Koala retrovirus diversity, transmissibility, and disease associations.

BACKGROUND: Koalas are infected with the koala retrovirus (KoRV) that exists as exogenous or endogenous viruses. KoRV is genetically diverse with co-infection with up to ten envelope subtypes (A-J) possible; KoRV-A is the prototype endogenous form. KoRV-B, first found in a small number of koalas with an increased leukemia prevalence at one US zoo, has been associated with other cancers and increased chlamydial disease. To better understand the molecular epidemiology of KoRV variants and the effect of increased viral loads (VLs) on transmissibility and pathogenicity we developed subtype-specific quantitative PCR (qPCR) assays and tested blood and tissue samples from koalas at US zoos (n = 78), two Australian zoos (n = 27) and wild-caught (n = 21) in Australia. We analyzed PCR results with available clinical, demographic, and pedigree data. RESULTS: All koalas were KoRV-A-infected. A small number of koalas (10.3%) at one US zoo were also infected with non-A subtypes, while a higher non-A subtype prevalence (59.3%) was found in koalas at Australian zoos. Wild koalas from one location were only infected with KoRV-A. We observed a significant association of infection and plasma VLs of non-A subtypes in koalas that died of leukemia/lymphoma and other neoplasias and report cancer diagnoses in KoRV-A-positive animals. Infection and VLs of non-A subtypes was not associated with age or sex. Transmission of non-A subtypes occurred from dam-to-offspring and likely following adult-to-adult contact, but associations with contact type were not evaluated. Brief antiretroviral treatment of one leukemic koala infected with high plasma levels of KoRV-A, -B, and -F did not affect VL or disease progression. CONCLUSIONS: Our results show a significant association of non-A KoRV infection and plasma VLs with leukemia and other cancers. Although we confirm dam-to-offspring transmission of these variants, we also show other routes are possible. Our validated qPCR assays will be useful to further understand KoRV epidemiology and its zoonotic transmission potential for humans exposed to koalas because KoRV can infect human cells.

Resolution of a Localized Granuloma Caused by Mycobacterium avium-intracellulare Complex on the Cere of a Bruce's Green Pigeon (Treron waalia).

A 3-year-old female Bruce's green pigeon (Treron waalia) was presented with granulomatous inflammation of the cere and underlying tissues with osteomyelitis and bone proliferation of the dorsal premaxilla. Biopsy and culture revealed the presence of Mycobacterium avium-intracellulare complex, and multi-antimicrobial treatment was initiated with clarithromycin, ethambutol, rifabutin, and enrofloxacin. The cere lesion improved and no evidence of systemic granulomas was observed over 4 months of treatment, although leukocytosis and monocytosis persisted. Five months after discontinuation of antibiotic therapy, the white blood cell count had normalized, but distal beak irregularities and partial recurrence of the mass were present. The bird died 15 months after discontinuation of antibiotic therapy and necropsy revealed no evidence of active mycobacteriosis of the beak or cere. This report documents an unusual clinical presentation of mycobacteriosis, in addition to its successful resolution.

Gastrointestinal torsions and intussusception in northern koalas (Phascolarctos cinereus) at San Diego Zoo (1976-2012).

The recent classification as threatened status of the northern koala (Phascolarctos cinereus) by the Australian Government highlights the importance of the conservation and health management of this iconic Australian marsupial. This case series describes gastrointestinal torsion and intussusception in six northern koalas (three males, three females, 2-11 yr old) at the San Diego Zoo from 1976 to 2012. Two koalas died shortly after presentation. Diagnoses of ileocecal intussusception, resulting from enteritis in one case and cecal torsion in the other, were made at postmortem examination. One koala died 4 days after an exploratory laparotomy, with negative findings, and an acute double colonic intussusception was diagnosed at postmortem examination. Two small intestinal mesenteric torsion and one proximal colon mesenteric torsion cases were successfully corrected surgically. In the case of colonic mesenteric torsion, the koala had recurrent clinical signs 2 wk later, and a second surgery requiring resection and anastomosis of ischemic jejunum was performed, with the koala dying shortly afterward. One koala with small intestinal torsion had a recurrence of torsion 22 mo later and subsequently died. The koala with the second case of small intestinal torsion remains alive 14 mo postsurgical correction. All six koalas presented with signs of colic that included anorexia, lethargy, depression, acute abdominal distension, abdominal stretching, decreased fecal output, open-mouth gasping, or a combination of symptoms. Abdominal radiographs may show stacked gastrointestinal linear gas patterns and contrast stasis. Prevalence of torsion and intussusception is low at this institution (2%), although recurrence in individuals is common (50%) and overall survival is poor (83%), which emphasizes the importance of timely recognition, surgical correction, and postoperative management. While inciting etiologies were unable to be determined in these cases, monitoring generalized gastrointestinal health and differing Eucalyptus sp. effects on individual koala's gastrointestinal function, parasite control, and stress minimization through standardized husbandry practices are likely important.

