Outbreak of avian mycobacteriosis in a flock of two-year-old domestic pigeons (Columba livia f. domestica)

This report describes an outbreak of avian mycobacteriosis in a flock of 100 two-yr-old pigeons. Over a 6-mo period, the sick pigeons showed cachexia followed by death. In Columbiformes classic tubercles rarely develop, but in these affected pigeons granulomatous nodular lesions of various sizes, containing numerous acid-fast bacilli, were found in the internal organs. The lesions were observed in the liver, spleen, intestine, bone marrow, ovary, and oviduct. Despite their breeding age, atrophy was also found in the ovary and oviduct. Microorganisms belonging to Mycobacterium avium complex were identified in the affected tissues by polymerase chain reaction.     

Disseminated Mycobacterium avium complex infection following renal transplantation in a cat

An 11-year-old cat with a history of renal transplantation and treatment with cyclosporine and prednisolone was examined because of vomiting, diarrhea, inappetence, lethargy, and weight loss. Abdominal ultrasonography revealed 2 large heteroechoic masses thought to be mesenteric lymph nodes. Ultrasound-guided biopsy was performed, and histologic examination of biopsy specimens revealed granulomatous inflammation of presumptive lymph node tissue. Examination of sections stained with acid-fast stains revealed innumerable acid-fast bacilli within histiocytes, and a presumptive diagnosis of mycobacteriosis was made. The cat's clinical condition deteriorated, and euthanasia was elected. At necropsy, granulomatous inflammation was present within the mesenteric lymph nodes, spleen, liver, small and large intestines, lungs, and bone marrow. Bacterial culture yielded Mycobacterium avium, a slow-growing, opportunistic, saprophytic mycobacterium that can cause tuberculous lesions that are clinically indistinguishable from those associated with classic tuberculosis. It is a rare cause of disseminated mycobacteriosis in human transplant recipients. To our knowledge, this is the first report of disseminated M avium complex infection in a feline transplant recipient.     

Humoral response to Mycobacterium avium subsp. avium in naturally infected ring-neck doves (Streptopelia risoria)

Creation of a reliable and easy to use serologic test would greatly improve ante mortem diagnosis of Mycobacterium avium subsp. avium and aid in the control of avian mycobacteriosis, particularly in captive birds. In order to determine whether serodiagnostics could be of value in testing ring-neck doves (Streptopelia risoria) for M. a. avium infection, Western blot analysis was used to assess the humoral response of ring-neck doves exposed to M. a. avium, and to evaluate whether an association could be made between the humoral response and necropsy findings, histopathology, culture, and PCR testing. Western blot results were examined for reactivity patterns associating humoral response with infection status, severity and type of lesions (diffuse vs. multifocal granulomatous inflammation) and phenotype (white vs. non-white). A sensitivity of 88.24% and a specificity of 100% were achieved utilizing Western blot analysis to detect M. a. avium infection in ring-neck doves, offering a negative predictive value of 93% and a positive predictive value of 100%. While Western blot analysis results did not reflect lesion severity, lesion type did partially correspond with the humoral response. The findings of the present study indicate that serologic testing can be used as a valuable ante mortem screening tool for identifying ring-neck doves infected with M. a. avium.     

Molecular epidemiology of Mycobacterium avium subsp. avium and Mycobacterium intracellulare in captive birds

Mycobacterium avium subsp. avium and Mycobacterium intracellulare are primary causes of mycobacteriosis in captive birds throughout the world, but little is known about how they are transmitted. To define the local epidemiology of infection, we strain-typed 70 M. avium subsp. avium and 15 M. intracellulare culture isolates obtained over a 4-year period from captive birds. Typing was performed using randomly amplified polymorphic DNA (RAPD) PCR, amplified fragment length polymorphic (AFLP) fragment analyses, and for a subset of isolates, DNA sequencing of a segment of the 16S-23S rRNA internal transcribed spacer region. Six strain clusters comprising 43 M. avium subsp. avium, isolates were identified; 42 isolates had unique typing patterns, including all M. intracellulare isolates. Phylo-geographical analyses using RAPD and AFLP fingerprints and animal confinement histories showed no correlation between housing of infected birds and mycobacterial strain-type, except for two animals. The diversity of M. avium subsp. avium and M. intracellulare isolates and minimal evidence for bird-to-bird transmission suggest that environmental reservoirs may be important sources of infection in captivity.     

