S. Carnaccini

Highly Pathogenic Eurasian H5N8 Avian Influenza Outbreaks in Two Commercial Poultry Flocks in California

SUMMARY In January 2015, a highly pathogenic Eurasian lineage H5N8 avian influenza (AI) virus (AIV) was detected in a commercial meat turkey flock in Stanislaus County, CA. Approximately 3 wk later, a similar case was diagnosed in commercial brown layers from a different company located in Kings County, CA. Five 14-wk-old turkey hens were submitted to the California Animal Health and Food Safety Laboratory System (CAHFS), Turlock, and eleven 12-wk-old chickens were submitted to CAHFS, Tulare laboratory due to an acute increase in flock mortality. Gross lesions included enlarged and mottled pale spleens and pancreas in turkeys and chickens. Histologically, the major lesions observed in turkeys and chickens were splenitis, pancreatitis, encephalitis, and pneumonia. In both cases, diagnosis was based on real-time reverse transcriptase PCR (RRT-PCR), sequencing, and virus isolation from oropharyngeal and cloacal swabs. Confirmatory diagnosis and AIV characterization was done at the National Veterinary Services Laboratory, Ames, IA. The sequence of the AIV from both cases was 99% identical to an H5N8 AI virus (A/gyrfalcon/Washington/41088-6/2014) isolated from a captive gyrfalcon (Falco rusticolus) from Washington State in December 2014. Immunohistochemistry (IHC) performed on various tissues from both cases indicated a widespread AIV tissue distribution. Except for minor variations, the tissue distribution of the AI antigen was similar in the chickens and turkeys. There was positive IHC staining in the brain, spleen, pancreas, larynx, trachea, and lungs in both chickens and turkeys. Hearts, ovaries, and air sacs from the turkeys were also positive for the AI antigen. The liver sections from the chickens had occasional AI-positive staining in mononuclear cells, but the IHC on liver sections from the turkeys were negative. The bursa of Fabricius, small intestine, kidney, and skeletal muscle sections were negative for the AI antigen in both chickens and turkeys.

Diagnosis and Control of a LPAI H5N8 Outbreak in a Japanese Quail (Coturnix coturnix japonica) Commercial Flock in the Central Valley of California

SUMMARY In April 2014 an outbreak of low pathogenic avian influenza H5N8 North American genetic lineage was diagnosed in a commercial quail operation in Stanislaus County, California. Sudden increase in mortality prompted the submission of 20 Japanese quail hens (Coturnix c. japonica) to the California Animal Health and Food Safety Laboratory, Turlock Branch. Oropharyngeal and cloacal swabs tested positive for influenza A virus H5N8 by real-time reverse transcription–polymerase chain reaction. The virus was subsequently isolated. In vivo assay and sequencing of the hemagglutinin protein cleavage site classified the virus as a North American genetic lineage of low pathogenicity for chickens. Following the diagnosis, a rapid and coordinated response took place to contain the outbreak. The affected premise was depopulated, cleaned, and disinfected. Three areas from the affected premises—a 3 kilometer (km) radius (High Risk Zone), a 3–10 km area (Buffer Zone), and a 10–20 km (Surveillance Zone)—were established for avian influenza testing of commercial and noncommercial poultry operations. Surveillance testing and rapid control measures were successful in the control and eradication of the outbreak and revealed no area of spread of the virus from the index flock. This report describes the history, diagnosis, surveillance, and control measures applied to manage this outbreak.

Ulcerative Dermatitis and Valvular Endocarditis Associated with Staphylococcus aureus in a Hyacinth Macaw (Anadorhynchus hyacinthinus)