Development of a case definition for clinical feline herpesvirus infection in cheetahs (Acinonyx jubatus) housed in zoos.

The identification of feline herpesvirus (FHV) infected cheetahs (Acinonyx jubatus) and characterization of shedding episodes is difficult due to nonspecific clinical signs and limitations of diagnostic tests. The goals of this study were to develop a case definition for clinical FHV and describe the distribution of signs. Medical records from six different zoologic institutions were reviewed to identify cheetahs with diagnostic test results confirming FHV. Published literature, expert opinion, and results of a multiple correspondence analysis (MCA) were used to develop a clinical case definition based on 69 episodes in FHV laboratory confirmed (LC) cheetahs. Four groups of signs were identified in the MCA: general ocular signs, serious ocular lesions, respiratory disease, and cutaneous lesions. Ocular disease occurred with respiratory signs alone, with skin lesions alone, and with both respiratory signs and skin lesions. Groups that did not occur together were respiratory signs and skin lesions. The resulting case definition included 1) LC cheetahs; and 2) clinically compatible (CC) cheetahs that exhibited a minimum of 7 day's duration of the clinical sign groupings identified in the MCA or the presence of corneal ulcers or keratitis that occurred alone or in concert with other ocular signs and skin lesions. Exclusion criteria were applied. Application of the case definition to the study population identified an additional 78 clinical episodes, which represented 58 CC cheetahs. In total, 28.8% (93/322) of the population was identified as LC or CC. The distribution of identified clinical signs was similar across LC and CC cheetahs. Corneal ulcers and/or keratitis, and skin lesions were more frequently reported in severe episodes; in mild episodes, there were significantly more cheetahs with ocular-only or respiratory-only disease. Our results provide a better understanding of the clinical presentation of FHV, while presenting a standardized case definition that can both contribute to earlier diagnoses and be used for population-level studies.

Mitochondrial Gene Diversity and Host Specificity of Isospora in Passerine Birds.

Isospora infections are common in both wild and captive passerine species. Many bird species have been shown to have co-evolved with a particular species of Isospora. Disease can range from subclinical to severe and fatal, making infection and transmission of this parasite a concern for birds under managed care, particularly in institutions housing endangered species for breeding and reintroduction purposes. Whether birds in mixed-species enclosures represent a risk factor for severe isosporiasis due to infection with non-host-adapted strains is of concern for institutions managing these populations. To begin answering this question, we sought to characterize the host-specificity of Isospora spp. in a large number of passerine birds via retrospective sequencing of mitochondrial gene cytochrome c oxidase subunit I (COI). Despite outliers, Isospora sequences largely grouped by host species and/or host family. Additional research is warranted into the degree of interspecies transmission and host-switching of Isospora parasites, and risk factors for the development of severe disease in passerine birds.

Longitudinal social network analysis of avian mycobacteriosis incidence in a large population of zoo birds.

