Pathogenicity, sequence and phylogenetic analysis of Malaysian Chicken anaemia virus obtained after low and high passages in MSB-1 cells.

Specific-pathogen-free (SPF) chickens inoculated with low passage Chicken anaemia virus (CAV), SMSC-1 and 3-1 isolates produced lesions suggestive of CAV infection. Repeated passages of the isolates in cell culture until passage 60 (P60) and passage 123 produced viruses that showed a significantly reduced level of pathogenicity in SPF chickens compared to the low passage isolates. Sequence comparison indicated that nucleotide changes in only the coding region of the P60 passage isolates were thought to contribute to virus attenuation. Phylogenetic analysis indicated that SMSC-1 and 3-1 were highly divergent, but their P60 passage derivatives shared significant homology to a Japanese isolate A2.

Isolation, identification and characterization of chicken anaemia virus in Malaysia.

A study was conducted to isolate and identify chicken anaemia virus (CAV) from field samples of clinically infected broiler chickens in Malaysia. A total of 125 samples were collected from chickens aged 2-6 weeks with clinically depressed and retarded growth, of which five samples were found positive to CAV directly by polymerase chain reaction (PCR). Later, five isolates of CAV from the respective five PCR positive samples were isolated in MDCC-MSB1 cells at passage 4 based on cytopathic effects, PCR and indirect immunofluorescent antibody test. The isolates were identified as BL-1, BL-2, BL-3, BL-4 and BL-5. These CAV isolates were found to resist treatment with chloroform and heat at 37 degrees C for 2 h, 56 degrees C for 30 min and 70 degrees C for 5 min. One of the isolates, BL-5 produced significant reduction (p < 0.001) of hematocrit values (9-19%), pale bone marrow, thymus atrophy and haemorrhages in skin/muscle when inoculated into 1-day old SPF chickens. Restriction enzyme digestion of 926 bp genomic fragments of all the isolates including Cux-1 isolate with HindIII exhibited a similar pattern of bands in 2% agarose gel. The present findings confirmed the presence of CAV in Malaysia.

Development of a modified method for sequencing high G:C regions in chicken anemia virus genome.

Two areas in the chicken anemia virus (CAV) genome have high G:C content with secondary structures. These two G:C rich areas could not be sequenced with Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit. Several modifications were carried out to solve the problem. Finally, a package of modified method was developed to sequence the high G:C areas. The result showed that the Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit with the normal procedures are not suitable for sequencing the high G:C regions of the CAV genome. The present developed method made the Perkin Elmer's Kit useful for the first time to sequence the G:C rich hairpin structures of the CAV genome. The system may be useful to sequence all other G:C rich DNA templates.

Economics of feeding pasteurized colostrum and pasteurized waste milk to dairy calves.

OBJECTIVE: To estimate the marginal contribution of pasteurization of waste milk and colostrum to gross margin per calf at weaning and to estimate the minimum number of cattle on a dairy farm for pasteurization to be profitable. DESIGN: Randomized, controlled, clinical trial. ANIMALS: 300 Holstein calves. PROCEDURE: The performance of calves fed pasteurized colostrum and waste milk was compared with the performance of calves fed nonpasteurized colostrum and waste milk. Costs, revenues, and gross margins for the 2 groups were compared. RESULTS: Calves fed pasteurized colostrum and waste milk were worth an extra $8.13 in gross margin/calf, compared with calves fed nonpasteurized colostrum and waste milk. The minimum number of cattle for which feeding pasteurized colostrum and waste milk was calculated to be economically feasible was 315 calves/d (1,260-cow dairy farm). CLINICAL IMPLICATIONS: An economic benefit was associated with feeding pasteurized colostrum and waste milk. Additional benefits that may accrue include higher mean weight gain and lower mortality rate of calves as well as calves that have fewer days in which they are affected with diarrhea and pneumonia.

Dairy cattle abortion in California: evaluation of diagnostic laboratory data.

A descriptive study was undertaken on 595 dairy cattle abortion submissions to the California Veterinary Diagnostic Laboratory System from July 1, 1987, to December 31, 1989, to determine the etiologic nature and distribution (seasonal and geographical) of dairy cattle abortion in California as reflected by laboratory submissions. Univariate analysis was performed to characterize abortion-related submissions by farm and laboratory variables, and logistic regression analysis was performed to determine factors that may influence success of abortion diagnosis in the laboratory. The proportions of dairies that submitted abortion-related specimens from northern, central, and southern milksheds during the 2.5-year period were 20.3%, 15.7%, and 13.1%, respectively, and 60% of submissions were from medium-sized (200-999 cows) dairies. Submissions consisted of fetus (58%), placenta (2%), fetus and placenta (12%), and fetus, placenta, and maternal blood (0.84%); fetal tissues and uterine fluid constituted the rest. An apparent pattern in abortion submissions was indicated by a peak in submissions during the winter and summer of 1988 and 1989. Infectious agents were associated with 37.1% of submissions; noninfectious causes, 5.5%, and undetermined etiology, 57.3%. Bacterial abortion accounted for 18% of etiologic diagnoses; protozoal, 14.6%; viral, 3.2%; and fungal, 1.3%. Submissions comprising fetus, placenta, maternal blood, or their combinations were associated with a higher likelihood of definitive diagnosis for abortion than tissues, as were fresher specimens and submissions associated with the second trimester of fetal gestation.