Experimental infection of brushtail possums (Trichosurus vulpecula) with Mycobacterium bovis by conjunctival instillation

In New Zealand, the brushtail possum (Trichosurus vulpecula) is the major wildlife reservoir of Mycobacterium bovis. Procedures for experimentally infecting possums are required to study the pathogenesis of the disease and to challenge possums in vaccine efficacy studies. Conjunctival instillation of a suspension of M. bovis was effective in producing bovine tuberculosis in captive possums. The experimental disease progressed slowly with the development of palpable lesions in superficial lymph node lesions, both characteristics of the disease in wild, naturally infected possums. At necropsy there was widespread distribution of macroscopic and microscopic lesions. The proportion of possums that became diseased, the rate of development and severity of lesions, the severity of clinical signs, all increased when the dose of M. bovis was increased. Of the three doses used, the medium dose (1000-2000 colony forming units) produced the disease with the most desired characteristics. As a procedure for exposing possums to infection with M. bovis the conjunctival route has advantages in that it is simple and safe to perform, and possums need only to be sedated for infection.     

Effects of sustained control of brushtail possums on levels of Mycobacterium bovis infection in cattle and brushtail possum populations from Hohotaka, New Zealand

AIMS: To examine the effect of reducing the abundance of brushtail possums (Trichosurus vulpecula) on the distribution and prevalence of bovine tuberculosis (Mycobacterium bovis, Tb) in possums and the incidence of Tb in domestic cattle on a group of farms in the central North Island, New Zealand. METHODS: The cumulative yearly incidence of Tb infection from 12 cattle herds was estimated from annual tuberculin testing and abattoir inspection data over the period 1983-98. Intensive control of possum populations began for six of the herds in 1988, five herds in 1994 and the remaining herd in 1996. The prevalence and distribution of macroscopic M. bovis infection in possums and an index of possum abundance was estimated during yearly cross-sectional surveys from 1988 to 1998. This enabled formal testing of the link between the abundance of tuberculous possums and the incidence of Tb in cattle. RESULTS: Before possum control, infected possums were clustered in foci on or adjacent to the farms with the highest annual incidence of tuberculosis in cattle, and had an overall prevalence of macroscopic M. bovis infection of 2.3%. The prevalence of disease declined to zero with ongoing possum control, although infected possums continued to be found during the first 5 years of control. Maintaining the possum population at an average of 22.1% of its pre-control density significantly reduced the odds of the cumulative yearly incidence of Tb in cattle by 77% during the first 5 years of possum control and a further 65% in the second 5-year period. Nine of 11 tuberculous possums identified since the start of possum control were found within the areas where infected possums were clustered during the pre-control survey, suggesting that the persistence of infection within these clusters rather than infected immigrants was the source of ongoing disease. Annual estimates of the prevalence of tuberculous possums broadly followed the predictions of Barlow's possum-Tb model for a controlled possum population. CONCLUSION: The results support the hypothesis that tuberculous possums transmit bovine tuberculosis to domestic cattle, and therefore that reducing the abundance of tuberculous possums reduces the incidence of Tb in cattle. If the level of possum culling is sufficient, it appears that M. bovis infection may be eradicated from possum populations. Better information on population density, rate of increase and annual culling rates would have been needed for a truly independent examination of the Barlow possum-Tb model.     

Do free-ranging common brushtail possums (Trichosurus vulpecula) play a role in the transmission of Toxoplasma gondii within a zoo environment?

To investigate the possible role of common brushtail possums (Trichosurus vulpecula) in the transmission of Toxoplasma gondii within a zoo environment, a serological survey of a free-ranging population resident within Taronga Zoo, Sydney, Australia was undertaken using the modified agglutination test (MAT). For comparison, the seroprevalence of T. gondii antibodies was also assessed in a possum population inhabiting a felid-free, non-urban woodland habitat. Six of 126 possums (4.8%) from the zoo population had antibodies to T. gondii with a MAT titre of 25 or higher, while in contrast, all of the 17 possums from woodland were seronegative. These observations suggest that possums were at a higher risk of exposure to the parasite as a consequence of co-existing with domestic, stray and captive felids associated with urbanisation. Screening of captive felids at the zoo indicated 16 of 23 individuals (67%) and all 6 species were seropositive for T. gondii, implicating them as a possible source of the parasite within the zoo setting. In addition captive, non-felid carnivores including the chimpanzee (Pan troglodytes), saltwater crocodile (Crocodylus porosus), dingo (Canis lupis) and leopard seal (Hydrurga leptonyx) were tested for the presence of T. gondii antibodies as these species predate and are a leading cause of death amongst zoo possums. In total, 5 of 23 individuals (22%) were seropositive, representing 2 of the 4 carnivorous species; the dingo and chimpanzee. These data suggest that carnivory was not a highly efficient pathway for the transmission of T. gondii and the free-ranging possum population posed minimal threat to the health of zoo animals.     

