (dogs microbial susceptibility testing was performed.
and cats visiting veterinary premises). The selected veterinary clinics
and hospitals were located in five provinces across different regions Identification of Escherichia coli
of Thailand, namely Nakhon Pathom (Central region), Nakhon
Ratchasima (Northeastern region), Chon Buri (Eastern region), For identification of E. coli, the specimens were inoculated on
Chumphon (Southern region), and Chiang Mai (Northern region), as MacConkey agar (Oxoid, UK) and incubated at 37 °C for 24 h. After
presented in Fig. 1. The study locations were purposively selected on incubation, the suspected bacterial colonies (lactose-fermenting
a voluntary basis with formal permission. The number of samples colonies) were identified. Three randomly selected from individual
was proportionally calculated based on population sizes and ratios of samples were subjected to Gram staining (Sigma-Aldrich, Germany).
dogs and cats in each province. Veterinary staff from each premise Conventional bacterial biochemical tests including triple sugar iron
and the animal owners were asked to participate in the study and agar (Oxoid, UK), ornithine decarboxylation/motility/indole (Oxoid,

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S. Buranasinsup, A. Wiratsudakul, B. Chantong et al. Journal of Infection and Public Health 16 (2023) 194–202

UK), and citrate utilization tests (Oxoid, UK) were used to identify Germany), and 1 µg of DNA template. The thermal cycler Flexcycler2
E. coli. (Analytik Jena, Germany) was used to performed the amplification .
The temperature profile was as follows: DNA denaturation at 95 °C
Antimicrobial susceptibility testing for 30 s, followed by annealing at the specific temperature for each
primer set for 30 s, extension at 72 °C for 60 s, and a final extension
Antimicrobial susceptibility testing and interpretation were at 72 °C for 10 min. Each run included both - positive and negative
performed using the minimum inhibitory concentration (MIC) controls. Positive control isolates for the detection of antimicrobial-
method according to the Clinical and Laboratory Standards Institute resistance genes were obtained from previous studies [30]. The
M100 [27] and VET01S [28] for samples isolated from humans and amplicons were analyzed using 1.5% agarose gel electrophoresis and
animals, respectively. The breakpoint for interpretation according to stained with SYBR-safe (Invitrogen, USA). The DNA bands were ob­
CLSI guidelines is shown in Supplement Table 1. served under the UV transilluminator (UVP Bioimaging system, In­
All E. coli isolates were examined for susceptibility to amikacin, vitrogen, USA).
gentamicin, ampicillin, amoxicillin–clavulanic acid, piperacillin/ta­
zobactam, cephalexin, cefazolin, cefpodoxime, cefovecin, ceftazi­ Network analysis
dime, imipenem, enrofloxacin, marbofloxacin, orbifloxacin,
pradofloxacin, trimethoprim/sulfamethoxazole, chloramphenicol, An undirected two-mode weighted network was constructed.
doxycycline, and tetracycline using the Sensititre™ Companion One mode was the antibiotics, and the other mode was the subjects
Animal Gram Negative COMPGN1F Vet AST Plate (Thermo Fisher involved, which included humans, cats, and dogs. However, in this
Scientific, USA). Antimicrobial susceptibility testing was performed analysis, both modes were treated equally, with a node representing
according to the manufacturer’s protocol. In brief, E. coli isolates either an antibiotic or a subject and an edge representing the re­
were diluted to 0.5 McFarland standard, inoculated on the AST plate, sistance between each pair of subjects and antibiotics. The node was
and incubated at 35 °C for 18–24 h. The MIC was determined after weighted based on degree centrality, which is defined as the number
adding resazurin dye (Sigma-Aldrich, Germany). Escherichia coli of immediate contacts of the node [31], and the edge was weighted
isolates from humans were further analyzed for susceptibility to based on the relative rate of resistance. All network analysis and
norfloxacin (Sigma-Aldrich, Germany) and ciprofloxacin (Sigma- visualization studies were performed using the package ‘igraph’ in
Aldrich, Germany). The susceptibility to imipenem, trimethoprim/ the R program.
sulfamethoxazole, chloramphenicol, doxycycline, and tetracycline in
animals was interpreted as per CLSI M100 [27]. Escherichia coli ATTC®
25922 was used as the control strain. Statistical analysis

