Post-mortem examination of a wild muntjac from Northern Ireland.
Dick, J. T. A., McKillen, J., Chapman, N., Collins, L., Provan, J., Freeman, M., Hogg, K., & Reid, N. (2012). Post-mortem examination of a wild muntjac from Northern Ireland. (12 ed.) Northern Ireland Environment Agency.https://www.doeni.gov.uk/publications/post-mortem-examination-wild-muntjac-northern-ireland
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Post-mortem examination of a wild
muntjac from Northern Ireland
www.doeni.gov.uk/niea Post-mortem of a wild muntjac in NI
Research and Development Series 12/12 A report commissioned by the Northern Ireland Environment Agency
Post-mortem examination of a wild muntjac from Northern Ireland
Authors: Jaimie Dick, John McKillen, Norma Chapman, Lisa Collins, Jim Provan, Marianne Freeman, Kayleigh Hogg & Neil Reid
Contractor: Quercus NHRP contract number: CON 2/1 (241) Quercus Project: QU11-05
ISSN 1751 – 7796 (Online)
This report should be cited as:
Dick, J.T.A., McKillen, J., Chapman, N., Collins, L., Provan, P. Freeman, M. Hogg, K. & Reid, N. (2012) Post-mortem examination of a wild muntjac from Northern Ireland. Report prepared by the Natural Heritage Research Partnership (NHRP) between Quercus, Queen’s University Belfast and the Northern Ireland Environment Agency (NIEA). Northern Ireland Environment Agency Research and Development Series No. 12/12, Belfast
For further information on this report please contact: John Early,
Northern Ireland Environment Agency,
Biodiversity Unit, Klondyke Building,
Gasworks Business Park,
Lower Ormeau Rd,
Belfast.
BT7 2JA.
The opinions expressed in this report do not necessarily reflect the current opinion or policy of
the Northern Ireland Environment Agency.
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EXECUTIVE SUMMARY
1. Deer are of major concern with regards to impacts on biodiversity, forestry and agriculture
as well as human health. The invasive Reeves’ muntjac deer Muntiacus reevesi, native to
Asia but established in Great Britain, has recently appeared in the wild in Ireland.
2. The first verified record in the wild in Northern Ireland was confirmed during June 2009 as
a result of a road traffic accident near Newtownards, Co. Down. The second record was a
culled animal shot in the grounds of Mount Stewart, Co. Down during June 2011.
3. The current report aimed to perform a detailed investigation of the most recently obtained
animal to establish a) its age, b) its genetic relationship with the first animal and c) the
threat it might pose in terms of carrying endo- or ecto-parasites or other micro-pathogens,
principally viruses and bacteria.
4. Analysis of its dentition suggested the culled animal was 56 weeks old (range 55-57
weeks).
5. Genetic analysis indicated that there was no possibility of a father-son relationship with the
buck killed in 2009. There was a 6.25% probability of both individuals being full-sibs and a
25% probability of a half-sib relationship.
6. The deer was free from all the major pathogens. Most significantly, a novel species of
Gammaherpesvirus was detected most closely matching type 2 ruminant rhadinovirus
(Gammaherpesvirinae) from mule deer. Further sequencing is required to provide a
definitive classification. There is no evidence suggesting this virus is pathogenic but its
detection is nonetheless of concern.
7. During late 2009 (after the first animal was recovered) and early 2010, there were a
number of sightings of muntjac within the vicinity of Mount Stewart. Our results indicate
that the animal shot in June 2011 could not have been this same animal, as the shot
animal must have been born in spring 2010. Moreover, genetic analysis indicates that the
two recovered individuals were highly unlikely to share the same mother and father,
suggesting they were the offspring of a minimum of three breeding adults (two fathers and
one mother or two mothers and one father). This brings the total number of known
individuals to 5 including the two offspring. The location of the breeding adults is unknown
and may be either in captivity with subsequent escapees, or deliberate releases, or be free
living in the wild. However, a further sighting during early 2012 may suggest a wild origin.
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INTRODUCTION
Deer, especially introduced species, are of major concern with regards to impacts on
biodiversity, forestry and agriculture (Putman & Moore 1998; Delahay, De Leeuw &
Claridge 2001). Deer also affect humans directly, causing fatalities and injury
through deer-vehicle collisions (Putman 1997). Consequently, expansion of
introduced and native deer populations will present considerable management
challenges in the future (Ward, 2005).