Resolution of Mycobacterium bovis infection in wild brushtail possums (Trichosurus vulpecula)

The brushtail possum (Trichosurus vulpecula) is the principal wildlife reservoir of Mycobacterium bovis in New Zealand (Morris and Pfeiffer 1995). Tuberculosis in possums is a progressive, fatal infection. The complete resolution of infection in possums has not been reported. The purpose of this communication is to present two cases of complete and spontaneous resolution of tuberculosis in wild, naturally infected possums. During an 11-year (1989-2000) longitudinal study of a possum population at Castlepoint in the Wairarapa, 90 bacteriologically-confirmed cases of M. bovis infection were observed, and 88 of the tuberculous possums were either found dead or were presumed to have died when they ceased to be trapped. As the home range of adult possums is stable (Efford et al 2000) it was assumed that if an adult possum could no longer be trapped, it had died. In many cases, the declining physical condition of a tuberculous possum prior to its disappearance supported the assumption. However, two cases survived and were euthanised, one 15 months and the other 30 months after they first showed clinical signs of infection. When examined post mortem, neither showed evidence of M. bovis infection. The case histories are described and the significance of the observations discussed.     

Longevity of Mycobacterium bovis in brushtail possum (Trichosurus vulpecula) carcasses,and contact rates between possums and carcasses

Abstract

AIM: To determine, for a variety of environmental conditions, how long Mycobacterium bovis might remain viable inside the carcass of a brushtail possum (Trichosurus vulpecula) that died of bovine tuberculosis (Tb), and to measure the rate of contact between free-ranging possums and possum carcasses.

METHODS: Lesions of M. bovis were simulated by inoculating excised spleens weighing 0.5–1 g with 0.2 mL liquid culture containing approximately 5 x 10⁷ cfu M. bovis/mL. Simulated lesions were inserted into possum carcasses (n=48) at the peripheral lymph nodes. Carcasses were placed in the field at two sites (a tussock grassland and a podocarp-broadleaved forest site) and in two seasons (summer and winter) for up to 62 days. Survival rates of M. bovis were estimated by sampling the simulated lesions over time, and culturing the recovered lesion to determine if any viable M. bovis bacteria were present.

The time taken for a free-ranging possum to first encounter a dead possum in its home range was estimated by live-trapping possums and fitting them with proximity loggers (n=13). A ‘contact’ was recorded if these possums came within 40–50 cm of proximity loggers fitted to possum carcasses.

RESULTS: There were strong seasonal and site effects in the survival rate of M. bovis in possum carcasses. In the grassland habitat, no viable bacilli were cultured from any carcass after 3 days in summer, whereas in winter all samples were culture-positive for the first 20 days, and some were still positive after 27 days. The survival rates for forest habitat were intermediate between the results for grassland, and there were no culture-positive carcasses after 9 days in summer or 27 days in winter.

In summer, infected carcasses (n=6) were first encountered by possums a mean 1.9 (range 0.4–6.7) days after placement.

CONCLUSIONS: Possum carcasses were contacted by free-ranging possums within the period that viable M. bovis were shown to survive in a carcass. The risk of such infection is likely to be most significant in winter or in areas with microhabitats where the survival of M. bovis is high. However, the generally low survival rate of M. bovis in possum carcasses and the low frequency of possum-to-carcass contacts indicate this route of transmission alone could not maintain Tb in a possum population.     