SUMMARY An 18-yr-old male hyacinth macaw (Anadorhynchus hyacinthinus) was found dead in his aviary with no preexisting signs. The bird had a chronic history of feather damaging behavior, with severe ulcerative dermatitis. Pathologic findings revealed a vegetative valvular endocarditis, myocarditis, septicemia, chronic severe glomerulonephritis, and thyroid dysplasia. Staphylococcus aureus was isolated from the valve, the liver, and the skin. Repeated trauma and low-rate bacteriemia may have contributed to the development of endocarditis. Translocation of S. aureus skin infection in the bloodstream may lead to subacute endocarditis in humans and such mechanism is suspected in this case. This case suggests that endocarditis associated with S. aureus septicemia is a potential complication of feather damaging behavior. This case also reports a systemic complication of ulcerative dermatitis secondary to feather damaging behavior. Endocarditis has been poorly reported in psittacine species, and such medical complication of feather damaging behavior has never been reported to our knowledge. Furthermore, S. aureus is a bacteria of public health concern and should be integrated into the differential when pet parrots with dermatitis are in proximity to owners. RESUMEN Dermatitis ulcerativa y endocarditis valvular asociada con Staphylococcus aureus en un guacamayo azul (Anadorhynchus hyacinthinus). Un guacamayo azul o jacinto macho de 18 años de edad, (Anadorhynchus hyacinthinus) fue encontrado muerto en su aviario sin signos preexistentes. El ave tenía una historia crónica de comportamiento caracterizado por autolesión de las plumas, con dermatitis ulcerativa grave. Los hallazgos patológicos revelaron una endocarditis valvular vegetativa, miocarditis, septicemia, glomerulonefritis crónica severa y displasia de la tiroides. Se aisló Staphylococcus aureus de la válvula cardiaca, del hígado y de la piel. El trauma repetido y la bacteriemia de bajo nivel pudieron haber contribuido al desarrollo de la endocarditis. La translocación de la infección cutánea por S. aureus en el torrente sanguíneo puede conducir a endocarditis subaguda en los seres humanos y se sospecha de este mecanismo en este caso. Este caso sugiere que la endocarditis asociada con la septicemia por S. aureus es una complicación potencial del comportamiento de autolesión de las plumas. Este caso reporta una complicación sistémica de dermatitis ulcerativa secundaria a un comportamiento de autolesión de las plumas. No se ha reportado frecuentemente endocarditis en especies de psitácidos y esta complicación médica del comportamiento de autolesión de las plumas nunca se ha reportado de acuerdo con nuestro conocimiento. Por otra parte, S. aureus es una bacteria de preocupación para la salud pública y debe integrarse en el diferencial cuando loros mascotas con dermatitis se encuentran en proximidad con sus propietarios.

Trichodinosis associated with pathology of the reproductive tract in waterfowl

ABSTRACT Trichodinid ciliophorans are opportunistic parasites of many species of fish, amphibians, and molluscs, but yet never reported in association with lesions in birds. Postmortem and histopathological evaluation of a commercial adult Toulouse gander and female goose, and a wild Mallard drake revealed the presence of severe pathological parasitic colonization of their reproductive tracts. Histopathological findings included moderate to severe granulocytic inflammation, acanthosis, accentuation of the rete pegs, and proliferative hyperplastic squamous metaplasia of the mucosa of the ejaculatory ducts and groove, sulcus spermaticus, glandular part of the phallus (cavum penis), and oviduct in association with large numbers of ciliated protozoa anchored to the tissues or free in the lumen. These protozoa had characteristic morphological features analogous to the family of Trichodinidae. The source of this parasitism could not be determined. To our knowledge, this is the first report of trichodinosis associated with pathology in birds.

Pathology and Tissue Distribution of an LPAI H5N8 of North American Lineage Isolated from an Outbreak in Commercial Japanese Quail (Coturnix c. japonica) in the Central Valley of California

SUMMARY This report describes the pathology and tissue distribution of avian influenza (AI) antigens by immunohistochemistry (IHC) in the tissues of commercial layer quail from a natural outbreak of low pathogenic avian influenza (LPAI) H5N8. LPAI virus H5N8 of North American lineage was diagnosed in commercial Japanese quail hens (Coturnix coturnix japonica) in California based on serology, reverse-transcriptase real-time polymerase chain reaction, virus isolation, and sequencing. The sudden increase in mortality in a flock of laying quail hens had prompted the submission of 15 live and 5 dead, 10- to 15-wk-old quail to the California Animal Health and Food Safety Laboratory System, Turlock branch in the beginning of April 2014. There was mild bilateral swelling of the eyelids and greenish diarrhea in 4/15 live quail submitted. On postmortem examination, there were severe, extensive hemorrhages and multifocal, confluent pale foci in the pancreas in 10/20 birds. Liver gross lesions in five birds ranged from a few pale areas to numerous disseminated foci. Histology revealed moderate to severe necrosis of acinar cells in the pancreas with little or no inflammation in most of the birds. Livers had acute multifocal coagulative necrosis of hepatocytes with fibrin exudation and infiltration of few to large numbers of heterophils and lymphocytes randomly scattered throughout. The AI virus was detected in the nucleus and cytoplasm of pancreatic acinar cells and hepatocytes by IHC targeting the nucleoprotein of the AI virus. A few birds had AI antigen in the reticuloendothelial cells of the spleen, endothelial cells of the lungs, epithelium of the respiratory mucosa, and lamina propria of the intestine. The severity of the lesions observed in this natural outbreak of LPAI in quail was higher than that expected for the pathotypic presentation in this species.

