FIVE YEARS OF PARTULA SNAIL PRE-EXPORT HEALTH SCREENING.

Between 2015 and 2019, a health screening was carried out annually on captive-bred Partula snails prior to export for reintroduction as part of an international effort to repopulate areas of French Polynesia, where the snails were extinct or critically endangered. In total, 129 separate tank populations of 12 different species were screened at ZSL London Zoo. Wet mounts and smears stained with modified Ziehl-Neelsen (MZN) of 535 fecal samples were examined, and 45% contained flagellated protozoa, and 35.5% had MZN-positive oocysts, measuring 3-5 µm in diameter. Smaller (2 µm) presumptive spores, MZN-positive bacilli, ciliated protozoa and nematodes were recorded less frequently. Fecal bacterial culture yielded mixed species, with a clear predominance of Myroides species (88.9% of samples). The MZN-positive oocysts (3-5 µm) were present in 6.5% of impression smears from the apices of 432 snails examined postmortem, plus acid-fast bacilli in a few cases, but no 2 µm spores. Mixed bacteria were cultured from coelomic swabs, with Myroides species again the most common (63.5%). Histologic examination was carried out on 292 snails. Autolysis affected almost 90% of those found dead but only 3.4% of euthanized snails. Histology commonly identified microsporidial sporocysts in the digestive gland and midgut epithelium of all but two species. Intracellular, extracytoplasmic Cryptosporidium-like organisms were also common in the midgut but were only observed when snails were fixed in 10% formalin (2017-2019), not ethanol. There were no clear pathologic changes associated with either organism. Pigmented hemocytic nodules were commonly observed, most frequently in the foot process; these were either age related or evidence of prior chronic inflammatory reaction and of low clinical significance. With no evidence of poor health and no significant organisms found, a total of 4,978 individuals representing 12 species were exported for reintroduction.

Identification and characterisation of Gamma-herpesviruses in zoo artiodactyla.

BACKGROUND: Viruses within the γ-herpesviruses subfamily include the causative agents of Malignant Catarrhal Fever (MCF) in several species of the order Artiodactyla. MCF is a usually fatal lymphoproliferative disease affecting non-adapted host species. In adapted host species these viruses become latent and recrudesce and transmit during times of stress or immunosuppression. The undetected presence of MCF-causing viruses (MCFVs) is a risk to non-adapted hosts, especially within non-sympatric zoological collections. This study investigated the presence of MCFVs in six different zoological collections in the UK, to evaluate the presence of subclinical/latent MCFVs in carrier animals. METHODS: One-hundred and thirty eight samples belonging to 54 different species of Artiodactyla were tested by Consensus Pan-herpes PCR. The positive samples were sequenced and subjected to phylogenetic analyses to understand their own evolutionary relationships and those with their hosts. RESULTS: Twenty-five samples from 18 different species tested positive. All viruses but one clustered in the γ-herpesvirus family and within the Macavirus as well as the non-Macavirus groups (caprinae and alcelaphinae/hippotraginae clusters, respectively). A strong association between virus and host species was evident in the Macavirus group and clustering within the caprinae group indicated potential pathogenicity. CONCLUSION: This study shows the presence of pathogenic and non-pathogenic MCFVs, as well as other γ-herpesviruses, in Artiodactyla species of conservation importance and allowed the identification of new herpesviruses in some non-adapted species.

Cancer Prevalence Across Vertebrates.

Cancer is pervasive across multicellular species, but what explains differences in cancer prevalence across species? Using 16,049 necropsy records for 292 species spanning three clades (amphibians, sauropsids and mammals) we found that neoplasia and malignancy prevalence increases with adult weight (contrary to Peto's Paradox) and somatic mutation rate, but decreases with gestation time. Evolution of cancer susceptibility appears to have undergone sudden shifts followed by stabilizing selection. Outliers for neoplasia prevalence include the common porpoise (<1.3%), the Rodrigues fruit bat (<1.6%) the black-footed penguin (<0.4%), ferrets (63%) and opossums (35%). Discovering why some species have particularly high or low levels of cancer may lead to a better understanding of cancer syndromes and novel strategies for the management and prevention of cancer.

Cancer Prevalence Across Vertebrates.

