Nonenteric Lesions of Necrotic Enteritis in Commercial Chickens in California: 25 Cases (2009-2018).

Necrotic enteritis (NE) is an important enteric disease affecting a wide variety of avian species, including poultry, caused by Clostridium perfringens type G and, rarely, type C. Significant economic losses can result from elevated mortality rates and poor performance, such as decreased weight gain associated with intestinal damage and impaired absorption of nutrients. Additional losses can result from elevated condemnation at the processing plant because of a high incidence of cholangiohepatitis. Nonenteric lesions associated with NE have been rarely reported. This paper describes uncommon presentations of NE in commercial chickens received by the California Animal Health and Food Safety Laboratory (Turlock and Tulare branches) between 2009 and 2018. Overall, extraintestinal lesions associated with C. perfringens were diagnosed in 25 cases of NE involving commercial broiler chickens. The extraintestinal sites most commonly affected included liver, followed by gizzard, bursa of Fabricius, gall bladder, and spleen. The etiology of these lesions, C. perfringens, was confirmed from a combination of gross, bacteriologic, microscopic, and immunohistochemical findings. The most common predisposing factors for NE identified were coccidiosis (56%, 14/25) and immunosuppressive disease agents, including infectious bursal disease virus (16%, 4/25) and fowl adenovirus group 1 (4%, 1/25). Additionally, four cases (16%) had microscopic lesions compatible with cystic enteritis, probably of viral etiology. This study describes the incidence of extraintestinal lesions of NE in chickens, underlying the role of enteric disorders and immunosuppression as major predisposing factors for the development of NE.

Partial Molecular Characterization and Pathogenicity Study of an Avian Reovirus Causing Tenosynovitis in Commercial Broilers.

This study describes the molecular characterization of avian reoviruses (ARVs) isolated during an outbreak in commercial chickens between 2015 and 2016. In addition, a pathogenicity study of a selected ARV strain isolated from a field case of viral tenosynovitis in commercial broiler chickens was performed. On the basis of phylogenetic analysis of a 1088-bp fragment of the ARV S1 gene, the investigated sequences were differentiated into five distinct genotypic clusters (GCs), namely GC1, GC2, GC3, GC4, and GC6. Specific-pathogen-free (SPF) and commercial broiler chickens were challenged with the GC1 genetic type MK247011, at 14 days of age via the interdigital toe web. No significant effects in body weight gain and feed conversion were detected in both chicken types. The Δ interdigital web thickness was most severe at 4 days postchallenge (DPC) in both the SPF and broiler subgroups. The inflammation in SPF birds was slightly more severe compared with broilers. Neither mortality nor clinical signs occurred in the infected groups for the duration of the experiment, despite the presence of significant microscopic lesions in challenged birds. Microscopic changes of tenosynovitis became evident at 3 DPC, with the highest incidence and severity detected at 14 and 21 DPC, respectively. Seroconversion against ARV occurred 3 wk postchallenge, and the microscopic lesions detected in tendon and heart sections were highly compatible with those described in the field. Increased severity of tenosynovitis and epicarditis lesions were noted in the ARV-challenged groups compared with the control groups. Although SPF and broiler chickens showed comparable responses to the challenge with an ARV genetic variant, detected lesions were subclinical, denoting the limitations of our challenge approach. The age selected in this experiment possibly influenced the course of the infection. Data from this study highlight the genotypic diversity of isolates in California, and the outcome of the pathogenicity study can be used as a basis to improve protocols for pathogenicity studies to characterize ARV variants causing clinical disease in the field.