Mycobacterium avium infection in an ostrich (Struthio camelus).

Acid-fast organisms were identified by histopathology of granulomatous lesions in an ostrich (Struthio camelus). The organisms were grown in Herrold's egg media with and without mycobactin and identified as Mycobacterium avium. An agar gel immunodiffusion (AGID) test for Mycobacterium avium paratuberculosis was performed for detection of antibody for M. avium in this infected ostrich and seven other ostriches that were in contact. The results of the AGID were consistent with the pathologic diagnosis of mycobacteriosis and the isolation of M. avium in the affected ostrich.     

Isolation and identification of Mycobacterium avium subspecies silvaticum from a horse

Routine cultivation methods are able to distinguish between isolates of the Mycobacterium avium and the Mycobacterium tuberculosis complex. However, molecular tools are needed to further identify the several subspecies in the M. avium complex, especially for the subspecies avium and silvaticum. A rapid technique using HhaI restriction digestion of a 349 bp amplification product of the 85B antigen (α-antigen) gene was used for the identification of M. avium subsp. silvaticum in a three-year-old gelding presenting with caseous, necrotizing, granulomatous lesions. The result was confirmed by sequencing of the 85B antigen gene.     

An outbreak of mycobacteriosis in Gouldian finches caused by Mycobacterium peregrinum.

An outbreak of mycobacteriosis was detected in an aviary containing Gouldian finches (Erythrura gouldiae) and golden shouldered parrots (Psephotus chrysopterygius). Affected birds developed granulomatous lesions, usually of the liver and intestine. Mycobacterium peregrinum, a species of the Mycobacterium fortuitum group, was cultured on pooled samples of intestinal tract from 31 euthanized finches. These rapid-growing mycobacteria are saprophytic organisms that are generally not associated with clinical disease in immunocomponenet hosts. This is the first report of mycobacteriosis in finches implicating M peregrinum as a causative agent.     

Mycobacterium avium abortion in a sow.

A 2-year-old sow aborted her entire near-term litter of 11. Gross and histologic examination of a fetus suggested a tuberculous infection, and a yellow-pigmented Mycobacterium avium serotype 1 was subsequently isolated from the fetal tissue. Efforts to rebreed the sow were unsuccessful. She was anergic to skin tests with purified protein derivative of M. avium on two occasions but had M. avium specific in vitro lymphocyte immunostimulation. Gross granulomatous lesions were found in the liver, kidneys, and endometrium when the sow was necropsied 5 months after the abortion. Histologic examination showed diffuse and focal non-encapsulated granulomas in lymph nodes, tonsils, kidney, liver, spleen, lung, and uterine and vaginal walls. There were a few encapsulated calcified foci in the endometrium. The centers of some granulomas in the tonsils, liver, kidneys, and some lymph nodes were caseated. The yellow-pigmented M. avium was isolated from the reproductive organs and from 11 of 12 other tissues cultured.     

Real-time polymerase chain reaction testing for the detection of Mycobacterium genavense and Mycobacterium avium complex species in avian samples

Diagnosis of avian mycobacteriosis, caused by Mycobacterium genavense or species belonging to the Mycobacterium avium complex (MAC), is problematic. Polymerase chain reaction (PCR) offers rapid and sensitive detection of minute quantities of DNA, and conventional protocols have been used for evaluating avian specimens. The recent development of real-time PCR offers several advantages over conventional PCR. In attempts to improve diagnosing avian mycobacteriosis, a real-time TaqMan PCR assay was developed targeting the 65-kD heat shock protein gene of M. genavense and MAC spp. Nineteen reference isolates, 16 clinical isolates, and 32 avian tissue samples were used to evaluate the assay. When sufficient amplicons were produced, the species of mycobacteria was determined by standard sequencing of TaqMan PCR products and compared with results from commercial mycobacteriology laboratories and/or standard sequencing of conventional PCR products. The TaqMan PCR detected DNA from reference isolates of M. genavense, MAC spp., and Mycobacterium tuberculosis complex spp. Of the clinical isolates, the TaqMan PCR detected DNA from 10 of 12 Mycobacterium avium avium isolates and two of three Mycobacterium avium intracellulare isolates. For the tissue samples, the TaqMan PCR amplified DNA in six of nine samples that were identified by sequencing of conventional PCR products and/or by commercial mycobacteriology laboratories as being MAC spp. positive and three of four samples that were positive for M. genavense. There was some disagreement between speciation results from the TaqMan PCR and those from commercial mycobacteriology laboratories or conventional PCR or both. This disagreement was suspected to be because of relatively small numbers of base pairs in the TaqMan PCR products. The TaqMan PCR may provide a useful tool for evaluating clinical samples for DNA from mycobacteria species that most commonly infect birds; however, further refinement is needed in order to improve sensitivity and provide more accurate speciation.     