The goal of this study was to evaluate longitudinal patterns of avian mycobacteriosis spread through a social network. Specifically, we wanted to determine whether the patterns of connectivity over time can predict future infections, and whether this pattern can distinguish between different sources of infection. The study population included 13,409 individuals nested in a larger population of birds that were closely monitored in zoological facilities for over 22 years (1992-2014). A retrospective cohort study design and social network connectivity were used to estimate the association between exposure to an infected bird, and development of mycobacteriosis. Avian mycobacteriosis was diagnosed from histopathology and network connectivity was defined by enclosure histories over discrete time periods. Single-variable and multivariable longitudinal, mixed effects logistic regression models examined whether exposure to infected birds, both directly- and indirectly-connected, was associated with development of mycobacteriosis at the next time step. Our adjusted model showed an increased odds of developing mycobacteriosis (odds ratio = 2.15; 95 % CI: 1.48-3.12; p < 0.001) for birds that were directly exposed (i.e., housed in the same aviary) to another infected bird, compared to those with no exposure. Exposure to a positive, indirectly-connected bird at a previous time step was independently associated with an increased risk of mycobacteriosis (odds ratio = 1.56; 95 % CI: 1.11-2.19). This association persisted in adjusted models even when the indirect contacts were housed in distinctly different aviaries and never had contact with the subject of interest or its environment. Adjusted, risk-stratified models further characterized the type of exposure that increased the risk of avian mycobacteriosis. Birds that were exposed in small aviaries were more likely to develop mycobacteriosis than those exposed in larger aviaries and those with no exposure. The lesion distribution and species of the contact (same species versus different species) were also significant predictors of disease risk. Some findings were sensitive to model variation of time divisions and initiation time. Our study shows avian mycobacteriosis spread through the social network in quantifiable and discernable patterns. We provide empirical evidence that a contagious process drives some of the observed infection, but we also show low transmissibility based on sustained patterns of low incidence over time even when large groups of birds are exposed. Targeted risk mitigation efforts based on the characteristics of the exposure may be effective at reducing risk of avian mycobacteriosis while enhancing population sustainability.

Herpesvirus surveillance and discovery in zoo-housed ruminants.

Gammaherpesvirus infections are ubiquitous in captive and free-ranging ruminants and are associated with a variety of clinical diseases ranging from subclinical or mild inflammatory syndromes to fatal diseases such as malignant catarrhal fever. Gammaherpesvirus infections have been fully characterized in only a few ruminant species, and the overall diversity, host range, and biologic effects of most are not known. This study investigated the presence and host distribution of gammaherpesviruses in ruminant species at two facilities, the San Diego Zoo and San Diego Zoo Safari Park. We tested antemortem (blood, nasal or oropharyngeal swabs) or postmortem (internal organs) samples from 715 healthy or diseased ruminants representing 96 species and subspecies, using a consensus-based herpesvirus PCR for a segment of the DNA polymerase (DPOL) gene. Among the 715 animals tested, 161 (22.5%) were PCR and sequencing positive for herpesvirus, while only 11 (6.83%) of the PCR positive animals showed clinical signs of malignant catarrhal fever. Forty-four DPOL genotypes were identified of which only 10 have been reported in GenBank. The data describe viral diversity within species and individuals, identify host ranges of potential new viruses, and address the proclivity and consequences of interspecies transmission during management practices in zoological parks. The discovery of new viruses with wide host ranges and presence of co-infection within individual animals also suggest that the evolutionary processes influencing Gammaherpesvirus diversity are more complex than previously recognized.

Social network analysis and whole-genome sequencing to evaluate disease transmission in a large, dynamic population: A study of avian mycobacteriosis in zoo birds.

This study combined a social network analysis and whole-genome sequencing (WGS) to test for general patterns of contagious spread of a mycobacterial infection for which pathways of disease acquisition are not well understood. Our population included 275 cases diagnosed with avian mycobacteriosis that were nested in a source population of 16,430 birds at San Diego Zoo Wildlife Alliance facilities from 1992 through mid-2014. Mycobacteria species were determined using conventional methods and whole genome sequencing (WGS). Mycobacterium avium avium (MAA) and Mycobacterium genavense were the most common species of mycobacteria identified and were present in different proportions across bird taxa. A social network for the birds was constructed from the source population to identify directly and indirectly connected cases during time periods relevant to disease transmission. Associations between network connectivity and genetic similarity of mycobacteria (as determined by clusters of genotypes separated by few single nucleotide polymorphisms, or SNPs) were then evaluated in observed and randomly generated network permutations. Findings showed that some genotypes clustered along pathways of bird connectivity, while others were dispersed throughout the network. The proportion of directly connected birds having a similar mycobacterial genotype was 0.36 and significant (p<0.05). This proportion was higher (0.58) and significant for MAA but not for M. genavense. Evaluations of SNP distributions also showed genotypes of MAA were more related in connected birds than expected by chance; however, no significant patterns of genetic relatedness were identified for M. genavense, although data were sparse. Integrating the WGS analysis of mycobacteria with a social network analysis of their host birds revealed significant genetic clustering along pathways of connectivity, namely for MAA. These findings are consistent with a contagious process occurring in some, but not all, case clusters.

