Natural Infection with H5N1 Highly Pathogenic Influenza (HPAI) Virus in 5- and 10-Day-Old Commercial Pekin Ducklings (Anas platyrhynchos domesticus).

Highly pathogenic avian influenza (HPAI) has resulted in catastrophic economic losses globally in poultry. This case report describes the diagnostic detection and pathology of HPAI H5N1 in 5-day-old commercial ducklings, which is an atypical age for detection of natural infection of HPAI in poultry. The pathology observed at 5 days of age was also compared to lesions observed in ducklings from the same flock evaluated at 10 days of age before depopulation. The California Animal Health and Food Safety (CAHFS) Laboratory, Tulare, received ten 5-day-old Pekin duckling (Anas platyrhynchos domesticus) carcasses for diagnostic evaluation due to mortality that started increasing at 3 days of age. The most common gross findings included bilateral pulmonary edema with congestion and enlarged, mottled livers and spleens. Microscopically, cerebral neuronophagia, pancreatic necrosis, and interstitial pneumonia with pulmonary edema were observed in the 5-day-old ducklings. Oropharyngeal and cloacal swabs were positive for avian influenza virus (AIV) by real-time reverse transcriptase PCR. The AIV was typed as HPAI, EA/AM 2.3.4.4b H5N1 goose/Guangdong clade lineage by the National Veterinary Services Laboratory. Ducks at the affected premises were depopulated 4 days after the 5-day-old ducklings were submitted to the CAHFS lab, at which time additional tissue samples were collected for comparison to 10-day-old ducklings on the same premises. Differences in microscopic lesions and AIV tissue distribution were observed between the 5-day and 10-day tissues collected. Notably, microscopic lesions were more severe in the brain and pancreas at 10 days of age. Findings in 10-day-old ducklings included cerebral lymphoplasmacytic perivascular cuffing, gliosis, neuronal degeneration, and pancreatic necrosis. AIV antigen distribution and intensity was greatest in the cerebral tissue of the brains at 10 days and in the lungs at 5 days of age. To the authors' knowledge, published studies are limited on AIV natural infection in domestic ducks less than 9 days of age.

Infección natural con el virus de la influenza altamente patógena (HPAI) H5N1 en patitos Pekín comerciales (Anas platyrhynchos domesticus) de 5 y 10 días de edad. La influenza aviar altamente patógena (HPAI) ha provocado pérdidas económicas catastróficas en todo el mundo entre las aves de corral. Este reporte de caso describe la detección diagnóstica y la patología de la infección por un virus de influenza aviar de alta patogenicidad H5N1 en patitos comerciales de 5 días de edad, que es una edad atípica para la detección de la infección natural del virus de la influenza aviar de alta patogenicidad en avicultura. La patología observada a los 5 días de edad también se comparó con las lesiones observadas en patitos de la misma parvada evaluados a los 10 días de edad, antes de la despoblación. El Laboratorio de Salud Animal y Seguridad Alimentaria de California (CAHFS), con sede Tulare, recibió 10 cadáveres de patito Pekín (Anas platyrhynchos domesticus) de 5 días de edad para su evaluación diagnóstica debido a que la mortalidad comenzó a aumentar a los 3 días de edad. Los hallazgos macroscópicos más comunes incluyeron edema pulmonar bilateral con congestión en hígado y bazos agrandados y moteados. Microscópicamente se observó neuronofagia cerebral, necrosis pancreática y neumonía intersticial con edema pulmonar en los patitos de 5 días de edad. Los hisopos orofaríngeos y cloacales fueron positivos para el virus de la influenza aviar (AIV) mediante transcripción reversa y PCR en tiempo real. El Laboratorio Nacional de Servicios Veterinarios clasificó al virus como de alta patogenicidad EA/AM 2.3.4.4b H5N1 clado de linaje de ganso/clado Guangdong. Los patos en las instalaciones afectadas fueron despoblados 4 días después de que los patitos de 5 días fueran enviados al laboratorio de CAHFS, momento en el cual se recolectaron muestras de tejido adicionales para compararlas con patitos de 10 días de las mismas instalaciones. Se observaron diferencias en las lesiones microscópicas y la distribución del tejido del AIV entre los tejidos recolectados de 5 y 10 días. En particular, las lesiones microscópicas fueron más severas en el cerebro y en el páncreas a los 10 días de edad. Los hallazgos en patitos de 10 días incluyeron infiltraciones linfoplasmocitarias perivasculares en el cerebro, gliosis, degeneración neuronal y necrosis pancreática. La distribución e intensidad del antígeno de influenza aviar fue mayor en el tejido cerebral de los cerebros a los 10 días y en los pulmones a los 5 días de edad. De acuerdo al conocimiento de los autores, los estudios publicados sobre la infección natural por el virus de la influenza aviar en patos domésticos de menos de 9 días de edad son limitados.

