Bacterial communities and signatures in the stomach and intestine of juvenile P enaeus (litopenaeus) vannamei shrimp affected by acute hepatopancreatic necrosis disease.

Acute hepatopancreatic necrosis (AHPND) is a severe bacterial disease affecting farmed shrimp. Although various pathogenic bacteria associated with AHPND-affected shrimp have been described, little is known about the bacterial signatures in the stomachs and intestines when the disease occurs naturally. In this study, we characterized the microbiome of P. vannamei by high-throughput sequencing (HTS). Shrimp samples were collected from a commercial farm and divided into two groups: healthy and affected by AHPND, confirmed by PCR. Stomach and intestine samples were subjected to microbiome analysis targeting the V3-V4 region of the 16S rRNA gene. PERMANOVA analysis revealed a significant disparity in the bacterial diversity between the stomach and intestine microbiomes of these two health conditions. Our results suggest that the significant abundance of Vibrio brasiliensis and V. sinaloensis in the intestines of affected shrimp plays a role in AHPND infection. This imbalance could be mitigated by the presence of Pseudoalteromonas, Gilvimarinus, and other members of the phylum Pseudomonadota such as Cellvibrionaceae, Psychromonadaceae, and Halieaceae, which showed significant abundance in healthy intestines. This study highlights the significance of the microbial community in the differentiation of specific microbial signatures in different organs of P. vannamei. These findings offer a deeper understanding of the intricate dynamics within the shrimp microbiome under these conditions, enriching our view of AHPND progression and paving the way toward future identification of probiotics tailored for more efficient management of this disease.

Molecular identification and characterizations of rhizobacterial isolates collected from lentil rhizosphere of Indo-gangetic plains.

Plant growth promoting rhizobacteria (PGPR) are also known to colonize in the soil rhizosphere and prevent the development of other soil borne pathogens residing in the root surface. These microorganisms play a vital role in growth and development of the plant and also enhances the soil fertility by enriching the soil with different beneficial nutrients. This study was aimed at isolation of different rhizobacteria and their molecular characterization in search of efficient bacterial strains with multiple growth regulating activities. A total 36 bacteria were isolated from lentil root nodule as well as soil from different lentil growing fields with a view to screen/evaluate their plant growth promoting potential. Morphological characterization of isolated rhizobacterial candidates were done by observing the colonies on YEMA and nutrient agar media. Determination of CFU, Congo red test and gram staining tests were done to further screen them according to their morphology. All the isolates were then undergone molecular phylogenetic analysis using the partial sequences of the 16 S rDNA. Based upon the Gram staining test, all the isolates were negative in gram reaction except six Bacillus isolates, PSB2 and AB3. Results of Ribosomal Database Project (RDP) and Basic Local Alignment Search Tool for Nucleotide Sequences (BLASTn) from 16 S rDNA gene sequences showed that these isolates are genetically diverse. A total of 15 isolates of Rhizobium, 6 isolates of Bacillus, 3 isolates of Pseudomonas, 2 isolates of Phosphate Solubilizing Bacteria, 4 isolates of actinomycetes were identified by molecular sequencing of their 16 S rDNA region and comparing them with the other isolates enlisted in the database of NCBI for the similarity percentage, query coverage. The purpose of the present study was to select native rhizosphere bacteria from the lentil nodule and soil of Lentil field and to evaluate their plant growth promoting potential as an alternative of chemical fertilizer for sustainable, environment friendly agriculture and assessment of their phylogenetic characterization.

Multi-omics data elucidate parasite-host-microbiota interactions and resistance to Haemonchus contortus in sheep.