Multiple antibiotic resistance (MAR) index calculation Descriptive statistics, including the percentage of isolations and
percentage of resistance, were used to describe the results. The an­
The MAR index was calculated and interpreted using the for­ timicrobial resistance of the bacteria was compared using the chi-
mula: a/b, where ‘a’ represents the number of antibiotics to which an square test to determine whether there were differences in regions
isolate was resistant, and ‘b’ represents the total number of anti­ and host categories. Host categories (veterinarian, veterinary assis­
biotics tested [29]. tant, pet owners, dog, and cat) that affected the occurrence of AMR
were also compared using Fisher’s exact test and chi-square test. The
cut-off for the statistical significance was set at a P-value of < 0.05.
Detection of antimicrobial resistance genes

Escherichia coli isolates with antibacterial resistance phenotypes Results

were cultured overnight in 2 ml of Luria-Bertani broth (HiMedia,
USA). The culture pellets were collected after centrifugation at Characteristics of the collected samples
6000 rpm for 10 min. The pellets were then resuspended in 200 µl of
DPEC-treated water. DNA was extracted by heating at 100 °C the In total, 955 samples were from five provinces. Samples were
suspension for 10 min and then centrifuging at 6000 rpm for 10 min most commonly collected from the Northern region (28.90%), fol­
to collect the supernatant. Antimicrobial-resistant genes, including lowed by the Northeastern (28.17%), Eastern (21.88%), Southern
blaCTX-M, blaTEM, blaSHV, aac(3)-IIa, aac(6’)-Ib, tetA, tetB, gyrA, dfrA5, (10.68%), and Central regions (10.37%). Rectal swabs were collected
sul1, sul2, and cmlA, were amplified via 30 cycles of PCR amplifica­ from 264 dogs and 170 cats. Fecal swabs were obtained from 67
tion using the specific primers listed in Supplement Table 2. The PCR owners, 19 veterinarians, and 41 veterinary assistants. Skin and ear
mixture (25 µl) contained 1 µM of each primer, 0.2 mM of dNTP, swabs were collected from 236 dogs and 158 cats with skin and ear
2 mM of MgCl2, 1 U of Taq DNA polymerase (Thermo-scientific, lesions of infection (Table 1).

Table 1
Characteristics and distribution of subjects across sampling locations.

Characteristics of samples Location

Central (Nakhon Northeastern (Nakhon Eastern Southern (Chumphon) Northern Total

Pathom) Ratchasima) (Chon Buri) (Chiang Mai)

Veterinarians (Fecal swab) 4 7 3 0 5 19

Veterinary assistants 6 13 2 0 20 41
(Fecal swab)
Owners (Fecal swab) 16 33 3 3 12 67
Dogs (Rectal swab) 24 81 52 36 71 264
Cats (Rectal swab) 13 42 51 15 49 170
Dogs (Ear or skin swab) 24 61 48 33 70 236
Cats (Ear or skin swab) 12 32 50 15 49 158
Total 99 269 209 102 276 955

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resistant to tetracycline (54.12%), and high rates of resistance to

ampicillin (43.53%) and trimethoprim/sulfamethoxazole (40.00%)
were detected. In dogs, resistance was commonly found for tetra­
cycline (42.33%), ampicillin (37.57%), and doxycycline (29.63%). In
cats, all isolates were resistant to ampicillin and amox­
icillin–clavulanic acid, whereas the resistance rates for the other
tested antimicrobials ranged from 2.27% to 43.94%.

