Infection of Eurasian badgers (Meles meles) with Mycobacterium bovis and Mycobacterium avium complex in Spain

The prevalence, distribution and pathology related to infection with Mycobacterium bovis and other mycobacteria were determined in trapped (n=36) and road-killed (n=121) badgers in Spain from 2006 to 2010. The prevalence of M. bovis based on bacteriological culture from road-killed badgers was 8/121 (6.6%) and from trapped badgers was 0/36 (0%). Tuberculosis/M. bovis infection was evident in 15/121 (12.4%) road-killed badgers when bacteriology and histopathology were combined. Mycobacterium avium complex was isolated by culture from the tracheal aspirate of 1/36 (2.8%) trapped badgers and from tissue pools from 8/121 (6.6%) road-killed badgers.     

Zoonotic aspects of rotaviruses

Rotaviruses are important enteric pathogens of humans and animals. Group A rotaviruses (GARVs) account for up to 1 million children deaths each year, chiefly in developing countries and human vaccines are now available in many countries. Rotavirus-associated enteritis is a major problem in livestock animals, notably in young calves and piglets. Early in the epidemiological GARV studies in humans, either sporadic cases or epidemics by atypical, animal-like GARV strains were described. Complete genome sequencing of human and animal GARV strains has revealed a striking genetic heterogeneity in the 11 double stranded RNA segments across different rotavirus strains and has provided evidence for frequent intersections between the evolution of human and animal rotaviruses, as a result of multiple, repeated events of interspecies transmission and subsequent adaptation.     

牛型提纯结核菌素国家标准品的研制

根据《中华人民共和国兽用生物制品规程》2000年版制备牛型提纯结核菌素(牛PPD)国家标准品候选物4批。根据多糖、核酸和氮含量测定结果,筛选200804批制备为待检国家标准品,其检验结果为:物理性状为乳白色疏松团块,加生理盐水后在5s内溶解;无菌检验合格。200804批准检国家标准品于稀释前各样品变异系数为1.4%,冻干后各样品变异系数为4.8%。效价测定:待检国家标准品4个稀释度引起的豚鼠皮肤红肿面积与国际标准品相对应的稀释度引起的皮肤红肿面积的差值平均值都在10%以内,2者的剂量反应曲线基本平行,即待检国家标准品效价与国际标准品基本相同。200804批待检国家标准品于50℃保存21d之内效价下降程度平均值小于10%,保存28d效价下降程度平均值大于10%。其特异性和安全性均符合要求。     

Experimental inoculation of pigeons (Columba livia) with Mycobacterium bovis

The purpose of this pilot study was to determine if pigeons (Columba livia) are susceptible to infection with Mycobacterium bovis by either oral or intratracheal inoculation and to assess their possible role in the lateral transmission of bovine tuberculosis. Six pigeons were orally inoculated with 1.3 x 10(5) colony-forming units of M. bovis, six pigeons were intratracheally inoculated with the same dose, and six pigeons served as noninoculated controls. The study continued for 90 days postinoculation (PI), with groups of birds necropsied at 30-day intervals, and fecal samples and tissues were collected for mycobacterial culture. Two pigeons, one intratracheally inoculated and one orally inoculated, shed M. bovis in their feces at 1 day PI, and one intratracheally inoculated bird shed M. bovis in its feces 60 days PI. Whereas no illness or weight loss was present during the course of the study, 2 of 12 inoculated birds exhibited microscopic lesions of mycobacteriosis, and the organism was isolated from tissues of three inoculated birds. Pigeons are susceptible to infection with M. bovis after high dose inoculation and can shed the organism in their feces for up to 60 days PI; intratracheally inoculated birds appear more likely to become active fecal shedders of M. bovis. Although these were high dose inoculations under experimental conditions, pigeons may potentially play a role in the lateral transmission of bovine tuberculosis between infected and uninfected mammalian hosts.     