Caracterización molecular parcial y estudio de patogenicidad de un reovirus aviar que causa tenosinovitis en pollos de engorde comerciales. Este estudio describe la caracterización molecular de reovirus aviares (ARV) aislados durante un brote en pollos comerciales entre los años 2015 y 2016. Además, se realizó un estudio de patogenicidad de una cepa de reovirus seleccionada que fue aislada de un caso de campo de tenosinovitis viral en pollos de engorde comerciales. Con base en el análisis filogenético de un fragmento de 1088 pb del gene S1 de reovirus, las secuencias investigadas se diferenciaron en cinco grupos genotípicos distintos (GCs), denominados, GC1, GC2, GC3, GC4 y GC6. Aves libres de patógenos específicos (SPF) y pollos de engorde comerciales se desafiaron con el tipo genético GC1 MK247011 a los 14 días de edad a través de la membrana interdigital. No se detectaron efectos significativos en el aumento de peso corporal ni en la conversión de alimento en ambos tipos de aves. El grosor de la banda interdigital diferencial fue más severa a los cuatro días posteriores al desafío en las aves libres de patógenos específicos y en los pollos de engorde. La inflamación en las aves libres de patógenos específicos fue ligeramente más severa en comparación con los pollos de engorde. No se presentó mortalidad ni signos clínicos en los grupos infectados durante la duración del experimento, a pesar de la presencia de lesiones microscópicas significativas en las aves desafiadas. Los cambios microscópicos de la tenosinovitis se hicieron evidentes a los tres días postinoculación, con la mayor incidencia y severidad detectadas a los 14 y 21días postinoculación, respectivamente. La seroconversión para reovirus ocurrió tres semanas después del desafío, y las lesiones microscópicas detectadas en secciones de tendón y corazón fueron altamente compatibles con las descritas en el campo. El aumento en la severidad de las lesiones de tenosinovitis y epicarditis se observó en los grupos expuestos a reovirus aviar en comparación con los grupos de control. Aunque las aves libres de patógenos específicos y los pollos de engorde mostraron respuestas comparables ante el desafío con una variante genética de reovirus, las lesiones detectadas fueron subclínicas, lo que denota las limitaciones de nuestro enfoque de desafío. La edad seleccionada en este experimento posiblemente influyó en el curso de la infección. Los datos de este estudio resaltan la diversidad genotípica de los aislamientos en California y el resultado del estudio de patogenicidad se puede usar como base para mejorar los protocolos de los estudios de patogenicidad para caracterizar las variantes de reovirus que causan enfermedades clínicas en el campo.

Otitis and meningoencephalitis associated with infectious coryza ( Avibacterium paragallinarum) in commercial broiler chickens.

Infectious coryza, caused by Avibacterium paragallinarum, is an acute respiratory disease of poultry that can result in substantial morbidity, mortality, and economic losses. In March 2017, the Turlock branch of the California Animal Health and Food Safety laboratory system encountered an unusual clinical and pathologic presentation of infectious coryza in 6 live, 29-d-old, commercial broiler chickens that were submitted for diagnostic investigation. Antemortem evaluation revealed severe neurologic signs, including disorientation, torticollis, and opisthotonos. Swollen head-like syndrome and sinusitis were also present. Histologically, severe sinusitis, cranial osteomyelitis, otitis media and interna, and meningoencephalitis were noted, explaining the clinical signs described. A. paragallinarum was readily isolated from the upper and lower respiratory tract, brain, and cranial bones. Infectious bronchitis virus (IBV) was also detected by PCR, and IBV was isolated in embryonated chicken eggs. Based on sequencing analysis, the IBV appeared 99% homologous to strain CA1737. A synergistic effect between A. paragallinarum and IBV, resulting in exacerbation of clinical signs and increased mortality, may have occurred in this case. A. paragallinarum should be considered among the possible causes of neurologic signs in chickens. Appropriate media should be used for bacterial isolation, and the role of additional contributing factors and/or complicating agents should be investigated in cases of infectious coryza.

Streptococcosis in Commercial and Noncommercial Avian Species in California: 95 Cases (2000-2017).

Streptococcal bacterial species represent common inhabitants of the intestinal tract of animals and humans with a potential for opportunistic infections. Streptococcosis has been identified in turkey poults ( Meleagris gallopavo), ducklings and goslings (Anatidae), broiler chickens, semimature-adult chickens ( Gallus gallus domesticus), and young and adult pigeons (Columbidae). However, the exact underlying factors that lead to bacterial invasion of the blood stream and tissue colonization have not been completely elucidated. The electronic database of the California Animal Health and Food Safety laboratory (Fresno, Tulare, and Turlock branches) was searched for necropsy cases in which streptococcosis was diagnosed in different avian species between January 2000 and August 2017. A total of 95 cases, involving both commercial operations and noncommercial premises, were analyzed. Streptococcus spp., Streptococcus bovis, and Streptococcus gallolyticus were identified from multiple organs, with macroscopic or histopathologic lesions (or both) indicative of septicemia in 23 (24%), 40 (42%), and 30 (32%) cases, respectively. Streptococcus pluranimalium and Streptococcus lutetiensis were also isolated from one (1%) and two (2%) cases, respectively. Turkey poults, broiler chickens, and ducklings were the most-commonly affected species with streptococcosis. Splenitis and hepatitis were the most-common lesions observed and these were the organs with the highest isolation rate. An overview of the clinical and pathologic presentation, and possible predisposing conditions associated with this bacterial infection, is provided.

