Invivo biocompatibility determination of acellular aortic matrix of buffalo origin.

In the present study, biocompatibility of native, acellular, 1,4-butanediol diglycidylether and 1-ethyl-3-(3-dimethyl aminopropyl carbodiimide (EDC) cross-linked acellular aortic grafts was evaluated following subcutaneous implantation in guinea pigs. Biocompatibility was evaluated based on macroscopic, histopathological observations and immune responses elicited by the implanted grafts. Results showed that macroscopically, no abnormal cellular reaction was observed at the host-graft junction in any of the implanted animals. Histopathological observations revealed that the inflammatory response was mild during first 15 days post-implantation and increased at 30 days post-implantation in acellular and cross-linked tissues. By day 60, marked ingrowth of host tissue was observed in EDC cross-linked acellular aortic grafts. ELISA and lymphocyte proliferation assay revealed that animals implanted with EDC grafts showed least immune response when compared to others. Therefore, it was concluded that EDC cross-linked acellular aortic grafts were more compatible and had better handling qualities than the other cross-linked grafts.

Determination of anti-inflammatory cytokine in periparturient cows for prediction of postpartum reproductive diseases.

Forty-one pregnant cows at 240 days of gestation were selected from the cattle herd at the Indian Veterinary Research Institute, Izatnagar, Bareilly, India. The cows were critically observed daily during the periparturient period until 45 days postpartum. The prepartum sampling day was fixed on the basis of AI date supported by the external signs of the animals approaching parturition. Blood sampling was done for each experimental animal on 15 days prepartum (-15 d), calving day (0 d), 15 days (15 d), and 30 days (30 d) postpartum, and thorough gynecological examinations were performed on 0 d, 15 d, 30 d, and 45 d for diagnosis of postpartum reproductive diseases like retained placenta (ROP), clinical metritis (CM), clinical endometritis, cervicitis, and delayed involution of the uterus. The blood serum was used for estimation of the anti-inflammatory cytokine interleukin 10 (IL-10). Determintaion of IL-10 concentration in serum was done by using a commercially available bovine-specific ELISA kit. The IL-10 concentration (pg/mL) was significantly (P < 0.01) higher for ROP (1956.96 ± 325.30) and CM (2283.01 ± 326.82) than normal cows (827.19 ± 127.11) at 15 days before calving. The values for IL-10 was also significantly (P < 0.05) higher for cows with ROP and CM than for normal cows at 0 d and 15 d postpartum. However, the values for IL-10 were significantly (P < 0.05) higher for cows with clinical endometritis (1847.83 ± 539.38) than for normal cows (770.75 ± 29.22) at 30 d postpartum. The concentration of IL-10 was lower (P > 0.05) in cows with delayed involution of uterus than other groups of cows at all days of the periparturient period. The cytokine level in cows with delayed involution of the uterus was 328.906 ± 107.19, 263.08 ± 84.92, 415.26 ± 102.14, and 386.28 ± 111.11 on -15 d, 0 d, 15 d, and 30 d, respectively. According to the results of this study it can be concluded that increased serum concentration of IL-10 at 15 d prepartum was observed in cows that developed ROP and CM and remained significantly higher until 15 d postpartum. Therefore, this cytokine might be used as a prognostic marker to identify the cows that are going to develop ROP and CM.

Isolation of bovine coronavirus (bcoV) in vero cell line and its confirmation by direct FAT and RT-PCR.

Bovine Coronavirus (BCoV) is widespread both in dairy and beef cattle throughout the world. The virus is one of the largest RNA virus and has specific tropism for intestinal and pulmonary epithelial cells. It is responsible for huge economic losses by causing winter dysentery in adult dairy cattle and respiratory and intestinal tract infections leading to pneumo-enteritis in young calves. Isolation of BCoV has been reported to be difficult. Studies regarding epidemiology, virus isolation and molecular detection from India are very few. In the present study Vero cell line was used for isolation of the BCoV from Enzyme Linked Immunosorbent Assay (ELISA) positive samples. Direct florescent antibody technique (dFAT) and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to confirm the isolated virus strains at antigenic and genomic levels, respectively. Out of the 15 positive fecal samples, virus from only seven was able to infect vero cell line. Subsequently BCoV got adapted to the vero cell line upto three passages, which was confirmed both at genomic and antigenic levels by dFAT and RT-PCR testing. It can be concluded that vero cell line can be used for isolation of BCoV, however due to the enormous stain diversity of the virus it is possible that many stains can't grow and get adapt in this cell line. Further studies are required for isolation of different viral strains, finding the susceptible cell lines and also to confirm the variations among the BCoV isolates at antigenic/genomic levels.

Use of RAPD-PCR for genetic analysis of Nicobari fowl of Andamans.

1. The present study was conducted to estimate genetic relatedness among Nicobari fowls (Brown, Black and White) and an exotic bird (White Leghorn) using random amplified polymorphic DNA (RAPD) polymorphism. 2. A total of 25 decamer primers were screened among all the breeds of which 24 primers amplified the genomic DNA, generating 2000 to 200 bp bands. Ten primers generated reproducible and distinct RAPD profiles and were used for further analysis. 3. A total of 94 bands were amplified and 30 polymorphic bands (32%) were produced. The number of polymorphic loci ranged from 1 to 5 with an average of 3.0. 4. Among the native breeds Brown Nicobari showed higher genetic similarity (0.85) than Black Nicobari (0.80) and White Nicobari fowl (0.82). 5. Brown Nicobari showed high genetic similarity with Black Nicobari (0.87 +/- 0.029); least similarity was between White Nicobari and White Leghorn (0.77 +/- 0.028). 6. The RAPD profile of all Nicobari fowls on amplification with the primers PBG5 and PBA12 showed specific bands of molecular size 1050 and 785 bp, respectively. 7. The native breeds showed the least genetic distance with each other while White Leghorn appeared to be most distant from the native breeds.