Zoonotic aspects of arenavirus infections

To date, the International Committee for Taxonomy of Viruses recognizes that the family Arenaviridae contains a unique genus Arenavirus that includes 22 viral species. There are nine additional arenaviruses that either have been discovered recently, or which taxonomic status remains pending. Arenaviruses have been classified according to their antigenic properties into two groups, the Lassa-Lymphocytic choriomeningitis (LCM) serocomplex and the Tacaribe serocomplex which has been further divided into four evolutionary lineages. Each arenavirus is more or less tightly associated with a mammal host. The distribution of the host dictates the distribution of the virus. Humans may become infected by arenaviruses through direct contact with infected rodents, including bites, or through inhalation of infectious rodent excreta and secreta. Lassa, Junin, Machupo, Guanarito, and Sabia viruses are known to cause a severe hemorrhagic fever, in western Africa, Argentina, Bolivia, Venezuela, and Brazil, respectively. Infection by LCM virus can result in acute central nervous system disease, congenital malformations, and infection in organ transplantation recipients. Detection of arenaviruses in their animal host can be achieved by virus isolation, and has recently taken advantage of PCR-based techniques. The approach based on consensus degenerate primers has shown efficient for both detection of known arenaviruses, and discovery of new arenaviruses.     

Mycobacterium bovis infection in New Zealand hedgehogs (Erinaceus europaeus)

Seventy-nine wild hedgehogs from the Wairarapa, an area of New Zealand in which bovine tuberculosis is endemic, were examined for lesions suggestive of tuberculosis. Sixteen animals with suspicious lesions had samples submitted for culture. Mycobacterium bovis was recovered from four animals with gross pulmonary lesions. The gross and histopathological appearance of the lesions are described and the significance discussed in the light of previously reported experimental and captive animal infections. It is likely that infection arose from the scavenging behaviour of hedgehogs. The moderate prevalence (5%) of tuberculosis in these animals combined with their small home ranges may allow them to be used in wildlife surveys to pinpoint the locality in which tuberculous possums have died.     

5-Bromodeoxyuridine (BUdR) and light inactivation of proliferating Mycobacterium bovis and Mycobacterium avium sensitized bovine lymphocytes

Maximum inactivation (83–98%) of proliferating Mycobacterium bovis and/or Mycobacterium avium sensitized bovine lymphocytes was obtained when the lymphocytes were treated with 10^?4 M of 5-bromodeoxyuridine (BUdR) and light. Inactivation was specific though not total whether the cultures were first stimulated with M. bovis PPD or M. avium PPD, though slightly higher inactivation resulted when cultures were first stimulated with M. bovis PPD. There was, however, a statistically significant suppression (P < 0.05) of the counts/min of cultures initially stimulated with either M. bovis PPD or M. avium PPD and later restimulated with either of the antigens after BUdR-light treatment. Thus experiments showed that inactivation occurs only to those cells responding by proliferation to a particular stimulant. There is a potential for the use of this assay for differentiating infection due to either M. bovis or M. avium if total inactivation of proliferating lymphocytes could be achieved.     

Experimental inoculation of wild turkeys (Meleagris gallopavo) with Mycobacterium bovis

Although avian species are known to be susceptible to infection with Mycobacterium spp. organisms, much remains unknown about the susceptibility of birds to infection with M. bovis. The objective of this current study was to determine if wild turkeys (Meleagris gallopavo) can be infected with M. bovis when inoculated by the oral or intratracheal route. Six turkeys were orally inoculated and another six were inoculated via the trachea with a high dose of M. bovis, 1 x 10(5) CFU/ml. Six turkeys were sham-inoculated controls. Two turkeys from each treatment group were sacrificed on days 30, 60, and 90 postinoculation. There were no gross or microscopic lesions consistent with mycobacteriosis in the 23 inoculated turkeys over the 90-day duration of this study. Fecal cultures were also consistently negative for M. bovis when sampled before inoculation and on days 1, 30, and 60 postinoculation. Two intratracheally inoculated turkeys were positive for M. bovis in visceral tissues at 30 days postinoculation. However, this finding was only indicative of passive persistence of mycobacteria in the tissues and not of infection, as there were no attendant lesions or clinical compromise to support infection. Thus, it can be concluded that young wild turkeys are resistant to infection with M. bovis and, therefore, pose minimal threat as reservoir or spillover hosts for this organism.     

