Use of cryopreserved semen has become an important tool in assisted reproduction but freezing and thawing cause sub-lethal damage to spermatozoa. This is detrimental to sperm because of the membrane damage including permeability and integrity. An excess generation of reactive oxygen species (ROS) creates oxidative stress due to reduced antioxidant status of the cryopreserved spermatozoa. In the present study fresh buffalo semen was collected and divided into two aliquots. One aliquot was used for fresh semen analysis and the other was cryopreserved in Tris-egg yolk-citrate extender. The semen samples were used to study different sperm quality parameters like motility, viability, membrane integrity and total antioxidant status. The DNA integrity in fresh and cryopreserved spermatozoa was also studied using comet assay. The sperm quality parameters like post-thaw sperm motility, viability, membrane integrity and total antioxidant status of cryopreserved spermatozoa were significantly lowered (P < 0.05) compared to fresh spermatozoa. The DNA fragmentation in cryopreserved spermatozoa was significantly higher (P < 0.01) as compared to fresh spermatozoa. The results show that the irreversible DNA damage occurs in spermatozoa during cryopreservation.
Heat shock protein 70 (HSP70) is one of the most abundant and best characterized heat shock protein family that consists of highly conserved stress proteins, expressed in response to stress, and plays crucial roles in environmental stress tolerance and adaptation. The present study was conducted to identify major types of genes under the HSP70 family and to quantify their expression pattern in heat-and cold-adapted Indian goats (Capra hircus ) with respect to different seasons. Five HSP70 gene homologues to HSPA8, HSPA6, HSPA1A, HSPA1L, and HSPA2 were identified by gene-specific primers. The cDNA sequences showed high similarity to other mammals, and proteins have an estimated molecular weight of around 70 kDa. The expression of HSP70 genes was observed during summer and winter. During summer, the higher expression of HSPA8, HSPA6, and HSPA1A was observed, whereas the expression levels of HSPA1L and HSPA2 were found to be lower. It was also observed that the expression of HSPA1A and HSPA8 was higher during winter in both heat-and cold-adapted goats but downregulates in case of other HSPs. Therefore, both heat and cold stress induced the overexpression of HSP70 genes. An interesting finding that emerged from the study is the higher expression of HSP70 genes in cold-adapted goats during summer and in heat-adapted goats during winter. Altogether, the results indicate that the expression pattern of HSP70 genes is species-and breed-specific, most likely due to variations in thermal tolerance and adaptation to different climatic conditions.
Background: Porcine reproductive and respiratory syndrome (PRRS) is an economically important viral disease of pigs worldwide. India has reported the first outbreak of PRRS in the pig population of Mizoram state to the Office International des Epizooties on the 26 June 2013. Hypothesis/Objectives: The aim of the present study was to determine the genotype and origin of porcine reproductive and respiratory syndrome virus (PRRSV) from the first outbreak in the pig population of Mizoram state, India, in 2013. Animals and methods: A total of 880 affected pigs from the outbreak were clinically examined, 51 animals were necropsied and tested by reverse transcription polymerase chain reaction (RT-PCR) to detect PRRSV and 148 serum samples were tested to detect PRRSV-specific antibodies. The full open reading frame 5 (ORF5) gene sequences from 12 and ORF7 gene sequences from three clinical cases were sequenced and analysed for genomic characterization, respectively. Results: The outbreak was confirmed by the detection of PRRSV-specific antibodies in 109 out of 148 serum samples (74%) and also by RT-PCR in 46 out of 51 necropsy samples (90%). Notably, ORF5 and ORF7 genes of Indian strain shares the same nucleotide positions i.e. 13,698À14,300 and 14,799À15,170, respectively, with the highly pathogenic (HP) strain of China and were grouped together in a phylogenetic tree. Conclusions and clinical importance: Sequence and phylogenetic analysis of ORF5 and ORF7 confirmed that the Indian strain has a close link with the HP-PRRSV of China. The current study forms an essential step for better understanding of the epidemiology as well as the movement and spread of the disease in India.
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