Effect of membrane stabilizers on semen quality and sperm membrane protein expression during cryopreservation of goat semen.

BACKGROUND: Semen cryopreservation is a complex process during which there is alteration in the expression of sperm and seminal plasma proteins, molecular weight of protein or loss of membrane proteins during the process. In order to compensate for these changes, different membrane stabilizers are used in freezing semen extenders. However, there is scarcity of such studies during cryopreservation of goat semen. OBJECTIVE: To investigate the effect of membrane stabilizers on sperm membrane protein expression during cryopreservation of goat semen. MATERIALS AND METHODS: A total of 36 semen ejaculates from nine Assam Hill Goat bucks aged 2 to 2.5 years was collected by artificial vagina method. Three membrane stabilizers, each at two different concentrations viz. 50 and 80 mM sucrose, 50 and 100 mM trehalose, and 100 and 150 ng per mL IGF-1 (insulin-like growth factor 1 protein) were added to Tris-citric acid fructose egg yolk glycerol (TCFEYG) extender and semen samples were cryopreserved. The sperm membrane protein profile was studied in fresh and cryopreserved semen by SDS-PAGE. RESULTS: SDS- PAGE of sperm membrane extract of fresh semen revealed the presence of 24 protein bands with molecular weights ranging from 10 kDa to 240 kDa. Samples supplemented with 50 mM sucrose and 80 mM sucrose revealed 21 protein bands with molecular weights ranging from 10 kDa to 240 kDa. All the 21 protein bands were same as those observed in the sperm membrane of fresh spermatozoa, except that the 23 kDa, 29 kDa and 42 kDa bands were absent in frozen semen. Similarly, frozen semen extended with 50 mM trehalose and 100 mM trehalose revealed 22 protein bands with molecular weights ranging from 10 kDa to 240 kDa, but lacking the 29 kDa and 42 kDa bands. Proteins with molecular weights of 29 kDa, 130 kDa and 240 kDa were absent in frozen semen supplemented with 100 ng per mL IGF-1 and 150 ng per mL IGF-1. CONCLUSION: The present study revealed that supplementation of tris basic extender with trehalose at 100 mM and or IGF-1 at 100 ng/mL or 150 ng per mL improves the post-thaw semen characteristics and protects certain fertility related sperm membrane proteins. Doi.org/10.54680/fr23510110612.

Development of an effective and efficient DNA isolation method for Cinnamomum species.

Different species of Cinnamomum are rich in polysaccharide's and secondary metabolites, which hinder the process of DNA extraction. High quality DNA is the pre-requisite for any molecular biology study. In this paper we report a modified method for high quality and quantity of DNA extraction from both lyophilized and non-lyophilized leaf samples. Protocol reported differs from the CTAB procedure by addition of higher concentration of salt and activated charcoal to remove the polysaccharides and polyphenols. Wide utility of the modified protocol was proved by DNA extraction from different woody species and 4 Cinnamomum species. Therefore, this protocol has also been validated in different species of plants containing high levels of polyphenols and polysaccharides. The extracted DNA showed perfect amplification when subjected to RAPD, restriction digestion and amplification with DNA barcoding primers. The DNA extraction protocol is reproducible and can be applied for any plant molecular biology study.

Serological evidence of coronavirus infection in mithuns (Bos frontalis) from India.

Studies conducted on mithuns at the National Research Centre on Mithun, and mithuns found in free-ranging conditions in Nagaland, India, revealed that the overall prevalence of antibodies to coronavirus was 80% (95% confidence interval [CI] 77-83) when detected by a commercially available enzyme-linked immunosorbent assay. The highest seroprevalence (92%; 95% CI 89-95) was found in mithuns above three years of age, and the lowest (53%; 95% CI 36-70) in mithuns of 7 to 12 months old (P = 0.00005). The study also revealed a higher degree of positivity (++++) in mithuns above three years old than in those below one year. No statistically significant difference in seroprevalence was observed between male (80%; 95% CI 75-85) and female mithuns (80%; 95% CI 76-84). The seroprevalence was found to be higher (92%; 95% CI 89-95) in mithuns reared in semi-intensive systems compared with those kept under free-ranging conditions (56%; 95% CI 49-63; P = 0.003). This is probably the first serological evidence of coronavirus infection in mithuns. The possible role of adults in the epidemiology of coronavirus infection in mithuns is also discussed.

Prevalence of Cryptosporidium parvum in mithuns (Bos frontalis) from India.

Systematic examination of faecal samples from mithuns maintained at the National Research Centre on Mithun, Indian Council of Agricultural Research (ICAR), Nagaland, India and mithuns found in free-ranging condition of Nagaland, by using a commercially available enzyme-linked immunosorbent assay kit, revealed that the overall prevalence of Cryptosporidium parvum in mithun was 56% (95% CI=48-64). Highest (prevalence rate=81, 95% CI=67-95) prevalence was found in mithuns of 1-6 months of age and lowest (prevalence rate=42, 95% CI=31-53) in mithuns above 2 years of age. The prevalence was found to be higher (94%) in diarrhoeic animals in comparison to non-diarrhoeic group (51%). No statistically significant difference was observed between male (prevalence rate=61, 95% CI=48-74) and female (prevalence rate=53, 95% CI=43-63) prevalences. The prevalence was found to be higher (prevalence rate=64, 95% CI=55-73) in mithuns kept in semi-intensive system than mithuns found in free-ranging condition (prevalence rate=40, 95% CI=27-53). The risk factor that may play a pivotal role in the epidemiology of cryptosporidiosis in mithun is also discussed in brief. The zoonotic implication of the disease is also discussed. This is probably the first report on prevalence of Cryptosporidium parvum in mithun.

Inheritance of heading date, plant height, ear length and spikelets per spike in an intervarietal cross of wheat.

A study to obtain information on early segregating generations of an intervarietal cross 'WG 357' X 'Tobari 66' in spring wheat on the genetics of days to heading, plant height, ear length and spikelets per spike was conducted. 'WG 357' has amber, hard and lustrous grains and is a well adapted high yielding variety of North India whereas 'Tobari 66' is red grained introduction from CIMMYT.The parental F1, F2, B1, B2, biparentals, F3 (parents of biparentals), F3 bulk and F4 bulk generations were studied in order to provide analysis of generations means (Mather 1949; Hayman 1958) and variance component analysis (Kearsay 1965; Perkins and Jinks 1970).There were highly significant differences among the generations for all the characters studied. There were significant differences among the F3 lines as well as among the biparental progenies. Only in case of ear length was the contrast between the two also significant. The mean value of most of the generations arising from the cross fell between the parental range.The three-parameter model failed to account for the variation in generation means in the case of days to heading. This character was concluded to be influenced by linkage and higher order interactions. For the other characters the three parameter model was adequate. For all characters, additive gene effects were most important as compared to dominance gene effects.The analysis of gene action as provided by the generation variance indicated that additive variance was much more pronounced as compared to dominance variance. The heritability was high for days to heading (71 per cent for narrow sense and 80 per cent in broad sense) and plant height (62 and 93 per cent in narrow and broad sense respectively.The implications of the results in breeding programmes have been discussed.