Genetic association of polymorphisms in bovine TLR2 and TLR4 genes with Mycobacterium avium subspecies paratuberculosis infection in Indian cattle population

Toll like receptors (TLRs) are pattern recognition molecules involved in cellular recognition of Mycobacterium avium subspecies paratuberculosis (MAP), the infectious agent causing Paratuberculosis (PTB), a notified disease of domestic and wild ruminants. The present study was undertaken to investigate the presence of single nucleotide polymorphisms (SNPs) in TLR2 and TLR4 gene and to evaluate association of these SNPs with occurrence of PTB in Indian cattle. A total of 213 cattle, were subjected to multiple diagnostic tests viz. Johnin PPD, ELISA test (Indigenous and Parachek kit method), fecal microscopy and fecal culture for detection of MAP infection. Based on screening results 51 animals each were assigned to case and control population. Two SNPs viz. rs55617172, rs41830058 in TLR2 gene and two SNPs viz. rs8193046, rs8193060 in TLR4 gene and were genotyped by PCR-RFLP method. All SNPs were found to be polymorphic except rs41830058 in the case-control population. Both SNPs in TLR4 gene but none in TLR2 genes were significantly associated with the occurrence of PTB in our population. The genotypes in SNP rs8193046 and SNP rs8193060 were significantly (P?<?0.01) different in case-control population. These findings suggest that SNPs rs8193046 and rs8193060 are likely a potential marker against MAP infection and a selection programme eliminating AG genotype for rs8193046 and CT genotype for rs8193060 might be beneficial in conferring resistance to MAP infection in Indian cattle population.

     

Identification of sperm motility markers in bovine transition protein genes

Transition proteins (TNPs) are essential in chromatin condensation during spermiogenesis, and hence, they are the candidate genes for identifying sperm motility markers. Coding and in silico predicted promoter regions of these genes were investigated in crossbred and purebred cattle, and also, their mRNA quantification was done to explore its use as a diagnostic tool of infertility. PCR‐SSCP analysis revealed two band patterns in fragment III of TNP1 and fragment II of TNP2 gene. Sequence analysis revealed a deletion of “G” nucleotide in 3′UTR region of TNP1 and C>T SNP in intronic region of TNP2 gene. Least square analysis of variance did not reveal any significant influence of nucleotide deletion on any sperm motility parameters in both crossbred and purebred cattle. However, C>T SNP had a significant effect on initial progressive motility (p < 0.05) in purebred cattle and post‐thaw motility in overall cattle population. RT‐qPCR analysis did not reveal any significant variation in TNP1 and TNP2 gene expression among poorly motile and good quality spermatozoa of Vrindavani bulls.     

Adrenal cortex reactivity in dairy cows differs between lactational stages and between different feeding levels

Changes in ACTH challenge test characteristics in dairy cows changing their physiological status at different lactational stages and different feeding levels were not investigated in terms of repeatability yet. In 23 multiparous Holstein cows (10 cows fed a sole fresh herbage diet without concentrate, 13 cows fed with concentrate), three ACTH challenge tests were performed: once during pregnancy shortly prior to drying off ( T1 ), and in week 3 ( T2 ) and 8 ( T3 ) after parturition. Test characteristics were correlated to performance and metabolic parameters: DMI, BW, energy balance (EB), plasma concentrations of free fatty acids (NEFA) and beta‐hydroxybutyrate (BHB). Basal plasma cortisol concentrations were higher at T1 compared with T2 and T3 (p < .05). The adrenal cortex sensitivity (expressed as total AUC (AUC_t) of cortisol response after ACTH application) was lowest at T2 compared with T1 and T3 (p < .05). Ranking of the individual animals’ responses was not repeatable between time points of the ACTH tests. Enhancing the energy deficiency during early lactation by omission of concentrate did not affect baseline cortisol concentrations in plasma, but decreased peak height at T2 (p < .05). Baseline plasma cortisol concentrations were positively correlated with cortisol peak values after ACTH application, previous lactation performance, milk yield and BW (p < .05). The AUC_t was positively correlated with baseline cortisol concentrations, EB and DMI. Cortisol release after ACTH injection was lower in animals with high plasma concentrations of NEFA, BHB and with higher contents of fat and free fatty acids in milk (p < .05). Cortisol peak height after ACTH administration was higher in cows with a more positive EB, higher DMI and lower plasma concentrations of NEFA and BHB. In summary, cortisol responses to ACTH challenges in this study were not repeatable in dairy cows changing their physiological status.     

Genetic studies of growth in domesticated rainbow trout

The heritability of body weight and the form of the growth curve was investigated in two crossbred strains of domesticated rainbow trout. Growth rate was nearly exponential at young ages but slowed to nearly linear at older ages. An empirical equation was derived that related exponential growth constants to the observed growth patterns. Heritabilities of body weight based on full-sib families were estimated at six ages for each stock. Estimates made at young ages were similar to other estimates of body weight in salmonid fish. However, non-additive genetic variance, culling bias and common environmental effects may have inflated estimates made at older ages, as the estimates were extremely high, and these possible sources of confounding were known to exist in the experimental design.     

Antibacterial activity of Punica granatum

Successive petroleum ether, chloroform, methanol and water extracts of Punica granatum were tested (in vitro) for their antibacterial activity. The methanolic extract was found to be most effective against all tested microorganisms.     

Mast cell stabilising activity of Bacopa monnieri

Successive petroleum ether, chloroform, methanol and water extracts of Bacopa monnieri were tested (in vitro) for mast cell stabilising effect. The methanolic fraction exhibited potent activity comparable to disodium cromoglycate, a known mast cell stabiliser.     

Drugs from bugs: the use of insects as a valuable source of transgenes with potential in modern plant protection strategies

Transgenic expression of antimicrobial peptides in crops has become a novel approach among the strategies to combat phytopathogens in modern plant protection measures. The first antimicrobial transgenes of insect origin, modified cecropins, have been demonstrated to confer resistance of several transgenic cultivars against both bacterial and fungal phytopathogens. Insects represent a promising reservoir for antimicrobial peptides to engineer disease resistant crops. The increasing knowledge about the potent insect innate immunity may help to develop a novel strategy in sustainable agriculture. Several approaches are presently under investigation to prevent evolution of phytopathogens that can overcome disease resistance in transgenic crops expressing an insect antimicrobial peptide. Pathogen-induced expression of insect antimicrobial peptides in crops and combined multiple expression of different antimicrobial peptides along with proteinase inhibitors from insects may prevent selection of resistant phytopathogens. The potential of insect antimicrobial peptides as transgenes to render disease resistant crops has just started to be explored and may provide tools to be ahead of the evolutionary adaptability of phytopathogens.     

Anthelmintic activity of essential oil of Ocimum sanctum and eugenol.

The essential oil of Ocimum sanctum and eugenol, tested in vitro, showed potent anthelmintic activity in the Caenorhabditis elegans model. Eugenol exhibited an ED(50) of 62.1 microg/ml. Eugenol being the predominant component of the essential oil, is suggested as the putative anthelmintic principle.