Whole-genome sequence of a genotype VIII infectious bronchitis virus isolated from California layer chickens in 2021.

Herein, we report the complete genome for an avian infectious bronchitis virus isolated from cecal tonsils of California layers in 2021. This whole-genome sequence belongings to genotype GVIII, previously classified as a unique variant.

Gastrointestinal Impaction and Necrotic Enteritis in a Backyard Chicken.

The carcass of a 4-mo-old, female, mixed-breed backyard chicken was submitted for postmortem evaluation and diagnostic workup. The bird was previously presented to a veterinary clinic because of chronic weight loss and loose stool, and was euthanized before submission to the California Animal Health and Food Safety, Turlock lab. On gross examination, the proventriculus, gizzard, and duodenum were markedly distended and impacted with a mixture of fibrous plant material, cereal grain, and litter material. The koilin layer of the gizzard was eroded. There were multifocal to coalescing, 0.2-1-cm diameter white nodules on the serosal surface of the duodenal loop and lesions extended into the distal jejunum. The duodenum had multifocal, transmural, umbilicated, and ulcerated mucosal lesions, which were covered with a white pseudomembrane. Microscopically, there was segmental, transmural necrosis of the intestinal wall with diffuse sloughing of villi epithelium and accumulation of fibrino-hemorrhagic exudate with numerous bacterial colonies in the lumen. The gross and microscopic findings were indicative of gastrointestinal impaction and necrotic enteritis. Proliferation of Clostridium perfringens within the intestine was demonstrated by anaerobic bacterial culture, intestinal gram stains, and immunohistochemistry. The C. perfringens isolate was type F (encoding the gene for alpha toxin -cpa- and for enterotoxin -cpe) by PCR toxinotyping. Overgrowth of C. perfringens was likely exacerbated by the rough fibrous forage and highly fermentable grain diet. To our knowledge, gastrointestinal impaction concurrent with necrotic enteritis has not been described in backyard chickens. In addition, to our knowledge, C. perfringens type F has not been associated with necrotic enteritis in chickens.

Reporte de caso- Impactación gastrointestinal y enteritis necrótica en un pollo de traspatio. Para realizar una evaluación post mortem y estudios de diagnóstico, se recibió un pollo de traspatio muerto, hembra, de raza mixta y de cuatro meses de edad. El ave fue presentada previamente a una clínica veterinaria debido a la pérdida de peso crónica y heces acuosas y fue sacrificada antes de ser enviada al Laboratorio de Salud Animal y Seguridad Alimentaria de California con sede en Turlock. En el examen macroscópico, el proventrículo, la molleja y el duodeno estaban marcadamente distendidos e impactados con una mezcla de material vegetal fibroso, granos de cereal y material de hojarasca. La capa de koilin de la molleja estaba erosionada. Había nódulos blancos de 0.2 a 1 cm de diámetro, multifocales a coalescentes, en la superficie serosa del asa duodenal y las lesiones se extendían hacia el yeyuno distal. El duodeno presentaba lesiones mucosas multifocales, transmurales, umbilicadas y ulceradas, las cuales estaban cubiertas por una pseudomembrana blanca. Microscópicamente, había necrosis transmural segmentaria de la pared intestinal con desprendimiento difuso del epitelio de las vellosidades y acumulación de exudado fibrino-hemorrágico con numerosas colonias bacterianas en el lumen. Los hallazgos macroscópicos y microscópicos fueron indicativos de impactación gastrointestinal y enteritis necrótica. La proliferación de Clostridium perfringens dentro del intestino se demostró mediante cultivo de bacterias anaerobias, tinciones de Gram intestinales e inmunohistoquímica. El aislado de C. perfringens fue tipo F (que codifica el gene de la toxina alfa ­cpa- y de la enterotoxina ­cpe) por tipificación de toxina mediante PCR. El crecimiento excesivo de C. perfringens probablemente fue exacerbado por el forraje áspero y fibroso y la dieta de granos altamente fermentables. Hasta donde se conoce, la impactación gastrointestinal concurrente con enteritis necrótica no se ha descrito en pollos de traspatio. Además, hasta donde se sabe, C. perfringens tipo F no se ha asociado con enteritis necrótica en pollos.

