PubMed Health⌕ Search

Biomedical subjects

A E Castro

Publications and source records attributed to A E Castro.

68 records · Page 4Linked to original sources

Comparison of electron microscopy and polyacrylamide gel electrophoresis in the diagnosis of avian reovirus and rotavirus infections.

Electron microscopy (EM) and genome electropherotyping by polyacrylamide gel electrophoresis (PAGE) for the detection of avian rotaviruses and reoviruses in intestinal specimens and cell cultures were compared. Fifty-eight field samples of intestine with intestinal contents, referred to as direct specimens, from turkey and chicken flocks located in different regions of California and submitted during 1989 for virus isolation were randomly selected as test samples. Also, 38 field intestinal specimens with suspected viral infection that had been passaged three times in primary chicken embryo kidney (CEK) cell cultures were used in their third passage. The percentage of agreement and the Kappa statistic of positive and negative results between these two tests were calculated. In the comparison, EM was considered the standard test. By statistical analysis, an agreement of 87% was observed in cell-culture samples analyzed by the two virus-detection methods, as contrasted with an agreement of 72% for direct specimens. The analysis of the number of segments and band migration profiles of reference and field virus strains indicated that only reoviruses replicated in CEK cell cultures and mainly rotaviruses were detected by both tests in direct specimens. The Kappa statistic analysis indicated substantial agreement (0.69) between the two tests for CEK samples, with moderate agreement (0.45) for the direct specimens examined.

Animals↗

Viral pathogenesis of a nephrotropic infectious bronchitis virus isolated from commercial pullets.

Infectious bronchitis was diagnosed in 3-to-4-week-old pullets from an outbreak in a commercial flock in California. The disease was characterized by head swelling, watery discharge from the eyes and nostrils, and urates in kidneys. Mortality ranged from 1.8% to 12.5% per week. The isolation of a coronavirus from a suspension of pooled kidneys from clinically ill chickens at the fifth passage in 10-day-old chicken embryos, gross and histologic renal lesions, and seroconversion by enzyme-linked immunosorbent assay in inoculated birds suggested that the virus isolated was a nephrotrophic strain of infectious bronchitis virus (IBV). The virus isolate was found to be a previously unrecognized serotype, based on virus neutralization tests performed in embryonated chicken eggs. Nephropathogenicity of the IBV isolate was confirmed by inoculation of the viral isolate into specific-pathogen-free chicks and demonstration of renal lesions. The isolation of nephrotropic strains of IBV has not been reported previously from poultry in California.

Animals↗

Experimental transmission of turkey viral hepatitis to day-old poults and identification of associated viral particles resembling picornaviruses.

Turkey viral hepatitis (TVH) was experimentally reproduced in two experiments in 1-day-old poults. In the first experiment, an infectious inoculum was prepared from filtered yolk materials harvested from dead embryonating chicken eggs (ECE) previously inoculated with suspensions of liver and pancreas tissues collected from TVH-affected birds in commercial turkey flocks. One-day-old poults given a yolk-sac inoculation or oral gavage with this preparation developed lesions in the liver and pancreas characteristic of TVH at 20 days postinoculation (PI) in 60% and 14% of the experimentally infected birds, respectively. With the identical inoculum, embryo mortality occurred at 8 and 10 days PI in embryonating turkey eggs (ETE) inoculated into the yolk sac. In the second experiment, an infectious inoculum was prepared from filtered yolk materials from dead ETE harvested in the first experiment. One-day-old poults given a yolk-sac inoculation with this filtered yolk material developed lesions in the liver and pancreas within 5 days PI. At 20 days PI, 67% of the experimentally infected birds had similar lesions. With the inoculum given to these poults, embryo mortality occurred at 6, 8, and 10 days PI in ETE inoculated into the yolk sac. Virus particles 26-28 nm in diameter with icosahedral morphology typical of picornaviruses were identified by EM in the yolk sacs of ETE that died in both experiments, and inoculated ETE that died following passage of filtered suspensions of pancreatic tissues collected from affected birds in the first experiment.

Animals↗

Herpes viral hepatitis in a toucan.