Comparison of antimicrobial resistance patterns

The patterns of AMR from different hosts were compared ac­

cording to the number of antimicrobials to which the isolates were
resistant. In this study, 29% of the E. coli isolates from dogs and 47%
from cats were resistant to ≥4 antimicrobials. Of the human groups,
Fig. 2. The percentage of human and animal samples classified according to
56% of the E. coli isolates from veterinarians were resistant to ≥ 4
sample type. medications. A comparable pattern was noted for multidrug re­
sistance (MDR), which describes resistance to ≥3 drug classes [32].
Occurrence of Escherichia coli Multidrug resistance phenotypes were found in 34.92% and 62.12%
of the E. coli isolates from dogs and cats, respectively. Veterinarian E.
Our findings indicated that E. coli was predominantly isolated coli isolates displayed high MDR phenotypic prevalence rates
from rectal swabs in dogs and cats, and from fecal swabs in humans. of 61.11%.
Fig. 2. illustrates the percentage of E. coli collected in each category. Multiple antibiotic resistance (MAR) indices of E. coli isolated
Table 2 reports the occurrence of E. coli among veterinarians, ve­ from dogs, cats, veterinarians, veterinary assistants, and pet owners
terinary assistants, owners, dogs, and cats from different locations in were also calculated. The results showed that 36.89% of the E. coli
Thailand. isolates had an index of > 0.2. High MAR index of > 0.2 in E. coli was
found in veterinarians, which was followed by samples obtained
from cats. MAR index is presented in Fig. 5.
Antimicrobial susceptibility testing
Antimicrobial resistance genes
Escherichia coli isolates from dogs were most frequently resistant
to tetracycline (42.33%), whereas those from cats were resistant to
Escherichia coli isolates identified as phenotypically resistant to
ampicillin (100%) and amoxicillin–clavulanic acid (100%). Regarding
antimicrobials were further investigated for the presence of re­
E. coli, isolates from owners were commonly resistant to tetracycline
sistance genes. Escherichia coli isolated from different hosts fre­
(61.9%); those from veterinarians were resistant to sulfamethox­
quently contained blaCTX-M, blaTEM, tetA, and tetB, which was
azole/trimethoprim (55.56%) and tetracycline (55.56%); and those
confirmed using the antimicrobial susceptibility test. High in­
from veterinary assistants were frequently resistant to ampicillin
cidences of blaCTX-M, blaTEM, and blaSHV genes encoding beta-lacta­
(36.36%), doxycycline (36.36%), and tetracycline (36.36%). The results
mases, enzymes that can inactivate beta-lactam antibiotics such as
of the antimicrobial susceptibility test are presented in Fig. 3.
penicillins and cephalosporins, were identified in E. coli isolated
from animals, veterinarians, and pet owners. This study identified a
Network analysis high incidence of tetA and tetB, which encode tetracycline efflux
pumps in E. coli in all groups consisting with the antimicrobial re­
Fig. 4. provides an overall picture of AMR in E. coli isolates from sistance phenotype. The results of the antimicrobial resistance genes
humans, dogs, and cats. We found that E. coli was resistant to 19 of are shown in Table 3.
21 tested antimicrobials in both dogs and cats, whereas it was re­
sistant to only 13 antimicrobials in humans (represented by degree Association between antibiotic resistance and hosts (humans and
centrality of the nodes). Escherichia coli was resistant to more than animals)
half of the tested antimicrobials (11/21) in all species and to eight
antimicrobials in two of the three species. Focusing on the rates of Table 4 shows the association between antibiotic resistance and
resistance, more than half of the E. coli isolates from humans were different categories of E. coli host. Only amoxicillin-clavulanic acid

Table 2
The occurrence of E. coli was analyzed among different subject groups and regions, and the results are reported as percentages.

Host category Location

Central (Nakhon Northeastern (Nakhon Eastern Southern (Chumphon) Northern Total

Pathom) Ratchasima) (Chon Buri) (Chiang Mai)

Veterinarians 40.00 18.75 14.29 4.76 0.00 12.50

(2/5) (3/16) (1/7) (1/21) (0/7) (7/56)
Veterinary assistants 33.33 0.00 17.65 2.13 0.00 6.25
(3/9) (0/32) (3/17) (1/47) (0/7) (7/112)
Owners 32.00 0.00 8.45 3.16 0.00 5.56
(8/25) (0/75) (6/71) (3/95) (0/40) (17/306)
Dogs 83.33 (20/24) 23.08 32.14 25.00 2.78 29.10
(12/52) (27/84) (18/72) (1/36) (78/268)
Cats 84.62 17.65 42.86 22.45 13.33 30.00
(11/13) (9/51) (18/42) (11/49) (2/15) (51/170)
Total 57.89 10.62 24.89 11.97 2.86 17.54
(44/76) (24/226) (55/221) (34/284) (3/105) (160/912)