Infectious Bronchitis Virus California Variant CA1737 Isolated from a Commercial Layer Flock with Cystic Oviducts and Poor External Egg Quality.

False layer syndrome is a condition in which the reproductive tract of chicks is infected with infectious bronchitis virus (IBV) strains that cause permanent damage to the oviduct. These chickens subsequently develop cystic oviducts and do not lay eggs, and affected flocks fail to reach expected egg production peaks. The California Animal Health and Food Safety laboratory, Turlock Branch, received four separate case submissions from a 25-to-28-wk-old commercial ISA Brown layer flock. Birds were submitted for diagnostic evaluation due to suboptimal egg production and vent pecking. Submissions totaled 31 birds and consisted of live layers, recent mortality, and a flat of eggs. No clinical signs were observed in the submitted live birds. The most common gross findings included cystic left oviducts, signs of vent pecking, ovarian regression, and yolk coelomitis. The eggs were abnormally shaped with irregular, white, gritty deposits on the surface of the shell. Microscopically, there was atrophy of the oviducts, glandular hypoplasia, and lymphocytic salpingitis. In addition, lymphoplasmacytic tracheitis was observed, and renal tubules were dilated with multifocal areas of mineralization. IBV was identified by reverse transcription quantitative PCR from cecal tonsil tissue pools and tracheal swab pools. Sequencing of the S1 hypervariable region of IBV and whole-genome IBV sequencing were 97% homologous to the California variant CA1737/04. Definitive proof of the CA1737 strain's causing reproductive abnormalities will require challenge studies with fulfillment of Koch's postulates and evaluation of confounding and risk factors.

Reporte de caso- Virus de la bronquitis infecciosa Variante de California CA1737 aislada de una parvada comercial de ponedoras con oviductos quísticos y mala calidad externa del huevo. El síndrome de la falsa capa es una condición en la cual el tracto reproductivo de las gallinas está infectado con cepas del virus de la bronquitis infecciosa (IBV) que causan daño permanente al oviducto. Posteriormente, estas gallinas desarrollan oviductos quísticos y bajas en la postura de huevo, las parvadas afectadas no alcanzan los picos de producción de huevos esperados. El laboratorio de Salud Animal y Seguridad Alimentaria de California, con sede en Turlock, recibió cuatro casos separados de una parvada comercial de ponedoras ISA Brown de 25 a 28 semanas de edad. Las aves se enviaron para evaluación diagnóstica debido a una producción de huevos subóptima y por presencia de picoteo en las cloacas. Se recibieron un total de 31 aves y consistieron en aves de postura vivas, mortalidad reciente y además una charola de huevos. No se observaron signos clínicos en las aves vivas enviadas. Los hallazgos macroscópicos más comunes incluyeron oviductos izquierdos quísticos, signos de picoteo en las cloacas, regresión ovárica y celomitis de la yema. Los huevos tenían una forma anormal con depósitos irregulares, blancos y arenosos en la superficie de la cáscara. Microscópicamente, había atrofia de los oviductos, hipoplasia glandular y salpingitis linfocítica. Además, se observó traqueítis linfoplasmocítica y túbulos renales dilatados con áreas multifocales de mineralización. El virus de la bronquitis infecciosa se identificó mediante PCR cuantitativa de transcripción inversa a partir de grupos de tejidos de tonsilas cecales y muestras agrupadas de hisopos traqueales. La secuenciación de la región hipervariable S1 de IBV y la secuenciación de IBV del genoma completo fueron homólogas en un 97 % a la variante de California CA1737/04. La prueba definitiva de las anomalías reproductivas causantes de la cepa CA1737 requerirá estudios de desafío con el cumplimiento de los postulados de Koch y la evaluación de los factores de riesgo y de confusión.

