Obtaining sufficient and high-quality genomic DNA from sludge samples is a fundamental issue of feasibility and comparability in genomic studies of microbial diversity. Commercial kits for soil are often used for the extraction of gDNA from sludge samples due to the lack of specific kits. However, the evaluation of the performance of commercial kits for sludge DNA extraction is scarce and optimization of these methods to obtain a high quantity and quality of DNA is necessary, especially for downstream genomic sequencing. Sequential batch reactors (SBRs) loaded with lignocellulosic biomass are used for the synthesis of renewable resources such as levulinic acid (LA), adipic acid (AA), and polyhydroxyalkanoates (PHAs), and the biochemical synthesis of these compounds is conducted through the inoculation of microbes present in the residual activated sludge (AS) obtained from a municipal wastewater treatment plant. To characterize these microbes, the extraction of DNA from residual sewage sludge was conducted with three different commercial kits: Nucleospin® Soil from Macherey-Nagel, DNEasy® PowerSoil® from Qiagen, and E.Z.N.A.® Plant DNA Kit from Omega BIO-TEK. Nevertheless, to obtain the highest load and quality of DNA for next-generation sequencing (NGS) analysis, different pretreatments and different combinations of these pretreatments were used. The pretreatments considered were an ultrasonic bath and a temperature of 80 °C, together and separately with different incubation time periods of 30, 60, and 90 min. The results obtained suggest a significant improvement in the efficiency and quality of DNA extraction with the three commercial extraction kits when used together with the ultrasonic bath and 80 °C for 60 min. Here, we were able to prove that physical pretreatments are a viable alternative to chemical lysis for DNA extraction from complex samples such as sludge.
Farmed salmonids show alterations in bone structure that result in skeletal deformities during formation, repair, and regeneration processes, with loss of mineralization at the level of the axial skeleton, mainly the head and spine, affecting their quality of life and even causing death. Despite improving factors, such as farming conditions, diets, and genetics, bone alterations appear more frequently in farmed fish than in wild fish. Thus, we used SEM-EDX, and TGA-DSC to study bone mineralization in farmed and wild rainbow trouts. As expected, we found significant differences in the nutritional parameters of farmed and wild fish (p < 0.05). Microstructural analyses indicated that farmed fish have a more robust mineral structure (p < 0.05), confirming the differences in mineralization and microstructure between both groups. However, the mechanisms regulating absorption and distribution in the organism and their effect on bone mineralization remain to be known. In our study, the combined use of techniques such as SEM-EDX and TGA-DSC allows a clearer assessment and detailed characterization beneficial to understanding the relationship between diet control and bone microstructure.
21 April 2022 in Microbiology Resource Announcements
Here, we announce the genome sequences of 408 strains of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) obtained from nasopharyngeal swabs in the Araucanía Region, Southern Chile. The genomes obtained are valuable to expand the availability of useful genomic data for future epidemiological studies of SARS-CoV-2 in Chile and worldwide.
Identifying the microbial taxa that structure assemblages in response to local disturbances along the Chilean Patagonian coast is critical for understanding community reorganization and ecological risk in this vulnerable marine environment. Here, we examined benthic microbial interactions across sixteen sites spanning 42–44°S in the Inner Sea of Chiloé and adjacent fjord systems. Using co-occurrence network analysis, we characterized spatial interaction patterns and identified keystone taxa whose relationships with environmental variables may serve as ecological indicators. The presence of keystone taxa supports the concept of microbial “seed banks,” which sustain community stability and ecosystem functionality under stress. Networks revealed structured communities dominated by niche-specialist taxa, consistent with ecological processes of niche differentiation and environmental filtering. The benthic environment was found to harbor niche-specialist taxa, including Flavobacteriaceae, Gemmataceae, Humimicrobiaceae, Clostridiaceae, and Sulfurovaceae, typically associated with organic enrichment and anthropogenic influence. These microbial families showed strong correlations with environmental gradients such as nitrate, phosphate, silicate, pH, salinity, and phaeopigments. Alpha-diversity analyses revealed greater microbial richness in FZ compared to the ISC, likely reflecting higher environmental heterogeneity and long-term adaptation to dynamic and fluctuating conditions. Beta-diversity analyses further supported the presence of distinct microbial assemblages between the two zones. Multivariate statistics and co-occurrence network analyses demonstrated that FZ communities display enhanced taxonomic interactions and greater network complexity, suggesting increased ecological stability. In contrast, microbial networks in the ISC exhibited reduced connectivity, potentially indicative of environmental stressors disrupting microbial spatial organization and network resilience. Moreover, the identification of indigenous bacterial isolates provided evidence for the presence of opportunistic host-associated pathogens, highlighting the utility of microbial biomarkers in detecting early ecological responses to human impacts. Microbial co-occurrence patterns in northern Patagonian sediments reveal shifts in community assembly and increased network complexity that may enhance resilience to perturbations. The detection of strains harboring opportunistic pathogen biomarkers highlights risks under intense anthropogenic pressures. Monitoring keystone taxa thus offers early warning of functional decline. Broader spatio-temporal assessments remain essential to clarify microbial dynamics and their implications for ecosystem health and antimicrobial resistance.