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S. Buranasinsup, A. Wiratsudakul, B. Chantong et al. Journal of Infection and Public Health 16 (2023) 194–202

Fig. 3. Profile of antimicrobial resistance in E. coli isolates from pets and humans. a) antimicrobial resistance in E. coli from pets. b) antimicrobial resistance in E. coli from humans.

was significantly different between humans and animals, and only transfer between humans and animals [9,33,34]. The emergence and
two antibiotics demonstrated a significant difference between dogs spread of E. coli strains carrying AMR genes in people, animals, and
and cats. No differences were identified among human groups. their environments are major global concerns [11,15,16,35]. Under­
standing the distribution of bacteria and their susceptibility to an­
Discussion timicrobial drugs is critical for antimicrobial resistance monitoring
[36]. The presence of resistant E. coli in pets and humans may be
The increasing proximity between humans and companion ani­ linked to the spread of drug-resistance genes within the same ha­
mals has raised concerns regarding the significant risk of AMR bitats [24,34,37]. Therefore, this study aimed to determine the

Ciprofloxacin Norfloxacin

Ampicillin
Cefazolin
Tetracycline

Ceftaxidime Imipenem

Doxycycline
Cepodoxime

Human

Gentamicin
Trimethoprim/Sulfamethoxazole
Dog Cat

Amoxicillin−clavulanic acid

Chloramphenicol

Orbifloxacin

Cephalexcin

Enrofloxacin
Amikacin Human
Pradofloxacin
Dog
Cefovecin
Cat

Piperacillin/Tazobactam Antibiotic
Marbofloxacin

Fig. 4. Networks of antimicrobial resistance in E. coli among humans, dogs, and cats in veterinary care settings. The size of the nodes denotes the proportional degree centrality of
the nodes, the width of the edges denotes the relative rate of resistance, and the color of the nodes and edges represents animal species and antimicrobials.

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S. Buranasinsup, A. Wiratsudakul, B. Chantong et al. Journal of Infection and Public Health 16 (2023) 194–202

Fig. 5. Multiple antibiotic resistance (MAR) index of E. coli isolated from dogs, cats, veterinarians, veterinary assistants, and pet owners.

pattern of antimicrobial susceptibility of E. coli in dogs and cats as resistance to ampicillin and tetracycline, and all isolates from cats
well as veterinarians, veterinarian assistants, and pet owners at were resistant to ampicillin [4,11,17,39,41]. In this study, the pro­
private veterinary hospitals and clinics and determine their anti­ portions of E. coli from dogs and cats with an MAR index of ≥ 0.2
microbial resistance genes. The minimum inhibitory concentrations were 34.92% and 62.12%, respectively, which were lower than those
of E. coli against 21 antimicrobial drugs were investigated. In addi­ observed in China [42]. The frequency of E. coli isolated from veter­
tion, the 12 antimicrobial resistance genes (blaCTX-M, blaTEM, blaSHV, inarians with an MAR index of ≥ 0.2 was relatively high, at 61.11%.
aac(3)-IIa, aac(6’)-Ib, tetA, tetB, gyrA, dfrA5, sul1, sul2 and cmlA) were According to previous reports from Austria and China, companion
determined. animals might act as overflow carriers for multidrug-resistant E. coli
In this study, E. coli was isolated from the rectal swabs of healthy in humans [43,44]. Our findings suggest a critical need for increased
dogs and cats, and also from skin and ear infection lesions. The awareness of the possibility of multidrug-resistant E. coli reservoirs
prevalence of E. coli isolated from pets was consistent with earlier in pets and veterinarians that may be shared during a visit or ad­
findings, in which it was 7–43% in dogs and 5–45% in cats mission to veterinary facilities. Our observations, although of con­
[11,17,38–40]. In this study, the prevalence of E. coli in people who cern, also provide an opportunity for proactive action. The need for
were in contact with pets was higher than that in previous studies. optimal infection control measures for veterinary practitioners in
According to Jung et al., the prevalence of E. coli was in the range of Thailand is clear, and the time to act is now. The absence of specific
0%-6% in veterinary staff, 41.11% in veterinarians, and 14.63% in ve­ infection control guidelines in small animal practice in Thailand
terinary assistants [39,40]. We found that E. coli isolates from cats should not stall the progress. Instead, it emphasizes the urgency for
were 100% resistant to ampicillin and amoxicillin–clavulanic acid, the country to develop and implement comprehensive infection
whereas those from dogs were most frequently resistant to tetra­ control protocols by leveraging international guidelines [45–47] and
cycline (42.33%). Escherichia coli from people was most frequently integrating them into the local context and existing national stra­
resistant to tetracycline, sulfamethoxazole/trimethoprim, and dox­ tegic plan on AMR [48].
ycycline. Similar to previous findings, E. coli isolated from humans The antimicrobial resistance genes found most commonly in this
and companion animals in this study exhibited high rates of study were bla and tet genes, which correlated with the results of the

