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.     

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.     

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.     

Mortality rate and movements of brushtail possums with clinical tuberculosis (Mycobacterium bovis) infection

AIM: To describe the mortality rate and movements of brushtail possums (Trichosurus vulpecula) with clinical tuberculosis (Mycobacterium bovis) from a site in native podocarp/hardwood forest, in the Orongorongo Valley, Wellington. METHODS: Possums with clinical tuberculosis (Tb) were identified through routine trapping on live capture grids and were fitted with radio-collars, and located at approximately monthly intervals for 2.5 years. The survival times of radio-collared tuberculous and non-diseased possums were used to estimate the instantaneous additional mortality rate due to clinical Tb. The spatial behaviour of clinically tuberculous possums was compared with that of non-diseased possums, using both den and trap locations. The frequency of long-distance movements of both tuberculous and non-diseased possums was also determined. A possum was defined as having undertaken along-distance movement when it moved at least 200 m from the centre of its activity range and then returned to its established activity range or died. RESULTS: The mean survival time of clinically tuberculous possums was 4.7 (SE 0.79) months. The instantaneous additional mortality rate due to clinical Tb was estimated to be between 1.08 year-1 and 2.38 year-1, with the lower figure including an unknown portion of the incubation period in the estimate. Estimates of denning and activity ranges of tuberculous possums were approximately 22-30% larger, on average, than the ranges of non-diseased possums, but the differences were not significant. Both tuberculous and non-diseased possums undertook long-distance movements up to 700 m from the centre of their activity range. However, the majority of tuberculous possums died within or near their established activity range. CONCLUSIONS: This study estimated the mortality rate during the clinical stages of Tb and, as such, did not represent a true estimate of the disease-induced mortality rate (alpha) used in most epidemiological models of possums and Tb. However, the lower estimate of the additional mortality rate due to clinical disease could be considered an approximation to alpha. The movement patterns of clinically tuberculous possums were, overall, not substantially different from non-diseased possums. The occurrence of long-distance movements by tuberculous possums sometimes coincided with the advanced stage of clinical disease. However, the majority of tuberculous possums died within their established activity area. Long-distance movements by resident possums during the terminal stages of disease represent a direct risk for transmission of Tb to livestock on farms within approximately 1 km of infected possum populations.     

Vaccination of the brushtail possum (Trichosurus vulpecula) against Mycobacterium bovis infection with bacille Calmette-Guérin: the response to multiple doses.

In New Zealand, the brushtail possum (Trichosurus vulpecula) is the principal wildlife vector of bovine tuberculosis. Control of infected possum populations contributes to the control of tuberculosis in domestic livestock. Vaccination is potentially a complementary strategy to population control, but to be cost-effective, administration of the vaccine to possums would need to be from an appropriately designed automatic vaccinator. Possums themselves would activate the vaccinator so that it would deliver an aerosol spray of vaccine. There would be no direct way to prevent possums receiving multiple doses of vaccine. This study examined the effect on protective immunity of repeated vaccination. Captive possums were vaccinated with BCG strain pasteur 1173P2 either 12 times at weekly intervals, twice at 6-weekly intervals, or once. Vaccination was by a combination of intranasal aerosol and conjunctival instillation. Eight weeks after the last dose of vaccine, all possums were challenged intratracheally with Mycobacterium bovis strain 83/6235. Vaccination induced a significant immune response as measured by the lymphocyte proliferation assay (LPA). A significant level of protection, as measured by the response to challenge, developed in all the vaccinated possum groups, but protection was greatest in the group vaccinated 12 times. It was concluded that protection would be enhanced if vaccinations were repeated at short intervals (weekly), but no benefit or detriment resulted from revaccination after longer intervals (1-2 months).     

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.     

The effect of fertility control on the transmission of bovine tuberculosis in wild brushtail possums

AIM: To determine the effect of fertility control on the rate of transmission of bovine tuberculosis (Tb), caused by Mycobacterium bovis, in brushtail possums (Trichosurus vulpecula). METHODS: At two study sites with a history of Tb infection in the resident possum population, a sample of adult male and female possums (n=50), determined by palpation to be Tb-free, was surgically sterilised by gonadectomy. A sample of untreated Tb-free male and female possums (n=54) served as controls. Each study site was trapped every 2 months over a 3-year period, and the Tb status of the individuals in the trial assessed. At the conclusion of the trial, all remaining experimental individuals were killed, necropsied and examined for characteristic lesions typical of Tb. The rate of transmission of Tb infection was estimated using the incidence of tuberculous cases in the experimental animals and comparing it between sites, sex and sterilisation treatment. RESULTS: Sterilisation by gonadectomy resulted in a reduction in the rate of transmission of Tb in male possums by 53%, but a corresponding increase in sterilised females for reasons that are still unclear. However, this interaction was statistically weak (p=0.10). When the sexes were combined, there was no overall effect of sterilisation on the rate of transmission of Tb (p=0.43). Sterility treatment notwithstanding, there was a highly significant difference in the rate of transmission between the sexes (p=0.01), being almost one order of magnitude higher in untreated males compared with untreated females. CONCLUSIONS: Although lacking strong statistical support, these results suggest that fertility control that targets endocrine control of fertility may provide some additional benefits for disease control over that achieved by reductions in population density, by reducing the rate of transmission of M. bovis in male possums. However, additional studies are needed to confirm the validity of these results. The large difference in the rate of transmission of M. bovis in untreated males compared with untreated females suggests that transmission of Tb among males may be an important driver of the dynamics of Tb infection in possums.     

