Avian Reoviruses of the Same Genotype Induce Different Pathology in Chickens.

The emergence of avian reovirus variant strains has caused negative effects in the poultry industry worldwide. Regardless of the efforts in molecular characterization and classification of these variants, information about the pathogenicity, transmissibility, and immunosuppression in chickens is limited. The genomes of two variant strains (A and B) and a classic S1133 strain (C) belonging to the same sigma C genotype 1 were compared. Additionally, these strains were used in a challenge experiment to evaluate inoculated and indirectly exposed specific-pathogen-free chickens. The whole-genome sequence analysis of the three strains revealed nucleotide identity differences in the L3, M2, and S1 genes. Strains A and B also showed homology differences in the S4 gene, despite having high homologies in all other genes. The in vivo challenge experiments showed that, whereas variant A induced high viral loads in tendons, hearts, and duodena of inoculated chickens, variant B induced high viral loads in indirectly exposed chickens. Likewise, histopathology reflected differences in the pathologic effects induced by these strains. For instance, the B and C strains induced more severe microscopic lesions compared with the A strain. Lymphoid depletion was more severe in bursas than in thymi, and inoculated birds were more affected than exposed birds. In conclusion, different pathologic outcomes in chickens were observed depending on the strain and transmission route. This study provides insights onto the relationship between pathogenicity and genomic composition of avian reoviruses.

Reovirus aviares del mismo genotipo inducen diferentes patologías en pollos. La aparición de cepas variantes del reovirus aviar ha causado efectos negativos en la industria avícola en todo el mundo. Independientemente de los esfuerzos en la caracterización molecular y clasificación de estas variantes, la información sobre la patogenicidad, transmisibilidad e inmunodepresión en pollos es limitada. Se compararon los genomas de dos cepas variantes (A y B) y una cepa S1133 clásica (C) perteneciente al mismo genotipo 1 del gene sigma C. Además, estas cepas se utilizaron en un experimento de desafío para evaluar pollos libres de patógenos específicos inoculados y expuestos indirectamente. El análisis de la secuencia del genoma completo de las tres cepas reveló diferencias de identidad de nucleótidos en los genes L3, M2 y S1. Las cepas A y B también mostraron diferencias de homología en el gene S4, a pesar de tener altas similitudes en todos los demás genes. Los experimentos de exposición in vivo mostraron que, mientras que la variante A inducía altas cargas virales en tendones, corazones y duodeno en los pollos inoculados, la variante B inducía altas cargas virales en pollos expuestos indirectamente. Asimismo, la histopatología reflejó diferencias en los efectos patológicos inducidos por estas cepas. Por ejemplo, las cepas B y C indujeron lesiones microscópicas más graves en comparación con la cepa A. La despoblación linfoide fue más severa en las bolsas que en el timo, y las aves inoculadas resultaron más afectadas que las expuestas. En conclusión, se observaron diferentes resultados patológicos en pollos según la cepa y la vía de transmisión. Este estudio proporciona información sobre la relación entre la patogenicidad y la composición genómica de los reovirus aviares.

Persistence of Highly Pathogenic and Low Pathogenic Avian Influenza Viruses in Footbaths and Poultry Manure.

A questionnaire was designed in order to gather information about bedding material and footbath preparation and maintenance in different productive units across the state of California.This information was used to plan two experiments. In the first experiment, we tested the effectiveness of footbaths in inactivating highly pathogenic (HP) and low pathogenic (LP) avian influenza viruses (AIVs) on rubber boots. Surprisingly, quaternary ammonia- and quaternary ammonia + glutaraldehyde-based footbaths were not able to eliminate live HPAIV (H5N8) and LPAIV (H6N2) particles on boots, while a chlorine-based granulated disinfectant was able to destroy the virus at contact. These results demonstrated the potential of AIV, particularly the HPAIV isolate, to persist even if exposed to disinfecting footbaths, and suggest that footbaths, as a single tool, are not capable of preventing pathogen introduction into commercial flocks. In the second experiment, we investigated the persistence of HPAIV (H5N8) and LPAIV (H6N2) in bedding material and feces obtained from turkey, broiler, and egg-layer commercial productive units. Samples were collected at different times after spiking the bedding materials and feces. Results showed that HPAIV (H5N8) was more persistent than LPAIV (H6N2) in layer feces and bedding material obtained from commercial broilers and turkeys. Live HPAIV particles persisted 96 hr, the last time point measured, in layer feces and less than 60 hr in broiler and turkey bedding. In contrast, LPAIV persisted less than 24 hr after being spiked in all the different substrates. Further research in biosecurity practices such as footbath preparation and maintenance and better understanding of the mechanism of the increased persistence of AIV is warranted in order to identify effective litter treatments that destroy live virus in bedding material.

Sarcocystis neurona encephalitis in a dog.

A 1.5-year-old male Feist dog was presented to a veterinarian for reluctance to stand on the hind legs. Treatment included dexamethasone and resulted in a favorable initial response, but posterior paresis returned and progressed to recumbency, hyperesthesia, and attempts to bite the owner. The dog was euthanized. The brain was negative for rabies by fluorescent antibody analysis. Multiple foci of encephalitis were found in the cerebrum and particularly in the cerebellum. Protozoa morphologically consistent with Sarcocystis sp. were identified at sites of intense inflammation and malacia. Additionally, multiple schizonts were identified in areas without inflammation. Immunohistochemistry using both polyclonal and monoclonal antibodies specific for Sarcocystis neurona was strongly positive. No reaction to polyclonal antisera for Toxoplasma gondii or Neospora caninum was found. Polymerase chain reaction confirmed that the protozoa were S. neurona. Additional aberrant hosts for S. neurona other than horses have been identified, but S. neurona encephalitis has not been documented previously in the dog.

Chronic polymyositis associated with disseminated Sarcocystosis in a captive-born rhesus macaque.

A 2-year-old, captive-born, clinically healthy male, rhesus macaque, was euthanatized as part of an experimental study. At necropsy, diffuse pale streaking of the trunk, lumbar, and limb muscles were noted macroscopically. On histology, numerous elongated cysts that contained crescent-shaped basophilic spores were found in the fibers of skeletal muscles. Scattered affected myofibers were degenerate and accompanied by eosinophilic-to-granulomatous inflammation. Sarcocysts had prominent villus-like projections with the morphology of a type 11 sarcocyst wall similar to Sarcocystis neurona but possessing many more villus microtubules than is reported for S. neurona. In addition, bradyzoites were very long, up to approximately 12 microm in length. The protozoa were consistent with a Sarcocystis sp., based on histology and ultrastructure, however, a definitive identification of the species was not possible. Nonspecific immunohistochemical crossreaction with Sarcocystis cruzi antisera was observed. The 18S ribosomal deoxyribonucleic acid sequence showed 91% similarity to Sarcocystis hominis, 90% similarity to Sarcocystis buffalonis, and 89% similarity to Sarcocystis hirsuta. Interestingly, the ITS1 sequence showed very little homology to any sequence in GenBank, suggesting that this is possibly a unique Sarcocystis sp. Sarcocystosis is often considered an incidental finding, particularly in wild-caught animals, with little clinical significance. However, as demonstrated in this report and others, disseminated sarcocystosis can occur in captive-born rhesus macaques with or without clinical signs. In some cases interference with research results can occur; including death in fulminant cases.