B. R. Charlton

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.

Investigation of problems associated with intramuscular breast injection of oil-adjuvanted killed vaccines in chickens.

Case submissions and a field investigation indicated that oil-adjuvanted killed vaccines may produce long-lived residual lesions in chickens. The lesions typically are yellow opaque cysts along the fascial planes separating the superficial and deep pectoral muscles. Microscopic evaluation shows that most lesions are cysts with thin fibrous capsules, sometimes associated with lymphocytic aggregates and, more rarely, a granulomatous reaction.

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.

Periodic recurrence of gangrenous dermatitis associated with Clostridium septicum in a broiler chicken operation

The association of gangrenous dermatitis (GD) of chickens with Clostridium septicum infection was well documented in the United States during the 1960s (Bickford AA: 1971, Proc 20th Western Poult Dis Conf, pp. 6, 7). At about the same time, GD was recognized as a source of considerable economic loss to broiler production operations in the United Kingdom and Australia. 1,3 Subsequently, the potentiating role of immunosuppression induced by infectious bursal disease virus (IBDV) and other viral agents was recognized. Reduction of losses caused by GD requires control procedures for IBDV and other immunosuppressive agents. Recent reports have suggested that there may be an increased incidence of the clostridial diseases of chickens in the United States. Among these diseases, GD associated with C. septicum is a major concern. At the California Veterinary Diagnostic Laboratory System (CVDLS), Turlock Branch, the diagnosis of this disease was once as frequent as 5% of all diagnostic cases (Bristow DL: 1971, Proc 20th Western Poult Dis Conf, p. 11). After producers implemented procedures to limit the effects of IBDV, the diagnosis of GD became a rare event in California. This report documents an unusual annual recurrence of C. septicum-associated GD over 3 consecutive years in a single house of a multiple-house broiler facility. The ranch involved in this report is situated on flat delta valley terrain and consists of 8 broiler houses. The houses are similar in design but differ in size. Three houses are 10,000 sq ft, 3 are 15,000 sq ft, and 2 are 17,500 sq ft. Feed sources, feed delivery systems, ventilation systems, and litter management procedures are uniform on the ranch. A single well serves as the water source for all houses, and hot-water brooding is practiced in all houses. The ranch is operated on a year-round all-in, all-out basis. On February 3, 1993, 7 dead 43-day-old broiler chickens were submitted from house 5 (15,000 sq ft) to CVDLS, Turlock Branch, for necropsy. Mortality in the house had increased suddenly to about 1% per day. Stocking density in all three 15,000-sq-ft buildings had been 19,000 chicks at placement. This density is approximately 8.5% higher than normal for these houses. Climatic conditions were normal for winter with characteristically high humidities and cool temperatures. Litter conditions in the house were described as old and wet. On January 18, 1994, a similar group of 6 dead broiler chickens was submitted from house 5. A sudden increase in mortality had occurred in this house only. The birds were

Mycoplasma bovis associated with decubital abscesses in Holstein calves.

Between April of 1990 and March of 1992, calves on a Holstein calf ranch experienced subcutaneous decubital abscesses involving the brisket region, dorsal aspect of the carpus, and lateral aspect of the stifle joints. Fifty out of 2,500 (2%) Holstein calves between the ages of 3 and 12 weeks were affected. Needle aspirates of brisket abscesses from 8 calves and 6 live or dead calves with 1 or more decubital abscesses were submitted for examination. Two of the 6 calves in addition had bronchopneumonia. Mycoplasma bovis was isolated from all abscesses and 1 lung. Formalin fixed tissues taken from the affected areas also revealed M. bovis by immunoperoxidase staining. No evidence of joint involvement was apparent, and no mycoplasma was isolated from the joints adjacent to affected areas. Attempts to isolate mycoplasma from milk and environmental samples were unsuccessful.

Identification and pathogenicity of a natural reassortant between a very virulent serotype 1 infectious bursal disease virus (IBDV) and a serotype 2 IBDV

Infectious bursal disease virus (IBDV) causes an economically important, immunosuppressive disease in chickens. There are two serotypes of the virus that contain a bi-segmented double-stranded RNA genome. In December 2008, the first very virulent (vv)IBDV was identified in California, USA and in 2009 we isolated reassortant viruses in two different locations. Genome segment A of these reassortants was typical of vvIBDV serotype 1 but genome segment B was most similar to IBDV serotype 2. The CA-K785 reassortant caused 20% mortality in chickens but no morbidity or mortality in commercial turkey poults despite being infectious. There have been previous reports of natural reassortants between vvIBDV and other serotype 1 strains, but a natural reassortant between IBDV serotypes 1 and 2 has not been described. The apparent reassorting of California vvIBDV with an endemic serotype 2 virus indicates a common host and suggests vvIBDV may have entered California earlier than originally thought.