ASSESSMENT OF DISEASE RISK ASSOCIATED WITH POTENTIAL REMOVAL OF ANTHROPOGENIC BARRIERS TO MOJAVE DESERT TORTOISE (GOPHERUS AGASSIZII) POPULATION CONNECTIVITY.

The Mojave Desert tortoise (Gopherus agassizii), federally listed as threatened, has suffered habitat loss and fragmentation due to human activities. Upper respiratory tract disease (URTD), a documented health threat to desert tortoises, has been detected at the Large-Scale Translocation Study Site (LSTS) in southwestern Nevada, US, a fenced recipient site for translocated animals. Our study aimed to 1) estimate prevalence of URTD and Mycoplasma infection at LSTS and three nearby unfenced sites; 2) assess whether Mycoplasma infection status was associated with developing clinical signs of URTD; and 3) determine whether such an association differed between LSTS and unfenced areas. We sampled 421 tortoises in 2016 to describe the current status of these populations. We evaluated three clinical signs of URTD (nasal discharge, ocular discharge, nasal erosions) and determined individual infection status for Mycoplasma agassizii and Mycoplasma testudineum by quantitative PCR and enzyme-linked immunosorbent assay. In 2016, LSTS had the highest prevalence of M. agassizii (25.0%; 33/132), M. testudineum (3.0%; 4/132), and URTD clinical signs (18.9%; 25/132). Controlling for other factors, clinical sign(s) were positively associated with M. agassizii infection (odds ratio [OR]=7.7, P=0.001), and this effect was similar among study sites (P>0.99). There was no association with M. testudineum status (P=0.360). Of the 196 tortoises in a longitudinal comparison of 2011-14 with 2016, an estimated 3.2% converted from M. agassizii-negative to positive during the study period, and incidence was greater at LSTS (P=0.002). Conversion to positive M. agassizii status was associated with increased incidence of clinical signs in subsequent years (OR=11.1, P=0.018). While M. agassizii and URTD are present outside the LSTS, there is a possibility that incidence of Mycoplasma infection and URTD would increase outside LSTS if these populations were to reconnect. Population-level significance of this risk appears low, and any risk must be evaluated against the potential long-term benefits to population viability through increased connectivity.

Molecular characterization of a novel gammaretrovirus in killer whales (Orcinus orca).

There are currently no published data documenting the presence of retroviruses in cetaceans, though the occurrences of cancers and immunodeficiency states suggest the potential. We examined tissues from adult killer whales and detected a novel gammaretrovirus by degenerate PCR. Reverse transcription-PCR also demonstrated tissue and serum expression of retroviral mRNA. The full-length sequence of the provirus was obtained by PCR, and a TaqMan-based copy number assay did not demonstrate evidence of productive infection. PCR on blood samples from 11 healthy captive killer whales and tissues from 3 free-ranging animals detected the proviral DNA in all tissues examined from all animals. A survey of multiple cetacean species by PCR for gag, pol, and env sequences showed homologs of this virus in the DNA of eight species of delphinids, pygmy and dwarf sperm whales, and harbor porpoises, but not in beluga or fin whales. Analysis of the bottlenose dolphin genome revealed two full-length proviral sequences with 97.4% and 96.9% nucleotide identity to the killer whale gammaretrovirus. The results of single-cell PCR on killer whale sperm and Southern blotting are also consistent with the conclusion that the provirus is endogenous. We suggest that this gammaretrovirus entered the delphinoid ancestor's genome before the divergence of modern dolphins or that an exogenous variant existed following divergence that was ultimately endogenized. However, the transcriptional activity demonstrated in tissues and the nearly intact viral genome suggest a more recent integration into the killer whale genome, favoring the latter hypothesis. The proposed name for this retrovirus is killer whale endogenous retrovirus.

