Trichodinosis associated with pathology of the reproductive tract in waterfowl.

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.

Atypical distribution of fowl pox lesions in broilers.

An unusual cutaneous fowl pox outbreak occurred in 8-wk-old broilers in California. Rounded and longitudinal, proliferative scratch-associated lesions were found only in feathered areas of the body. Both sides of the hip, the lower abdomen, pericloacal area, and lateral lower neck area were involved. The head, legs, feet, and toes did not have lesions. Birds in only one section of one of five houses were affected. Fifteen percent condemnations occurred in birds from the affected house due to the skin lesions. A diagnosis of fowl pox was achieved by histopathology, viral isolation, and direct electron microscopy. The unusual distribution of pox lesions was assumed to be associated with skin scratches. There was no evidence that mosquitoes or other types of insects were involved in this outbreak. To the knowledge of the authors, this is the first report of this kind of unusual fowl pox in the United States.

Respiratory tract trichomoniasis in breeder squab candidates in Northern California.

Breeder squab candidates between the ages of 6 and 16 wk were submitted to the California Animal Health and Food Safety Laboratory, Turlock branch, as a result of respiratory distress and increased mortality. These cases were submitted from one Northern California commercial squab operation on three separate occasions occurring between December 2007 and March 2008. Severe trichomoniasis was identified, primarily in the tracheal epithelium and lung of squabs, with few or no lesions in the oral cavity, crop, esophagus, and livers, where the organism commonly infiltrates. Infiltration of the trachea and lung sections with trichomonads was associated with a severe inflammatory response in the surrounding tissue. Diagnosis was confirmed with the use of histopathology and an immunoperoxidase special stain. Oxytetracycline supportive antibiotic therapy to prevent secondary bacterial infections was administered to remaining squabs on the farm, but no specific treatment regimen was instituted. This novel respiratory presentation of trichomoniasis continued over a period of 3 mo, until mortality gradually returned to normal.

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.

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.

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

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.

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.

Time course for neurotoxic esterase inhibition in hens given multiple diisopropyl fluorophosphate injections.

Neurotoxic esterase has been useful for predicting delayed neurotoxicity after acute administration of organophosphorus esters. The present study determined how it could be used to predict neurotoxicity after 20 daily sc injections of 0.05 mg/kg diisopropyl fluorophosphate (DFP) in adult hens. Brain neurotoxic esterase activities were measured during the course of intoxication. No significant occurred after a single DFP injection but the level of inhibition gradually increased to a maximum of 54.3% after 20 injections. The time courses for inhibition of plasma cholinesterase, brain pseudocholinesterase and brain acetylcholinesterase were distinctly different from that for neurotoxic esterase and the former 3 enzymes reached a plateau of inhibition after 1 or 5 DFP injections. Walking behavior was regularly measured in treated hens and distinct motor impairment was first noted after 5 daily DFP injections. Brains, spinal cords and sciatic nerves were removed 24 hr after the last injection and examined microscopically. All hens exhibited central-peripheral distal axonopathy that was characteristic for organophosphate delayed neurotoxicity. Neurotoxicity developed in the absence of a high level of inhibition of neurotoxic esterase following multiple injections of DFP and the "critical" level neurotoxic esterase inhibition used to predict subchronic neurotoxicity may be lower than that used after single treatments.

Complementary randomly amplified polymorphic DNA (RAPD) analysis patterns and primer sets to differentiate Mycoplasma gallisepticum strains.

Randomly amplified polymorphic DNA (RAPD) analysis was used to differentiate 7 strains of Mycoplasma gallisepticum. Six commercially available primers or primer combinations were screened for their ability to differentiate vaccine and type strains. Although major and minor bands were produced with each primer, many of the primers were unsuitable for strain differentiation. The use of primer 6 and combined primers 3 and 4 resulted in complementary RAPD banding patterns for each M. gallisepticum strain. Eleven different isolates representing 7 different strains were segregated into 7 different patterns, corresponding to the 7 strains.

Randomly amplified polymorphic DNA (RAPD) analysis of Mycoplasma gallisepticum isolates from turkeys from the central valley of California.