Cancer is pervasive across multicellular species. Are there any patterns that can explain differences in cancer prevalence across species? Using 16,049 necropsy records for 292 species spanning three clades (amphibians, sauropsids and mammals) we found that neoplasia and malignancy prevalence increases with adult weight and decreases with gestation time, contrary to Peto’s Paradox. Evolution of cancer susceptibility appears to have undergone sudden shifts followed by stabilizing selection. Outliers for neoplasia prevalence include the common porpoise (<1.3%), the Rodrigues fruit bat (<1.6%) the black-footed penguin (<0.4%), ferrets (63%) and opossums (35%). Discovering why some species have particularly high or low levels of cancer may lead to a better understanding of cancer syndromes and novel strategies for the management and prevention of cancer.

Somatic mutation rates scale with lifespan across mammals.

The rates and patterns of somatic mutation in normal tissues are largely unknown outside of humans1-7. Comparative analyses can shed light on the diversity of mutagenesis across species, and on long-standing hypotheses about the evolution of somatic mutation rates and their role in cancer and ageing. Here we performed whole-genome sequencing of 208 intestinal crypts from 56 individuals to study the landscape of somatic mutation across 16 mammalian species. We found that somatic mutagenesis was dominated by seemingly endogenous mutational processes in all species, including 5-methylcytosine deamination and oxidative damage. With some differences, mutational signatures in other species resembled those described in humans8, although the relative contribution of each signature varied across species. Notably, the somatic mutation rate per year varied greatly across species and exhibited a strong inverse relationship with species lifespan, with no other life-history trait studied showing a comparable association. Despite widely different life histories among the species we examined-including variation of around 30-fold in lifespan and around 40,000-fold in body mass-the somatic mutation burden at the end of lifespan varied only by a factor of around 3. These data unveil common mutational processes across mammals, and suggest that somatic mutation rates are evolutionarily constrained and may be a contributing factor in ageing.

SYSTEMIC ISOSPORIASIS (ATOXOPLASMOSIS) IN PASSERINE BIRDS AT THE ZOOLOGICAL SOCIETY OF LONDON, LONDON ZOO.

Infection with systemic Isospora species (systemic isosporiasis [SI]) is common in passerine birds and may cause substantial mortality in zoological collections. Ten years of postmortem records of 26 species of captive, nonnative passerine birds maintained at the Zoological Society of London, London Zoo, plus seven free-ranging species found dead within the zoo, were reviewed to assess cause of death and occurrence of SI (presence of merozoites in tissue impression smears and/or polymerase chain reaction [PCR] testing for Isospora DNA). The records of 287 juveniles and adults were reviewed, of which 161 had SI test results. The most common cause of death was physical (trauma, predation, drowning, and hypothermia), diagnosed in 39.0% of cases. Virulent SI was considered the cause of death in only nine individuals from five species (3.1% of all cases, 5.6% of tested birds). However, merozoites were recorded in 36.0% of the 150 individuals examined cytologically (representing 18 of the 33 species), while 45.3% of 53 spleen samples (14 species) were positive for Isospora DNA. Test agreement for the 42 birds tested by both methods was 69.0%. Assuming that the PCR result was correct in these, 37.9% of the 161 birds (21 species) were positive for SI at the time of death. These figures might underestimate prevalence because of poor DNA preservation and low numbers of individuals of some species tested. Eight new 28S rDNA sequences and 12 new internal transcriber spacer 1/2 sequences were amplified. Sequences from individuals of the same host species clustered together, suggesting a single Isospora species, and there was no evidence of overlap among hosts. These results confirm that systemic infection with Isospora species in zoo passerines is generally of low pathogenicity and most likely coevolved with their hosts. Severe disease may occur, however, with overwhelming exposure, secondary to immunosuppression, or following coinfection with another pathogen.

Cortical Morphology and White Matter Tractography of Three Phylogenetically Distant Primates: Evidence for a Simian Elaboration.

Comparative neuroimaging has been used to identify changes in white matter architecture across primate species phylogenetically close to humans, but few have compared the phylogenetically distant species. Here, we acquired postmortem diffusion imaging data from ring-tailed lemurs (Lemur catta), black-capped squirrel monkeys (Saimiri boliviensis), and rhesus macaques (Macaca mulatta). We were able to establish templates and surfaces allowing us to investigate sulcal, cortical, and white matter anatomy. The results demonstrate an expansion of the frontal projections of the superior longitudinal fasciculus complex in squirrel monkeys and rhesus macaques compared to ring-tailed lemurs, which correlates with sulcal anatomy and the lemur's smaller prefrontal granular cortex. The connectivity of the ventral pathway in the parietal region is also comparatively reduced in ring-tailed lemurs, with the posterior projections of the inferior longitudinal fasciculus not extending toward parietal cortical areas as in the other species. In the squirrel monkeys we note a very specific occipito-parietal anatomy that is apparent in their surface anatomy and the expansion of the posterior projections of the optical radiation. Our study supports the hypothesis that the connectivity of the prefrontal-parietal regions became relatively elaborated in the simian lineage after divergence from the prosimian lineage.

