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New Zealand Veterinary Journal
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Mycobacterium avium subsp. paratuberculosis infection
in wildlife on three deer farms with a history of Johne's
disease
G Nugent
a
, EJ Whitford
a
, JC Hunnam
b
, PR Wilson
c
, ML Cross
a
& GW de Lisle
d
a
Landcare Research Manaaki Whenua, PO Box 40, Lincoln, 7640, New Zealand
b
Johne's Management Limited, PO Box 161, Ashburton, New Zealand
c
Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Private Bag 11222,
Palmerston North, 4442, New Zealand
d
AgResearch, National Centre for Biosecurity and Infectious Disease Wallaceville, PO Box 40063,
Upper Hutt, New Zealand
Accepted author version posted online: 28 Jul 2011.Published online: 31 Oct 2011.
To cite this article: G Nugent , EJ Whitford , JC Hunnam , PR Wilson , ML Cross & GW de Lisle (2011): Mycobacterium avium subsp.
paratuberculosis infection in wildlife on three deer farms with a history of Johne's disease, New Zealand Veterinary Journal, 59:6,
293-298
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Scientic Article
Mycobacterium avium subsp. paratuberculosis infection in wildlife
on three deer farms with a history of Johnes disease
G Nugent*
x
, EJ Whitford*, JC Hunnam
{
, PR Wilson
{
, ML Cross* and GW de Lisle
#
Abstract
AIM: To determine the prevalence of Mycobacterium avium
subsp. paratuberculosis (Map) infection in wildlife, in pastoral
landscapes with a recent history of clinical Johnes disease in
livestock.
METHODS: A total of 449 wild mammals and birds from
three farms in the South Island of New Zealand with recent
histories of clinical Johnes disease in their deer herds were
trapped and examined for gross pathological changes in the
gastrointestinal tract. Additionally, individual mesenteric lymph
nodes from 380 mammals, and segments of gastrointestinal
tract from 32 birds were excised, homogenised and cultured for
viable Map bacilli. The prevalence of Map infection was then
calculated for the various species. Faecal samples from those
mammals which had culture-positive tissues were further
cultured for the presence of Map.
RESULTS: Gross pathological changes were identied in the
gastrointestinal tract of four brushtail possums, one cat, six
ferrets, 12 hares, six hedgehogs, three rabbits, one stoat, and
one paradise shelduck. Infection with Map in the gastrointest-
inal tract was conrmed in only three of these cases, one each of
brushtail possums, hares and hedgehogs. In contrast, Map
infection in the absence of gross pathological changes was
frequently recorded in enteric tract tissues of mammals and
birds. Among mammals, Map infection was recorded in 18/73
(25%) brushtail possums, 4/23 (17%) cats, 15/42 (36%)
hedgehogs and 29/113 (26%) rabbits. Among birds, intestinal
tract tissue Map infection was recorded in 3/17 (18%) paradise
shelducks. Among 64 of the 74 mammals which had Map
culture-positive tissues, 38% (n5) of hedgehogs and 11%
(n3) of rabbits also had culture-positive faecal samples.
CONCLUSIONS: This study is the rst to identify that Map
infection can be prevalent in wildlife in New Zealand. There
was a high prevalence of Map infection among both scavenging
and grazing wild animals. Both mammals and birds are capable
of harbouring viable Map organisms in their gastrointestinal
tract; further, viable Map was excreted into the environment via
faeces by hedgehogs and rabbits.
CLINICAL RELEVANCE: Previous studies overseas have
postulated a role of wildlife as reservoirs of Map infection and
possible vectors of Johnes disease to livestock. Here, brushtail
possums, hedgehogs and rabbits and in particular were
identied as potential wildlife hosts for Map infection in
New Zealand. This suggests that several wildlife species could
contribute to the persistence of Map infection within a wildlife/
livestock complex, and potentially, perhaps more importantly,
to the spread of infection between farms.
KEY WORDS: Mycobacterium avium subsp. paratuberculosis,
Johnes disease, infection, wildlife, infection reservoir
Introduction
Johnes disease is an enteric inammatory disease, caused by
chronic infection with the bacterium Mycobacterium avium
subsp. paratuberculosis (Map). Johnes disease has long been
present in farmed cattle and sheep in New Zealand and since the
mid-1980s has also emerged as a potentially major problem on
deer farms (de Lisle et al. 1993; Mackintosh and de Lisle 1998).