A model of avian mycobacteriosis: clinical and histopathologic findings in Japanese quail (Coturnix coturnix japonica) intravenously inoculated with Mycobacterium avium

Mycobacterial infections are an important cause of morbidity and mortality in birds and a considerable diagnostic challenge until the disease is advanced. In order to develop more clinically useful antemortem tests, a biological model was created that replicated naturally occurring disease. Japanese quail (Coturnix coturnix japonica; n = 8) were inoculated intravenously with Mycobacterium avium. Two additional birds served as uninoculated controls. Mean survival time of the inoculated birds was 68 +/- 13 days postinoculation (PI). Seven of the eight inoculated birds died naturally. Clinical and postmortem abnormalities in inoculated birds were characteristic of naturally occurring mycobacteriosis. Abnormal clinical findings included decreased activity, feather erection, and sudden death. Mean body weight and packed cell volume declined and mean total white blood cells (primarily heterophils, bands, and monocytes) increased from 28 days PI onward. Similar to birds that are naturally infected with mycobacteriosis, the inoculated birds were thin and had severe hepatosplenomegaly on postmortem examination. All eight birds had lesions in the liver, spleen, intestine, lung, gonads, and serosa. Less commonly affected tissues included bone marrow, thymus, gizzard, heart, pancreas, and brain. Lesions were invariably severe in the liver and spleen. These gross postmortem findings were consistent with natural infections of avian mycobacteriosis. Mycobacterium avium was isolated from the liver, spleen, and intestine of all inoculated birds. Both control birds remained disease free and culture negative. This inoculation protocol is a reliable and practical means of inducing avian mycobacteriosis for further study.     

A report of mycobacteriosis caused by Mycobacterium marinum in bullfrogs (Rana catesbeiana)

The occurrence of mycobacteriosis caused by Mycobacterium marinum in a commercial breeding farm of bullfrogs (Rana catesbeiana) in Rio de Janeiro, Brazil is described. Ten animals presented skin lesions on the head and extremities. These and 38 other asymptomatic adult animals from various tanks were killed and at necropsy disseminated granulomatous lesions were observed in the 10 clinically affected animals and in 16 (42.1%) of the asymptomatic frogs. Acid-fast bacilli were observed in all smears of the 10 symptomatic frogs and in all but one from the 16 asymptomatic animals with visceral lesions. Ten samples from the 25 positive animals were randomly selected for culture which yielded four isolates of fast-growing (<7 days) mycobacteria. Those purified isolates were characterised by biochemical traditional means as M. marinum. Identification of the strains was confirmed using reverse-phase high-performance liquid chromatography and a polymerase chain reaction (PCR) restriction enzyme analysis assay. It is suggested that M. marinum is an important agent of granulomatous disease in bullfrogs and that infected animals, even when asymptomatic, could act as reservoirs spreading the disease and contaminating other frogs in the farm.     

Polymerase chain reaction detection of Mycobacterium tuberculosis complex and Mycobacterium avium organisms in formalin-fixed tissues from culture-negative ruminants