RISK FACTORS ASSOCIATED WITH YOLK SAC RETENTION IN CAPTIVE-BRED HUMBOLDT PENGUIN (SPHENISCUS HUMBOLDTI) CHICKS.

Multiple occurrences of yolk sac retention prompted a retrospective investigation in a recently formed colony of captive Humboldt penguins (Spheniscus humboldti). Necropsy reports of 141 parent-reared penguin chicks that died between January 2014 and December 2018 were reviewed for evidence of yolk sac retention, defined as the presence of a yolk sac at postmortem examination of a chick aged 7 d or greater, and analyzed by demographic and pathological variables for identification of risk factors. Fifty-nine (65%) chicks that died at age 7 d or greater had a retained yolk sac at postmortem examination, revealing that this was a common condition in penguins in this population. Chicks that retained their yolk sac were also more likely to present with minimal gut contents (P = 0.02), have a prominent bursa of Fabricius (P < 0.01), and be the first chick hatched of their clutch (P = 0.02). Parental experience and age were not predictive of yolk sac retention, but there was a trend for chicks with retained yolk sacs to present with a poorer body condition, reduced weight, and reduced crown-rump length compared to chicks without a retained yolk sac. Histopathological and bacteriological findings of retained yolk sacs were not significantly different from those of chicks under 7 d of age. Although likely to be multifactorial, the association between yolk sac retention and indicators of suboptimal feed intake and growth (empty gastrointestinal tract, poor body condition score, decreased crown-rump length, and decreased weight at death) is hypothesized to be a result of parental neglect, leading to starvation and absorption arrest of the yolk, as previously indicated in broiler chicks.

Diagnosis and Treatment of Heavy Metal Toxicosis in Six Waldrapp Ibis (Geronticus eremita).

Six adult Waldrapp ibis (Geronticus eremita) were presented with heavy metal toxicosis secondary to paint chip ingestion after being placed in an indoor confinement enclosure because of a regional highly pathogenic avian influenza epornitic and government restrictions. Four of the ibis developed clinical disease signs between 45 and 64 days of confinement, including weakness, lethargy, inability to fly, low body condition, and bilateral wing droop while 2 birds appeared to remain normal during that period. Common clinicopathological findings included elevated plasma aspartate aminotransferase, creatine kinase, glutamate dehydrogenase, and bile acids values. Erythrocyte morphological changes included prominent hypochromasia, decreased size of polychromatophils with occasional cytoplasmic stippling, abnormal shapes, and D cells. Whole-body radiographic imaging revealed particulate radio-opaque material in the ventriculus of all affected birds. One bird died before chelation therapy could be instituted and heavy metal testing of the liver revealed increased concentrations of lead and zinc. Lead toxicosis was diagnosed in the 5 remaining birds by plasma lead analysis. These 5 birds were hospitalized for an average of 40 days and treated with sodium calcium edetate and fluid therapy, which resulted in resolution of the toxicosis. Plasma biochemistry, complete blood counts, and blood lead values were performed repeatedly on all 5 birds and were used to guide therapy and monitor treatment response. This case series emphasizes the importance of early contingency planning and reviews the treatment of lead toxicosis in birds with a focus on long-term clinical monitoring and hematology.

Longitudinal connections and the organization of the temporal cortex in macaques, great apes, and humans.

The temporal association cortex is considered a primate specialization and is involved in complex behaviors, with some, such as language, particularly characteristic of humans. The emergence of these behaviors has been linked to major differences in temporal lobe white matter in humans compared with monkeys. It is unknown, however, how the organization of the temporal lobe differs across several anthropoid primates. Therefore, we systematically compared the organization of the major temporal lobe white matter tracts in the human, gorilla, and chimpanzee great apes and in the macaque monkey. We show that humans and great apes, in particular the chimpanzee, exhibit an expanded and more complex occipital-temporal white matter system; additionally, in humans, the invasion of dorsal tracts into the temporal lobe provides a further specialization. We demonstrate the reorganization of different tracts along the primate evolutionary tree, including distinctive connectivity of human temporal gray matter.