Randomly amplified polymorphic DNA (RAPD) analysis was used to investigate the molecular epidemiology of 26 Mycoplasma gallisepticum (MG) isolates obtained from turkeys located in the central valley of California. The MG isolates were recovered from 5 different companies and 13 ranches. Each company had unique MG strains. No evidence of spread of MG between companies was detected. RAPD analysis of MG isolates within a ranch during an outbreak revealed only a single strain involved in each outbreak. RAPD analysis identified an isolate from 1 ranch with a banding pattern identical to that of the 6/85 vaccine strain, which had been used on that particular ranch. Similar RAPD banding patterns of isolates from different ranches within the same company suggested horizontal spread of MG between ranches. The use of 2 primer sets in RAPD analysis was critical to prevent misinterpretation of relationships between different isolates.

Postural and neurological deficits in broiler chicks after cervical vaccination with live vaccine.

A disease characterized by paresis and paralysis was seen in 7-9-day-old broiler chicks after vaccination in the neck area at day-of-age with a live virus vaccine containing viruses of Marek's disease, fowl pox, and infectious bursal disease. Affected birds presented with variable signs of ataxia, lateral recumbency, leg paralysis, and twisting or S-shaped flexure of the neck. Gross lesions noted at necropsy included swelling and edema of the subcutaneous tissues and muscles of the neck at the injection site area. A heavy mononuclear inflammatory cell infiltration was seen in the subcutaneous tissues, connective tissues, and muscles of the neck at the injection site. In some cases, the inflammatory process extended along fascial planes to involve the epidural spaces surrounding the spinal cord. Fatty changes with possible demyelination of nerve fibers were noted in some sections of the spinal cord adjacent to the inflammatory lesions. Clusters of poxviruses were found within some inflammatory lesions on transmission electron photomicrographs.

Association of Reoviridae particles in an enteric syndrome of poults observed in turkey flocks during 1988.

An enteric syndrome of turkey poults, characterized by enteritis, crop mycosis, intestinal changes (pale, thin-walled ballooning with watery contents), and rickets, occurred during 1988 in 74 turkey flocks from different farms belonging to 9 California turkey growers. The flocks ranged in size from 9,000 to 120,000 birds. Pools of intestine sections from 618 birds, representing 78 field cases, were examined. Histopathological examination of the intestines showed a mild to severe atrophy with a reduced depth of crypts, which was more prominent in the distal part of the small intestine. Viral isolation attempts with primary cell cultures of chicken embryo kidney cells were negative. Examination by electron microscopy of negatively stained intestinal specimens revealed the presence of Reoviridae particles of 58.8 to 80 nm in diameter. Enzyme-linked immunosorbent assay results on the intestinal pools for mammalian and group A avian rotaviruses were negative. A statistically significant relationship was found for the presence of Reoviridae particles in the intestines of 10-21-day-old birds. Of the 7 most common pathological conditions analyzed, 2, rickets and intestinal changes (thin-walled ballooning intestine with watery contents), showed a statistically significant association with the presence of Reoviridae particles.

Preliminary characterization of a pleomorphic gram-negative rod associated with avian respiratory disease.

An unidentified, pleomorphic, gram-negative rod (PGNR) bacterium has been isolated from domestic fowl with respiratory disease. The PGNR was isolated in 5% of turkey accessions and 3% of chicken accessions, primarily from the respiratory tract. Preliminary characterization of this organism included reviewing accession records, conducting cultural and biochemical tests, and analyzing cellular fatty acids. The PGNR was also compared with other bacteria capable of inhabiting the avian respiratory system. Biochemical and cellular fatty acid analysis failed to identify the organism, however all 14 isolates were similar.

Spirochetosis in California game chickens.

An outbreak of spirochetosis occurred in a flock of 75 game chickens in California during fall 1991. Affected birds were weak and anemic. Many had greenish diarrhea. Spirochetes were seen on Giemsa-stained blood smears and in silver-stained tissue sections of kidney, liver, and spleen. Splenomegaly, which is reported to be characteristic of fowl spirochetosis, was not observed in two acutely infected chickens.

Wry necks associated with Mycoplasma meleagridis infection in a backyard flock of turkeys.

Two 5-week-old bronze turkeys were submitted with crooked necks that could not be straightened. Both birds were serologically and culturally positive for Mycoplasma meleagridis (MM). Histologically, there were lymphoid follicles in the cranial thoracic and cervicoclavicular air sacs, and inflammatory changes in the cartilage of some cervical vertebrae. This appeared to be a case of MM-associated wry necks.

An outbreak of coccidiosis and necrotic enteritis in 16-week-old cage-reared layer replacement pullets.