Necrotic enteritis in a commercial turkey flock coinfected with hemorrhagic enteritis virus.

Four turkeys from a commercial flock with acutely elevated mortality levels were submitted for postmortem examination and diagnostic workup. No clinical signs had been observed before death. On gross examination, hemorrhage and necrosis were present throughout the intestinal tracts, and the spleens were markedly enlarged and speckled. Microscopically, numerous, large basophilic-to-amphophilic intranuclear inclusion bodies were observed in mononuclear cells of the spleen and the lamina propria of the small intestine. In addition, there were lesions of diffuse villus blunting and necrosis of the small intestine, with large numbers of rod-shaped bacteria adhered to the epithelium and in the intestinal lumen. Hemorrhagic enteritis virus (HEV) infection was confirmed via PCR on the spleen. Clostridium perfringens was demonstrated in the small intestine by anaerobic culture and immunohistochemistry. The C. perfringens isolate was type F by PCR and, to our knowledge, necrotic enteritis in turkeys has not been described in association with C. perfringens type F infection.

An Uncommon Case of Trueperella pyogenes Infection in an Adult Backyard Rooster and a Retrospective Study; 2000-20.

Trueperella pyogenes is an opportunistic Gram-positive bacterium that induces purulent lesions and abscesses in cattle, small ruminants, and swine. In birds, T. pyogenes infections have been linked to lameness and osteomyelitis in turkeys (Phasianidae) and hepatic fibriscess in turkeys and pigeons (Columbidae). An 18-mo-old backyard rooster with a history of progressive emaciation was submitted to the California Animal Health and Food Safety (CAHFS) laboratory system. At necropsy, unusual numerous miliary granulomas were identified, primarily in the spleen, but granulomas were also observed in air sacs and lungs. Microscopically, few to moderate numbers of granulomas with giant cells were observed in the spleen, lung, air sacs, and crop composed of necrosis and mixed inflammatory cell inflammation including multinucleated giant cells, fibrin deposition, and fibrosis. Trueperella pyogenes was isolated from the air sacs and trachea. Avibacterium paragallinarum PCR was positive from the tracheal swab. A retrospective analysis of CAHFS data on T. pyogenes between 2000 and 2020 identified 24 cases in avian species: chickens (Gallus gallus domesticus; 16/24), turkeys (5/24), Pekin duck (Anas platyrhynchos domesticus; 1/24), parrot (Psittaciformes; 1/24), and pheasant (Phasianidae; 1/24). Although T. pyogenes infection in birds is rare, the clinical signs and gross lesions might be indistinguishable from avian mycobacteriosis in some cases and should be considered in the differential diagnosis.

Reporte de caso­Un caso no común de infección por Trueperella pyogenes en un gallo adulto de traspatio y un estudio retrospectivo; entre los años 2000-20. Trueperella pyogenes es una bacteria grampositiva oportunista que induce lesiones purulentas y abscesos en bovinos, pequeños rumiantes y porcinos. En las aves, las infecciones por T. pyogenes se han relacionado con cojera y osteomielitis en pavos (Phasianidae) y fibrosis hepática en pavos y palomas (Columbidae). Un gallo de traspatio de 18 meses de edad con antecedentes de emaciación progresiva fue enviado al sistema de Laboratorios de Salud Animal y Seguridad Alimentaria de California (CAHFS). En la necropsia, se identificaron numerosos granulomas miliares inusuales, principalmente en el bazo, pero también se observaron granulomas en los sacos aéreos y los pulmones. Microscópicamente, se observaron pocos a moderados granulomas con células gigantes en el bazo, pulmón, sacos aéreos y buche compuesto por necrosis e inflamación celular inflamatoria mixta, incluidas células gigantes multinucleadas, depósito de fibrina y fibrosis. Trueperella pyogenes se aisló de los sacos aéreos y la tráquea. Un método de PCR para Avibacterium paragallinarum fue positivo realizado a partir de hisopos traqueales. Un análisis retrospectivo de los datos de CAHFS sobre T. pyogenes entre los años 2000 y 2020 identificó 24 casos en especies aviares: pollos (Gallus gallus domesticus; 16/24), pavos (5/24), pato Pekín (Anas platyrhynchos domesticus; 1/24), loro (Psittaciformes; 1/24) y faisán (Phasianidae; 1/24). Aunque la infección por T. pyogenes en aves es poco común, los signos clínicos y las lesiones macroscópicas pueden ser indistinguibles de micobacteriosis aviar en algunos casos y debe considerarse como diagnóstico diferencial.