The rapidity of the spread of introduced Reeves’ muntjac Muntiacus reevesi (Ogilby
1839) is of great concern currently due to deliberate translocations and natural
dispersal from their initial inoculation site(s) in the south-east of Great Britain (Ward
2005).
The first verified record of a muntjac in the wild in Northern Ireland was confirmed on
the 3rd June 2009 as a result of a road traffic accident near Newtownards, Co. Down
(J554764). It was confirmed by a veterinary surgeon that the injuries presented were
consistent with a recent deer-vehicle collision, including broken limbs, blunt trauma
and scrape marks. The animal was a young buck. Identification was further verified
by two independent muntjac experts (Dick et al. 2011). This is the first record of a
new large mammal species in Northern Ireland since the introduction of Sika deer
Cervus nippon (Temminck 1838) in 1870.
This record followed the first reports of muntjac in the Republic of Ireland during
2007 when a free-living adult buck was shot near Avoca, Co. Wicklow, with a second
animal sighted alive shortly thereafter near Trooperstown (Dick et al. 2011). Muntjac
are held and bred in captivity throughout Ireland (Jaimie Dick pers. obs.) and
anecdotal sightings of the species in the wild in both Northern Ireland and Republic
of Ireland have increased in recent years. These include aggregations of sightings
near Mount Stewart on the Ards Peninsula, Co. Down (Freeman, Reid & Dick, 2010)
as recently as February 2012 (Nigel Lafferty pers. obs., Ballyhaft Road,
Newtownards).
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Reeves’ muntjac was listed as one of the ‘most unwanted’ non-native species by
Invasive Species Ireland (www.invasivespeciesireland.com). The Government in the
Republic of Ireland have taken action to facilitate the eradication of any establishing
population(s) by placing the species on the Wildlife (Wild Mammals) (Open Seasons)
Amendment Order 2008. Dick, Provan & Reid (2009) reviewed candidate control
procedures for muntjac and outlined five ‘Options’ for response should the species
be confirmed in the wild. In Northern Ireland, muntjac has been added to Schedule 9
(Part I) of the Wildlife Order (NI) 1985 (as amended) which makes it an offence ‘if
any person releases or allows to escape into the wild any animal which is included in
Part I of Schedule 9’. In addition, to help facilitate control of muntjac, changes have
been made to Article 20 (Section 8a) in relation to the minimum calibre of gun
suitable for deer culling to account for the species-specific characteristics of muntjac.
In common with the Republic of Ireland, there is no closed season for muntjac in
Northern Ireland. Moreover, there is now a cross-border co-ordinated approach to
the species with the publication of an All-Ireland ‘Invasive Species Action Plan’
(ISAP) to further bolster recommendations for action if the species is confirmed in
the wild.
A second confirmed record of muntjac was made in Northern Ireland when a young
buck was shot at Mount Stewart, Co. Down on 11th June 2011 (Jaimie Dick, pers.
obs.). The current report aimed to perform a detailed investigation of this carcass to
establish:
1. The age of the culled animal
2. Whether this animal was related to the previous animal killed in a
road traffic accident
3. Whether the animal carried any endo- or ecto-parasites or other
micro-pathogens, principally viruses/bacteria e.g. bovine tuberculosis
(bTB)
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METHODS
Ageing
The jaw of the animal was examined by Norma Chapman to describe dentition for
the purposes of ageing. The conformation of the teeth was recorded and compared
to a series of reference specimens of known age. The age was estimated in weeks
and is accurate to within 2 weeks.
Genetics
Genetic analysis was performed on both the original buck killed in a road-traffic
accident during 2009 and the second buck shot at Mount Stewart during 2011.
Microsatellite markers developed for M. crinifrons were used (Wu et al. 2008).
Post-mortem examination
A standard post-mortem examination was conducted of the animal culled in June
2011 by the Veterinary Science Division, Agri-food and Biosciences Institute (AFBI).
Routine bacteriology and mycobacterial analysis was performed. Parasitological
examination was conducted for a range of common infections. Virology PCR was
conducted on a range of tissues including for a large range of standard bovine
viruses and using degenerate ‘broad spectrum’ assays for other/novel viruses.
Sequencing was used for confirmation of PCR positives.