Isolation and characterisation of Vibrio cholerae from fish examined postmortem at ZSL London Zoo between 2014 and 2018.

BACKGROUND: When suspect Vibrio cholerae were cultured from fish at ZSL London Zoo, investigations were carried out to determine whether they were possible causes of cholera. METHODS: Bacterial culture was carried out on fish examined postmortem and colonies were identified using standard techniques including the API 20NE biochemical test kits. Suspect isolates were submitted to the Public Health England laboratory for additional testing. Separately, a number of fish were submitted for routine histopathology. RESULTS: On 13 occasions between 2014 and 2018, suspected V cholerae were cultured from individuals of eight different freshwater fish species. Archived cultures for eight of these (from six different fish species) were investigated and seven isolates (from five fish species) were confirmed as V cholerae, but all were non-O1, non-O139 strains. Whole-genome sequencing showed that the five fish species had unique V cholerae multilocus sequence types (three isolates from Aphanius danfordii were identical), all of which were genetically distant from human isolates. CONCLUSIONS: There was no evidence that these isolates could cause cholera. Histopathological changes consistent with vibriosis were seen in several fish, suggesting that V cholerae were causing the disease, but there were also concurrent infections or predisposing stress factors.

POSTMORTEM FINDINGS IN EIGHT SPECIES OF CAPTIVE CAECILIAN (AMPHIBIA: GYMNOPHIONA) OVER A TEN-YEAR PERIOD.

Between July 2007 and June 2017 there were 86 deaths in the populations of eight caecilian species at the Zoological Society of London (ZSL) London Zoo. The mortality rate (deaths per animal-year at risk) ranged from 0.03 in the Congo caecilian (Herpele squalostoma) to 0.85 in Kaup's caecilian (Potomotyphlus kaupii). Among the 73 individuals examined post mortem, no cause of death or primary diagnosis could be established in 35 cases, but of the others the most common cause of death was dermatitis (22 cases). When all significant pathological findings were considered, skin lesions of varying types were again the commonest (56 cases), particularly among the aquatic species: Typhlonectes compressicauda (18 out of 21 cases), T. natans (8/10) and P. kaupii (12/14). Other common findings were poor gut-fill (35 cases), kidney and gastrointestinal lesions (10 cases each), generalized congestion (8 cases) and poor body condition (6 cases). This review adds to the growing body of knowledge regarding the presentations and causes of disease in captive caecilians.

Digestive anatomy, physiology, resting metabolism and methane production of captive maras (Dolichotis patagonum).

The digestive physiology of maras (Dolichotis patagonum) has not been investigated in detail. Maras have a particular limb anatomy facilitating a unique cursoriality among rodents. This may also have led to additional adaptations such as a reduced volume of the gastrointestinal tract. We performed macroanatomical measurements of, and determined mean particle size along, the digestive tract of 10 semi-free-ranging animals (7.04 ±â€¯1.05 kg). Additionally, we measured CH4 emission in five captive animals (7.67 ±â€¯0.98 kg) fed a diet of pelleted lucerne, and measured food intake, digestibility, and digesta mean retention time (MRT) of a solute and three particle markers (fed at <2, 10 and 20 mm particle size). The digestive tract contents represented 11.1 ±â€¯1.4% of body mass, similar to other mammals and rodents, and there was slight indication of selective small particle retention in the caecum. Secondary peaks in marker elimination patterns suggested the possibility of caecotrophy. The MRTs were 15.4 h for the solute and 13.6 h, 13.3 h and 13.3 h for the three particle markers, respectively. At a dry matter intake of 61 ±â€¯12 g kg body mass-0.75 d-1, the maras digested organic matter and neutral detergent fibre to 48 ±â€¯8% and 34 ±â€¯10%, respectively, which is in the lower range of results from horses fed on a diet with a similar fibre content. The respiratory quotient (CO2/O2) was 0.93 ±â€¯0.03, the resting metabolic rate 346 ±â€¯35 kJ kg body mass-0.75 d-1, and CH4 emissions averaged at 3.85 ±â€¯0.47 L d-1 and 14.5 ±â€¯5.2 L per kg dry matter intake; this at a CH4/CO2 ratio of 0.042 ±â€¯0.004. Thus, the methane yield was of a magnitude expected for a hypothetical ruminant of this body mass. The results are consistent with the general understanding of hystricomorph rodent digestive physiology, including caecotrophy, but do not indicate a reduction of digestive capacity to support cursoriality. These results, and those obtained from other hystricomorph rodents, suggest that CH4 production may be more prominent in rodents than previously thought.

