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Kadirvel G.,Indian Veterinary Research Institute | Kadirvel G.,ICAR Research Complex for NEH Region | Kathiravan P.,Animal Genetics and Breeding | Kumar S.,Animal Reproduction
Theriogenology | Year: 2011

Many similarities between the changes associated with normal capacitation and cryocapacitation have been demonstrated. The present study was undertaken to determine whether similarities exist in the protein tyrosine phosphorylation pattern and zona binding ability between in vitro capacitated (heparin induced; 20 μg/ml) and frozen-thawed (cryocapacitated) buffalo spermatozoa. Semen from seven buffalo bulls (eight ejaculates each) was divided into two parts. Part I was used as fresh semen and part II was extended in Tris-egg yolk extender, equilibrated and frozen in liquid nitrogen. Localization of phosphotyrosine-containing protein was determined using an indirect immunoflourescence assay with anti-phosphotyrosine antibody. For zona binding assay, good quality oocytes collected by aspiration technique from fresh buffalo ovaries were used. The bound spermatozoa were stained with Hoechst 33342 dye and observed under fluorescent microscope. The results revealed sperm head associated protein tyrosine phosphorylation in both in vitro capacitated and frozen-thawed spermatozoa. In the zona binding assay, the mean number of bound spermatozoa was 90.6 ± 1.9 and 104.7 ± 2.2 in fresh semen after incubation in non capacitating media at 0 h and 3 h, respectively. But after incubation in capacitating media with heparin for 3 h, the mean number of spermatozoa attached to zona pellucida was 138.4 ± 2.6. The in vitro capacitated spermatozoa had significantly (P < 0.05) higher binding ability than that of fresh spermatozoa. After freezing and thawing, 2.5 fold reductions in the zona binding ability of cryopreserved spermatozoa was observed compared to in vitro capacitated spermatozoa. The binding ability of in vitro capacitated spermatozoa was significantly (P < 0.01) higher than that of frozen-thawed (cryocapacitated) spermatozoa. The study concluded that both in vitro capacitated and frozen-thawed (cryocapacitated) spermatozoa had similar immune-localization of tyrosine phosphorylated protein pattern, however, differed in the zona binding ability. © 2011 Elsevier Inc. Source

Sen S.,Animal Genetics and Breeding | Shukla R.,Animal Genetics and Breeding | Ranjan R.,Animal Genetics and Breeding | Parmar S.N.R.,Animal Genetics and Breeding
Indian Journal of Animal Research | Year: 2015

A total of 60 crossbred cattle were screened for subclinical and clinical mastitis in and around Jabalpur using California mastitis test (CMT) followed by somatic cell count (SCC). The DNA was isolated and 311 bp fragments of IL8R gene was amplified using PCR. PCR-SSCP (Single Strand conformational polymorphism) method was used to identify genetic polymorphism of 311 bp amplicon of IL8R gene. PCR-SSCP revealed five SSCP patterns, SSCP pattern 1 (P1) was present in highest percentage 63.6% in healthy group, SSCP pattern 3 (P3) was present in highest percentage 64.2% in subclinical mastitis group while SSCP patterns (P2 and P5) were present in highest frequency 100% and 100% respectively in clinical mastitis group. Cattles exhibiting pattern P2, P5 and P3 were significantly (p<0.05) associated with susceptibility to clinical and subclinical mastitis in crossbred cattle while those exhibiting pattern P1 were significantly (p<0.05) associated with mastitis resistance. The animals exhibiting SSCP pattern P1 may have IL-8R haplotypes that are good genetic markers for increased mastitis resistance while animals exhibiting pattern P2, P5 and P3 may have IL-8R haplotypes as good genetic markers for clinical and subclinical mastitis susceptibility. Thus, animals exhibiting these patterns can be selected or culled at an early age for future breeding. © 2015, Agricultural Research Communication Centre. All rights reserved. Source

Shukla R.,Animal Genetics and Breeding | Sen S.,Animal Genetics and Breeding | Ranjan R.K.,Animal Genetics and Breeding | Parmar S.N.S.,Animal Genetics and Breeding
Indian Journal of Animal Research | Year: 2015

Toll-like receptors (TLRs) are essential in the host defense against microbial pathogens. Individual TLRs recognize distinct structural components of pathogens and evoke inflammatory responses. In the present study, the amplicon of 316 bp of TLR4 gene was amplified in the thermal cycler (Gene Amp 6700, Applied Biosystem) using the primers of TLR4 gene and the genetic polymorphism was detected by Single Strand Conformation Ploymorphism (SSCP). The SSCP analysis of 316 bp region of TLR4 gene revealed similar type of SSCP patterns in healthy, subclinical and clinical mastitis cases suggesting that Frieswal cattle is monomorphic for TLR4 gene. © 2014, Indian Journal of Animal Research. All Rights Reserved. Source

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