Genetic Evaluation of TLR4 and CaLA-DRB Genes for Mastitis Susceptibility in Dairy Goats and the Response of Mastitis-Bacteria to Garlic and Black pepper Extracts
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2012Author
Petlane, Molefe
Noor, Ronny Rachman
Maheswari, Rarah.R.A.
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Mastitis is a bacterial disease that affects mammary glands of nursing animals. It results in serious production and economic losses due to costs attached to its treatment and loss of animals through culling. This study was aimed at evaluating TLR4 and CaLA-DRB genes for mastitis susceptibility in Saanen, PE and PESA using PCR-RFLP and also to investigate the antibacterial effects of garlic and black pepper extracts on mastitis causing bacteria. TLR4 is a transmembrane protein that recognizes PAMPs, convey signals into nucleus thus causing transcription factors that inhibit primary infection while DRB is a cell surface protein that recognizes antigens and present such antigens to T-cells and CD4. Allicin is an active ingredient in garlic; it confers antibiotic properties by interfering with the bacterial polymerase enzyme system thus inhibiting the formation of nucleic acids whereas piperine is an alkaloid that forms active ingredient in black pepper. Piperine alters the permeability of bacterial membrane and inhibits NorA efflux pump mechanism thus causing leakage and accumulation of extracellular substances into the cell which may cause cell death. A total of 68 lactating dairy goats; consisting of animals from Balitnak, Caprito farm, Cordero dairy farm and Bangun dairy farm were used in this study, the population consisted of both mastitic and healthy animals. Genetic polymorphism was detected by digesting TLR4 PCR product with AluI while DRB amplicon was digested with PstI and TaqI in two separate reactions. Antimicrobial tests were performed using agar well diffusion technique. The study found that TLR4|AluI locus was monomorphic and fixed with allele “T” in all breeds analyzed. Allele P had high frequency in PE (0.7069) followed by PESA (0.6900) then Saanen (0.6705) while allele T had high frequency in PESA (0.9205), followed by PE (0.8976) then Saanen (0.7188). DRB|PstI and DRB|TaqI loci were found polymorphic where Saanen exhibited most heterozygosity. DRB|PstI had moderate to high PIC in all breeds while DRB|TaqI had low PIC in PE and PESA. χ2 tests indicated that with the exception of DRB|TaqI locus in PE and in Saanen, all breeds were still in Hardy-Weinberg equilibrium. Staphylococcus aureus was the most frequently (80%) isolated bacteria in mastitic animals while Escherichia coli was less frequent (<30%), thus leading to decision that S.aureus is the main etiological agent in caprine mastitis. The study also found that all genotypes are susceptible to mastitis but genotype PP and TT are comparatively more resistant as compared to other genotypes since they displayed comparatively higher frequency of healthy animals against other genotypes. Antibacterial tests revealed that E.coli was more sensitive to garlic while S.aureus was more sensitive to black pepper.
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