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.     

The effect of fertility control on the transmission of bovine tuberculosis in wild brushtail possums

AIM: To determine the effect of fertility control on the rate of transmission of bovine tuberculosis (Tb), caused by Mycobacterium bovis, in brushtail possums (Trichosurus vulpecula).

METHODS: At two study sites with a history of Tb infection in the resident possum population, a sample of adult male and female possums (n=50), determined by palpation to be Tb-free, was surgically sterilised by gonadectomy. A sample of untreated Tb-free male and female possums (n=54) served as controls. Each study site was trapped every 2 months over a 3-year period, and the Tb status of the individuals in the trial assessed. At the conclusion of the trial, all remaining experimental individuals were killed, necropsied and examined for characteristic lesions typical of Tb. The rate of transmission of Tb infection was estimated using the incidence of tuberculous cases in the experimental animals and comparing it between sites, sex and sterilisation treatment.

RESULTS: Sterilisation by gonadectomy resulted in a reduction in the rate of transmission of Tb in male possums by 53%, but a corresponding increase in sterilised females for reasons that are still unclear. However, this interaction was statistically weak (p=0.10). When the sexes were combined, there was no overall effect of sterilisation on the rate of transmission of Tb (p=0.43). Sterility treatment notwithstanding, there was a highly significant difference in the rate of transmission between the sexes (p=0.01), being almost one order of magnitude higher in untreated males compared with untreated females.

CONCLUSIONS: Although lacking strong statistical support, these results suggest that fertility control that targets endocrine control of fertility may provide some additional benefits for disease control over that achieved by reductions in population density, by reducing the rate of transmission of M. bovis in male possums. However, additional studies are needed to confirm the validity of these results. The large difference in the rate of transmission of M. bovis in untreated males compared with untreated females suggests that transmission of Tb among males may be an important driver of the dynamics of Tb infection in possums.     

Dominance hierarchies in cattle and red deer (Cervus elaphus): their possible relationship to the transmission of bovine tuberculosis

A behavioural study was conducted to assess the dominance structure of cattle and deer herds and to assess the possible relationship of dominance to the risk of becoming infected with bovine tuberculosis. Five groups of cattle containing newly identified intradermal tuberculin test reactors were evaluated to determine the dominance hierarchy, and then exposed to sedated possums to assess the response of reactors and non-reactors. Eighty-six percent of the tuberculin test-positive cattle were among the 20% most dominant animals in their herds. In four of the five herds, the dominant animals investigated the sedated possum most actively, and in three of these four the reactors were in the investigating group. Six deer were exposed to a naturally tuberculosis-infected possum population, and the four highest animals in the dominance hierarchy (which also showed strong investigative behaviour when exposed to simulated terminally ill tuberculous possums) all subsequently became infected with tuberculosis. The fifth animal in the hierarchy became test-positive for tuberculosis later than the first four, but was subsequently also shown to be culture-positive for M. bovis. The lowest animal in the hierarchy, which showed no active interest in simulated tuberculous possums, did not become infected. This study strongly suggests a central role for terminally ill tuberculous possums in the transmission of tuberculosis to cattle and farmed deer. Management techniques designed to reduce contact between these few possums and farmed livestock may be expected to reduce the incidence of tuberculosis.     

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.     

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.     

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.     

Endoparasites of brushtail possums (Trichosurus vulpecula) from the South Island, New Zealand

As part of a study to assess whether endoparasites could assist in the biological control of brushtail possums in New Zealand, we investigated the composition and distribution of possum endoparasites in the South Island. Possums were collected near five of the original release areas in the South Island : Banks Peninsula, Hokitika, Nelson, Dunedin and Invercargill. Among the nematodes, those most frequently encountered were Trichostrongylus spp., which were present in possums from all five study areas. Trichostrongylus species from possums in the Invercargill area comprised 4.5% T. colubriformis, 0.9% T. vitrinus and 11.3% T. retortaeformis. Paraustrostnmgylus trichosuri and Parastrongyloides trichosuri were found only in the Invercargill area, where they infected 1.4% and 14% of possums respectively. The cestode Bertiella trichosuri was present in possums from all locations except Dunedin. The protozoan Eimeria spp. occurred in all areas. These are the first records of Parastrongyloides trichosuri, Paraustrostrongylus trichosuri, T. vitrinus, T. retortaeformis and Eimeria spp. in South Island possums. The prevalence of endoparasites and the intensity of infection was very low compared to the lower North Island of New Zealand. Endoparasites at the existing levels in the South Island probably have very little effect on possum populations.     