In the US eradication program for bovine tuberculosis, a definitive diagnosis depends on the isolation of Mycobacterium bovis. However, in some cases bacterial culture is unsuccessful, even though the tissue is considered suspicious by histopathology because granulomatous lesions and acid-fast organisms are present. The purpose of this study was to determine if polymerase chain reaction (PCR) tests on formalin-fixed tissue would successfully identify the organisms observed in suspect lesions from culture-negative animals. Diagnostic laboratory records were used to select paraffin blocks of tissue from 102 ruminants that had suspect microscopic lesions but no bacterial isolation. Sections from these blocks were examined with PCR primers for IS6110 to detect Mycobacterium tuberculosis complex infection, or with 16S ribosomal RNA and IS900 primers for detection of Mycobacterium avium. The PCR tests successfully identified a mycobacterial infection in 58 of 102 tissues, including 41 M. tuberculosis complex and 17 M. avium (11 subspecies paratuberculosis). These results demonstrate that PCR testing of formalin-fixed tissue, in combination with bacterial culture, may increase the effectiveness of laboratory diagnostic efforts to detect and identify the most common mycobacterial diseases of ruminants.     

Study of Mycobacterium avium complex strains isolated from cattle in the Czech Republic between 1996 and 2000

This study surveys 2,593,348 cattle slaughtered between 1996 and 2000, and further investigates 571 (0.02%) animals found to have tuberculous lesions. Culture of 346 randomly selected tissue samples from animals younger (n = 215) and older (n = 131) than 2 years, isolated mycobacteria from 91 animals (26.3%). These included 74 Mycobacterium avium subsp. avium isolates of IS901+ and IS1245+ genotype and serotype 2, 13M. avium subsp. hominissuis isolates of IS901- and IS1245+ genotype and serotypes 8 (n = 7) and 4 (n = 6), two M. chelonae, one M. avium subsp. paratuberculosis (RFLP type B-C1), and one M. terrae. Culture of mesenteric lymph node samples obtained 66 isolates of M. avium complex (MAC) and four isolates of other mycobacterial species. M. bovis was significantly absent from all samples. Mycobacteria were more frequently (P = 0.01) isolated from tissues of animals under 2 years (34.4%) than animals over 2 years (13.0%). IS901 and IS1245 RFLP methods were used to type 17 randomly selected MAC isolates, virulent after intramuscular inoculation of pullets, from 17 different cattle herds. These revealed 11 distinct IS901 RFLP types and three IS1245 RFLP profiles. Polyclonal infection of individual animals was detected by IS901/IS1245 typing in 2 of the 17 selected isolates.     

Pathogenesis of systemic Mycobacterium avium infection in pigs through histological analysis of hepatic lesions

Mycobacterium avium causes systemic infections through primary intestinal lesions in pigs. However, its pathogenesis is not well understood. The aim of this study was to confirm the effects on swine after enteral infection. One hundred and twelve pigs with hepatic lesions infected with M. avium were used in this study. We investigated the involvement of other organs and the distribution of hepatic lesions in the lobular structure. Most lesions involved the mesenteric lymph nodes. Hepatic lymph nodes were the secondary nodes involved. In 74 cases (66.1%), the hepatic lesions were predominantly distributed in the portal tract of the affected livers. The other 38 cases (33.9%) showed granulomatous lesions in the hepatic lobule. Many cases showed interface hepatitis. There was a significant relationship between focal lesions within hepatic lobule and splenic lesions. These findings suggest that granulomatous lesions formed in hepatic lobules upon establishment of bacteremia in pigs systemically infected with M. avium.     

Development of a Johne's disease infection model in laboratory rabbits following oral administration of Mycobacterium avium subspecies paratuberculosis

To assess the rabbit as a model for the study of Johne's disease pathogenesis, a breeding group of adult and juvenile New Zealand white rabbits were orally challenged with three doses of the Mycobacterium avium subspecies paratuberculosis wildtype bovine strain, CLIJ623, on three occasions. Faecal culture, post-mortem tissue bacteriological culture and histopathology were used to monitor the disease progression in the rabbits for more than 2 years. Of 4 adult and 16 juvenile orally dosed rabbits M. paratuberculosis organisms were recovered bacteriologically from two and three animals, respectively, using the BACTECtrade mark radiometric culture system. Tissue sites from which the bacteria were recovered included the mesenteric lymph nodes, ileocaecal valve, vermiform appendix, caecum, proximal colon and jejunum. Body weight loss, reduced abdominal fat and mild lesions were observed at necropsy in four infected rabbits. Diarrhoea and persistent faecal shedding of bacteria were not observed. Faecal culture did not yield any cultivable mycobacterial organisms on solid media.     