Behavioural studies on the potential for direct transmission of tuberculosis from feral ferrets (Mustela furo) and possums (Trichosurus vulpecula) to farmed livestock

Studies were conducted to evaluate the response of cattle and deer to ferrets which were sedated so they behaved like terminally tuberculous animals, and to compare this with the response of cattle, deer and sheep to sedated possums. Six groups of deer and two groups of cattle were exposed to a sedated ferret and to a sedated possum. Both livestock species showed interest in the possum by sniffing and licking it, but they only briefly touched the ferret and no licking or extended investigation was observed. The proportion of available time spent in physical contact with the possum by cattle was 7.7 times as high as for the ferret, and for the deer was 5.7 times as high. The behavioural response of three groups of sheep to a sedated possum was investigated, and sheep showed limited interest beyond viewing the possum from a distance. The amount of time spent by sheep investigating the possum was very low and the intensity of exploration was also low. For possums, at least one deer was within 1.5 m (an estimate of the distance that tuberculosis can be transmitted by aerosol) for 50.9% of observation time, and in physical contact with the possum for 9.5% of time. The figures for cattle were 69.3% and 17.3%, while those for sheep were 6.9% and 0.3%. In interactions with ferrets, the equivalent figures were 29.8% within 1.5 m and 2.2% in physical contact for cattle, and 20.8% and 1.1% for deer. Tuberculous possums commonly and tuberculous ferrets less commonly have lung lesions and/or discharging sinuses, and may excrete Mycobacterium bovis intermittently or continuously in aerosols or discharges. The exploratory behaviour of deer and cattle in this study would provide opportunities for them to become infected with M. bovis if they had contact with infectious possums, and less probably with ferrets. The response of sheep to possums suggests that they would be much less likely to contract the disease.     

Ranging behaviour and duration of survival of wild brushtail possums (Trichosurus vulpecula) infected with Mycobacterium bovis

AIM: To quantify the duration of survival of possums (Trichosurus vulpecula) infected with Mycobacterium bovis, and identify aspects of their behaviour which may influence the likelihood of disease transmission to domestic stock or wildlife. METHODS: Capture and den locations of 14 naturally infected tuberculous possums, eight possums experimentally infected with M. bovis and eight non-infected possums were recorded between May 1998 and February 2000 at a study site near Castlepoint on the Wairarapa coast of the North Island in New Zealand. Denning behaviour was observed weekly using radiotelemetry, and possums were captured, examined and released bi-monthly. Data were used to estimate survival period; create denning, activity, and total ranges; and to identify extended forays by possums as individuals and groups. RESULTS: Seventeen tuberculous possum carcasses were recovered, of which 14 (82%) were close to or within their activity range. Denning ranges were known for 10/17 possums that died. Four tuberculous possums were found dead within their denning range. Three possums made extended forays in the 3 weeks before death. Twelve possums were found dead in dense scrub, three in long grass in open woodland and two on pasture. Mean duration of survival of naturally infected possums following detection of clinical signs was 3.4 months (95% CI=2.1-5.4) and the instantaneous mortality rate was 0.293 per month (95% CI=0.184-0.470). Signs of disease were obvious for about 3 weeks prior to death. Tuberculous possums were commonly trapped on only part of the area where the total non-infected population was trapped. CONCLUSION: Most tuberculous possums died within their activity range and in scrub, representing a risk of transmission of M. bovis to wildlife and livestock that forage in scrub. Smaller proportions dying on pasture represent a less frequent, but highly visible risk. Tuberculous possums were clustered on the study site, and localised possum control operations would be more effective if focussed on such areas.     

Contact patterns as a risk factor for bovine tuberculosis infection in a free-living adult brushtail possum Trichosurus vulpecula population

The aim of this study was to identify risk factors for bovine tuberculosis (TB) in a free-roaming, capture-mark-recapture monitored possum Trichosurus vulpecula population in a 22-ha study site at Castlepoint, New Zealand from 1 April 1989 to 31 March 1994. A matched case-control design was used to evaluate the influence of sex, habitat and contact opportunities on TB risk. Cases comprised possums identified as TB-positive throughout the study period. Controls were selected from the group of possums that were captured and showed no clinical signs of TB throughout the study period. Measures derived from a social network analysis of possum capture locations such as degree, clustering coefficient (CC) and betweenness were used to represent potential contact opportunities among possums. Network analysis measures recorded for individual possums in the 12-month period before a diagnosis of TB were evaluated in a conditional logistic regression model. We found no evidence of an association between case status and the total number of possums with which there was potential contact (degree) (P=0.5). The odds of cases being exposed to unit increases in the number of TB-positive contacts was 2.50 (95% CI 1.24-5.05; P<0.01) times that of controls. This effect was conditional on the total number of potential contacts made, with a negative interaction with increasing degree. These findings indicate that potential contact with TB-positive possums increases the odds of disease whereas potential contact with large numbers of possums does not. This suggests that multiple contacts with TB-positive possum(s) are necessary for transmission of TB and this is more likely to occur in networks that are smaller. We challenge the hypothesis that contact with large numbers of individuals increases the probability of becoming TB infected and argue that individual contact behaviour is a determinant of the creation of TB foci within free-living possum populations.     

