Biotelemetry, ichthyoplankton and genetic data can provide detailed information about the migratory dynamics and reproductive cycle of freshwater fishes. However, few studies have combined these techniques in Neotropical systems. The objective of this study was to examine the migratory and reproductive dynamics of Prochilodus costatus in the São Francisco River watershed, south‐east Brazil, by comparing the ecological importance of two rivers to the species, an undammed segment of the São Francisco River and a dammed segment of one of its main tributaries, the Pará River. In total, 215 fish were radio‐tagged over three years (2014–16). Eggs and larvae were sampled at seven locations and analysed by PCR to identify Prochilodus spp. ichthyoplankton. Most radio‐tagged individuals (97%) used the undammed segment of the São Francisco River as spawning migration route, even those captured and released in the Pará River. Fish migrated to spawn from late September to late November with the arrival of the rains and returned to feeding sites from December to May after spawning. The highest densities of fish eggs and larvae were recorded in the upper reaches of the São Francisco River during months of peak river discharge. Returning fish showed high fidelity to sites occupied before spawning migration. Fish spent roughly 71% of the year at feeding sites, 25% at spawning sites and 4% moving between them. This study provides novel information about the migratory dynamics of Neotropical fishes and underscores the key role of undammed river segments for the conservation of Neotropical migratory fish species.
Genetic diversity and population studies are essential for conservation and wildlife management programs. However, monitoring requires the analysis of multiple loci from many samples. These processes can be laborious and expensive. The choice of microsatellites and PCR calibration for genotyping are particularly daunting. Here we optimized a low-cost genotyping method using multiple microsatellite loci for simultaneous genotyping of up to 384 samples using next-generation sequencing (NGS). We designed primers with adapters to the combinatorial barcoding amplicon library and sequenced samples by MiSeq. Next, we adapted a bioinformatics pipeline for genotyping microsatellites based on read-length and sequence content. Using primer pairs for eight microsatellite loci from the fish Prochilodus costatus, we amplified, sequenced, and analyzed the DNA of 96, 288, or 384 individuals for allele detection. The most cost-effective methodology was a pseudo-multiplex reaction using a low-throughput kit of 1 M reads (Nano) for 384 DNA samples. We observed an average of 325 reads per individual per locus when genotyping eight loci. Assuming a minimum requirement of 10 reads per loci, two to four times more loci could be tested in each run, depending on the quality of the PCR reaction of each locus. In conclusion, we present a novel method for microsatellite genotyping using Illumina combinatorial barcoding that dispenses exhaustive PCR calibrations, since non-specific amplicons can be eliminated by bioinformatics analyses. This methodology rapidly provides genotyping data and is therefore a promising development for large-scale conservation-genetics studies.
The main organic contaminants in municipal wastewater are proteins, polysaccharides, and lipids, which must be hydrolyzed to smaller units. A high concentration of oil and grease in wastewater affects biological wastewater treatment processes by forming a layer on the water surface, which decreased the oxygen transfer rate into the aerobic process. Microbial proteases, lipases, amylases, and celullases should play essential roles in the biological wastewater treatment process. The present study aimed to isolate lipase- and other hydrolytic enzyme-producing microorganisms and assess their degradation capabilities of fat and oil wastewater in the laboratory. We also evaluated microbial interactions as an approach to enhance lipolytic activity. We place emphasis on lipase activity because oil and grease are not only environmental pollutants, but also form an undesirable tough crust on pipes of sewage treatment plants. Thirty-five lipolytic microorganisms from sewage were identified and assessed for hydrolytic enzyme profiles. Lipases were characterized in detail by quantification, chain length affinity, and optimal conditions for activity. The good stability of isolated lipases in the presence of chemical agents, thermal stability, wide range of pH activity and tolerance, and affinity for different lengths of ester chains indicates that some of these enzymes may be good candidates for the hydrolysis of organic compounds present in wastewater. A combination of enzymes and fermenting bacteria may facilitate the complete hydrolysis of triglycerides, proteins, and lingo-cellulose that normally occur in the wastes of industrial processes. This study identifies enzymes and microbial mixtures capable of digesting natural polymeric materials for facilitating the sewage cleaning process.
Detection of microbial enzymes in natural environments is important to understand biochemical activities and to verify the biotechnological potential of the microorganisms. In the present report, 346 isolates from soil, water, and plants were screened for enzyme production (caseinase, gelatinase, amylase, carboxymethyl cellulase, and esterase). Our results showed that 89.6% of isolates produced at least one tested enzyme. A predominance of amylase in soil samples, carboxymethyl cellulase in plants, as well as esterase and gelatinase in water was observed. Interesting enzymatic profiles were found in some microenvironments, suggesting specificity of available nutrients and/or natural selection. This study revealed the potential of microorganisms present in water, soil, and plant to produce important enzymes for biotechnological exploration. A predominance of certain enzymes was found, depending on the type of environmental sample. The distribution of microbial enzymes in soil, water and plants has been little exploited in previous reports.
Introduction Freshwater ecosystems provide propitious conditions for the acquisition and spread of antibiotic resistance genes (ARGs), and integrons play an important role in this process. Material and methods In the present study, the diversity of putative environmental integron-cassettes, as well as their potential bacterial hosts in the Velhas River (Brazil), was explored through intI-attC and 16S rRNA amplicons deep sequencing. Results and discussion ORFs related to different biological processes were observed, from DNA integration to oxidationreduction. ARGs-cassettes were mainly associated with class 1 mobile integrons carried by pathogenic Gammaproteobacteria, and possibly sedentary chromosomal integrons hosted by Proteobacteria and Actinobacteria. Two putative novel ARG-cassettes homologs to fosB3 and novA were detected. Regarding 16SrRNA gene analysis, taxonomic and functional profiles unveiled Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria as dominant phyla. Betaproteobacteria, Alphaproteobacteria, and Actinobacteria classes were the main contributors for KEGG orthologs associated with resistance. Conclusions Overall, these results provide new information about environmental integrons as a source of resistance determinants outside clinical settings and the bacterial community in the Velhas River.
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