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RESULTS
Ageing
Muntjac have lost their first premolar during their recent evolution, thus the first tooth
in the row is called second premolar, in both upper and lower deciduous and
permanent dentitions.
1. The right mandible was in 2 pieces and had a loose molar.
Conformation of the teeth were as follows: I1 I2 I3 c pm2 pm3 broken, pm4 (3 cusps confirms it is deciduous), M1 M2 and separate from bone M3 (partly erupted). The tip of permanent replacement of pm2 was just
appearing from the inner (lingual).
2. In the portion of right maxilla there were as follows: pieces of pm2 pm3
pm4 M1 M2 not fully erupted and M3 erupting. Above pm4 there was its
permanent replacement developing. One upper canine of which 14mm
protruded below gum.
Given the conformation of the teeth, this specimen was judged to be 56 weeks of
age, falling within the variation typically seen from reference specimens that are 55
and 57 weeks of age.
Genetics
Genetic analysis indicated that there was no possibility of a father-son relationship
between the original buck killed in a road-traffic accident during 2009 and the second
buck shot at Mount Stewart during 2011. There was a 6.25% probability of both
individuals being full-sibs and a 25% probability of a half-sib relationship.
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Post-mortem examination
The deer was free from gross signs of disease and all major pathogens. No gross
lesions present in oeseophagus, trachea, lungs, heart, rumen, reticulum, omasum,
abomasum, small and large bowel, liver, kidney, spleen, mesenteric lymph node,
pancreas or thyroid gland. Alimentary tract contents normal.
Routine bacteriology found low levels of Streptoccocus spp. and Bacillus
licheniforms but this is what is expected in a healthy animal. Results indicate that
there was no E. coli which is highly unusual for an ungulate species.
Mycobacterial analysis was negative in mesenteric lymph node and heart. Faeces
negative for Johne’s disease.
The parasite count was zero for worms, Fasciola eggs, paramphistome eggs,
Coccdia and worm eggs.
Virological PCR and sequencing was carried out on kidney, heart, spleen, lung,
bronchial lymph node thyroid, caecum, faeces, oesophogus, colon, small intestine
regions 1-6, pancreas, liver, abomasun, medistinal lymph node, mesenteric lymph
node, trachea, rumen contents, blood clot and blood.
The carcass was negative for bovine herpesvirus 1 (infectious broncial rhinitis virus),
bovine parainfluenza virus 3, bovine viral diarrhoea virus, bovine respiratory syncytial
virus, bluetongue virus, kobuvirus, calicivirus (norovirus and sapovirus), anellovirus,
adenovirus, coronavirus, reovirus, orthoreovirus, rotavirus and hepatitis E virus.
However, colon, mediastinal lymph node, lung and blood were weakly positive for
herpesvirus. The band amplified from the colon was sequenced and identified as
matching rhadinovirus of mule deer (gammaherpesvirus). A number of tissues were
also positive for picornavirus and further sequencing is needed determine the correct
classification.
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Other PCR or RT-PCRs tests were carried out for astrovirus, bocavirus, circovirus,
paramyxovirus, bovine hokovirus and bovine parvovirus type 2. Assays were
negative, but results were inconclusive due to lack of fully and properly validated
assays/controls.
DISCUSSION
The chronology of events surrounding muntjac in the Ards peninsula, Co. Down,
Northern Ireland is thus:
1. From May 2009, anecdotal muntjac sightings began to increase in frequency near
Mount Stewart, with a photograph nearby at Spring Lane (25th May 2009).
2. On 3rd June 2009, a buck muntjac was killed in a road-traffic accident on the Killaughy
Road approximately 3.5km from the photographed animal the preceding month.
3. However, anecdotal sightings continued in the area throughout late 2009 and early 2010
(Freeman et al. 2010).
4. On 11th June 2011, a 55-57 week old buck was shot at Mount Stewart (the subject of
this report). This animal could not have been responsible for earlier sightings during late
2009 or early 2010 (subsequent to the first animal being killed in a road-traffic accident)
as it must have been born during spring 2010.
5. Genetic analysis indicates that the two individuals were highly unlikely to share the
same mother and father, suggesting parentage from a minimum of three breeding adults
(two fathers and one mother or two mothers and one father). This brings the total
number of known individuals to 5 including the two offspring. The location of the
breeding adults at present is unknown and they may be either held in captivity or may
represent an establishing feral population.