Age-Associated Changes in Recombinant H5 Highly Pathogenic and Low Pathogenic Avian Influenza Hemagglutinin Tissue Binding in Domestic Poultry Species.

The 2014 outbreak of clade 2.3.4.4A highly pathogenic avian influenza (HPAI) led to the culling of millions of commercial chickens and turkeys and death of various wild bird species. In this outbreak, older chickens and turkeys were commonly infected, and succumbed to clinical disease compared to younger aged birds such chicken broilers. Some experimental studies using waterfowl species have shown age-related differences in susceptibility to clinical disease with HPAI viruses. Here, we evaluate differences in H5 Hemagglutinin (HA) tissue binding across age groups, using recombinant H5 HA (rHA) proteins generated using gene sequences from low pathogenic (A/mallard/MN/410/2000(H5N2 (LPAIV)) and a HPAIV (A/Northern pintail/Washington/40964/2014(H5N2)) influenza A virus (IAV). Respiratory and intestinal tracts from chickens, ducks (Mallard, Pekin, Muscovy) and turkeys of different age groups were used to detect rHA binding with protein histochemistry, which was quantified as the median area of binding (MAB) used for statistical analysis. There were species and tissue specific differences in the rHA binding among the age groups; however, turkeys had significant differences in the HPAIV rHA binding in the respiratory tract, with younger turkeys having higher levels of binding in the lung compared to the older group. In addition, in the intestinal tract, younger turkeys had higher levels of binding compared to the older birds. Using LPAIV, similar species and tissues, specific differences were seen among the age groups; however, only turkeys had overall significant differences in the respiratory tract MAB, with the older birds having higher levels of binding compared to the younger group. No age-related differences were seen in the overall intestinal tract rHA binding. Age-related differences in rHA binding of the LPAIV and HPAIV demonstrated in this study may partially, but not completely, explain differences in host susceptibility to infection observed during avian influenza outbreaks and in experimental infection studies.

A Case Report of Avian Malaria (Plasmodium spp.) in Pen-Reared Pigeons (Columba livia).

One dead 6-wk-old male racing pigeon (Columba livia) was submitted for postmortem evaluation after presenting with weight loss, anorexia, dry shanks, dehydration, and lethargy. The bird belonged to a confined flock with 12 other pigeons raised by a hobbyist. Two pigeons in the flock reportedly had died with a history of similar clinical signs. On gross examination, the liver and the spleen were diffusely dark brown to black. Histopathology revealed moderate to large amounts of anisotropic, intracytoplasmic black pigment, compatible with hemozoin, in the spleen, liver, lung, and kidneys, with small amounts in the heart and meninges of the brain. Marked plasmacytic infiltrates were observed in liver, lungs, heart, and kidneys. Blood smears from a clinically affected concomitant pigeon from the flock revealed numerous light-blue, round to oval, intraerythrocytic trophozoites and meronts suggestive of Plasmodium spp. PCR and sequencing tests were performed from spleen and ceca with fragments of the 18S ribosomal RNA and the mitochondrial cytochrome b (cytB) genes. Sequencing results confirmed the presence of Plasmodium in the affected pigeon. Although an exact genetic match could not be determined, the most similar species to the isolate from this study are Plasmodium relictum, Plasmodium matutinum, Plasmodium lutzi, and Plasmodium homocircumflexum.