FERLAVIRUS-RELATED DEATHS IN A COLLECTION OF VIPERID SNAKES.

Between June and October 2013, 26 snakes of six viperid species kept in two adjoining rooms died ( n = 16) or were euthanized on medical (1) or welfare grounds (9). Two were from the main zoo collection, but the other 24 had been imported and quarantined for a minimum of 6 mo. Four of those that died and the single snake euthanized on medical grounds showed minor signs of respiratory disease prior to death, and five were weak, lethargic, and/or poor feeders. Frequent postmortem findings among all snakes were poor body condition (18) and respiratory disease (13). Seventeen cases were examined histologically, and pneumonia, sometimes with air sacculitis and/or tracheitis, was present in 15 individuals. Lung samples from 24 snakes were ferlavirus polymerase chain reaction (PCR) positive, and one of the two snakes for which only liver was available was also positive. The negative liver sample was from a snake that died of sepsis following anesthesia for surgical removal of a spindle cell sarcoma. Correlation with antemortem PCR testing of glottal and cloacal swabs in five cases was poor (sensitivity = 40%). Immunohistochemistry (IHC) for ferlaviruses on the tissues of 13 PCR-positive cases showed positive labeling in 7 only. Tissues samples from 22 ferlavirus PCR-positive snakes were examined for Chlamydia species by PCR, and 9 were positive, although DNA sequencing only confirmed two of three tested as Chlamydia pneumoniae. Immunohistochemistry for Chlamydia pneumoniae of seven cases (two Chlamydiales PCR positive, one of which was sequenced as C. pneumoniae, plus five negative) confirmed the Chlamydia PCR results. These two Chlamydiales PCR and IHC positive snakes were ferlavirus PCR positive, but IHC negative suggesting that, even though a ferlavirus was the predominant cause of the outbreak, in a few cases death may have been due to chlamydiosis with ferlavirus present, but not acting as the primary pathogen.

Development of a body condition score for the mountain chicken frog (Leptodactylus fallax).

The Critically Endangered mountain chicken frog (Leptodactylus fallax) has undergone drastic population decline due to habitat loss, hunting, invasive species, and chytridiomycosis. In response, several partner institutions initiated a conservation breeding program. It is important to maintain the captive population in good health. Therefore the program partners have recommended establishment of protocols for health examination of the species, including body condition assessment. Visual body condition scoring is a useful means to assess body condition in zoo animals for which regular bodyweight measurements are impractical or associated with capture-related stress. In this study, the authors developed a visual body condition score for the mountain chicken frog based on an ordinal categorical scale from 1 to 5 (1 = lowest body condition, 5 = highest body condition) using anatomical features that vary with total body energy reserves. Veterinary staff, animal managers, keepers, researchers, and students subsequently used the body condition score to assign scores to 98 mountain chicken frogs (41 male, 57 female) aged between 8 months and 12 years housed in five zoos in the UK and Jersey between February and March 2016. Body condition scores showed moderate (rho = 0.54; males) to strong (rho = 0.6; females) correlation with the scaled mass index, an objective measure of total energy reserves. The majority of pairwise comparisons between scores showed slight to substantial intra-observer agreement (93.8%) and slight to almost perfect inter-observer agreement (97.2%). Cases of poor agreement were likely due to limited observer experience working with the species.

OSTEOCHONDROSIS IN THE DISTAL FEMURS OF AN ADULT RETICULATED GIRAFFE (GIRAFFA CAMELOPARDALIS RETICULATA): MACROSCOPIC, RADIOLOGIC, AND HISTOLOGIC FINDINGS.