Vaccination with Mycobacterium bovis BCG Strains Danish and Pasteur in White‐tailed Deer (Odocoileus virginianus) Experimentally Challenged with Mycobacterium bovis

Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis in domestic livestock and the cause for many faltering bovine tuberculosis eradication programmes. One approach in dealing with wildlife reservoirs of disease is to interrupt inter‐species and intraspecies transmission through vaccination of deer or cattle. To evaluate the efficacy of BCG vaccination in white‐tailed deer, 35 deer were assigned to one of three groups; one s.c. dose of 10⁷ CFU of M. bovis BCG Pasteur (n = 12); 1 s.c. dose of 10⁷ CFU of M. bovis BCG Danish (n = 11); or unvaccinated deer (n = 12). After vaccination, deer were inoculated intratonsilarly with virulent M. bovis. Lesion severity scores of the medial retropharyngeal lymph node, as well as all lymph nodes combined, were reduced in vaccinated deer compared to unvaccinated deer. BCG Danish vaccinated deer had no late stage granulomas characterized by coalescent caseonecrotic granulomas containing numerous acid‐fast bacilli compared to BCG Pasteur vaccinated or unvaccinated deer where such lesions were present. Both BCG strains were isolated as late as 250 days after vaccination from deer that were vaccinated but not challenged. In white‐tailed deer, BCG provides protection against challenge with virulent M. bovis. Issues related to vaccine persistence, safety and shedding remain to be further investigated.     

Comparison of Mycobacterium avium complex (MAC) strains from pigs and humans in Sweden by random amplified polymorphic DNA (RAPD) using standardized reagents

Infections with atypical mycobacteria belonging to the Mycobacterium avium/intracellulare complex (MAC) can cause infection in both animals and humans. Using a standardized reagents commercial kit for random amplified polymorphic DNA (RAPD) analysis, 49 MAC strains isolated from 32 slaughter pigs and 17 humans in Sweden were identified and sorted out, yielding 6 RAPD types. By combining the results of RAPD primers 4 and 5 and the primer IS1245A, we found that pigs and humans may be infected with the same types of MAC strains, since 14 strains from humans and 8 strains from pigs were essentially identical and together, comprised RAPD type 2, the largest group of strains (44.8% of strains). With respect to grouping of strains, serotype and RAPD type were uncorrelated, except for serotype 20 and RAPD type 6. Using standardized beads, RAPD analysis is a reproducible technique for typing MAC strains, as the indistinguishable banding patterns obtained with repeated analyses of two isolates from each strain in this study demonstrate. However, primer selection and DNA purity were crucial for differentiating closely related strains.     

An outbreak of tuberculosis by Mycobacterium bovis in coatis (Nasua nasua)

Mycobacterium tuberculosis complex, which includes Mycobacterium bovis, infrequently causes severe or lethal disease in captive wildlife populations. A dead coati from a wildlife triage center showing pulmonary lesions compatible with tuberculosis had raised suspicion of a potential disease caused by mycobacteria species and was further investigated. Four native coatis (Nasua nasua) with suspected mycobacterial infection were sedated, and bronchoalveolar lavages and tuberculin skin tests (TSTs) were performed. All animals tested positive upon TST. Mycobacterial culturing, Ziehl-Neelsen staining, and genetic testing were performed on postmortem samples and the etiologic agent was identified as M. bovis. Molecular genetic identification using a polymerase chain reaction panel was crucial to achieving a definitive diagnosis.     

Experimentally induced infection of reindeer (Rangifer tarandus) with Mycobacterium bovis.

