The dissemination of antibiotic resistant bacteria from anthropogenic sources into the environment poses an emerging public health threat. Antibiotic resistance genes (ARGs) and gene-capturing systems such as integron-associated integrase genes (intI) play a key role in alterations of microbial communities and the spread of antibiotic resistant bacteria into the environment. In order to assess the effect of anthropogenic activities on watersheds in southwestern British Columbia, the presence of putative antibiotic resistance and integrase genes was analyzed in the microbiome of agricultural, urban influenced, and protected watersheds. A metagenomics approach and high-throughput quantitative PCR (HT qPCR) were used to screen for elements of resistance including ARGs and intI. Metagenomic sequencing of bacterial genomic DNA was used to characterize the resistome of microbial communities present in watersheds over a 1-year period. There was a low prevalence of ARGs relative to the microbial population (<1%). Analysis of the metagenomic sequences detected a total of 60 elements of resistance including 46 ARGs, intI1, and groEL/intI1 genes and 12 quaternary ammonium compounds (qac) resistance genes across all watershed locations. The relative abundance and richness of ARGs was found to be highest in agriculture impacted watersheds compared to urban and protected watersheds. A downstream transport pattern was observed in the impacted watersheds (urban and agricultural) during dry months. Similar to other reports, this study found a strong association between intI1 and ARGs (e.g., sul1), an association which may be used as a proxy for anthropogenic activities. Chemical analysis of water samples for three major groups of antibiotics was below the detection limit. However, the high richness and gene copy numbers (GCNs) of ARGs in impacted sites suggest that the effects of effluents on microbial communities are occurring even at low concentrations of antimicrobials in the water column. Antibiotic resistance and integrase genes in a year-long metagenomic study showed that ARGs were driven mainly by environmental factors from anthropogenized sites in agriculture and urban watersheds. Environmental factors such as land-use and water quality parameters accounted for 45% of the variability observed in watershed locations.

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The study investigated the occurrence of trypanosomosis in water buffaloes using parasitological and molecular methods. Buffaloes of different ages and both sexes comprising Bulgarian Murrah and some crosses were used in the study. Two of the 145 samples (0.13%) were positive for Trypanosoma evansi in both, blood parasite examination and PCR. One female buffalo (1/54 samples; 1.86%) from a lactating herd in the city of Muñoz, Nueva Ecija (Region 3) and another (1/51 samples; 1.96%) non-pregnant female belonging to caraheifer population from Los Baños, Laguna (Region 4) were found infected by T. evansi. The positive animals were in the age group of 3-4 years. Blood samples obtained from 40 buffaloes from Cagayan (Region 2) were found negative for trypanosome infection.

ABSTRACT. During the attempt to seek T. congolense species-specific diagnostic antigens, we discovered one cDNA clone (P74) encoding 74 kDa putative abc1 protein (p74) from T. congolense PCF cDNA library. It has been suggested that members of the abc1 family are novel chaperonins and essential for both the mitochondrial electron transfer in the bc 1 complex and the coenzyme Q biosynthesis. Although abc1 protein in yeast has a nuclear or mitochondrial subcellular location, neither nuclear localization signal nor mitochondrial targeting signal was found within p74. Northern blot analysis revealed that the transcription level of P74 mRNA in bloodstream form (BSF) cells were 4 times higher than that in procyclic form cells. Western blot analysis also indicated that p74 was only expressed in T. congolense BSF cells, and revealed that molecular mass of native p74 was not 74 kDa but 56 kDa. This indicates extensive posttranslational modification in p74. Although further characterization of p74 will be required, our findings provide implications for CoQ biosynthesis pathway in T. congolense.