An adult male reticulated giraffe (Giraffa camelopardalis reticulata) was presented for postmortem examination. During radiologic examination of the hindlimbs, osseous cyst-like lesions were detected in both medial femoral condyles. These lesions were subsequently examined macroscopically and histologically. The gross appearance suggested a diagnosis of bilateral osteochondrosis that was confirmed with histopathologic examination. This finding has not previously been reported in giraffes. Macroscopic visualization of the major limb joints, including the femorotibial joints, is therefore encouraged in future postmortem examinations of giraffes (Giraffa camelopardalis), and further assessment of clinical significance is required.

A novel investigation of a blister-like syndrome in aquarium Echinopora lamellosa.

This study investigates potential causes of a novel blister-like syndrome in the plating coral Echinopora lamellosa. Visual inspections of this novel coral syndrome showed no obvious signs of macroparasites and the blisters themselves manifested as fluid-filled sacs on the surface of the coral, which rose from the coenosarc between the coral polyps. Histological analysis of the blisters showed that there was no associated necrosis with the epidermal or gastrodermal tissues. The only difference between blistered areas and apparently healthy tissues was the presence of proliferated growth (possible mucosal cell hyperplasia) directly at the blister interface (area between where the edge of the blister joined apparently healthy tissue). No bacterial aggregates were identified in any histological samples, nor any sign of tissue necrosis identified. We conclude, that the blister formations are not apparently caused by a specific microbial infection, but instead may be the result of irritation following growth anomalies of the epidermis. However, future work should be conducted to search for other potential casual agents, including viruses.

Batrachochytrium dendrobatidis infection and lethal chytridiomycosis in caecilian amphibians (Gymnophiona).

Batrachochytrium dendrobatidis (Bd) is commonly termed the 'amphibian chytrid fungus' but thus far has been documented to be a pathogen of only batrachian amphibians (anurans and caudatans). It is not proven to infect the limbless, generally poorly known, and mostly soil-dwelling caecilians (Gymnophiona). We conducted the largest qPCR survey of Bd in caecilians to date, for more than 200 field-swabbed specimens from five countries in Africa and South America, representing nearly 20 species, 12 genera, and 8 families. Positive results were recovered for 58 specimens from Tanzania and Cameroon (4 families, 6 genera, 6+ species). Quantities of Bd were not exceptionally high, with genomic equivalent (GE) values of 0.052-17.339. In addition, we report the first evidence of lethal chytridiomycosis in caecilians. Mortality in captive (wild-caught, commercial pet trade) Geotrypetes seraphini was associated with GE scores similar to those we detected for field-swabbed, wild animals.

Positive skin and serologic test results of diagnostic assays for bovine tuberculosis and subsequent isolation of Mycobacterium interjectum in a pygmy hippopotamus (Hexaprotodon liberiensis).

A 20-yr-old male pygmy hippopotamus (Hexaprotodon liberiensis), weighing 250 kg, arrived at Zoological Society London Whipsnade Zoo (United Kingdom) from a captive collection in Portugal. A quarantine health check was performed including a comparative intradermal tuberculosis (IDTB) test. Assessment of the comparative IDTB test at 72 hr revealed a strong positive reaction at the bovine site. Serum was tested with a rapid immunochromatographic assay (TB STAT-PAK) and was positive for tuberculosis antibodies. The tuberculosis tests were repeated 6 wk later with the same positive test outcome. In addition, a broncho-alveolar lavage (BAL) was submitted for mycobacterial culture. The positive IDTB test and TB STAT-PAK results were supported by multiantigen print immunoassay (MAPIA). Based on these results, the animal was suspected to be infected with Mycobacterium tuberculosis complex organisms and was euthanized. No gross or histologic signs of tuberculosis were found at postmortem examination. Mycobacterium interjectum was cultured from the BAL but not from necropsy samples. The antigens used in the TB STAT-PAK and MAPIA tests are reportedly specific for the M. tuberculosis complex, and so it is possible this animal presented with a latent case of tuberculosis or had a previous tuberculosis infection that resolved prior to testing. Cross-reactions with nontuberculous mycobacteria have been described with TB STAT-PAK and MAPIA tests. However, Western blotting analysis using serum from this animal did not recognize M. interjectum proteins of equivalent size to the M. tuberculosis-Mycobacterium bovis proteins recognized in the MAPIA. Thus, antigenic cross-reactivity with M. interjectum can be deemed less likely, but other nontuberculous mycobacterial proteins cannot be ruled out. It is therefore possible that false-positive reactions were obtained. These results highlight the difficulty of diagnosing tuberculosis in the absence of pathology and the presence of nontuberculous mycobacteria.