The annual cost from loss of production caused by the disease has
been estimated at between NZ$3.8 million and NZ$31.7 million
for dairy farmers (Burton and Voges 2002), and is also a serious
economic burden to some deer farmers (Glossop et al. 2005).
There is also a possibility that Map may have human health
implications as there is an association between Map infection and
Crohns disease, an enteric inammatory condition in humans,
although the causality of this association remains unknown
(Mendoza et al. 2009). This drives concerns about the potential
presence of Map in dairy and meat products (Rossiter and
Henning 2001), its occurrence in river systems and water supplies
(Pickup et al. 2005), and the increasing prevalence of Map
infection in farmed deer in New Zealand (de Lisle et al. 2003). As
a consequence, the control of Map infection and Johnes disease
in livestock in New Zealand, particularly deer, is assuming greater
importance than in the past.
Different strains of Map show different host species afnities. In
New Zealand, Type S strains of the bacillus infect sheep and less
commonly deer and cattle, whereas Type C strains frequently
infected cattle and deer and less commonly sheep (OBrien et al.
2006; de Lisle et al. 2006). However, other animal species,
particularly wildlife, may also serve as permissive hosts for Map.
* Landcare Research Manaaki Whenua, PO Box 40, Lincoln 7640, New Zealand.
{
Johnes Management Limited, PO Box 161, Ashburton, New Zealand.
{
Institute of Veterinary, Animal and Biomedical Sciences, Massey University,
Private Bag 11222, Palmerston North 4442, New Zealand.
#
AgResearch, National Centre for Biosecurity and Infectious Disease
Wallaceville, PO Box 40063, Upper Hutt, New Zealand.
x
Author for correspondence. Email: [email protected]
CPC Cetyl pyridinium chloride
Map Mycobacterium avium subsp. paratuberculosis
New Zealand Veterinary Journal 59(6), 293298, 2011 293
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Studies in Europe and North America have identied a range of
wildlife species including lagomorphs, wild ruminants and wild
and feral carnivores as hosts for Map infection (Beard et al. 2001;
Marco et al. 2002; Palmer et al. 2005; Raizman et al. 2005;
Pedersen et al. 2008). One study in Europe suggested that Map
infection could be maintained in a wild lagomorph population
(Judge et al. 2006). Thus wild animal populations could act as
reservoirs of Map infection and could, potentially, contribute to
the spread and/or persistence of Johnes disease in livestock.
New Zealand has a diverse exotic mammalian fauna, among
which at least brushtail possums (Trichosurus vulpecula) and
ferrets (Mustela furo) are known to maintain infections of another
mycobacterial pathogen, Mycobacterium bovis, the causative agent
of bovine tuberculosis, and can serve as wildlife reservoirs of that
disease (Caley and Hone 2004). Unlike M. bovis infection, the
status of Map infection in wildlife in New Zealand is currently
uncertain, and the potential for inter-species transmission in
pastoral environments is unknown. If wild animals in New
Zealand can act as maintenance hosts for Map infection and are
capable of transmitting the disease back to livestock, this might
well undermine monitoring and control programmes and farm
husbandry practices being used to control Johnes disease.
In the present study, the prevalence of Map infection in wildlife
on three farms in the South Island of New Zealand in a pastoral
setting was surveyed in 20042005, focussing mostly on the
introduced mammalian fauna, but including some of the avian
species common in pastoral lands. Because deer seem
particularly susceptible to Map infection and Johnes disease
(OBrien et al. 2006), the surveys were conducted on deer farms
that had had recent and ongoing reports of outbreaks of clinical
Johnes disease that had been conrmed as Map infection by
culture.
Materials and methods
Farm proles and Johnes disease history
Cross-sectional surveys for the prevalence of Map infection in
wildlife were conducted on three separate farms in the mid-lower
South Island. The deer herds that comprised the exclusive or
predominant stock on each of these farms had experienced
signicant outbreaks of clinical Johnes disease shortly before the
farms were surveyed in 20042005.