The Diagnosis of Very Virulent Infectious Bursal Disease in California Pullets

Abstract 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, Peyers 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 105.5 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.

Infectious coryza in meat chickens in the San Joaquin Valley of California.

Two cases of infectious coryza in meat chickens are reported. The first case involved 6-week-old broiler chickens in which only Haemophilus paragallinarum was isolated. The second case involved 11-week-old roaster chickens in which H. paragallinarum and Mycoplasma synoviae were isolated. Both farms were in close proximity to layer-chicken farms where infectious coryza had been previously diagnosed. In both cases, only certain houses on the farm were affected, and mortality in these houses increased slightly. At processing, the condemnation rates for affected houses were considerably higher than rates for unaffected houses. Condemnations for affected houses were mostly due to airsacculitis. A dissecting fibronopurulent cellulitis was a prominent lesion in the second case. This lesion could lead to confusion with chronic fowl cholera and swollen-head syndrome.

Characteristics of a Very Virulent Infectious Bursal Disease Virus from California

Abstract An outbreak of infectious bursal disease (IBD) in two California layer flocks resulted in the isolation of two infectious bursal disease viruses designated rA and rB. Increased mortality plus gross and histopathology in the layer flocks suggested rA and rB could be very virulent infectious bursal disease virus (vvIBDV). Preliminary studies indicated that high mortality resulted when bursa homogenates from the layer farms were used to inoculate specific-pathogen-free (SPF) chicks. In addition, rA and rB contained VP2 amino acid sequences typically seen in vvIBDV. Molecular and in vivo studies were conducted to more thoroughly identify and characterize the rA and rB viruses. Nucleotide sequence analysis demonstrated that rA and rB had identical sequences across the hypervariable VP2 (hvVP2) and segment B regions examined, and their amino acid sequences in the hvVP2 region were identical to the vvIBDV type strains UK 661, OKYM, and Harbin. Furthermore, the genome segment B nucleotide sequences examined for rA and rB were a 98.1% match with vvIBDV and only an 88.0% match with classic IBDV strains. Phylogenetic analysis placed the rA and rB viruses with other vvIBDV and confirmed these viruses were close genetic descendants of vvIBDV seen around the world. Pathogenicity studies in 4-wk-old SPF chicks demonstrated that at a high dose (105.5 50% egg infective dose [EID50]) and a low dose (102.0 EID50) of rA and rB, mortality ranged from 91% to 100%. A pathogenic classic virus, standard challenge (STC), at similar doses did not cause mortality in the SPF chicks. In addition, mortality occurred in three out of four SPF birds exposed by direct contact to rA and rB inoculated chicks. Serum from convalescent birds inoculated with rA had high titers to IBDV but were negative for antibodies to infectious bronchitis virus, avian influenza virus, chicken anemia virus, Newcastle disease virus, Mycoplasma gallisepticum, and Mycoplasma synoviae. Virus isolation attempts on the rA and rB bursa homogenate inocula also indicated that no contaminating microorganisms contributed to the high mortality and pathology observed in the SPF chicks. In one experiment, broilers with maternal immunity to IBDV were protected from infection and disease when they were challenged with 102 EID50 and 105 EID50 of the STC virus. When challenged with 102 EID50 of the rA virus, the maternally immune broilers were protected from disease but not infection as evidenced by a positive reverse transcription–polymerase chain reaction (RT-PCR) assay for the virus. When the broilers were challenged with 105 EID50 of the rA virus, they had typical gross and histopathologic signs of IBD but no mortality by 7 days postinoculation. It was concluded that the rA and rB viruses meet the genotypic and phenotypic characteristics of a vvIBDV.

Avian Paramyxovirus Type 1 Infections in Racing Pigeons in California. I. Clinical Signs, Pathology, and Serology

An outbreak of diarrhea and neurological disease in California racing pigeons caused by avian paramyxovirus type 1 (PMV-1) is documented. Predominant clinical signs were polydipsia, ataxia, poor balance, torticollis, head tremors, inability to fly, and diarrhea that was unresponsive to therapy. Gross pathologic findings were often unremarkable or non-specific. The predominant histologic lesions were interstitial nephritis, chronic tubular necrosis, lymphoplasmacytic infiltration within the kidney, liver, and pancreas, and focal non-suppurative encephalitis. Pigeons from 20 submissions demonstrated characteristic clinical signs of PMV-1 infection. Pigeons from 17 submissions exhibited typical histopathology. Serologic evidence of PMV-1 infection was present in pigeons from 13 submissions, and PMV-1 was isolated from pigeons received in six submissions. None of these pigeons had been vaccinated against PMV-1.