Herpesvirus infection was diagnosed in a toucan. The herpesvirus was isolated from the liver and identified by electron microscopy in the liver and in cell culture. A negative immunofluorescent reaction was obtained when virus-infected cell cultures were reacted with a conjugate to the herpesvirus of Pacheco's disease. The main pathologic finding in the toucan consisted of a severe necrotizing hepatitis with intranuclear inclusions in the liver and spleen. A presumptive diagnosis of chlamydiosis was also made, based on a positive direct fluorescent monoclonal antibody reaction to chlamydial antigens in impression smears of liver and spleen. Chlamydial isolation attempts were unsuccessful. The toucan had been in contact with two macaws that had died 5 days before the toucan died and were diagnosed by histology as having herpesvirus hepatitis.

Animals↗

Hypoglycemia, enteritis, and spiking mortality in Georgia broiler chickens: experimental reproduction in broiler breeder chicks.

The clinical signs, hypoglycemia, and mortality of "spiking mortality syndrome" were experimentally reproduced. Seven groups of day-old male primary broiler breeder chicks were orally inoculated with tissue and/or fecal-urate homogenates taken from field broilers with spiking mortality syndrome and from field broilers with enteritis and/or runting-stunting syndrome. All homogenates used as inocula were shown by transmission electron microscopy and negative staining to contain arenavirus-like particles. Inocula produced from field broilers with spiking mortality syndrome contained the highest numbers of the arena-virus-like particles and produced the highest percentage of hypoglycemic chicks 13-15 days postinoculation after a 5-to-9-hour fast. These homogenates also produced the most significant differences in mean plasma growth hormone and insulin-like growth factor-1 levels. The significance of the arenavirus-like particles is unknown but is currently being investigated.

Animals↗

Experimental reproduction of severe hypoglycemia and spiking mortality syndrome using field-derived and embryo-passaged preparations.

The clinical signs, enteritis, weight depression, and hypoglycemia of spiking mortality syndrome were experimentally reproduced in broiler breeders and broiler chicks. Inocula included 1) virus-like particles from intestines of chicks with spiking mortality syndrome that had been banded in a discontinuous Renograffin gradient, 2) homogenized darkling beetles collected from litter of farms where spiking mortality syndrome had occurred repeatedly, and 3) homogenized embryos which had been inoculated with the Renograffin-banded material. Arkansas variant infectious bronchitis virus and arenavirus-like particles were identified in the inocula. Serology on samples from surviving chicks suggested the presence of an avian encephalomyelitis virus in one of the inocula. One-day-old (n = 172) and 2.5-day-old (n = 30) chicks were inoculated orally, and some were also injected intraperitoneally or subcutaneously, with 0.5 ml of the inocula. Twelve to fourteen days postinoculation, chicks were fasted for 4-6 hours, then briefly stressed with a cool water spray. Within 1.5 hours, inoculated chicks began dying with severe hypoglycemia and clinical signs of spiking mortality syndrome. Body weights were significantly depressed. Uninoculated controls (n = 130) from the same hatches, also fasted and stressed, were unaffected clinically and were not hypoglycemic. One group (n = 52) of inoculated chicks exposed to a controlled lighting program was unaffected clinically, had significantly higher mean plasma glucose levels, and had significantly less body weight depression than chicks exposed to continuous lighting. We concluded that exposure to controlled amounts of light/darkness can ameliorate much of the hypoglycemia, mortality, and runting-stunting associated with spiking mortality syndrome of chickens. The significance of the viruses and virus-like particles detected in the inocula is currently under investigation.

Animals↗

Experimental reproduction of hypoglycemia-spiking mortality syndrome in broiler chickens with the use of homogenized brains containing arenaviruslike particles.

Severe hypoglycemia-spiking mortality syndrome was experimentally reproduced in broiler chicks. Inoculum was homogenized brains from 28-day-old commercial broiler chicks with central nervous system signs (50% [v/v] in phosphate-buffered saline with 2% fetal calf serum). Oral inoculations of 1.2 ml of the homogenate were given at 1 day of age to broiler chicks (n = 15). Fourteen days later, chicks were fasted and stressed with a 2-sec cool water spray. Six chicks (40%) developed clinical signs of spiking mortality syndrome and were severely hypoglycemic. Uninoculated control chicks (n = 15) from the same hatch, also fasted and stressed simultaneously, were unaffected. Examination of a banded fraction produced from the inoculum with the use of transmission electron microscopy with negative staining revealed viruslike particles indistinguishable from arenavirus particles stained and examined simultaneously. Avian encephalomyelitis virus was isolated by one of three laboratories attempting virus isolation with the use of embryonating chicken eggs.

Animals↗