Farm A, South Canterbury
Farm A was 4,200-ha sheep, cattle, and deer farm comprising
*1,000 ha of alluvial at land and rolling downs with bushy
gullies and woodlots, and *3,200 ha of steeper hill country with
rougher tussocky pasture and large areas of native bush. It was
stocked with approximately 3,000 deer, mostly breeding stock for
venison production, and *9,000 sheep. Clinical Johnes disease
was rst conrmed by necropsy and histological examination in
eight adult red deer (Cervus elaphus) in 1995, and up until the
time of the survey annual losses in deer attributed to Johnes
disease had averaged *5% of the adult stock. Wildlife were
surveyed around this farm site in December 2004, through most
of the 1,340 ha that were deer-fenced (*1,000 ha, 340 ha at
and downs).
Farm B, South Canterbury
This farm comprised 220 ha of ats and downs, with interspersed
willow trees (Salix spp.), pine trees (Pinus spp.) and gorse (Ulex
europaeus). About 1,000 deer were farmed on the property,
mainly for velvet production, but with some on-farm breeding of
replacement stags. Some sheep were also farmed. Clinical Johnes
disease had been recorded on this farm since autumn 2003, when
49/350 (14%) weaners, 20/100 (20%) rising 2-year-old stags and
several adult hinds died. Fewer cases were reported in 2004, but
occasional deaths in adult deer, particularly older stags, still
occurred. The wildlife survey was conducted in April 2005, and
covered the entire farm.
Farm C, North Otago
This 2,024-ha farm consisted of 500 ha of ats and lower terraces
and 1,500 ha of hill country. At the time of survey only deer were
farmed; sheep and cattle had last been grazed on the pastures in
2001, prior to it being deer-fenced. Deer on this site were farmed
primarily for venison, and were apparently free from Johnes
disease until 2001, when the herd was supplemented by external
stock. In 2002, six animals were diagnosed with clinical Johnes
disease by post-mortem examination and bacteriological culture.
In 2003, serological tests for Johnes disease were positive or
suspicious for 22/201 (11%) yearling hinds. Further animals
were diagnosed with clinical Johnes disease in 2004 and 2005
but in lesser numbers than in 2002 (Grifn et al. 2007). The
wildlife survey was conducted in autumn 2005, and was conned
to the intensively farmed area of ats and terraces.
Collection of samples
Each survey was conducted over a period of about a week, and
involved shooting or trapping of most of the wild mammal
species present other than mice (Mus musculus) and, largely
incidentally, a few of the bird species. The mammals included
brushtail possums, feral cats (Felis catus), ferrets, hares (Lepus
europaeus occidentalis), hedgehogs (Erinaceus europaeus occidenta-
lis), rabbits (Oryctolagus cuniculus), rats (Rattus norvegicus) and
stoats (Mustela erminea). The birds surveyed included Australa-
sian harriers (Circus approximans), black-backed gulls (Larus
dominicanus), magpies (Gymnorhina tibicen), paradise shelducks
(Tadorna variegata), spur-winged plovers (Vanellus miles), and
starlings (Sturnus vulgaris). The aim was to obtain at least 30
individuals of each of the common mammals on each farm, but
the numbers actually collected depended on the combination of
local density and the ease with which they could be trapped or
shot. The hunting effort was spread over the widest possible area
within each farm over which Johnes disease-infected deer were
present.
Trapping and shooting techniques were approved by Landcare
Researchs Animal Ethics Committee, Lincoln, New Zealand.
Brushtail possums and cats were captured using leg-hold traps
(Victor No.1; Philproof Pest Control Products, Hamilton, NZ)
lured with our or meat, and placed approximately 20 m apart
along bush edge margins, forested gullies or woodlots. Ferrets
were captured in Holden kill traps (Philproof Pest Control
Products) baited with rabbit meat, set *100 m apart along fence
lines and other habitat boundaries favoured by ferrets. Hedgehogs
were mostly shot, although some were caught incidentally in leg-
hold traps set for brushtail possums, cats and stoats. Hares,
rabbits, and the various bird species were shot using either a .22-
calibre rie or 12-gauge shotgun. Rats were trapped using snap-
traps lured with peanut butter, placed around farm buildings and
haysheds.