Table 3
Antimicrobial resistance genes of E. coli isolated from dogs, cats, veterinarians, veterinary assistants, and pet owners.

Drugs classes Genes Dogs N = 189 (%) Cats N = 132 (%) Veterinarians N = 18 (%) Veterinary assistants N = 11 (%) Pet owners N = 42 (%)

Beta-lactams blaCTX-M 21.69 71.97 27.78 - 21.43

blaTEM 24.87 43.18 44.44 36.36 42.86
blaSHV 8.99 20.45 11.11 - 4.76
Aminoglycoside aac(3)-IIa 5.29 8.33 - - 7.14
aac(6’)-Ib 2.65 7.58 - - -
Tetracycline tetA 26.46 37.88 44.44 27.27 42.86
tetB 26.46 40.91 33.33 36.36 54.76
Quinolone gyrA 8.99 14.39 5.56 - 19.05
Trimethoprim dfrA5 6.88 15.15 22.22 - 14.29
Sulfonamide sul1 5.29 2.27 22.22 18.18 14.29
sul2 12.7 10.61 27.78 9.09 21.43
Chloramphenicol cmlA 3.7 13.64 11.11 - 4.76

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Table 4 Companion animals can serve as a reservoir for E. coli strains

Association between antibiotic resistance and different categories of hosts (humans carrying genes associated with antimicrobial resistance, which have
vs. animals, veterinarians vs. veterinary assistants vs. pet owners, and dogs vs. cats) of
the potential to be transmitted to humans via regular contact
Escherichia coli.
[24,55,59,60]. Consequently, the findings of this research, together
Antibiotics P-value with good veterinary practice, can be applied to reduce the transfer
Humans vs Human groups Animal of AMR bacteria between pets and humans [61]. Optimizing anti­
Animals (veterinarians vs groups (dogs microbial use in veterinary clinics will limit the selection and spread
veterinary vs cats)
of resistant bacteria among both pets and humans [11]. The pre­
assistants vs pet
owners) vention of AMR transmission among pets and their owners should
be prioritized via the promotion of improved personal and profes­
Amikacin 1.00 NA# 1.00
Amoxicillin- < 0.001*** NA# < 0.001***
sional hygiene, as well as client education. Moreover, the connection
clavulanic acid between AMR in animals and people at veterinary facilities, in­
Ampicillin 0.15 0.86 < 0.001*** cluding both direct and indirect paths, must be explored further.
Cefazolin 0.17 0.73 0.19 Nevertheless, we acknowledge that the environmental dimension is
Cefovecin NA NA 0.25
missing from the One Health concept. Indeed, environmental con­
Cefpodoxime 0.25 0.77 0.98
Ceftazidime 1.00 0.17 0.71 tamination with AMR bacteria should not be overlooked in hospital
Cephalexin NA NA 0.13 settings. Several factors influence the pattern and frequency of AMR
Chloramphenicol 0.81 0.81 0.08 among veterinary staff, pet owners, and pets at veterinary clinics/
Ciprofloxacin NA 0.26 NA hospitals, including antimicrobial types and dosages, the exposure
Doxycycline 0.39 0.25 0.84
Enrofloxacin NA NA 0.05
frequency of different animals, and personal hygiene [62]. The po­
Imipenem 1.00 NA# 1.00 tential for AMR in owners and pets has also been documented, in­
Gentamicin 0.12 0.77 0.22 cluding human–companion animal interactions, knowledge
Marbofloxacin NA NA 0.05 regarding the causes of AMR, and owner perspectives of anti­
Norfloxacin NA 0.26 NA
microbial usage. Further studies are warranted to demonstrate the
Orbifloxacin NA NA 0.13
Pradofloxacin NA NA 0.05 link between AMR in pets and humans using molecular techniques,
Piperacillin/ NA# NA# NA# such as random amplification of polymorphic DNA–PCR, multilocus
Tazobactam sequence typing, and whole-genome sequencing. Additionally, fur­
Tetracycline 0.42 0.77 0.89 ther studies are needed to explore strategies to reduce AMR occur­
Trimethoprim/ 0.06 0.91 0.37
Sulfamethoxazole
rence in veterinary practices, including establishing an antimicrobial
stewardship policy, enhancing personal hygiene, and improving
***indicates statistically significant relationships for p < 0.001
biosafety in occupational health.
vs: versus
NA: No association between groups of hosts available
NA#: No statistics were computed. Conclusions