BACKGROUND: The integration of molecular data from hosts, parasites, and microbiota can enhance our understanding of the complex biological interactions underlying the resistance of hosts to parasites. Haemonchus contortus, the predominant sheep gastrointestinal parasite species in the tropics, causes significant production and economic losses, which are further compounded by the diminishing efficiency of chemical control owing to anthelmintic resistance. Knowledge of how the host responds to infection and how the parasite, in combination with microbiota, modulates host immunity can guide selection decisions to breed animals with improved parasite resistance. This understanding will help refine management practices and advance the development of new therapeutics for long-term helminth control. METHODS: Eggs per gram (EPG) of feces were obtained from Morada Nova sheep subjected to two artificial infections with H. contortus and used as a proxy to select animals with high resistance or susceptibility for transcriptome sequencing (RNA-seq) of the abomasum and 50 K single-nucleotide genotyping. Additionally, RNA-seq data for H. contortus were generated, and amplicon sequence variants (ASV) were obtained using polymerase chain reaction amplification and sequencing of bacterial and archaeal 16S ribosomal RNA genes from sheep feces and rumen content. RESULTS: The heritability estimate for EPG was 0.12. GAST, GNLY, IL13, MGRN1, FGF14, and RORC genes and transcripts were differentially expressed between resistant and susceptible animals. A genome-wide association study identified regions on chromosomes 2 and 11 that harbor candidate genes for resistance, immune response, body weight, and adaptation. Trans-expression quantitative trait loci were found between significant variants and differentially expressed transcripts. Functional co-expression modules based on sheep genes and ASVs correlated with resistance to H. contortus, showing enrichment in pathways of response to bacteria, immune and inflammatory responses, and hub features of the Christensenellaceae, Bacteroides, and Methanobrevibacter genera; Prevotellaceae family; and Verrucomicrobiota phylum. In H. contortus, some mitochondrial, collagen-, and cuticle-related genes were expressed only in parasites isolated from susceptible sheep. CONCLUSIONS: The present study identified chromosome regions, genes, transcripts, and pathways involved in the elaborate interactions between the sheep host, its gastrointestinal microbiota, and the H. contortus parasite. These findings will assist in the development of animal selection strategies for parasite resistance and interdisciplinary approaches to control H. contortus infection in sheep.

Bacteriomic Profiles of Rock-Dwelling Lichens from the Venezuelan Guiana Shield and the South African Highveld Plateau.

Lichens are not only fungal-algal symbiotic associations but also matrices for association with bacteria, and the bacterial diversity linked to lichens has been receiving more attention in studies. This study compares the diversity and possible metabolism of lichen-associated bacteria from saxicolous foliose and fruticose taxa Alectoria, Canoparmelia, Crocodia, Menegazzia, Usnea, and Xanthoparmelia from the Venezuelan Guiana Shield and the South African Highveld Plateau. We used DNA extractions from the lichen thalli to amplify the eukaryotic 18S rRNA gene (rDNA) and the V3-V4 region of the bacterial 16S rDNA, of which amplicons were then Sanger- and MiSeq-sequenced, respectively. The V3-V4 sequences of the associated bacteria were grouped into operational taxonomic units (OTUs) ascribed to twelve bacterial phyla previously found in the rock tripe Umbilicaria lichens. The bacterial OTUs emphasized the uniqueness of each region, while, at the species and higher ranks, the regional microbiomes were shown to be somewhat similar. Nevertheless, regional biomarker OTUs were screened to predict relevant metabolic pathways, which implicated different regional metabolic features.

Different microbial communities in paddy soils under organic and nonorganic farming.

Organic agriculture is a farming method that provides healthy food and is friendly to the environment, and it is developing rapidly worldwide. This study compared microbial communities in organic farming (Or) paddy fields to those in nonorganic farming (Nr) paddy fields based on 16S rDNA sequencing and analysis. The predominant microorganisms in both soils were Proteobacteria, Chloroflexi, Acidobacteria, Actinobacteria, and Nitrospirota. The alpha diversity of the paddy soil microbial communities was not different between the nonorganic and organic farming systems. The beta diversity of nonmetric multidimensional scaling (NMDS) revealed that the two groups were significantly separated. Distance-based redundancy analysis (db-RDA) suggested that soil pH and electrical conductivity (EC) had a positive relationship with the microbes in organic paddy soils. There were 23 amplicon sequence variants (ASVs) that showed differential abundance. Among them, g_B1-7BS (Proteobacteria), s_Sulfuricaulis limicola (Proteobacteria), g_GAL15 (p_GAL15), c_Thermodesulfovibrionia (Nitrospirota), two of f_Anaerolineaceae (Chloroflexi), and two of g_S085 (Chloroflexi) showed that they were more abundant in organic soils, whereas g_11-24 (Acidobacteriota), g__Subgroup_7 (Acidobacteriota), and g_Bacillus (Firmicutes) showed differential abundance in nonorganic paddy soils. Functional prediction of microbial communities in paddy soils showed that functions related to carbohydrate metabolism could be the major metabolic activities. Our work indicates that organic farming differs from nonorganic farming in terms of microbial composition in paddy soils and provides specific microbes that might be helpful for understanding soil fertility.

Minuca panema (Coelho, 1972): Resurrection of a Fiddler Crab Species from Brazil Closely Related to Minuca burgersi (Holthuis, 1967) (Crustacea, Decapoda, Brachyura, Ocypodidae).

