Emerging Pathogenic Gammaproteobacteria Wohlfahrtiimonas chitiniclastica and Ignatzschineria Species in a Turkey Vulture (Cathartes aura).

New World vultures, such as turkey vultures (Cathartes aura), are obligate scavengers with large geographic ranges. In a preliminary characterization of the turkey vulture (TV) gastrointestinal microbiome in Southern California, we identified 2 recently described emerging bacterial pathogens not previously known to be associated with this avian species. High-throughput sequencing of broad-range 16S rRNA gene amplicons revealed sequences from TV cloacal swabs that were related closest to Wohlfahrtiimonas chitiniclastica and Ignatzschineria species, both Gammaproteobacteria considered by the United States Centers for Disease Control and Prevention as emerging zoonotic pathogens. None of these bacterial sequence types have been previously identified from samples obtained from the turkey vulture gastrointestinal microbiome. With the use of bioinformatics workflows previously established by our research group, we designed specific and sensitive polymerase chain reaction primer sets that represent novel diagnostic assays for the genera Wohlfahrtiimonas and Ignatzschineria. These primer sets were validated by Sanger sequence confirmation from complex TV samples. Because the genera Wohlfahrtiimonas and Ignatzschineria are both known to have dipteran hosts, the molecular diagnostic tools we present here should be useful for better understanding the role of flies, vultures, and other scavengers in the ecology and epidemiology of the genera Wohlfahrtiimonas and Ignatzschineria from a One Health perspective.

Evaluation of Multidrug Therapy With Azithromycin, Rifampin, and Ethambutol for the Treatment of Mycobacterium avium subsp avium in Ring-neck Doves (Streptopelia risoria): An Uncontrolled Clinical Study.

An uncontrolled clinical study was conducted to evaluate the efficacy of a multidrug protocol in 16 ring-neck doves ( Streptopelia risoria ) from a flock naturally infected with Mycobacterium avium subsp avium. The doves were considered infected on the basis of a high prevalence of infection in a group of 29 birds that were euthanatized from the same flock and clinical signs and pathologic results consistent with infection in the remaining birds. All birds were treated with azithromycin (43 mg/kg), rifampin (45 mg/kg), and ethambutol (30 mg/kg) administered orally once daily for 180 days. Five birds died during treatment and were confirmed positive for mycobacteriosis on postmortem examination. Of the remaining 11 birds, infection and disease were present in 9 (81.8%) at the end of the treatment. Postmortem investigation showed that 2 mycobacterial isolates were resistant to ethambutol, intermediately sensitive to rifampin, and sensitive to azithromycin. Microscopic examination of liver sections equivalent of those that would be taken for biopsy showed that biopsy as a method of monitoring birds for treatment success had poor sensitivity. Toxicity associated with drug therapy was not observed in these doves nor in 6 outwardly healthy ring-neck doves exposed to the same treatment. The results of this study showed that this protocol of azithromycin, rifampin, and ethambutol has poor efficacy when administered for 180 days for treatment of doves infected with M avium subsp avium.

A DNA vaccine expressing ENV and GAG offers partial protection against reticuloendotheliosis virus in the prairie chicken (Tympanicus cupido).

Recurring infection of reticuloendotheliosis virus (REV), an avian oncogenic gammaretrovirus, has been a major obstacle in attempts to breed and release the endangered Attwater's prairie chicken (Tympanicus cupido attwateri). The aim of this study was to develop a DNA vaccine that protects the birds against REV infection. A plasmid was constructed expressing fusion proteins of REV envelope (env) and VP22 of Gallid herpesvirus 2 or REV gag and VP22. Birds vaccinated with these recombinant plasmids developed neutralizing antibodies; showed delayed replication of virus; and had significantly less infection of lymphocytes, specifically CD4+ lymphocytes. Although the vaccine did not prevent infection, it offered partial protection. Birds in field conditions and breeding facilities could potentially benefit from increased immunity when vaccinated.

Comparison of sampling methods, culture, acid-fast stain, and polymerase chain reaction assay for the diagnosis of mycobacteriosis in ring-neck doves (Streptopelia risoria).