Yersinia pseudotuberculosis in Eurasian collared doves (Streptopelia decaocto) and retrospective study of avian yersiniosis at the California animal health and food safety laboratory system (1990-2015)

SUMMARY In February 2015, two Eurasian collared doves (Streptopelia decaocto) were submitted dead to the California Animal Health and Food Safety (CAHFS) Laboratory, Turlock branch, from a private aviary experiencing sudden, high mortality (4/9) in adult doves. In both doves, the gross and histologic lesions were indicative of acute, fatal septicemia. Grossly, there were numerous pale yellow foci, 1 to 2 mm in diameter, in the liver and spleen. Microscopically, these foci were composed of acute severe multifocal coagulative necrosis of hepatocytes and splenic pulp with infiltration of heterophils mixed with fibrin and dense colonies of gram-negative bacteria. Yersinia pseudotuberculosis was isolated from the lung, liver, spleen, heart, ovary, kidney, and trachea. The organism was susceptible to most antibiotics it was tested against, except erythromycin. Based on a retrospective study of necropsy submissions to CAHFS between 1990 and 2015, there were 77 avian case submissions of Y. pseudotuberculosis. There were 75/77 cases identified from a wide range of captive avian species from both zoo and private facilities and 2/77 cases from two backyard turkeys submitted from one premise. The largest number of cases originated from psittacine species (31/77). The lesions most commonly described were hepatitis (63/77), splenitis (49/77), pneumonia (30/77), nephritis (16/77), and enteritis (12/77). From 1990 to 2015, there was an average of three cases of avian pseudotuberculosis per year at CAHFS. Although there were no cases diagnosed in 1993 and 1994, in all other years, there were between one and eight cases of Y. pseudotuberculosis detected from avian diagnostic submissions.

Spatial and temporal epidemiology of infectious laryngotracheitis in central California: 2000-2012.

