The quick diagnosis of Newcastle disease requires known serum against the disease. In this study, an attempt was made to raise anti- Newcastle disease virus hyperimmune serum in rabbits. Three different inoculum were prepared to inoculate in the rabbits; (i) Fresh harvested allantoic fluid containing Newcastle disease virus (NDV) Mukteswar vaccine strain; (ii) Freshly harvested ND virus pelletted through centrifuging at 40,000 rpm for two hours and resuspended in normal saline and (iii) Pelletted virus (centrifuged and suspended as in ii) with addition of incomplete Freund's adjuvant. It was observed during the monitoring of haemagglutination inhibition (HI) titre that the serum collected after series of inoculation of 1st and 2nd inoculum provided maximum titre upto 1:256. However, the serum collected after series of injection of 3rd inoculum gave maximum HI titre 1:1024. This study suggested that antigen containing incomplete Freund's adjuvant provided better immune response against Newcastle disease virus in rabbits.

This study was carried out for investigation of an outbreak in Karachi. A disease outbreak in poultry was reported during April, 2000 in Karachi. The main symptoms included respiratory distress, sneezing and gasping. Autopsy of dead birds showed tracheitis, air-sacculitis and involvement of lungs. The morbid material was collected for processing in the laboratory. During the investigation for isolation of pathogenic bacteria or virus, Ornithobacterium rhinotracheale was isolated on 5% sheep blood agar plates in an atmosphere containing 5-10% CO2 at 37 degree C and identified through biochemical and fermentation tests. A Lasota like virus was also isolated from the same material which showed HA activity but was found negative to Avian Influenza virus against known Avian Influenza serum. The isolated virus was sent to Central Veterinary Laboratory, Weybridge, U.K. which confirmed it to be Lasota virus having comparatively high Intracerebral Pathogenicity Index (ICPI) as 0.90. Attempts for the transmission of the disease in susceptible healthy broiler chicks was successful through aerosol route using combination of inoculum of the isolated bacteria and virus. On post-mortem of infected birds air- sacculitis, tracheitis and unilateral pneumonia were noticed. Again ORT and NDV were isolated from the material harvested from these chicks. According to the observations of these experiments, it was concluded that the isolated ORT had a triggering effect on Lasota virus or vice versa. The environmental temperature and other stress factors might have aggravated the disease problem.

This study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells (fetal fibroblasts, adult skin and muscle cells, and cumulus cells) after culture under a variety of conditions ; 1 ) growth to 60- 70% confluency (cycling) , 2) serum starvation, 3) culture to confluency. Cell -cycle phases were determined by flow cytometry with propidium iodide staining enabling the calculation of percentages of cells in GO /G1, S and G2 /M. The majority was in GO /GI regardless of cell type and treatment. Serumstarved or confluent cultures contained higher percentages of cells in GO /G1 (89.5-95.4% ; P <0.05) . Percentages of cells in GO /G1 increased as cell size decreased regardless of the cell type and treatment. In the serum-starved and confluent cultures, about 98% of small cells were in GO /G1. Serum-starved cultures contained higher percentages of small cells (38.5-66.9%) than cycling and confluent cultures regardless of cell type (P < 0.05) . After trypsinization of fetal fibroblasts and adult skin cells that were serum-starved and cultured to confluency, the percentages of cells in GO /G1 increased (P < 0.05) on incubation for 1.5 (95.7-99.5%) or 3 hr (95.9-98.6%). These results verify that serum starvation and culture to confluency are efficient means of synchronizing bovine somatic cells in GO /G1, and indicate that a more efficient synchronization of the cells in GO /G1 can be established by incubation for a limited time period after trypsinization of serum-starved or confluent cells.

Bovine chondrocytes were cultured in monolayers and alginate beads with or without ascorbic acid (Asc) for 16 days. Cell proliferation was examined every 4 days by staining with Hoechst 33258 dye. The gene expression of aggrecan, and collagen type I and II was analyzed at 16 days by reverse transcription and polymerase chain reaction.Cell morphology and the production of extracellular matrix (ECM) were evaluated by cytochemical, immunocytochemical and electron microscopical methods.Cells were continuously cultured in alginate beads with Asc for 2 months, and the cell morphology and ECM were examined. The proliferation of chondrocytes was significantly stimulated with Asc in both monolayers and alginate beads at 16 days. Expression of the collagen type I gene in both cultures was increased, and that of the collagen type II gene in alginate beads was decreased, by Asc. There were no significant cytochemical and immunocytochemical differences between the cultures in alginate beads with or without Asc at 16 days.In alginate beads cultured with Asc for 2 months, proliferating cells were observed mainly at the periphery of the beads, and glycosaminoglycan and collagen type II were found around the cells. These results suggest that Asc stimulated the proliferation of chondrocytes and maintained the chondrogenic properties of the cells in an alginate beads culture.

Endothelial cells are important for maintenance of vascular integrity by producing a variety of bioactive molecules such as nitric oxide (NO) . Recent evidence has suggested that there are some differences in characteristics between endothelial cells from different origins. Here we examined responses of two typical endothelial cells to hepatocyte growth factor (HGF), which induces endothelium-dependent relaxation of microvessels. Stimulation of human umbilical vein endothelial cells (HUVEC) and bovine aortic endothelial cells (BAEC) with HGF increased endothelial NO synthase activity, accompanied with an increase of activity-related site-specific phosphorylation of protein kinase B/Akt. However, HGF stimulated phosphorylation of p38 mitogenactivated protein kinase (MAPK) only in HUVEC, but not in BAEC, while it induced phosphorylation of p44 /p42 MAPK in both cells. These results suggest that HGF transduces different intracellular signals between aortic and umbilical venous endothelial cells, and that the differences might represent divergent endothelial responses to growth factors, especially those that activate receptor-tyrosine kinases.

In this study ELISA was standardized for detecting antibodies against Hydropericardium Syndrome Virus. Factors like Antigen, Conjugate and incubation time were studied. Standardization was carried out by checker board titration method. Optimum incubation time for colour development was found to be 15 minutes while 50 mug/ml of antigen gave better results than 100 mug/ml. The conjugate dilution of 1:500 was observed to give satisfactory results as determined by regression analysis. Cut off value was determined by adding 2SDs and 3SDs in the mean optical density (OD) values of negative serum samples. It was found that mean OD+2SD (0.0684) resulted in better diagnostic sensitivity) and diagnostic specificity than mean OD+3SDs (0.0741).

In this study five samples each of serum glucose, serum cholesterol and uric acid contents were measured in Fayoumi, Rhode Island Red, Parent desi, desi Fl and desi F2 strains of chickens. Cholesterol contents were found to be lowest in desi F2 chickens whereas in other strains there was least significant difference, which can be attributed to reduced lipogenic activity in desi F2 generation. When five samples each of meat from two locations i.e. breast and thigh was subjected to proximate analysis it was found that there was no difference (P greater than 0.05) in crude protein, crude fat, total ash and moisture contents regardless of sex and strains. Dry matter contents differed (P less than 0.05) in breast meat and found to be lowest in Parent desi stocks.