Bovine tuberculosis in a possum from the Orongorongo Valley, Wellington

The first confirmed case of a tubercular possum in the Orongorongo Valley forest is reported from a population intensively studied for 20 years. The victim was a five-year-old male recaptured 20 times over 3% years. Its annual home range was about 1.8 hectares but this dropped to 0.4 ha during its last year of life. Its range overlapped with at least 15 others. The animal appeared normal and healthy in July 1985 but died of advanced, generalised bovine tuberculosis 12 weeks later. The nearest confirmed cases of tubercular possums were reported 1.3 km away in 1976 and 4 km away in 1979. The Orongorongo possum was unlikely to have contacted any cattle for over three years. The infection was probably transmitted from the Wainuiomata Valley by possums or pigs. The Orongorongo River was no barrier to the spread of the infection. Although most of the neighbouring possums were trapped and examined several times over the following year, no tubercular animals were detected.     

Isolation of Mycobacterium bovis from brushtail possums with non-visible lesions

AIM: To determine the prevalence of Mycobacterium bovis infection in brushtail possums (Trichosurus vulpecula) that did not have macroscopic lesions of bovine tuberculosis, and to evaluate culture of pooled tissues from multiple possums as a method for determining the M. bovis-infection status of wildlife populations in New Zealand.

METHODS: Pools of selected tissues were collected from possums from four different populations known to be infected with M. bovis. Tissue pools from individual animals, and combined pools from multiple animals, were cultured for M. bovis.

RESULTS: In the four populations investigated, the prevalence of possums with macroscopic lesions confirmed by culture to be infected with M. bovis ranged from 1 to 19 (mean 31/283; 10.9)%. The prevalence of possums with non-visible lesions that were culture positive for M. bovis in the same populations ranged from 4 to 10 (mean 24/283; 8.5)%. The mean of the log10 cfu of M. bovis of the macroscopic lesions and of the culture-positive samples that did not have visible lesions was 3.85 (SE 0.26) and 1.46 (SE 0.26) log₁₀ cfu, respectively (p<0.01). Mycobacterium bovis was cultured from pools of 30–50 animals in the four populations studied.

CONCLUSIONS: The finding of M. bovis infection in possums with non-visible lesions identified a potential deficiency of declaring possum populations free of M. bovis on the basis of absence of macroscopic lesions. The culturing of pools of selected tissues from multiple animals without visible lesions can be used to reduce laboratory costs of possum surveys without a major reduction in the ability to detect M. bovis infection.     

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.     

Aerosol vaccination of the brushtail possum (Trichosurus vulpecula) with bacille Calmette-Guérin: the duration of protection

Bovine tuberculosis is endemic in wild brushtail possums (Trichosurus vulpecula) in New Zealand. The disease is controlled by reducing or eliminating infected possum populations, but control methods do not kill all possums in the targeted area, leaving some tuberculous possums to maintain the disease. Vaccination with bacille Calmette-Guérin (BCG) has been shown to provide significant levels of protection. Vaccination is a potential alternative or complementary control strategy if protection is long lasting. Captive possums were vaccinated with a single dose of BCG by intranasal aerosol and challenged by intratracheal instillation of Mycobacterium bovis 2, 6 or 12 months after vaccination. Vaccination produced significant immunity as measured by the lymphocyte proliferative response to bovine PPD and protection in response to challenge. The protective response was seen as a decrease in the mass of pulmonary lesions and decreased dissemination to the abdominal organs and body lymph nodes. The protective effect was strongest at 2 months after vaccination but was still present at a lower level at 12 months. Delivery of an aerosol vaccine to possums in the wild using a self-delivery system could contribute substantially to wildlife tuberculosis control.     

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.     

A descriptive spatial analysis of bovine tuberculosis in intensively controlled cattle farms in New Zealand

We describe the temporal and geographical distribution of confirmed cases of bovine tuberculosis (TB) in a population of cattle in the south-east of the North Island of New Zealand. Data were derived from routine TB testing conducted between 1980 and 2003 and included details for 69 farms. Four six-year periods were defined to coincide with changes in depopulation strategies against the wildlife TB reservoir, the brushtail possum Trichosurus vulpecula. For the periods 1980 to 1985 and 1986 to 1991 the median annual incidence rate of TB was 0.4 and 4.7 cases per 1000 cattle-years at risk, respectively. For the period 1992 to 2003 the median annual incidence rate of TB decreased to 1.8 cases per 1000 cattle-years at risk, coincident with the use of poisoning to control possums in the surrounding forest park (a major possum habitat area). We identified clusters of TB cases adjacent to the forest park and found no evidence of spatio-temporal interaction of TB risk among farms. Our findings support the hypothesis that possums living in the forest park are a source of bovine TB in this area and that farm-to-farm spread of disease was not an important infection mechanism.