The efficacy of bacille Calmette-Guérin vaccine in wild brushtail possums (Trichosurus vulpecula)

A population of wild brushtail possums (Trichosurus vulpecula) in which bovine tuberculosis was endemic was vaccinated with live bacille Calmette-Guérin (BCG) to determine the efficacy of vaccination. The population on the 56 hectare site was monitored bimonthly over 2 years using a capture-release regime. During the study tuberculosis was diagnosed by clinical and post mortem examination. Possums were vaccinated with BCG by both intranasal aerosol and conjunctival instillation. Possums were revaccinated on average every 5 months. Over the 2 years, 300 possums were recruited to the study with 149 being allocated to the vaccination group. There were significantly fewer cases of tuberculosis in the vaccinated (4 cases) than in the unvaccinated group (13 cases; P=0.023). The vaccine efficacy was 69%. An attempt was made to increase the incidence of disease by releasing onto the site possums that had been experimentally infected with a strain of M. bovis unknown in the area. However, this did not result in any additional cases. BCG vaccine was shown to have a level of efficacy which could be of assistance in controlling tuberculosis in wild possum populations. The future use of vaccination for the control of tuberculosis in wild possum populations is discussed.     

The relationship between prevalence of Mycobacterium bovis infection in feral ferrets and possum abundance

AIM: To determine the relationship between the prevalence of macroscopic Mycobacterium bovis infection in feral ferrets (Mustela furo) and the abundance of brushtail possums (Trichosurus vulpecula). METHODS: The predictive power of a previously reported positive association between the prevalence of macroscopic M. bovis infection in ferrets and possum abundance was examined by undertaking surveys of M. bovis infection in ferrets at sites of low and high possum abundance. The association was then tested by a manipulative experiment that measured changes in the prevalence of macroscopic M. bovis infection in feral ferrets after reducing possum abundance. RESULTS: The positive relationship between the prevalence of macroscopic M. bovis infection in ferrets and possum abundance remained valid for new survey data, although the goodness of fit of the relationship was reduced. Experimentally reducing possum abundance reduced the odds of macroscopic M. bovis infection in ferrets by 80% in the years immediately following possum control (Odds Ratio=0.23, p=0.003). CONCLUSIONS: There is a causal link between possum abundance and the prevalence of macroscopic M. bovis infection in feral ferrets in areas in which M. bovis infection is known to occur in ferret populations. This suggests that possum-to-ferret transmission of disease occurs and accounts for most of the disease evident in ferret populations, though does not determine whether ferrets are spillover or maintenance hosts of M. bovis. Management to reduce the prevalence of M. bovis infection in ferrets should consider reducing possum abundance as a control tactic. KEY WORDS: Mustela furo, ferret, Trichosurus vulpecula, brushtail possum, Mycobacterium bovis, bovine tuberculosis.     

Broad-scale possum and ferret correlates of macroscopic Mycobacterium bovis infection in feral ferret populations

AIMS: To examine the relationships between the prevalence of macroscopic Mycobacterium bovis infection (bovine tuberculosis) in feral ferrets (Mustela furo), the abundance of ferrets, and the abundance of brushtail possums (Trichosurus vulpecula) . METHODS: Data on the prevalence of macroscopic M. bovis infection in ferrets, abundance of ferrets, and abundance of possums were analysed from 12 comparable independent broad-scale surveys. RESULTS: The prevalence of macroscopic M. bovis infection in ferrets was positively correlated with possum abundance but unrelated to ferret abundance, suggesting that possums are an important source of M. bovis infection in ferrets. The lack of any positive relationship between the prevalence of M. bovis infection in ferrets and ferret abundance does not support the hypothesis that per capita transmission rates, and therefore disease prevalence, should be higher at higher ferret abundance. CONCLUSION: The results support the hypothesis that tuberculous possums are the major underlying source of M. bovis infection for feral ferrets in New Zealand.     