SUMMARY In October of 2005 an outbreak of a vaccine-like strain of infectious laryngotracheitis (ILT), indistinguishable from the chicken embryo origin (CEO)-like vaccine strains, was detected by routine passive surveillance in the Central Valley of California, U. S. A. In response, a highly coordinated industry effort by two companies led to a significant decrease in the incidence of ILT over the same geographic region between 2008–2012. In order to understand the geographic and temporal spread of ILT in California before and after the outbreak, Global Information Systems (GIS) mapping coupled with spatial, temporal, and spatial-temporal statistics were used to identify retrospective and prospective low-rate clustering (i.e., less ILT than statistically expected) and high-rate clustering (i.e., more ILT than statistically expected) of ILT spatially and temporally. Results showed two high-rate retrospective spatial-temporal clusters and one low-rate prospective spatial-temporal cluster which were all statistically significant (P < 0.05). Overall, spatial-temporal clustering accounted for 36.9% of the positive ILT cases, while temporal clustering and spatial clustering done separately each accounted for 0% of the ILT cases, respectively. This demonstrates the utility of combining spatial and temporal clustering for ILT surveillance. Due to the risk of reversion to virulence and spread to immunologically naive broilers, future application of the CEO-based vaccine in the identified high rate spatial-temporal clusters should be avoided and other vaccine alternatives considered in order to avoid repeat outbreaks in those areas. This should especially be followed during the winter months of December, January, and February, which were found to have the highest prevalence of ILT (P < 0.05). Analysis of GIS data within the high-rate clusters showed that wind direction and farm density were minor factors in the spread of ILT. Shared roads may have played a role in the spread of ILT in one of the two high rate spatial-temporal clusters. RESUMEN Epidemiología espacial y temporal de laringotraqueitis infecciosa en el centro de California: 2000–2012. En octubre del 2005 un brote de una cepa similar a la vacuna contra la laringotraqueítis infecciosa (ILT), indistinguible de las cepas vacunales originadas en embrión de pollo (CEO), fue detectada mediante vigilancia pasiva de rutina en el Valle Central de California, en los Estados Unidos. En respuesta, un esfuerzo altamente coordinado de la industria por dos empresas dio lugar a una disminución significativa en la incidencia de laringotraqueítis durante la misma región geográfica entre los años 2008–2012. A fin de comprender la distribución geográfica y temporal de la laringotraqueítis en California antes y después del brote, se utilizaron Sistemas de Información Global (GIS), junto con las estadísticas espaciales, temporales y espacio-temporales para identificar la agrupación espacial y temporal de tasa baja retrospectiva y prospectiva (es decir, menor que la tasa esperada estadísticamente para la laringotraqueítis) y de agrupamiento de tasa alta (es decir, mayor que la tasa esperada estadísticamente para laringotraqueítis) para laringotraqueítis. Los resultados mostraron dos agrupamientos espaciales-temporales retrospectivos de tasa alta y una agrupación espacial-temporal prospectiva de tasa baja que fueron estadísticamente significativas (P <0.05). En general, la agrupación espacio-temporal representó el 36.9% de los casos positivos de laringotraqueítis, mientras que la agrupación temporal y la agrupación espacial por separado representó el 0% de los casos de laringotraqueítis, respectivamente. Esto demuestra la utilidad de la combinación de la agrupación espacial y temporal para la vigilancia de laringotraqueítis. Debido al riesgo de reversión a la virulencia y la diseminación a los pollos de engorde inmunológicamente susceptibles, la aplicación futura de vacunas basadas en CEO en agrupaciones espacio-temporales identificadas de tasa alta deben ser evitados y se deben considerar otras alternativas de vacunas a fin de evitar brotes repetidos en esas áreas. Esto sobre todo se debe seguir durante los meses de diciembre, enero y febrero, que se encontró que tenían la mayor prevalencia de laringotraqueítis (P <0.05). Los análisis de datos de los sistemas de información global dentro de los grupos de alta tasa mostraron que la dirección del viento y la densidad de las granjas fueron factores de menor importancia en la difusión de la laringotraqueítis. Los caminos compartidos pudieron haber desempeñado un papel en la propagación de la laringotraqueítis en uno de los dos grupos espaciales-temporales de alta tasa.

Isolation of Avipoxvirus from Tongue of Canaries (Serinus canaria) Show Severe Localized Proliferative Glossitis

SUMMARY Poxvirus was the causative agent of two unusual outbreaks of proliferative glossitis in canary (Serinus canaria forma domestica) breeders in the Northern Italy. A total of 45, 7–9-mo-old canaries were submitted in fair postmortem conditions to the Istituto Zooprofilattico Sperimentale delle Venezie at the beginning of November 2005 for diagnostic investigation. Birds belonged to two unrelated and geographically distant aviaries in northern Italy, herein identified as Aviary A and Aviary B. The two breeder flocks had both attended the same bird exposition held at the beginning of October and started experiencing an onset of high mortality 3 wk after the show. Twelve red factor-melanin canaries from Aviary A (Mantua) and 33 dominant white and recessive white canaries from Aviary B (Vicenza) were submitted for laboratory investigations. Clinical signs were unspecific and consisted of depression, ruffling of the feathers, epistaxis, and anorexia due to decreased feed and water intake. Postmortem findings revealed a severe increase in volume, thickening, and hardening of the tongue, which had turned pinkish to dark brown. No apparent gross lesions were noticed in integumentary, respiratory, and digestive systems or other internal organs. Histopathologic evaluation of the tongue revealed severe proliferation of the stratified squamous epithelium containing numerous large eosinophilic intracytoplasmic inclusion bodies (Bollinger bodies) displacing the nuclei of the cells peripherally. Severe ulceration of the surface epithelium, fibrinoheterophilic plaque formation, and moderate basal lymphoplasmacytic infiltrations were also associated with the proliferative lesion. Poxvirus was successfully isolated from the lesions in tissue cultures but not in specific-pathogen-free chicken embryonated eggs. Typical large, brick-shaped viral particles of 300–450 nm were also observed in affected tongues by transmission electron microscopy. This is the first report of multiple outbreaks of “poxvirus glossitis” in canaries.