6. A further muntjac sighting was made during February 2012 near Mount Stewart on the
Ballyhaft Road (Nigel Lafferty pers. obs.) supporting the latter hypothesis that there may
be multiple males and females in the wild raising the possibility of an establishing feral
population.
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The post-mortem carried out here suggests that the animal recovered poses little
threat in terms of parasites or pathogens (i.e. no risk from bovine TB, Johne’s
disease or salmonella). Notably, there were low levels of bacteria in the animal
digestive tract (specifically, no E. coli) and no parasite fauna which may be
characteristic of an animal that had been treated recently with antibiotics and
anthelmintics. However, there may be natural reasons for this lack bacterial and
parasite communities so it is not certain that this can be taken as an indication that
the animal had been in captivity prior to being shot.
However, the animal was positive for picornavirus and herpesvirus, the latter of
which closely matched a sequence of type 2 ruminant rhadinovirus
(Gammaherpesvirinae) from mule deer. Other Rhadinovirus members include
bovine herpesvirus-4, implicated in post-partum metritis (Donofrio et al., 2009) and a
number of primate viruses associated with cancer in immunosuppressed individuals
such as Kaposi sarcoma herpesvirus i.e. human herpesvirus 8 (Cesarman, 2011).
The sequence matched with lower homology to a wide range of Gammaherpesvirus
from the Rhadinovirus and Macavirus genera, including ovine herpesvirus 2, genus
Macavirus, which causes sheep-associated malignant catarrhal fever (Russell et al.,
2009), and caprine herpesvirus 2, genus Macavirus, which is known to cause
malignant catarrhal fever in some species of deer. However, there are a number of
ruminant Macaviruses that are known to cause no clinical disease (Li et al. 2005).
The discovery of a novel species of Gammaherpesvirus is notable but its
significance as a pathogen of deer is unknown at this time. While there is no
evidence to suggest that this virus is pathogenic, and while this taxonomic group
contains non-pathogenic viruses of ruminants, the relatedness of the virus to known
pathogens of ruminants is of some concern as there is always a possibility, however
small, of a benign virus evolving to become more pathogenic or to expand its host
range. In addition it is well known for apparently harmless viruses proving pathogenic
when introduced to immunologically naive or susceptible populations. So while the
risks associated with this virus are probably small the fact that DNA from a
completely novel virus has been identified in such a small sample would suggest that
there is significant value in future virological work involving invasive species,
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particularly muntjac deer. Further sequencing of the viral genome through standard
molecular methods would allow definitive classification of the virus and a
metagenomic analysis of other samples using next generation sequencing would be
a powerful method of further virus discovery.
RECOMMENDATIONS
We make 3 recommendations for action:
1. Integrated and co-ordinated species surveillance and monitoring to document
the establishment, spread and impact of muntjac on the Ards Peninsula.
Specifically, proactive detection involving baited camera trap stations in Mount
Stewart estate and surrounding area coordinated by the ‘Mount Stewart
Eradication Plan’ members (principally National Trust, Forest Service, Local
Shooting Syndicate, Invasive Species Ireland, QUB and NIEA) in order to
detect further individuals in the wild and initiate culling.
2. Proactive detection and reaction throughout Northern Ireland at sites of other
possible innocula i.e. locations of escapees or releases (see Freeman, Reid &
Dick, 2010). Data on anecdotal sightings are held by Queen’s University Belfast
and the National Biodiversity Data Centre, Waterford.
3. Any future carcasses should be subjected to similar analyses as presented
here in order to estimate population sizes and further assess disease profiles.
Specifically, further sequencing of the novel Gammaherpesvirus genome
through standard molecular methods to facilitate definitive classification of the
virus and a metagenomic analysis of future muntjac samples using next
generation sequencing would be a powerful method of further virus discovery.
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ACKNOWLEDGEMENTS
Thanks to Jason Barle and Jim McCarthy, Disease Surveillance and Investigation
Branch, Agri-Food and Biosciences Institute for carrying out the post-mortem
examination (2011-20287). Conrad Watson (Disease Surveillance and Investigation
Branch) and Eric Walton (Bacteriology) carried out routine bacteriology and
mycobacterial analyses respectively, Hillary Edgar (Disease Surveillance and
Investigation Branch) preformed parasitiological tests and Paula Lagan and Michael
McMenamy (Virology) carried out virology PCR and sequencing. John Early acted as
NIEA Client Officer.
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