Reporte de caso­Reporte de un caso de malaria aviar (Plasmodium spp.) en palomas criadas en corrales (Columba livia) Una paloma mensajera macho de 6 semanas muerta (Columba livia) fue remitido a evaluación post mortem después de presentar pérdida de peso, anorexia, patas secas, deshidrataciœn y letargo. El pájaro pertenecía a una parvada confinada con otras 12 palomas criadas por un criador aficionado. Dos palomas de la parvada habían muerto con antecedentes de signos clínicos similares. En el examen macroscópico, el hígado y el bazo se observaron de color marrón oscuro a negro. La histopatología reveló cantidades moderadas a abundantes de pigmento negro intracitoplasmático y anisotrópico, compatible con hemozoína, en el bazo, hígado, pulmón y riñones, con pequeñas cantidades en el corazón y en las meninges del cerebro. Se observaron marcados infiltrados plasmocíticos en hígado, pulmones, corazón y riñones. Los frotis de sangre de otra paloma clínicamente afectada de la parvada revelaron numerosos trofozoítos intraeritrocíticos y esquizontes de color azul claro, redondos a ovalados, que sugerían Plasmodium spp. Se realizaron pruebas de PCR y secuenciación a partir del bazo y el ciego con fragmentos de los genes de ARN ribosómico 18S y del citocromo b mitocondrial (cytB). Los resultados de la secuenciación confirmaron la presencia de Plasmodium en la paloma afectada. Aunque no se pudo determinar una identidad genética exacta, las especies más similares al aislado de este estudio son Plasmodium relictum, Plasmodium matutinum, Plasmodium lutzi y Plasmodium homocircumflexum.

Thymoma in an aged backyard Leghorn chicken, with reviews of a database and literature.

A 7-y-old backyard Leghorn chicken (Gallus domesticus) was submitted to the California Animal Health and Food Safety Laboratory System (CAHFS)-Turlock branch for postmortem examination, with a history of unexpected death. At postmortem examination, a hemorrhagic soft tissue mass was observed in the cervical region. Microscopically, a densely cellular neoplasm of polygonal epithelial cells and small lymphocytes was observed. The microscopic features of the neoplasm in combination with positive immunohistochemistry for pancytokeratin and CD3 were used to classify the lesion as a thymoma. Thymoma was diagnosed in only 5 birds submitted to CAHFS from 1990 to 2019. Thymoma has been described only rarely in birds, and is an unusual diagnosis in backyard chickens.

Case Report - A Case Report of Avian Malaria (Plasmodium spp.) in Pen-Reared Pigeons (Columbia livia).

One dead 6-week-old, male racing pigeon ( Columbia livia ) was submitted for postmortem evaluation after presenting with weight loss, anorexia, dry shanks, dehydration and lethargy. The bird belonged to a confined flock with 12 other pigeons raised by a hobbyist. Two pigeons in the flock reportedly had died with a history of similar clinical signs. On gross examination, the liver and the spleen were diffusely dark brown to black. Histopathology revealed moderate to large amounts of anisotropic, intracytoplasmic black pigment, compatible with hemozoin, in the spleen, liver, lung and kidneys, with small amounts in the heart and meninges of the brain. Marked plasmacytic infiltrates were observed in liver, lungs, heart and kidneys. Blood smears from a clinically affected concomitant pigeon from the flock revealed numerous light-blue, round to oval, intraerythrocytic trophozoites and meronts suggestive of Plasmodium spp. PCR and sequencing tests were performed from spleen and ceca using fragments of the 18S ribosomal RNA and the mitochondrial cytochrome B (cytB) genes. Sequencing results confirmed the presence of Plasmodium in the affected pigeon. Although an exact genetic match could not be determined, the most similar species to the isolate from this study are P. relictum , P. matutinum, P. lutzi and P. homocircumflexum .

Infectious Bronchitis Virus Associated with Nephropathy Lesions in Diagnostic Cases from Commercial Broiler Chickens in California.

In March 2019, the California Animal Health and Food Safety Laboratory (CAHFS), Turlock branch, received two submissions of broiler chickens from commercial flocks reporting increased mortality. Submissions consisted of either white or brown broilers. Submitted chickens appeared depressed with ruffled feathers. At necropsy, moderate to severely enlarged and pale kidneys were observed, with gross lesions indicative of dehydration. Microscopically, renal tubules were degenerated and distended with necrotic debris and tubular casts. The kidney parenchyma contained mononuclear inflammatory cell infiltrates and interstitial edema. Infectious bronchitis virus (IBV) was isolated and identified by reverse transcription quantitative PCR from kidney tissue pools and tracheal swab pools from both cases. Partial sequencing of the S1 hypervariable region was most similar to a local California variant, CA1737. The outbreak lasted roughly 1 wk in both flocks, with 2% total mortality in the brown broilers and 20% total mortality in the white broilers. Final proof of the IBV strains causing nephropathy will require fulfillment of Koch postulates. IBV associated with nephropathy has been sporadically reported in California chicken flocks and represents a significant pathogen due to its potential for inducing high flock mortality. The incidence of IBV associated with a nephropathy diagnosis in chicken necropsy submissions to the CAHFS system-wide from 1998 to 2019 is also reviewed.