Genetic characterization and epidemiology of Helicobacters in non-domestic animals.

BACKGROUND: Novel helicobacter infections and associated disease are being recognized with increasing frequency in animals and people. Yet, the pervasiveness of infection in distantly related animal taxa, genetic diversity of helicobacters, and their transmissability are not known. AIM: To better understand the ecology of helicobacters, we did a PCR survey and epidemiologic analysis of 154 captive or wild vertebrate taxa originating from 6 continents. MATERIALS AND METHODS: One hundred twenty nine helicobacter 16s rRNA gene segments were amplified by PCR and sequenced from ninety-three mammalian, reptilian, avian, or amphibian host species. Prevalence estimates were generated, and univariate logistic regression analyses were used to explore relationships between infection status and the health and characteristics of the 220 individual animals. RESULTS: One hundred and nineteen novel helicobacter DNA sequences were found. No significant relationship between infection and host health was found; however, multi-infection or infections with particular genotypes were associated with mild clinical signs. Phylogenetic and genetic comparisons of helicobacters suggested prolonged co-adaptation and niche-associated divergence as well as periodic inter-species transmission. CONCLUSION: The genus Helicobacter should accordingly be viewed as a collection of hundreds of organisms that have colonized most tetrapod taxa and have the potential to expand into new hosts as contact among animals and between animals and people increases.

Diclofenac residues as the cause of vulture population decline in Pakistan.

The Oriental white-backed vulture (OWBV; Gyps bengalensis) was once one of the most common raptors in the Indian subcontinent. A population decline of >95%, starting in the 1990s, was first noted at Keoladeo National Park, India. Since then, catastrophic declines, also involving Gyps indicus and Gyps tenuirostris, have continued to be reported across the subcontinent. Consequently these vultures are now listed as critically endangered by BirdLife International. In 2000, the Peregrine Fund initiated its Asian Vulture Crisis Project with the Ornithological Society of Pakistan, establishing study sites at 16 OWBV colonies in the Kasur, Khanewal and Muzaffargarh-Layyah Districts of Pakistan to measure mortality at over 2,400 active nest sites. Between 2000 and 2003, high annual adult and subadult mortality (5-86%) and resulting population declines (34-95%) (ref. 5 and M.G., manuscript in preparation) were associated with renal failure and visceral gout. Here, we provide results that directly correlate residues of the anti-inflammatory drug diclofenac with renal failure. Diclofenac residues and renal disease were reproduced experimentally in OWBVs by direct oral exposure and through feeding vultures diclofenac-treated livestock. We propose that residues of veterinary diclofenac are responsible for the OWBV decline.

Investigation of factors predicting disease among zoo birds exposed to avian mycobacteriosis.

OBJECTIVE: To characterize infection patterns and identify factors associated with avian mycobacteriosis among zoo birds that were housed with infected enclosure mates. DESIGN: Matched case-control study. ANIMALS: 79 birds with avian mycobacteriosis (cases) and 316 nondiseased birds (controls) of similar age and taxonomic group that were present in the bird collection of the Zoological Society of San Diego from 1991 through 2005. PROCEDURES: Inventory and necropsy records from all eligible, exposed birds (n = 2,413) were examined to determine disease incidence and prevalence in the exposed cohort. Cases were matched in a 1:4 ratio to randomly selected controls of similar age and taxonomic grouping. Risk factors for mycobacteriosis (demographic, temporal, enclosure, and exposure characteristics as well as translocation history) were evaluated with univariate and multivariable conditional logistic regression analyses. RESULTS: Disease prevalence and incidence were estimated at 3.5% and 8 cases/1,000 bird-years at risk, respectively. In the multivariable model, cases were more likely to have been imported into the collection, exposed to mycobacteriosis at a young age, exposed to the same bird species, and exposed in small enclosures than were controls. Odds for disease increased with an increasing amount of time spent with other disease-positive birds. CONCLUSIONS AND CLINICAL RELEVANCE: The low incidence of mycobacteriosis and the risk factors identified suggested that mycobacteria may not be easily transmitted through direct contact with infected enclosure mates. Identification of risk factors for avian mycobacteriosis will help guide future management of this disease in zoo bird populations.