A flock of 16-week-old layer replacement pullets housed in cages since 1 day old experienced an outbreak of necrotic enteritis with concurrent coccidiosis 3 weeks after debeaking.

Pyogranulomatous typhlitis and hepatitis of market turkeys.

Pyogranulomatous lesions in the liver and cecum were observed in groups of 7-to-8-week-old market turkeys. Necropsy revealed multifocal lesions in the liver and the presence of cecal cores that, in some cases, distended the cecum to the point of rupture. Mortality was associated with the ruptured ceca. Two successive flocks also succumbed to these lesions. An epizootiological study was performed to determine the cause and/or initiation of lesions leading to the formation of cecal cores.

Critical neurohistopathologic evaluation of clinically healthy commercial single-comb White Leghorn hens.

A neurohistopathologic study was conducted to determine the types and incidence of spontaneous lesions of the nervous system in healthy adult hens. Tissues examined were brain, three levels of spinal cord, and sciatic nerves from 92 healthy commercial hens from a single commercial egg-production flock judged above average in health. These hens represented negative controls (untreated or given corn oil) in 9 separate toxicologic studies. Minimal to mild focal proliferative or degenerative microscopic changes were detected at all levels of the nervous system. More than 50% of the hens had lymphocytic perivascular cuffing and focal gliosis in the central nervous system. Similarly, lymphocytic foci (in nerve sheaths or within fasciculi) and Schwann cell hyperplasia were commonly seen in sciatic nerves. The spinal cord exhibited neuronal swelling and chromatolysis frequently; axonal degeneration (in brain and spinal cord) and swelling or degeneration of nerve fibers in sciatic nerves were observed sporadically. Most changes were rare or mild. A few chickens had mild or moderately severe perivascular cuffing and focal gliosis. The significance of these findings is discussed in relation to the use of the adult hen in neurotoxicologic evaluations.

Local reaction and serological response in commercial layer chickens injected intramuscularly in the leg with oil-adjuvanted Mycoplasma gallisepticum bacterin.

The incidence of local vaccine reaction was investigated in three flocks of commercial layer chickens after they were vaccinated intramuscularly in the leg with oil-adjuvanted Mycoplasma gallisepticum (MG) bacterin by commercial crews. Local vaccine reaction typically involved swelling due to a granulomatous cellulitis in the connective tissue above the tibiotarsal joint of the vaccinated leg. Histopathology revealed that the inflammatory reaction was confined primarily to subcutis and muscle fascia and rarely extended into muscle fibers, tendons, or tendon sheaths. Movement of the vaccinated flocks from growing to laying houses was associated with a significant increase in the incidence and severity of leg swelling. Antibodies against MG as measured by the hemagglutination-inhibition test were significantly higher in chickens with severe leg swelling than in chickens without leg swelling.

Scoliosis and tibiotarsal deformities in broiler chickens.

The incidence and degree of scoliosis were investigated in broiler chickens with and without intertarsal deformities associated with slipped gastrocnemius tendons. In both groups, the incidence of scoliosis was similar and there was a significant tendency for scoliosis to be convex on the right side. However, scoliosis was significantly greater in birds with intertarsal deformities, and in a significant proportion of these the joint with a slipped tendon was on the convex side of scoliosis. In birds with deformities, inequalities between right and left tibiotarsi were significantly greater, and tibiotarsi with greater length, narrower condyles and trochleae, and shallower trochlear grooves were significantly more often on the convex side of scoliosis. Significant positive correlations were found between scoliosis and rotational and bending deformities of the distal tibiotarsus on the convex side of scoliosis. These findings suggest a cause-and-effect relationship between scoliosis and tibiotarsal deformities associated with slipped tendons.

Relationship of sex, age, and concurrent intestinal disease to necrotic enteritis in turkeys.

A retrospective case-control study of necrotic enteritis in turkeys was done. Male flocks were involved in a significantly greater proportion of necrotic enteritis cases compared with control cases. Necrotic enteritis cases occurred most frequently between 6 and 11 weeks of age. Within this age range, the probability of a necrotic enteritis case having a concurrent intestinal disease diagnosis was significantly higher than the probability in control cases. Coccidiosis was the most frequently diagnosed concurrent intestinal disease with necrotic enteritis, followed by clinical hemorrhagic enteritis and ascaridiasis.