Most captures were necropsied within 12 h of death, but 22
(mostly ferrets) were stored frozen for some weeks before
examination. The sex, weight, and age class (juvenile/adult) were
294 New Zealand Veterinary Journal 59(6), 2011 Nugent et al.
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recorded post mortem. At necropsy, the abdominal organs of
each carcass were then examined for any pathological changes
suggestive of mycobacterial lesions. Samples routinely collected
included any anomalous gastrointestinal tract, e.g. discoloured or
thickened intestinal walls, enlarged mesenteric nodes, or any gross
lesions seen in the gastrointestinal tract and/or associated lymph
nodes; mesenteric lymph nodes (mammals); 510-mm sections
of the duodenum, ileum, caecum and colon in mammals, or of
the post-caecal gastrointestinal tract for birds; and faecal samples
from the lower gastrointestinal tract.
New gloves, and freshly sterilised instruments and work surfaces
(cutting boards) were used for each animal to prevent cross
contamination between animals. The above tissues were frozen
(7208C) within 2 h of collection, and later prepared for culture
and identication of Map, as outlined below.
Identication of viable Map bacilli
The presence of viable Map organisms in tissues was identied
using mycobacterial culture, based on methods described
previously (de Lisle et al. 2003). Tissues were homogenised
using a Coleworth Stomacher (Seaward, Norfold, UK), deconta-
minated in 0.35% cetyl pyridinium chloride (CPC) for
40 minutes, and then centrifuged at 3,500g for 20 minutes.
The supernatant was discarded, and the pellet reconstituted in
1 mL sterile distilled water, 0.5 mL of which was inoculated into
a BACTEC 12B vial (Becton Dickinson, Sparks MO, USA)
supplemented with egg yolk, mycobactin and antibiotics
(PANTA; Becton Dickinson). The vials were incubated at
378C, and examined weekly for the release of
14
CO
2
, for a
minimum of 8 weeks. Faecal samples were decontaminated using
the procedure based on the double-incubation method described
by Whitlock and Rosenberger (1990). Approximately 2 g of
faeces was added to 40 mL sterile distilled water, vigorously
shaken, and allowed to stand for 30 minutes. A 5-mL aliquot
off the top of the liquid was added to 1% CPC. After
decontaminating overnight, the samples were centrifuged, the
supernatants discarded, and the pellets reconstituted in 1 mL of
an antibiotic cocktail (Whittington et al. 1998). After a further
incubation period of 3 days, the sample was inoculated into a
supplemented BACTEC 12B vial, as described for the tissues.
Mycobacteria were identied as Map based on criteria described
previously (de Lisle et al. 2003).
The mesenteric lymph nodes were excised, processed and
cultured from each mammal for Map. Additionally, any
anomalous tissues from the gastrointestinal tract or gross lesions
were cultured. Culture of faecal samples from an individual
was only conducted if either the mesenteric lymph node or a
lesion proved culture-positive. For birds, a homogenate of the
gastrointestinal tract was cultured.
Statistical analysis
All analyses were performed using GLM in the statistical
package GenStat (GenStat for Windows, Release version 8.1,
8th Edition 2002; VSN International Ltd, Oxford, UK).
Binomial regression was used to analyse data, with each
individual animal being characterised as infected with Map or
not infected. The proportion of animals with Map was
modelled using the independent variables of farm (three levels),
sex (two levels), age class (two levels, juvenile and adult), and
sex-by-age class interaction. Initially, this model was tted to
the data, and any non-signicant variables were removed by
backward elimination (Crawley 2002), until a minimum
adequate model was obtained; this analysis was undertaken
to provide data on the prevalence of infection for each species
individually. A separate but similar analysis was carried out to
investigate whether any differences in the prevalence of
infection of individual species was dependent on the farm
where the animal was killed, i.e. location, using the variables of
farm (three levels) and species (six levels). No analyses were
performed on rats, stoats and any of the bird species as sample
sizes were too small.
Results
Gross pathological ndings
In total, 449 wild animals were trapped, killed and necropsied.
Tissues from 412/449 (92%) of them were submitted for culture
and provided a diagnostic result, as outlined in Table 1. Tissues
could not be cultured from 37 animals due to errors in processing
and collection of samples.
