RESUMEN
The aim of this study was to evaluate the in vitro anthelmintic effect of crude aqueous, methanol, ethanol, hydro alcohol and acetone extracts of Vitex negundo leaves against Haemonchus contortus eggs and larvae. Phytochemical analysis to identify the number of compounds in extracts was done by chemical tests and gas chromatography coupled to a mass spectrophotometer detector (GC-MS). First off all the effectiveness of dried plant materials was evaluated on larval development by mixing powdered material (no nano particles) to faecal cultures from donor sheep. Adding powder to the faecal culture resulted into 100% inhibition in larval development at 200 and 300 mg/g of faeces. The anthelmintic activity was assessed using the egg hatch assay (EHA) and the larval mortality assay (LMA). Comparison of mean inhibition percentage of egg embryonation, mean inhibition percentage of egg hatching and mean percentage of larval mortality at different concentrations with control was performed by one-way ANOVA. The means were compared for statistical significance using DMRT at P < 0.05. For both the assays, 50% inhibitory concentration (IC50) and lethal concentration (LC50) were calculated by probit analysis. Chemical test revealed presence of high concentration of saponin and flavoinoids and moderate concentration of total phenols in leaves. The antioxidant activity (radical scavenging activity, RSA %) measured was 35.47%. On GC-MS, the methanolic leaves extract revealed 30 phyto-compounds. On EHA, there was marked effect on inhibition of egg hatching by aqueous, hydro alcohol and acetone extracts. On LMA all the five extracts showed excellent larvicidal activity. V. negundo leaves methanol extract mediated silver nanoparticles were found very effective at much lower concentrations as compared to crude methanol extract. The results indicated that the V. negundo leaves crude extracts possessed excellent in vitro ovicidal and larvicidal properties against H. contortus which needs more investigation, especially in vivo trials for the control of parasite.
RESUMEN
The genetic diversity of MHC class II DQ genes was investigated in riverine buffalo (Bubalus bubalis) by PCR-RFLP and sequencing. Highly variable regions (exons 2-3) of DQ genes were amplified from 152 buffaloes and genotyped by PCR-RFLP. Alleles identified by differential restriction patterns were sequenced for the characterization. PCR-RFLP was a rapid method to discriminate between DQA1 and duplicated DQA2 genes in buffalo, however, the method appeared to be inadequate for determining the more complicated DQB genotypes. A total of 7 and 10 alleles were identified for DQA and DQB loci, respectively. Nucleotide as well as amino acid variations among DQ alleles particularly at peptide binding regions were high. Such variations were as expected higher in DQB than DQA alleles. The phylogenetic analysis for both genes revealed the grouping of alleles into two major sub-groups with higher genetic divergence. High divergence among DQ allelic families and the isolation of two diverse DQA and DQB sequences from individual samples indicated duplication of DQ loci was similar in buffalo to other ruminants.