A behaviour study on the potential for direct transmission of tuberculosis from possums (Trichosurus vulpecula) to alpacas (Lama pacos), and the converse from alpacas to possums

AIMS: To evaluate the potential for the direct transmission of tuberculosis from possums to alpacas, and vice versa. METHODS: A field study was conducted on an alpaca farm in Northland, New Zealand on 7 January 1999. Observations were recorded on the interaction of one group of male alpacas with a simulated dead possum, one male and one female alpaca group with a simulated terminally tuberculous possum, and one group of male alpacas with a normal possum in an enclosure from which the animals could not escape. The possum was sedated with ketamine as hydrochloride to simulate death (inactive; no movement), and terminal illness (active, inco-ordinated movement around the paddock). The observations were based on the focal animal sampling technique, they were un-replicated, and recorded visually and manually with intermittent still photography. RESULTS: Both male and female alpacas showed strongly inquisitive interaction with the possum. They clustered around the possum (focal animal) very soon after it was observed by the first member of the group. The interest of the majority of both sex groups remained high for the observation periods of approximately 30 minutes, and most individuals remained within 5 metres of the possum for that time. Approximately 50% of the alpacas were within possible aerosol transmission distance of 2 metres from the sedated, erratically mobile possum with their heads towards it for approximately 50% of the two observation periods. Aggressive behaviour was recorded for a young male with stamping on the moving possum. Similar, but more vigorous and prolonged stamping behaviour was recorded for a female with a young (<1 week) offspring (cria). The stamping behaviour was accompanied by very close nose to nose contact of the alpaca and possum. At one point the female threw the possum approximately 1.5 metres in the air with her teeth. The group of male alpacas placed in an enclosure with an unsedated normal possum generally moved away from the possum during its rapid active attempts to escape. When it became inactive their approaches were cautious and only once elicited a defence reaction from the possum, from which they recoiled. One male made one attempt to stamp on the active possum. Soon after the possum became inactive in a small loose hay pile, the alpacas lost interest in it. CONCLUSIONS: The alpaca / possum behavioural interactions show there is potential for direct aerosol transmission of tuberculosis from possums to alpacas, but probably not from alpacas to possums.     

Interactions between beef cattle and simulated tuberculous possums on pasture

Brushtail possums (Trichosurus vulpecula) were sedated with ketamine and placed within a fenced observation area with 17 Hereford cross steers. The behaviour of the sedated possums simulated that of terminally ill possums. Behavioural analysis was carried out during eight observation periods in which sedated possums were successfully released and interactions occurred. Cattle were attracted from over 50 m away by the movements of the possums, and for 34% of observation time cattle investigated the possums. Inactive animals engendered far less interest, and cattle investigated them for 3% of observation time. An average of seven to eight cattle came in physical contact with the possum in each of the observation periods. Cattle actively investigated the possum while following it as it moved, commonly close enough to be within aerosol transmission distance, and some sniffed the possum, touched it with their noses, and in many cases licked it extensively. All of these activities would expose the cattle to high risk of receiving an infective dose of Mycobacterium bovis, especially via the respiratory route. The type and duration of contact between individual steers and the possum varied between animals and observation period. The cattle exposed themselves to greater risk, overall, during the first week of observations, and there was evidence of habituation with increased contact. Although all 17 steers came within 5 metres of the possum at some point during the observation periods, some individual cattle behaved in ways which would put them at greater risk of contracting tuberculosis than did others. Cattle showed little interest in possum carcasses placed on pasture. These were unlikely to be a significant source of infection for cattle, although they could be for scavenger species.     

Toxoplasmosis as a cause of ovine perinatal mortality

Aims. To determine temporal and spatial patterns of bovine tuberculosis (Tb) in a population of brushtail possums (Trichosurus vulpeculu) free from commercial and recreational hunting and living contiguously with livestock, and to relate these patterns to measures of possum health and density.

Methods. Possums were trapped on 11 lines located on a forest-rough grazing margin in the Ahaura Valley in Westland in 1979-80 and each August from 1992 to 1996, and were examined post mortem for gross lesions indicative of Tb. Infection levels in possums were compared with Th test data from cattle grazing in the same area.

Results. Tuberculosis was identified from the trapped sample of possums in 1980. Trapping of further possums on the same trap lines confirmed the presence of the disease each year from 1992 to 1996, and suggested a clustering of infection in time and space. The prevalence of Th in possums declined with increasing possum population size and well being, and decreasing age. The prevalence of Tb in possums was highest in 1980 and 1992, and appeared to roughly coincide with similar upsurges in Tb in nearby cattle.