At a macroscopic level, abnormal gastrointestinal tract tissue
suggestive of inammatory enteritis was identied in four
brushtail possums, one cat, six ferrets, 12 hares, six hedgehogs,
three rabbits, one stoat, and one paradise shelduck. The most
common gross pathological changes were foci of tissue
discolouration and thickening in the intestinal wall, and
enlargement of the mesenteric lymph nodes (in mammals). In
two cases, one hare and one rabbit, there were noticeable lesions
in a small area of the mesenteric lymph nodes; however, neither
case was conrmed by culture as Map infection. Additionally, one
hedgehog had a single 20-mm diameter lesion in its mesenteric
lymph node, which was conrmed by culture as Map infection.
The only other two cases of gross pathological changes in the
gastrointestinal tract which were conrmed by culture as Map
infection involved a brushtail possum which had noticeable
diarrhoea on capture, and a hare with vascularisation and
thickening of the intestinal submucosa.
Prevalence of Map infection in tissues
Culture of gastrointestinal tract tissues identied Map infection
in 78/412 (19%) animals from which tissues were cultured
(Table 1). Infection was detected in six mammal and two bird
species, viz brushtail possums, cats, ferrets, hares, hedgehogs,
rabbits, black-backed gulls and paradise shelducks. The raw
prevalence of infection varied between species, and, overall, was
highest in brushtail possums, cats, hedgehogs, and rabbits (Table
1). In total, one quarter of the samples from these four species
were culture-positive for viable Map organisms. Prevalence also
varied between farms, with about one-tenth of the animals on
Farm C infected compared with a quarter of those from the other
two farms. There was an interaction between farm and species
(w
2
8
29.0; p50.001; Figure 1), indicating that the different
prevalence of Map infection among wildlife species was not
consistent between farms. For example, a low prevalence (2%)
was recorded from samples from rabbits on Farm A, whereas
almost half of the rabbits on Farm C were infected, while the
reverse pattern applied to feral cats, and for hedgehogs the highest
prevalence was on Farm B.
The prevalence of Map infection was higher in adult animals than
in juveniles for both hedgehogs (14/30 adults infected vs 1/12
juveniles; w
2
1
6.53; p0.011) and rabbits (28/85 adults infected
vs 1/28 juveniles; w
2
1
5.27; p0.022). The prevalence in
brushtail possums, cats, ferrets and hares did not differ
Nugent et al. New Zealand Veterinary Journal 59(6), 2011 295
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statistically between age classes, but the number of infected
animals was low in these species.
Prevalence of Map infection in faeces
Faecal samples were collected from 64 individual mammals which
returned culture-positive results for their mesenteric lymph nodes
and/or gastrointestinal tract, comprising four cats, 13 brushtail
possums, two ferrets, ve hares, 13 hedgehogs and 27 rabbits. Of
these samples, eight (13%) animals were Map culture-positive,
comprising 5/13 (38%) hedgehogs and 3/27 (11%) rabbits. All
ve hedgehogs were from Farm A, whereas two rabbits were from
Farm B and one from Farm C. The single brushtail possum
which had noticeable diarrhoea upon capture, and which had
Map culture-positive mesenteric lymph nodes, was Map culture-
negative for its faeces.
Discussion
This case study represents the rst survey for the prevalence of
Map infection in wildlife in any landscape in New Zealand. Of
the eight mammalian and six avian species surveyed, Map
infection was recorded in eight, with a prevalence of up to 57%.
The surveys were conducted in pastoral landscapes where there
was believed to be a substantial source of Map bacilli in the form
of farmed deer with clinical Johnes disease. Thus, the results
possibly represent a higher than average prevalence in wildlife
than may be found from farms with a lower or zero prevalence of
Johnes disease among livestock; further case-controlled studies
would be required to clarify this situation.