Our findings emphasize the issue of antibiotic resistance in E. coli

antimicrobial susceptibility test. The detection of any one of the isolates from pets, pet owners, and veterinarians in Thailand that
variants, including CTX-M, TEM, and SHV gene products, confirmed had a high frequency of E. coli with the MAR index and AMR-related
the presence of ESBL-producing enterobacteria. BlaCTX-M and blaTEM genes. Furthermore, E. coli with ESBL genotypes (blaCTX-M, blaTEM, and
were more predominant than blaSHV in the E. coli isolated from all blaSHV) and tetracycline resistance genes (tetA and tetB) are pre­
species (Table 3). The observed presence of blaCTX-M and blaTEM in valent in veterinarians and pets. The results suggest that preventing
E.coli isolates corresponded to previous findings, indicating a high AMR transmission among veterinary professionals and pets during
level of detection that identified the presence of these genes in physical examination, clinical procedures, and admission should be
gram-negative isolates from healthcare and community settings prioritized. The results also indicate the need to promote client
[49,50]. Both patients and healthy people are increasingly carrying education for improving the personal hygiene of pet owners. Our
ESBL-producing E. coli in their intestinal tract, which is a global findings also demonstrate the importance of persistence in mon­
concern in community and healthcare settings [51,52]. Additionally, itoring the presence of MDR and AMR genes among E. coli in pet
studies have suggested that companion animals potentially serve as animals.
ESBL reservoirs [16,53]. The high prevalence of ESBL genotypes
(specifically blaCTX-M, blaTEM, and blaSHV) observed in our study might
Funding
have significant implications on the health of pet owners, veterinary
personnel, and pets, as well as on the surrounding environment. The
This research was funded by Mahidol University (Grant No. 11569
study identified additional AMR genes of significance, particularly
and Grant No. 23039).
tetA and tetB, which correlated with tetracycline resistance in E. coli.
Efflux mechanisms encoded by tetA and/or tetB genes have been
Consent for publication
reported to be the common pathway of tetracycline resistance in E.
coli isolated from humans and animals in several countries
Not applicable.
[34,54–57]. The occurrence of tetA, tetB, and tetC genes was observed
in most tetracycline-resistant E. coli isolates obtained from both
healthy and sick pets in the United States, Mexico and Portugal Data availability
[55,56,58]. In addition, the fecal E. coli isolates in the present study
exhibited comparable patterns of virulence and antimicrobial re­ The data used to support the findings of this study are available
sistance genes among both pets and their owners whichagrees with from the corresponding author upon request.
a previous study [34]. Therefore, it is possible that E. coli strains
obtained from both human and pet sources, which harbor genes Declaration of Competing Interest
linked to antimicrobial resistance, can disseminate enteric patho­
gens and antibiotic resistance determinants. The authors have no conflict of interest to declare.

200
S. Buranasinsup, A. Wiratsudakul, B. Chantong et al. Journal of Infection and Public Health 16 (2023) 194–202

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