Even when different diagnostic modalities are available, mycobacteriosis is difficult to diagnose in a live bird. To investigate the diagnostic value of sampling different tissues and using different diagnostic methods, we evaluated results of mycobacterial culture, Ziehl-Neelsen (ZN) staining, and single-amplification polymerase chain reaction assay (PCR) of 18 ring-neck doves (Streptopelia risoria) with confirmed natural infection with Mycobacterium avium avium. Results of testing liver biopsy, duodenal aspirate, and bone marrow aspirate samples and liver and spleen samples collected at necropsy were compared. Results showed the use of one single technique did not allow identification of all infected birds. In liver biopsy and bone marrow aspirate samples, culture had the highest sensitivity, whereas PCR assay and ZN staining had low sensitivity, and their combination was less sensitive than culture alone. Examination of ZN staining of the intestinal aspirate samples failed to detect infection in most birds. More splenic lesions contained acid-fast organisms than did liver lesions, suggesting that splenic biopsy may have the greatest potential for diagnosis of mycobacterial infection antemortem. Sensitivity was higher for postmortem examination of multiple liver sections than of a single biopsy section; therefore, obtaining multiple liver biopsy sections may increase detection of mycobacteria. Examination of multiple tissues and the use of several different diagnostic techniques significantly increases the probability of diagnosis of mycobacteriosis.

Mycobacteriosis in naturally infected ring-neck doves (Streptopelia risoria): investigation of the association between feather colour and susceptibility to infection, disease and lesions type.

Prevalence of infection and disease, the degree of organ involvement and the nature of the lesions were investigated in 11 white and 18 non-white ring-neck doves coming from a flock naturally infected with Mycobacterium avium subsp. avium. Lesions were common in the liver, spleen, lung, kidney, intestines, ovary and bone marrow. Overall, 18 out of 29 (62%) birds were considered infected with a sequevar of M. avium subsp. avium that contains serotypes 2, 3, 4 and 9. The prevalence of infection in the white doves (36.4%) was significantly lower than in the non-white morphs (77.7%). White doves had on average fewer organs affected (mean =3.1) than the non-white doves (mean =5.9). A diffuse pattern of inflammation in the liver and spleen was observed mainly in non-white doves. Focal or multifocal granulomatous inflammation of the liver and spleen was predominant in white doves. Genetic mechanisms of immunity to mycobacteriosis may be contributing or determining these differences. There are three basic colour morphs in ring-neck doves--dark or wild type, blond and white--and the alleles coding for colour are sex-linked and located on the sex (Z) chromosome. Female's single sexual chromosomed (ZW) and homozygous males (ZZ) can be white if they carry the white alleles. It is very probable that the gene or genes modulating the immune response to M. avium subsp. avium infection in these doves could be associated to these loci or at least located in the same (Z) chromosome, as the association with white colour suggests.

Gross and microscopic findings and investigation of the aetiopathogenesis of mycobacteriosis in a captive population of white-winged ducks (Cairina scutulata).

The white-winged duck (Cairina scutulata) is critically endangered. Breeding collections of this duck are established in the United Kingdom and the USA. Infection with Mycobacterium avium avium serotype 1 is a major cause of mortality in the UK collection. In this study, the aetiopathogenesis of deaths occurring in the US collection was studied. All ducks (n=21) that died over a 21-month period were examined. Mycobacteriosis was diagnosed in 20 ducks, killing 19 of them. Multifocal to diffuse granulomatous lesions, often with abundant intralesional organisms, were seen in all 20 ducks. Unusual manifestations of this disease were the extensive involvement of the respiratory system and the absence of multinucleated giant cells. Sequence analysis showed that the ducks were infected with a sequevar of M. a. avium that contains serotypes 2, 3, 4, and 9. Given that the long-term ingestion of metals affects immune function, we measured an array of such elements in the liver of six ducks. Concentrations were undetectable or low. The disseminated nature of the disease, high concentration of mycobacteria and absence of multinucleated giant cells within lesions suggest that these ducks were unable to effectively kill the mycobacteria and point to a possible defect or inhibition in cell mediated immunity. Taken together with previously reported UK data, these results suggest that captive white-winged ducks are highly susceptible to at least two sequevars of M. a. avium and that mycobacteriosis is a major threat to ex situ breeding. We hypothesize that the minimal heterozygosis previously shown in these ducks could be contributing to an apparently ineffective immune response.