Many previous surveys have been conducted in New Zealand
to determine the prevalence of the related mycobacterial
pathogen M. bovis (see overview by Coleman and Cooke
2001). There is substantial overlap in the host range of these
pathogens in New Zealand, and a key question for both is
whether the various wildlife species are true maintenance hosts
capable of forming independent reservoirs or, conversely, whether
the observed prevalences result from spillover from infected
livestock or other species, and whether the infected wildlife can
then pass the pathogen back to livestock, i.e. spillback (Nugent
2011). For Map infection, single cross-sectional surveys such as
this are of little use in characterising host status, but some
inferences are possible. A faecaloral transmission pathway, with
spillover infection arising from the ingestion of Map-infected deer
faeces, or feed contaminated with infected faecal material, could
explain the high prevalence of infection in the predominantly
herbivorous species, the brushtail possums, hares, rabbits and
paradise shelducks. Further, the increased prevalence of Map with
age in rabbits is consistent with an accumulation of spillover
infection through life. However, spillover transmission via
contaminated pasture seems unlikely for the carnivores (feral
cats and ferrets) or omnivores (hedgehogs and black-backed
Table 1. Prevalence (%) of Mycobacterium avium subsp. paratuberculosis infection in wildlife species, with number infected and total number of
individuals from which tissue samples were cultured, as confirmed by bacteriological culture of samples from mesenteric nodes and/or
gastrointestinal tract lesions of mammals, and gastrointestinal tracts of birds, from three farms in the South Island of New Zealand with histories of
Johnes disease among their farmed deer.
Species Farm A Farm B Farm C Total
Mammal
Brushtail possum 26% (8/31) 24% (10/42) 25% (18/73)
Feral cat 43% (3/7) 6% (1/16) 17% (4/23)
Ferret 0% (0/2) 0% (0/6) 8% (3/36) 7% (3/44)
Hare 6% (3/52) 0% (0/10) 14% (2/14) 7% (5/76)
Hedgehog 12% (2/17) 57% (12/21) 25% (1/4) 36% (15/42)
Rabbit 2% (1/42) 24% (6/25) 48% (22/46) 25.7% (29/113)
Rat 0% (0/4) 0% (0/4)
Stoat 0% (0/1) 0% (0/1) 0% (0/3) 0% (0/5)
All mammals 11.2% (17/152) 25.7% (28/109) 24.4% (29/119) 19.5% (74/380)
Bird
Australasian Harrier 0% (0/1) 0% (0/2) 0% (0/3)
Black-backed gull 25% (1/4) 0% (0/1) 20% (1/5)
Magpie 0% (0/3) 0% (0/1) 0% (0/4)
Paradise Shelduck 18% (3/17) 18% (3/17)
Spur-winged plover 0% (0/1) 0% (0/1)
Starling 0% (0/2) 0% (0/2)
All birds 20% (1/5) 0% (0/4) 13% (3/23) 12% (4/32)
0
10
20
30
40
50
60
70
80
Feral
cat
Ferret Hare Hedgehog Rabbit
Species
Brushtail
possum
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(
%
)
Figure 1. Mean (with back-transformed SE) prevalence of Mycobacter-
ium avium subsp. paratuberculosis infection in a total 371 individuals
of six mammalian wildlife species from Farm A (&), Farm B (&) and
Farm C () in the South Island of New Zealand, cultured from samples
collected between December 2004 and April 2005.
296 New Zealand Veterinary Journal 59(6), 2011 Nugent et al.
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gulls). For the carnivores, feeding on Map-infected carcasses of
deer was possible on at least one of the farms, and both cats and
ferrets were likely feeding on brushtail possums, hares, and
rabbits. Overall, regardless of how wildlife become infected, the
presence of Map in wildlife clearly creates a potential risk of
spillback to livestock, as has been suggested from eld studies of
rabbits overseas (Judge et al. 2005; 2006) Further detailed
studies, aimed specically at understanding the modes of
transmission that are operative for Map in wildlife in New
Zealand, will be required to address how each species acquires
infection.
Despite an overall prevalence of Map infection of 19% among
wild mammals in the present study, very few macroscopic lesions
indicative of overt pathological changes in the gastrointestinal
tract were recorded here in wildlife. This nding matches the
results of wildlife surveys for Map in rabbits in Scotland (Beard
et al. 2001; Judge et al. 2006) and white-tailed deer (Odocoileus
virginianus) in Florida (Pedersen et al. 2008), which have
similarly shown a low prevalence of pathological changes in the
gastrointestinal tract among wild mammals that have a high
prevalence of Map infection. It is uncertain whether a proportion
of the Map-infected brushtail possums, hedgehogs and rabbits
here would go on to develop macroscopic lesions in the
gastrointestinal tract, but assuming that the animals surveyed
represented an accurate cross-sectional sample of animal ages, it
seems that few do. In line with that, results from a eld survey
indicated that Map-infected wild rabbits frequently exhibited
low-grade but not macroscopic pathological changes (Beard et al.