Recombinant hemagglutinin glycoproteins provide insight into binding to host cells by H5 influenza viruses in wild and domestic birds.

Clade 2.3.4.4, H5 subtype highly pathogenic avian influenza viruses (HPAIVs) have caused devastating effects across wild and domestic bird populations. We investigated differences in the intensity and distribution of the hemagglutinin (HA) glycoprotein binding of a clade 2.3.4.4 H5 HPAIV compared to a H5 low pathogenic avian influenza virus (LPAIV). Recombinant HA from gene sequences from a HPAIV, A/Northern pintail/Washington/40964/2014(H5N2) and a LPAIV, A/mallard/MN/410/2000(H5N2) were generated and, via protein histochemistry, HA binding in respiratory, intestinal and cloacal bursal tissue was quantified as median area of binding (MAB). Poultry species, shorebirds, ducks and terrestrial birds were used. Differences in MAB were observed between the HPAIV and LPAIV H5 HAs. We demonstrate that clade 2.3.4.4 HPAIV H5 HA has a broader host cell binding across a variety of bird species compared to the LPAIV H5 HA. These findings support published results from experimental trials, and outcomes of natural disease outbreaks with these viruses.

MYONECROSIS AND DEATH DUE TO PRESUMED MICROCYSTIN TOXICOSIS IN AMERICAN WHITE PELICANS (PELECANUS ERYTHRORHYNCOS).

Over a period of 5 mo, seven out of eight American white pelicans (Pelecanus erythrorhynchos) housed on a spring-fed pond at a zoo died or were euthanized. Clinical signs included inability to stand, anorexia, and weight loss. Clinicopathologic findings included heterophilic leukocytosis and elevated creatine kinase and aspartate aminotransferase. Histopathologic findings on all pelicans demonstrated severe, chronic, diffuse rhabdomyofiber degeneration and necrosis, making vitamin E deficiency a differential diagnosis despite routine supplementation. Based on tissue and pond water assays for the cyanobacterial toxin, microcystin, toxicosis is suspected as the inciting cause of death in these cases. We hypothesize that vitamin E exhaustion and resultant rhabdomyodegeneration and cardiomyopathy were sequelae to this toxicosis.

Focal duodenal necrosis in chickens: attempts to reproduce the disease experimentally and diagnostic considerations.

Focal duodenal necrosis (FDN) is an intestinal disease of egg-layer chickens characterized by multifocal necrosis of the duodenal loop and proximal jejunum. Affected flocks usually have decreased egg weights and drops in egg production. Previous studies have associated this condition with Clostridium perfringens infection. We tried to reproduce FDN by experimental infection of egg-laying chickens using different netB-positive and netB-negative C. perfringens strains, and duodenal homogenate obtained from FDN lesions. Chickens challenged with C. perfringens and/or duodenal homogenate developed duodenitis after challenge. Gross lesions included mucosal erosions, hyperemia, mucosal hemorrhages, and watery intestinal content. Microscopic lesions included mild enterocyte degeneration and necrosis, and mild-to-moderate hemorrhage and lymphoplasmacytic and heterophilic infiltration of the lamina propria. Two netB-positive C. perfringens strains closely related to necrotic enteritis pathogenic strains, by genomic composition, were re-isolated from lesions. Necrosis of intestinal crypts was observed in chickens challenged with duodenal homogenate with or without C. perfringens coinfection. Characteristic microscopic FDN lesions with significant necrosis and loss of villus enterocytes were not reproduced.

Retrospective Summary of Erysipelothrix rhusiopathiae Diagnosed in Avian Species in California (2000-19).