In the USA, all species of Cervidae are included in the USDA's uniform methods and rules for the eradication of bovine tuberculosis and, therefore, are subject to regulations regarding intradermal tuberculin testing. In reindeer (Rangifer tarandus), infection with Mycobacterium bovis is exceedingly rare and the response of reindeer to infection with M. bovis in pathologic and immunologic terms is unknown. The objectives of the study reported here were to describe the pathologic changes associated with M. bovis infection in reindeer and evaluate the effectiveness of intradermal tuberculin testing as a means of diagnosis of tuberculosis in reindeer. Thirteen reindeer were inoculated intratonsilarly with 10(5) colony-forming units (CFU) of M. bovis, and 4 noninoculated reindeer served as negative controls. The comparative cervical test (CCT) was done on all reindeer 90 and 240 days after inoculation. Thirteen months after inoculation, all reindeer were euthanized and examined. All experimentally inoculated reindeer developed lesions in the medial retropharyngeal lymph nodes. The CCT accurately identified all M. bovis-inoculated reindeer, but false-positive results were common among negative-control reindeer. Modifications of the method for interpretation of the CCT decreased false-positive results without increasing false-negative results. Reindeer are susceptible to infection with M. bovis; however, lesions are fewer in number, less severe in nature, and less widely disseminated than are those seen in white-tailed deer (Odocoileus virginianus). Comparative cervical skin testing of reindeer can be highly sensitive, but has low specificity. Specificity can be improved by modification of criteria for interpretation of the CCT.     

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.     

An outbreak of Mycobacterium bovis infection in fallow deer (Dama dama)

In an outbreak of Mycobacterium bovis infection in fallow deer in South Australia, 3 herds related by recent movement of deer were infected. From these 3 infected herds, 47 of 51 animals were tuberculosis at necropsy. A range of lesions was seen most of which differed from classical bovine tuberculosis in that pus was a white liquid, fibrous encapsulation was not marked and calcification was rare. Histopathology was of classical tuberculosis. M. bovis was cultured from lesions and M. avium-intracellulare was cultured from one deer with no visible lesions. The source of M. bovis infection has not been determined.     

Zoonotic aspects of infections with noroviruses and sapoviruses

The close genetic relationship of noroviruses and sapoviruses found in animals and humans has raised the question whether these viruses have a zoonotic potential. Transmission from animals to humans and vice versa would have far-reaching consequences for epidemiology and food safety. So far animal noro- and sapoviruses have not been found in humans. However detection of human noroviruses in animals as well as simultaneous presence of animal and human viruses in bivalve molluscs suggest a risk of transmission. Furthermore, antibodies against animal noroviruses were detected in humans as well as antibodies against human noroviruses in swine. Experimental infection of gnotobiotic calves and pigs with human noroviruses demonstrated that virus replication and seroconversion can occur. Accordingly the possible role of noro- and sapoviruses as zoonotic agents needs to be further investigated.     

Spoligotype diversity of Mycobacterium bovis and Mycobacterium caprae animal isolates

The genetic diversity of 283 Mycobacterium bovis (M. bovis) and 10 Mycobacterium caprae (M. caprae) strains, isolated between 2002 and 2007 from cattle, goat, red deer and wild boar from six different geographical regions of Portugal was investigated by spoligotyping. The technique showed a good discriminatory power (Hunter-Gaston Index, h=0.9) for the strains, revealing 29 different patterns. One pattern (SB0121) was clearly predominant, accounting for 26.3% of the isolates; ten patterns, representing 20.7% of the isolates, had never been reported previously. Multiple spoligotypes were detected in thirteen cattle and one goat herd, most of which were found in beef cattle and extensive management regions, suggesting different infection sources. With the exception of two spoligotypes, those in wildlife species were also found in domestic species.     

Vaccinating badgers (Meles meles) against Mycobacterium bovis: the ecological considerations

Bovine tuberculosis (TB) is a serious zoonotic disease, which despite a largely successful test and slaughter programme has persisted in cattle herds in parts of the UK. The badger (Meles meles) is widely considered to represent a significant wildlife reservoir for the transmission of Mycobacterium bovis to cattle, and has been the subject of a variety of culling strategies since the mid 1970s. Nevertheless, the incidence of herd breakdowns has continued to rise, and the efficacy of culling is currently the subject of a large-scale field trial. One potential alternative tool for the management of disease in wildlife populations is vaccination. However, the successful development of an effective vaccine and a strategy for its delivery will require careful consideration of the practical constraints imposed by ecological factors. In the current paper, we discuss relevant ecological and epidemiological characteristics of badger populations and practical aspects of vaccine delivery in the field.     

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.