Conclusions. Our results suggest that the exceptional levels of Tb recorded in possums in 1992 had a direct effect on possum numbers, and gave rise to very low trap catches in 1993-94. They also suggest thatTb can persist for several years in possum populations existing well below the 5% trap catch targeted by regipnal councils forTb possum control.     

Temporal patterns in bovine tuberculosis in a brushtail possum population contiguous with infected cattle in the Ahaura Valley, Westland

AIMS: To determine temporal and spatial patterns of bovine tuberculosis (Tb) in a population of brushtail possums (Trichosurus vulpecula) free from commercial and recreational hunting and living contiguously with livestock, and to relate these patterns to measures of possum health and density. METHODS: Possums were trapped on 11 lines located on a forest-rough grazing margin in the Ahaura Valley in Westland in 1979-80 and each August from 1992 to 1996, and were examined post mortem for gross lesions indicative of Tb. Infection levels in possums were compared with Th test data from cattle grazing in the same area. RESULTS: Tuberculosis was identified from the trapped sample of possums in 1980. Trapping of further possums on the same trap lines confirmed the presence of the disease each year from 1992 to 1996, and suggested a clustering of infection in time and space. The prevalence of Tb in possums declined with increasing possum population size and well being, and decreasing age. The prevalence of Tb in possums was highest in 1980 and 1992, and appeared to roughly coincide with similar upsurges in Tb in nearby cattle. CONCLUSIONS: Our results suggest that the exceptional levels of Tb recorded in possums in 1992 had a direct effect on possum numbers, and gave rise to very low trap catches in 1993-94. They also suggest that Tb can persist for several years in possum populations existing well below the 5% trap catch targeted by regional councils for Tb possum control.     

Trends in the incidence of tuberculosis in possums and livestock, associated with differing control intensities applied to possum populations

AIM: To determine the trap-catch index (an estimate of abundance) of brushtail possum (Trichosurus vulpecula) populations infected with bovine tuberculosis (Tb; Mycobacterium bovis) that must be achieved, and the length of time such an index must be maintained, for Tb to be eliminated from possum populations and adjacent livestock. METHODS: Between 1997-1998 and 2000-2001, trap-catch surveys of possum populations naturally infected with Tb and subjected to population-control measures were undertaken at four forest sites and two farmland sites. At the same time, possum carcasses were collected at these sites and their Tb status determined, and all contiguous cattle and deer herds were Tb tested and abattoir slaughter data for these herds were interrogated. RESULTS: Trap-catch surveys indicated that numbers of possums on the farmland sites surveyed were usually very low and well below the control targets set (i.e. a 5% trap catch or approximately 0.5-1 possum/ha) for the study. In contrast, trap-catch surveys undertaken in forest sites indicated possum numbers were more variable, and often recovered rapidly from control operations to exceed control targets within 1-3 years. The annual rate of recovery of possum populations in half of the forest population surveys undertaken exceeded published intrinsic rates of increase for possums. The overall prevalence of Tb in possum populations was < or =1.9% at 5/6 sites, and was 6.5% at the sixth site. Juvenile possums infected with Tb were trapped within but near the edge of control zones and appeared to represent an immigrant source of infection. Mature infected possums survived control operations apparently by having home ranges in uncontrolled patches within control areas. Infection in possums appeared to be eliminated from one study site by the intensive control undertaken, but elimination at other sites appeared less likely. Levels of Tb in livestock on or adjacent to the study sites fell by at least 50% during the study, and cattle in one area tested clear for the first time in 20 years. CONCLUSIONS: Initial control of possums in forest appeared to achieve national control targets set by the Animal Health Board (AHB), despite trap-catch data often providing misleading population estimates. Such targets were often exceeded within 1-3 years. By comparison, possum control on farmland appeared to maintain populations at very low levels, while control on forest margins maintained populations at intermediate levels. Control was least effective in deep forest where human access was most difficult. Intensive population control measures appeared to have led to a reduced incidence of Tb in livestock at 3/4 sites, and elimination of Tb in livestock at one site. This result supports modelling studies that predict the eradication of Tb from possums through ongoing intensive control and may explain the lower success achieved with earlier less-intensive possum control.     

