Revista Argentina de microbiologia最新文献

Understanding the gut mycobiota composition and its impact on health requires reliable methods for fungal community assessment. This study explores the influence of DNA extraction methods in GM analysis. Three protocols were evaluated: Qiagen DNeasy blood and tissue kit with mechanical glass bead lysis (DNgb), Thermofisher MagMax Microbiome ultra-nucleic isolation kit automated method (MM), and MM combined with glass beads lysis (MMgb). Fecal samples from healthy volunteers were collected, DNA extracted and ITS2 amplicon library preparation and sequencing performed. Results showed that DNA yields did not significantly differ among methods and the addition of glass bead beating favored the recovery of DNA more appropriate for fungal analysis. Beta diversity revealed distinct clusters, with MMgb showing the most pronounced variation in mycobiota composition, exposing particularly the low abundance taxa. LEfSe analysis identified significant differences in the abundance of fungal species among the extraction methods. Samples extracted with bead beating were enriched in filamentous species, while those without this step showed higher relative abundance of yeast fungi. This study underscores the importance of selecting appropriate DNA extraction methods for accurate characterization of the gut mycobiota, emphasizing the need for standardized methodologies to ensure reproducibility and reliability in microbial data acquisition.

Bovine viral diarrhea virus (BVDV) causes significant economic losses in the international livestock industry and in Argentina, where it circulates at high prevalence. Under high prevalence conditions, BVDV infections are controlled through vaccination once persistently infected animals are identified and segregated. This study evaluated the feasibility of controlling BVDV circulation under field conditions by combining diagnosis, management measures, and vaccination in 2 dairy farms in the province of Santa Fe. Commercial ELISAs were used for the detection of the NS3 (P80) protein or antibodies against this protein as well as an RT-nested PCR for the detection of the viral genome, and viral seroneutralization to assess vaccine efficacy. The average seroprevalence of the farms was 58.4%, with a persistently infected animal rate of 2.4%. After segregating the persistently infected animals and vaccinating them with a commercial combined vaccine containing inactivated BVDV, abortion rates significantly decreased (p<0.05) in farm 1 (from 20.5 to 11.6%) and in farm 2 (from 34 to 23.4%) during the second year of the control strategy. Conception rates increased from 29 to 33% in farm 1 during the first year, while in farm 2, the increase was 7 points during the second year. This methodology achieved conditions in which BVDV ceased to circulate, constituting the first controlled report on BVDV management in dairy farms using tools available to producers in Argentina.

DNA extraction is crucial for conducting procedures, such as whole-genome sequencing, which demand methods that are reproducible and cost-effective. Lysing Staphylococcus aureus cells is particularly challenging due to their peptidoglycan layer that is resistant to common treatments. Traditional methods involve costly enzymatic lysis using lysostaphin. Here, we developed a novel approach for lysis utilizing liquid nitrogen and mechanical disruption in a mortar. DNA from S. aureus USA300 and related clinical isolates were purified using phenol-chloroform extraction followed by precipitation. The integrity and purity of DNA were confirmed, obtaining suitable concentration and purity for various molecular biology techniques. The quality of the employed DNA was validated by amplifying fragments of different genes using PCR. This method circumvents lysostaphin, yielding DNA that is suitable for use in other techniques.

Acute respiratory infection (ARI) is one of the principal causes of morbidity worldwide, with respiratory viruses being common etiological agents. Among them, endemic human coronaviruses (hCoVs) including CoV-229E, CoV-OC43, CoV-NL63, and CoV-HKU1 can cause mild ARI but are usually not evaluated in the clinical setting. The aim of this work was to determine the prevalence of all respiratory pathogens, with the focus placed on endemic hCoVs in the pre-pandemic period. Circulating species, clinical associations and coinfections with other respiratory pathogens were evaluated in 510 immunocompetent patients (children and adults) with ARI using the FilmArray® Respiratory Panel (BioFire/bioMérieux). A total of 399 children (252 outpatients and 147 hospitalized) and 111 adult outpatients were enrolled in the pre-pandemic period (2008-2010 and 2016). Endemic hCoVs were the third and fifth more frequently detected viruses among adults and outpatient children, respectively, with an overall frequency close to 10%. The most prevalent species were CoV-OC43 (42.8%) and CoV-HKU1 (40.5%), followed by CoV-NL63 (19.0%) and CoV-229E (4.8%). Tachypnea, wheezing and chest indrawing were more frequent in hospitalized children compared to outpatients. All adult patients presented with symptoms of a common cold. Endemic hCoVs were detected year-round, primarily between June and November. Our results highlight their clinical relevance, and the need to include endemic hCoVs in routine screening. In the post-pandemic period, further long-term surveillance is needed for understanding the epidemiology of endemic hCoVs and their evolution, as a tool to anticipate the possible emergence of new species.