Preventive medicine success: thymoma removal in an African spot-necked otter (Lutra maculicollis).

During a preventive medicine examination on a 13-yr-old intact female African spot-necked otter (Lutra maculicollis), radiographs were obtained and a cranial thoracic mass was noted. Cytology from an ultrasound-guided fine-needle aspirate was suggestive of a thymoma. Surgical removal was performed and this diagnosis was confirmed by histopathology. At a recheck examination 6 mo postsurgery, there was no evidence of recurrence. This case highlights the value of a comprehensive preventive medicine program that resulted in the early detection of a subclinical thymoma and its successful removal.

Spatiotemporal network structure among "friends of friends" reveals contagious disease process.

Disease transmission can be identified in a social network from the structural patterns of contact. However, it is difficult to separate contagious processes from those driven by homophily, and multiple pathways of transmission or inexact information on the timing of infection can obscure the detection of true transmission events. Here, we analyze the dynamic social network of a large, and near-complete population of 16,430 zoo birds tracked daily over 22 years to test a novel "friends-of-friends" strategy for detecting contagion in a social network. The results show that cases of avian mycobacteriosis were significantly clustered among pairs of birds that had been in direct contact. However, since these clusters might result due to correlated traits or a shared environment, we also analyzed pairs of birds that had never been in direct contact but were indirectly connected in the network via other birds. The disease was also significantly clustered among these friends of friends and a reverse-time placebo test shows that homophily could not be causing the clustering. These results provide empirical evidence that at least some avian mycobacteriosis infections are transmitted between birds, and provide new methods for detecting contagious processes in large-scale global network structures with indirect contacts, even when transmission pathways, timing of cases, or etiologic agents are unknown.

Association between Mycobacterium avium subsp. paratuberculosis infection among offspring and their dams in nondomestic ruminant species housed in a zoo.

The objective of the present study was to determine whether offspring of dams infected with Mycobacterium avium subsp. paratuberculosis (Map) have an increased risk for Map infection. Antemortem and postmortem disease surveillance data were used to identify positive and test-negative ruminants born at the Zoological Society of San Diego (ZSSD) between 1991 and 2007 and to estimate cumulative lifetime incidence. A matched case-control study, nested within the population, was conducted and conditional logistic regression analyses were used to quantify the association between infection status of offspring and their dams. Cases (infected ruminants, n = 47) were matched to controls (test-negative ruminants, n = 152) by species, birth date, birth enclosure, and follow-up time to control for confounding factors. The overall cumulative lifetime incidence of infection was estimated at 2.2%, but it decreased over time and varied by species. There was a significant association between infection status of offspring and their dams (odds ratio [OR] = 6.8, P < 0.01), which is consistent with studies in domestic livestock species. The association was stronger for animals whose dam was diagnosed within 2 years of their birth (OR = 9.0, P < 0.01) than for animals whose dam was diagnosed more than 2 years after their birth (OR = 6.0, P < 0.01) compared to animals with test-negative dams. For positive animals born to a positive dam, 85.3% of the Map infections were attributable to having a positive dam. For the entire population of ZSSD ruminants, 28.8% [corrected] of the cases were attributable to having a positive dam. Findings will help guide future management of Map infection in zoo ruminant populations.

Myxozoan parasitism in waterfowl.

Myxozoans are spore-forming, metazoan parasites common in cold-blooded aquatic vertebrates, especially fishes, with alternate life cycle stages developing in invertebrates. We report nine cases of infection in free-flying native and captive exotic ducks (Anseriformes: Anatidae) from locations across the United States and describe the first myxozoan in birds, Myxidium anatidum n. sp. We found developmental stages and mature spores in the bile ducts of a Pekin duck (domesticated Anas platyrhynchos). Spores are lens-shaped in sutural view, slightly sigmoidal in valvular view, with two polar capsules, and each valve cell has 14-16 longitudinal surface ridges. Spore dimensions are 23.1 microm x 10.8 microm x 11.2 microm. Phylogenetic analysis of the ssrRNA gene revealed closest affinity with Myxidium species described from chelonids (tortoises). Our novel finding broadens the definition of the Myxozoa to include birds as hosts and has implications for understanding myxozoan evolution, and mechanisms of geographical and host range extension. The number of infection records indicates this is not an incidental occurrence, and the detection of such widely dispersed cases suggests more myxozoans in birds will be encountered with increased surveillance of these hosts for pathogens.