A neurological syndrome in a free-living population of possums (Trichosurus vulpecula)

AIMS: To determine the clinical and pathological features of a neurological disease syndrome in a free-living possum population in New Zealand and to compare this syndrome with wobbly possum disease. METHODS: An outbreak of a neurological disease in possums in the Rotorua district was investigated in 1994. A variety of tissues was collected and investigated microbiologically and histopathologically. Tissues stored from clinically affected possums were homogenised, clarified and inoculated into healthy possums by the intra-peritoneal route. The clinical signs and histopathological lesions in naturally-infected and in experimentally-inoculated possums were assessed and compared with those of possums affected with wobbly possum disease. RESULTS: Histopathological investigation of three of the naturally-affected possums revealed non-suppurative encephalitis with perivascular cuffing, diffuse non-suppurative meningitis and focal non-suppurative myocarditis. These lesions were suggestive of a viral infection. No pathogenic bacteria were recovered and no viruses were isolated in tissue culture. A neurological disease, indistinguishable from wobbly possum disease, was reproduced in five out of the eight experimentally inoculated possums. In two experimental cases the clinical signs were very mild and, in most cases of the natural and experimental disease, histopathological lesions in the central nervous system were mild in comparison with wobbly possum disease. Possums which did not develop clinical signs of neurological disease or have lesions in the central nervous system did have infiltrations of mononuclear inflammatory cells in the liver and kidney. CONCLUSIONS: This neurological disease, reported for the first time in a free-living population, closely resembles and may be the same as wobbly possum disease. The milder nature of this disease could suggest there may be more than one strain of the aetiological agent.     

A behaviour study on the potential for direct transmission of tuberculosis from possums (Trichosura vulpecula) to alpacas (Lama pacos), and the converse from alpacas to possums

Aims. To evaluate the potential for the direct transmission of tuberculosis from possums to alpacas, and vice versa.

Methods. A field study was conducted on an alpaca farm in Northland, New Zealand on 7 January 1999. Observations were recorded on the interaction of one group of male alpacas with a simulated dead possum, one male and one female alpaca group with a simulated terminally tuberculous possum, and one group of male alpacas with a normal possum in an enclosure from which the animals could not escape. The possum was sedated with ketamine as hydrochloride to simulate death (inactive; no movement), and terminal illness (active, into-ordinated movement around the paddock). The observations were based on the focal animal sampling technique, they were un-replicated, and recorded visually and manually with intermittent still photography.

Results. Both male and female alpacas showed strongly inquisitive interaction with the possum. They clustered around the possum (focal animal) very soon after it was observed by the first member of the group. The interest of the majority of both sex groups remained high for the observation periods of approximately 30 minutes, and most individuals remained within 5 metres of the possum for that time. Approximately 50% of the alpacas were within possible aerosol transmission distance of 2 metres from the sedated, erratically mobile possum with their heads towards it for approximately 50% of the two observation periods. Aggressive behaviour was recorded for a young male with stamping on the moving possum. Similar, but more vigorous and prolonged stamping behaviour was recorded for a female with a young (<1 week) offspring (cria).The stamping behaviour was accompanied by very close nose to nose contact of the alpaca and possum. At one point the female threw the possum approximately 1.5 metres in the air with her teeth. The group of male alpacas placed in an enclosure with an unsedated normal possum generally moved away from the possum during its rapid active attempts to escape. When it became inactive their approaches were cautious and only once elicited a defence reaction from the possum, from which they recoiled. One male made one attempt to stamp on the active possum. Soon after the possum became inactive in a small loose hay pile, the alpacas lost interest in it.

Conclusions. The alpaca / possum behavioural interactions show there is potential for direct aerosol transmission of tuberculosis from possums to alpacas, but probably not from alpacas to possums.     

Mycobacterium bovis infection in the brush-tailed possum (Trichosurus vulpecula): II. Comparison of experimental infections with an Australian cattle strain and a New Zealand possum strain

Experimentally induced M. bovis infection in brush-tailed possums (Trichosurus vulpecula) with a possum-adapted strain from New Zealand, produced a more rapidly progressive, severe disease than did that induced by an Australian cattle strain. Possums infected with either strain excreted large numbers of mycobacteria in urine, faeces and discharges from abscesses. Aerosol transmission of infection occurred only with the New Zealand possum strain. The infectious aerosol most probably was of respiratory origin but urine may have been the source.All possums had focal interstitial nephritis consistent with Leptospira infection. Histopathological evidence of severe stress was observed in only one possum; the remainder had either mild or no stress-associated lesions.