Avian Encephalomyelitis in Layer Pullets Associated with Vaccination.

Avian encephalomyelitis (AE) was diagnosed in three flocks of leghorn layer pullets following AE vaccination. Ages of the birds were 11, 12, and 14 wk. The submissions came from three different companies located in two geographic areas of the Central Valley of California. The clinical signs included birds down on their legs, unilateral recumbency or sitting on their hocks, lethargy, reluctance to move, dehydration, unevenness in size, low weight, tremors of the head in a few birds, and mildly to moderately elevated mortality. The flocks had been vaccinated against fowl pox and AE with a combined product in the wing-web 2 wk prior to the onset of AE clinical signs. Histopathologic examination revealed lesions consistent with AE, including lymphocytic perivascular infiltration and neuronal central chromatolysis in the brain and spinal cord, as well as gliosis in the cerebellar molecular layer. The AE virus was detected by reverse-transcriptase PCR in the brain homogenate from three cases and peripheral nerves in one case. Additionally, the AE virus was isolated in specific-pathogen-free (SPF) embryonated eggs from brain tissue pool samples. Other avian viral infections capable of causing encephalitis, including avian paramyxoviruses, avian influenza virus (AIV), West Nile virus (WNV), eastern equine encephalitis virus (EEEV), and western equine encephalitis virus (WEEV), were ruled out by attempting virus isolation and molecular procedures.

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).

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.

The diagnosis of very virulent infectious bursal disease in California pullets.

This report documents the occurrence of a very virulent infectious bursal disease virus (vvIBDV) in Northern California commercial brown pullets. Diagnosis was made from multiple accessions from two neighboring and epidemiologically related ranches submitted to the California Animal Health and Food Safety (CAHFS) laboratory. Pullets, 11 and 14 wk of age from ranch A (rA) and ranch B (rB) respectively, were submitted from infectious bursal disease virus vaccinated flocks experiencing a drastic increase in mortality. The December 2008 outbreak resulted in 26% and 34% mortality on rA and rB respectively. Gross and histologic lesions characteristic of acute vvIBDV were observed. Gross lesions included edematous bursas, hemorrhages at the junction of the proventriculus and gizzard as well as hemorrhages on skeletal muscles. Microscopic lesions included severe lymphoid necrosis and inflammation in edematous bursas, lymphoid necrosis in thymus, spleen, Peyer's patches and cecal tonsils. Diagnosis of vvIBDV was confirmed by molecular characterization of the IBDV from bursas as well as viral pathogenicity in specific-pathogen-free birds. RT-PCR and nucleotide sequencing of the hypervariable region of the VP2 (vVP2) gene segment of the IBDV genome was performed on rA, rB and embryo passaged rA virions.The amino acids compatible with vvIBDV isolates: 222(Ala), 242(Ile), 256(Ile), 294(Ile) and 299(Ser) were reported from both ranches. In addition, nucleotide sequencing of a fragment of the VP1 gene demonstrated the viruses have the segment B genotype associated with highly pathogenic vvIBDV. Inocula of 10(5.5) 50% egg infective dose of vvIBDV virus from rA and rB were introduced orally into two groups (g1 and g2 respectively) of 4 wk 2-day-old SPF leghorns. At 4 days postinoculation, there was 100% (22/22) morbidity in g1 and g2; 91% (20/22) mortality in g1; 100% (22/22) mortality for g2; 0% (0/20) morbidity and 0% (0/ 20) mortality was reported in the control group. This is the first occurrence of vvIBDV reported from birds in the United States.