Investigation of characteristics and factors associated with avian mycobacteriosis in zoo birds.

The objective of the current study was to identify factors associated with avian mycobacteriosis in zoo birds. Inventory data, population health records, and necropsy data from eligible birds in the Zoological Society of San Diego's (ZSSD) collection from 1991-2005 (n = 13,976) were used to describe disease incidence, prevalence, and postmortem findings. A matched case-control study was then conducted to identify factors describing demographic, temporal, and enclosure characteristics, along with move and exposure histories. Cases (disease-positive birds; n = 167) were matched in a 17 ratio with controls (disease-negative birds; n = 1169) of similar age and taxonomic grouping. Potential risk factors were evaluated using univariate and multivariable conditional logistic regression. Disease prevalence and incidence were estimated for the study period at 1.2% and 3 cases/(1,000 bird-years at risk), respectively. Lesion characteristics and order prevalence are described. In the multivariable model, case birds were more likely to have been previously housed with a bird with mycobacterial disease involving the intestinal tract (odds ratio [OR] = 5.6, P < 0.01) or involving only nonintestinal sites (OR = 2.0, P < 0.01). Cases were more likely to have been imported into the collection than hatched at the ZSSD (OR = 4.2, P < 0.01). Cases were moved among ZSSD enclosures more than controls (OR = 1.1 for each additional move, P < 0.01). Findings will help guide future management of this disease for zoo bird populations.

Early-life exposures and Johne's disease risk in zoo ruminants.

Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic, progressive bacterial enteritis of ruminants that can cause serious losses in both livestock and exotic species. Infection risk in exotic ruminants is associated with maternal infection status, but the effect of other herdmates on risk of infection has not been reported, to our knowledge. We conducted a retrospective cohort study to evaluate the association between MAP infection status and early-life contact with infected herdmates. The study population included 3,234 individuals representing 128 species at San Diego Zoo Global facilities between 1991 and 2010. Animal movement, health, and pathology records were used to trace enclosure-sharing contacts between members of the study population and any MAP-infected animal. Contact-days were counted by age of the reference animal and the number of unique infected individuals contacted. Herdmate infection status was stratified by stage of infection (180 d prior to diagnosis), age, and whether relevant lesions were found at autopsy. Having an infected herdmate was a strong risk factor for infection (OR = 4.4; 95% CI: 1.9-10.3), and each method of defining herdmate infection status showed significant differences in infection risk. The best predictor was number of contact-days within the first week of life, with a 2-fold increase in risk associated with each doubling in the number of contact-days (OR = 2.1; 95% CI: 1.1-4.0). We conclude that early contact with infected animals is an important predictor of MAP infection risk, although the effect size is smaller than that previously described for maternal infection status.

Oral eosinophilic granulomas in tigers (Panthera tigris)--a collection of 16 cases.

Oral eosinophilic granulomas were diagnosed in 16 tigers (Panthera tigris). All lesions were located on the hard or soft palate and typically consisted of flat or slightly raised circular ulcers. Histologic features of these lesions were essentially identical to those seen in oral eosinophilic granulomas of domestic cats and dogs. No clinical signs were noted in eight cases, though various degrees of inappetence, excessive salivation, and dysphagia were noted in the other eight tigers. Six cases were not treated. Treatment for the remaining 10 cases centered on corticosteroids and additional treatments included surgical removal, cryotherapy, antibiotics, and chlorpheniramine. Treatment with corticosteroids did appear to be effective in some cases, though lesions would worsen after cessation of therapy and no cases were cured. In addition, three cases developed complications possibly related to this corticosteroid therapy. The etiology of these lesions remains unknown, though an underlying allergic condition is likely.