2001), while a laboratory infection study of rabbits (Vaughan
et al. 2005) has shown that even two years after oral infection
with Map bacilli, rabbits developed only low-grade lesions in the
gastrointestinal tract. Overall, those observations suggest that
while rabbits may be permissive for Map infection, they are quite
refractory to clinical disease.
Further, in the present study, evidence of Map infection was
found in the gastrointestinal tract of two carnivore species, feral
cats and ferrets, but there was no evidence of concurrent gross
pathological changes. Similarly, other studies have reported Map-
positive tissues in the absence of pathological changes for wild
carnivores from the United States of America, including feral cats
(Palmer et al. 2008) and raccoons (Procyon lotor) (Corn et al.
2005; Pedersen et al. 2008). In contrast, Map-positive lesions in
the gastrointestinal tract have been recorded in carnivores in
Europe, namely stoats and weasels (Mustela nivalis) captured
from a Johnes disease-endemic region in Scotland (Beard et al.
2001), while among wild foxes (Vulpes vulpes) captured in the
same study, half of the animals which had lesions in mesenteric
lymph nodes were culture-positive for Map. In the present study,
Map infection was also recorded in the gastrointestinal tract of
two bird species, namely black-backed gulls and paradise
shelducks. An 18% prevalence of Map infection was recorded
in paradise shelducks, a bird which grazes and which shows a
preference for pastoral habitats, indicating a potential for this
species to be involved in the acquisition and/or transmission of
Map infection. Results from studies overseas have also included
Map infection in wild birds (Corn et al. 2005; Gaukler et al.
2009; Miranda et al. 2009), although again, and in common with
this study, Map infection in birds was recorded largely in the
absence of gross pathological changes in the gastrointestinal tract.
In contrast, infection of birds with Mycobacterium avium subsp.
avium can lead to tuberculous lesions in somatic tissues
and granuloma-type pathological changes (Biet et al. 2005).
Nevertheless, despite a paucity of Map-induced pathological
changes in birds in the present study, the presence of viable Map
in the gastrointestinal tract presents an obvious route by which
viable mycobacteria may be shed into the environment. Thus,
although we have no evidence here that infected birds play any
role in either maintenance or inter-species transmission of Map
infection, there is clearly potential for both them, and wide-
ranging mammalian hosts, to spread the pathogen over large
areas and to provide a route of farm-to-farm transmission of
infection.
In summary, this survey is the rst study to report Map infection
in a number of wildlife species associated with pastoral land in
New Zealand, at least in the case of areas which had a recorded
history of clinical Johnes disease in livestock. Further studies will
be required to determine the prevalence of infection in other
parts of the country, including those pastoral landscapes largely
free of clinical Johnes disease in farmed livestock, and to
determine how widespread Map infection in wildlife is through-
out the country. Additionally, further studies are planned to
DNA type the Map isolates from wildlife and compare them with
isolates obtained from farmed deer from the same properties.
If spillback transmission is operative among wildlife in New
Zealand, then this could confound current methods for
controlling infection within livestock herds, and could present a
risk of spread of Map between properties. Thus, this study had
provided data to justify further investigation of the prevalence of
Map infection in wildlife in New Zealand, and to determine the
role that wildlife plays in the epidemiology of Johnes disease in
livestock.
Acknowledgements
We acknowledge and thank the cooperation of farm owners
during conduction of this study. We thank Gary Yates
(AgResearch, Wallaceville) for mycobacterial culture, and
Fernanda Castillo-Alcala who participated in the post-mortem
examinations and collection of samples. We also thank Dr Colin
Mackintosh (AgResearch, Invermay) for guidance in conducting
this study. This research was nancially supported in part by a
research grant awarded by the Foundation for Research Science
and Technology of New Zealand, and in part by Landcare
Research Capability Funding (Emerging Diseases project), with
additional support from the Johnes Research Group (Massey
University, Palmerston North, New Zealand) and DEEResearch
Ltd (Wellington, New Zealand).
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