Mycotic pododermatitis and mycotic pneumonia in commercial turkey poults in northern California.

Seven 5-week-old broad-breasted white commercial meat turkeys were submitted to the California Animal Health and Food Safety laboratory in Turlock with a history of respiratory illness. The primary diagnostic findings were mycotic pododermatitis and mycotic pneumonia. The unique feature of this case was the colonization of footpad epidermis and subcutis by fungal hyphae in commercial turkey species. No fungal cultures were undertaken at the time of the necropsy; therefore, paraffin-embedded tissue sections of lung and footpads were used to extract, amplify, and sequence mycotic DNA. A mixed population of fungi was identified in both lung and footpads by polymerase chain reaction amplification of part of the large subunit ribosomal RNA gene using broad-range fungal primers and DNA sequencing. In footpads, sequences matching Cryptococcus saitoi and Cladosporium and Cudoniella species were identified. It is believed that these fungi were opportunistic pathogens originating from the litter. The fungi identified from lungs were Aspergillus species, most closely matching Aspergillus flavus and Arxiozyma telluris (most likely a contaminant). Mycotic pododermatitis in avian species is considered a rare pathologic finding, and few documented reports are available. The on-farm prevalence of footpad lesions was estimated at 3%, and there was no associated increase in the incidence of lameness or weight depression in affected birds. Microscopically, a granulomatous inflammatory reaction associated with fungal hyphae was observed in lung parenchyma. Disruption of keratinized epidermis, encrustations, and acute inflammation were also noted in footpads invaded with fungal hyphae.

Host inflammatory response governs fitness in an avian ectoparasite, the northern fowl mite (Ornithonyssus sylviarum).

Vertebrate immune responses to ectoparasites influence pathogen transmission and host fitness costs. Few studies have characterized natural immune responses to ectoparasites and resultant fitness effects on the ectoparasite. These are critical gaps in understanding vertebrate-ectoparasite interaction, disease ecology and host-parasite co-adaptation. This study focused on an ectoparasite of birds--the northern fowl mite (NFM) (Ornithonyssus sylviarum). Based on prior evidence that chickens develop resistance to NFM, these experiments tested two hypotheses: (i) skin inflammation blocks mite access to blood,impairing development, reproduction and survival; and (ii) host immunogenetic variation influences the inflammatory response and subsequent effects on the ectoparasite. On infested hosts, histology of skin inflammation revealed increased epidermal cell number and size, immigration of leukocytes and deposition of serous exudates on the skin surface. Survival of adult mites and their offspring decreased as the area of skin inflammation increased during an infestation. Inflammation increased the distance to blood vessels beyond the length of mite mouthparts (100-160 lm) and prevented protonymphs and adults from reaching a blood source. Consequently, protonymphs could not complete development, evidenced by a significant inverse relationship between inflammation and protonymph feeding success, as well as an increasing protonymph/adult ratio. Adult females were unable to feed and reproduce, indicated by an inverse relationship between inflammation and egg production, and decreasing female/juvenile ratio. These combined impacts of host inflammation reversed NFM population growth. Intensity of inflammation was influenced by the genotype of the major histocompatibility complex(MHC), supporting previous research that linked these immunological loci with NFM resistance. Overall, these data provide a model for a mechanism of avian resistance to an ectoparasitic arthropod and the fitness costs to the parasite of that host defense.

Acute depigmentation of fertile brown eggs in a commercial layer operation.

Rapid depigmentation of brown eggs is an infrequent but startling event in the commercial egg industry that can result in significant economic losses. Loss of shell pigment in brown-shelled eggs is caused by various factors. In many cases, the exact cause of flock-wide pigment loss remains undetermined. A rapid decline in shell pigmentation was observed in 2 flocks of Hyline brown layers. The lack of evidence of an infectious disease process suggested a feed or management problem. On the basis of a small-scale, "in-house" feeding trial, the feed was identified as the cause of depigmentation. Feed analysis by liquid chromatography with mass spectrometry confirmed the presence of 4,4'-dinitrocarbanilide, a major component of nicarbazin (NCZ). There was no evidence of increased mortality, and only a slight but transient drop in the egg production was observed. Depigmentation effects were rapidly reversed after replacing the feed with NCZ-free feed.