Mycobacterium avium infection in a farmed deer herd

Tuberculous lesions were identified over a 2-year period in 36 clinically normal red deer from a single herd. The lesions were only present in the retropharyngeal lymph nodes and lymph nodes draining the intestinal tract, indicating infection by the oral route. Mycobacterium avium was isolated from 27 of 29 lesions examined by bacterial culture. Grossly and histologically, the lesions were indistinguishable from those caused by Mycobacterium bovis. DNA restriction endonuclease analysis revealed that all the 26 M. avium isolates available for examination had identical cleavage patterns. These patterns were identical to a New Zealand M. avium serotype 2 isolate from a pig and were very similar to a reference strain of M. avium serotype 2. The DNA examinations indicated that the deer were infected from a common source that was not identified.     

Disseminated mycobacteriosis due to Mycobacterium avium in captive Bengal tiger (Panthera tigris).

A 2-year-old captive female Bengal Tiger (Panthera tigris) died after prolonged anorexia in the Gwangju Uchi Park Zoo, Gwangju, Republic of Korea. Necropsy revealed multiple nodules of varying sizes in the lung, liver, kidney, and spleen. Histopathologic examination revealed a typical granuloma composed of caseous necrotic areas surrounded by lymphocytes with a few giant cells and foamy macrophages. Periodic acid-Schiff stain and Gomori methenamine silver stain did not reveal any fungal bodies. The Ziehl-Neelsen acid-fast stain revealed few acid-fast organisms in the lung, liver, kidney, and spleen. A polymerase chain reaction assay of the lung, liver, kidney, and spleen yielded a positive result for Mycobacterium avium subsp. avium. This is an unusual case of disseminated infection of a wild mammal with avian mycobacteriosis, and is believed to be most likely associated with the feeding of tigers with culled chickens infected with M. avium.     

Mycobacterial disease in cats in Great Britain: I. Culture results, geographical distribution and clinical presentation of 339 cases

This study investigated 339 cases of feline mycobacterial disease from cats with cutaneous lesions or masses found at exploratory laparotomy. Tissue samples were submitted to the Veterinary Laboratories Agency for mycobacterial culture over a 4-year period to December 2008. The study assessed which species of culturable mycobacteria were involved, where the cats lived, and their clinical presentation (physical findings, serum biochemistry, radiography, feline leukaemia virus and feline immunodeficiency virus status). Mycobacterium microti was cultured from 19%, Mycobacterium bovis 15%, Mycobacterium avium 7%, non-M avium non-tuberculous mycobacteria 6%, with no growth in 53% of samples. M microti, M bovis and M avium were found in almost mutually exclusive clusters within Great Britain (GB) (ie, M bovis in South-West England/Wales/Welsh Border, M avium in eastern England and M microti south of London and in South-West Scotland). While differences were seen in the clinical presentation and distribution of lesions caused by the different infections, these were not sufficiently different to be diagnostic. Cats commonly presented with single or multiple cutaneous lesions (74%), which were sometimes ulcerated or discharging, located most frequently on the head (54%). Lymph nodes were usually involved (47%); typically the submandibular nodes. Systemic or pulmonary signs were rarely seen (10-16%). When a cat is suspected of having mycobacteriosis, accurate identification of the species involved helps to determine appropriate action. Our findings show that knowing the cat's geographic location can be helpful, while the nature of the clinical presentation is less useful. Most cases of feline mycobacterial disease in GB are cutaneous.     

Disseminated Mycobacterium avium subsp. avium infection in a pet Korean squirrel (Sciuris vulgaris coreae)

A disseminated Mycobacterium avium subsp. avium infection was diagnosed in a pet Korean squirrel. Grossly, multiple small nodules in the lung, liver, spleen, and skin were observed. Adrenal glands were very enlarged. The only tissue exhibiting necrosis and calcification was a very enlarged bronchial lymph node. The remaining lymph nodes were slightly enlarged. Moderate ascites was also observed. Microscopically, a disseminated granulomatous inflammation with numerous lymphocytes was seen. Acid-fast bacilli were detected in macrophages, in giant cells, free in the interstitium, and in some lymphatic vessels, both within cells and free in the lumen. M. avium subsp. avium was isolated and identified by polymerase chain reaction-restriction endonuclease analysis.