A Novel Mycoplasma sp. Associated with Phallus Disease in Goose Breeders: Pathological and Bacteriological Findings

SUMMARY In April 2014, poor fertility in a major commercial goose breeder operation in California triggered the submission of six live affected Toulouse ganders (Anser anser) to the California Animal Health and Food Safety Laboratory, Turlock branch (University of California–Davis). Toulouse were principally affected among all breeds, and their egg fertility dropped from 65.7% to less than 33.9% in the first 40 days of the 2014 breeding season. The flock consisted of 410 adult birds, 90 males and 320 females, between 2 and 5 yr of age. Inspection of the flock revealed that 44.4% of the Toulouse ganders had severe phallic deformities that prevented them from mating. At postmortem examination, severe yellowish fibrocaseous exudate disrupted the architecture of the phallus and occasionally produced fistulating tracts through the wall of the organ. Microscopically, multifocal lymphoid nodules were noted in the mucosa and submucosa of the phallus and were associated with extensive granulomatous reaction, intralesional bacteria, and spermatozoa. Mycoplasma spp. were isolated from the phallus of affected and nonaffected birds, and PCR protocols targeting the 16S–23S ribosomal RNA intergenic spacer regions and the RNA polymerase beta subunit gene were performed to identify the isolates. Three distinct species were identified on sequencing and analysis using the National Center for Biotechnology Information basic local alignment search tool: Mycoplasma cloacale, Mycoplasma anseris, and an unknown novel Mycoplasma sp. Additionally, Pasteurella multocida, in combination with other bacteria, was also isolated from the phallic lesions and identified as serotype 3 with a DNA profile of 1511 (National Veterinary Service Laboratory). This is the first report of these Mycoplasma spp. and other bacteria associated with reproductive disease in ganders in the United States.

Characterization of Seven Outbreaks of Hemorrhagic Hepatopathy Syndrome in Commercial Pullets Following the Administration of a Salmonella Enteritidis Bacterin in California

SUMMARY Between April 2013 and April 2015, seven flocks belonging to three different major commercial egg producers inCalifornia experienced a mild increase in mortality 2 to 3 wk after administration of Salmonella Enteritidis bacterins. Strains of chickens involved were H&N (flock A1, A2, B2, C1, C2, and C3) and Lohmann white (flock B1). Vaccination was administered individually through injection either in the breast muscles or subcutis in the legs between 11 and 18 wk of age in all flocks. Clinical signs ranged from inapparent to lameness, reluctance to walk, greenish diarrhea, and retching-like symptoms. The mortality ranged from 0.16% to 1.38% per week, with the highest peaks occurring usually 2 to 3 wk postvaccination, and then declined rapidly. Postmortem examinations revealed enlarged livers with disseminated hemorrhages and pale foci of necrosis. Also, severe extensive hemorrhages in the intestine, heart, and proventriculus were observed in a few birds. Various degrees of productive, exudative giant cell granulomatous myositis were observed invading deeply the muscles and subcutis at the site of vaccination. The myositis was always associated with optically empty vacuoles positive for neutral lipids by Oil Red O stain. Droplets of Oil Red O material were also noticed in the affected livers and intestines. Congo red stain highlighted the presence of amyloid in moderate to severe amounts in the breast muscles and moderate amounts in livers, spleens, and intestines. Salmonella antigens were detected in the injection sites and livers by immunohistochemical staining. No viruses or toxic substances were recovered from the liver, spleen, intestine, and pectoral muscles, and the few bacteria isolated were interpreted as secondary postmortem invaders. In addition, livers and bile tested for hepatitis E virus were negative by reverse-transcriptase polymerase chain reaction.