Erysipelas is a bacterial disease caused by Erysipelothrix rhusiopathiae that affects multiple mammalian and avian species. In poultry, the disease is of sporadic prevalence and more often observed in older birds, leading to decreased egg production and mortality. Among avian species, turkey breeders seem to be the most affected, but outbreaks have been reported in ducks, layer chickens, quails, geese, and various captive and free-range birds. Sixty-seven cases of erysipelas have been diagnosed in animals submitted for necropsy evaluation at the California Animal Health and Food Safety Laboratory System from January 2000 to December 2019. Of these, 38 cases (56.72%) were in avian species, and a retrospective analysis of these avian cases was performed. The majority of the avian cases were in turkeys (17/38, 44.74%). Most of the turkey breeder cases reported performing artificial insemination prior to the increase in mortality. In other birds, mortality was often observed without observing previous clinical signs. The majority of cases presented with coinfections with other pathogens (23/38, 60.53%), which might have affected the clinical outcome. Despite the occasional occurrence in avian species, erysipelas is an important pathogen in poultry and should be considered as a differential diagnosis in other avian species when acute septicemia is suspected as the cause of mortality.

Immune Responses in the Eye-Associated Lymphoid Tissues of Chickens after Ocular Inoculation with Vaccine and Virulent Strains of the Respiratory Infectious Laryngotracheitis Virus (ILTV).

Infectious laryngotracheitis (ILT) is an acute respiratory disease of poultry caused by infectious laryngotracheitis virus (ILTV). Control of the disease with live attenuated vaccines administered via eye drop build upon immune responses generated by the eye-associated lymphoid tissues. The aim of this study was to assess cytokine and lymphocyte changes in the conjunctiva-associated lymphoid tissues (CALT) and Harderian gland (HG) stimulated by the ocular inoculation of the ILTV chicken embryo origin (CEO) vaccine strain and virulent strain 63140. This study offers strong evidence to support the roles that the CALT and HG play in the development of protective ILTV immune responses. It supports the premise that ILTV-mediated immunomodulation favors the B cell response over those of T cells. Further, it provides evidence that expansions of CD8α+ cells, with the concomitant expression of the Granzyme A gene, are key to reducing viral genomes in the CALT and halting ILTV cytolytic replication in the conjunctiva. Ultimately, this study revealed that the early upregulation of interleukin (IL)-12p40 and Interferon (IFN)-γ cytokine genes, which shape the antigen-specific cell-mediated immune responses, retarded the decline of virus replication, and enhanced the development of lesions in the conjunctiva epithelium.

Tacrolimus therapy for dermal arteritis of the nasal philtrum refractory to surgery and anti-inflammatory therapy (doxycycline/niacinamide and topical fluocinolone) in a dog.

BACKGROUND: Dermal arteritis of the nasal philtrum is a rarely reported condition commonly affecting large breed dogs. OBJECTIVE: To describe the effective treatment of nasal philtrum arteritis with topical tacrolimus in one dog. ANIMAL: A 9-year-old, intact male German shorthair pointer dog was presented with well-demarcated deep erythematous ulcers targeting exclusively the skin of the nasal philtrum, accompanied by frequent series of haemorrhage. METHODS: Complete blood count, serum chemistry profile, urinalysis, histopathological examination and immunohistochemistry of skin biopsies. RESULTS: The presence of a V-shaped ulcer with subendothelial spindle cell proliferation resulting in stenosis of dermal arteries and arterioles on histological evaluation, together with a well-demarcated deep nasal philtrum ulcer was consistent with arteritis of the nasal philtrum. Treatment was initiated with twice daily oral doxycycline and niacinamide in conjunction with topical fluocinolone cream. Over the course of two years, the lesions progressed with frequent bleeding episodes. A novel surgical approach provided deep resection of all grossly affected tissue; four months later a recurrence of fissures and occasional mild bleeding from the original site was noted and there was no improvement after another two months of oral doxycycline/niacinamide and topical fluocinolone treatment. Topical application of 0.1% tacrolimus twice daily resulted in complete healing of the ulceration and normalization of the epidermis. Over the subsequent 15 months, the dog's lesions remained in remission with topical tacrolimus application twice daily. CONCLUSIONS AND CLINICAL IMPORTANCE: Topical tacrolimus ointment appeared effective at inducing and maintaining lesion remission in this dog with nasal philtrum arteritis.