We redescribe a species of fiddler crab, Minuca panema (Coelho, 1972), from the Atlantic coast of South America. It is closely related to M. mordax (Smith, 1870), and until now, the taxon has been considered to be synonymous with another closely related species Minuca burgersi (Holthuis, 1967). However, we found that two clades of M. burgersi sensu lato were restricted to the Caribbean Basin. This distribution differs from than that of M. panema, which occurs primarily along the eastern coast of South America, ranging from the island of Trinidad to Praia da Armação, Santa Catarina, Brazil. Based on our field studies, the geographical boundary between M. burgersi sensu stricto and M. panema is the Tobago Basin, north of Trinidad. Since the two species diverged only 3 to 4 million years ago, as dated from the phylogeny of the genus Minuca Bott 1954, there are few reliable morphological features that can be used to distinguish them clearly. In live crabs, there is a striking difference in coloration between the cherryred South American M. panema and the rusty-red Caribbean M. burgersi sensu lato. In males, the pattern of tubercles on the inner surface of the major cheliped varies between the two species. In females, the vulva is slightly larger in M. burgersi sensu stricto. Ocean tides and currents together with siltation owing to freshwater outflow from the Amazon and Orinoco rivers most likely have driven the divergence of these species. In the Caribbean, small tidal amplitudes have minimized long-distance gene flow in M. burgersi sensu stricto from isolated insular lagoons. In contrast, large tidal amplitudes and exposed habitats on riverbanks along the eastern Atlantic coast of South America have enabled long-distance dispersal in M. panema. DNA analysis reveals that haplotypes of cytochrome c oxidase subunit 1 are not shared between the species. Since natural selection and/or genetic drift have yet to produce extensive morphological divergences between M. panema and M. burgersi sensu stricto, we speculate that changes in the genes regulating mitochondrial DNA functions have led to speciation at the molecular level. According to the mitonuclear compatibility concept, we propose that mitochondrial DNA may be at the forefront of speciation events and that co-evolved mitonuclear interactions are responsible for some of the earliest genetic incompatibilities arising among isolated populations.

Spatial distribution of sediment bacterial communities from São Francisco River headwaters is influenced by human land-use activities and seasonal climate shifts.

Riverbed sediments are dynamic freshwater environments colonized by a great diversity of microorganisms which play important roles in supporting freshwater ecosystem by performing a vast array of metabolic functions. Recent evidence generated by HTS approaches has revealed that the structure of sediment microbial communities is influenced by natural seasonal variations in water such as temperature or streamflow as well by disturbances caused by local human activities. Here, a spatiotemporal analysis of sediment microbial distribution from São Francisco River headwaters section was conducted using Illumina 16S rRNA-V4 region amplicon sequencing in order to accomplish three major goals: (i) to investigate whether the diversity and composition of bacterial communities accessed in riverbed sediments vary in response to distinct land-use activities; (ii) to estimate whether the diversity patterns vary between the dry and wet seasons; and (iii) to evaluate whether the diversity of bacterial metabolic functions, predicted by PICRUSt2 approach, varies similarly to the estimated taxonomic diversity. Our findings revealed that bacterial communities in the sediment show differences in diversity and taxonomic composition according to the anthropic activities performed in the local environment. However, the patterns in which this taxonomic diversity is spatially structured show differences between the dry and wet seasons. On the other hand, the most changes in predicted bacterial metabolic functions were verified between sediment samples accessed in portions of the river located in protected and unprotected areas. Our findings contributed with new evidence about the impact of typical land-use practices conducted in countryside landscapes from developing countries on riverbed bacterial communities, both in their taxonomic and functional structure.

Identification of bacteria associated with canine otitis externa based on 16S rDNA high-throughput sequencing.

Bacteria are regarded as predisposing and perpetuating factors causing otitis externa (OE), whereas auricular anatomy is a predisposing factor. This study aims to investigate bacterial populations in the external auditory canals of healthy dogs and dogs with OE. Four categories of ear swabs included healthy erect-ear dogs, erect-ear dogs with OE, healthy pendulous-ear dogs and pendulous-ear dogs with OE. After bacterial DNA extraction, 16S rDNAs were amplified using specific primers within a V3/V4 region. Following DNA library construction, high-throughput sequencing was performed on MiSeq (Illumina). CLC Microbial Genomics Module was used to determine the rarefaction curve, bacterial classification, relative abundance, richness and diversity index. The results demonstrated that healthy dogs had higher bacterial richness and diversity than the dogs with OE. Comparable with culture-dependent methods described previously, this study revealed predominant Corynebacterium spp., Pseudomonas spp., Staphylococcus spp., and Proteus spp. in OE cases. Furthermore, high-throughput sequencing might disclose some potential emerging pathogens including Tissierella spp., Acinetobacter spp., and Achromobacter spp., which have not been reported in previous canine OE cases. Nevertheless, larger sample sizes are further required for an extensive evidence-based investigation.

Morphological and molecular identification of the brown dog tick in Mexico.

Ticks of the Rhipicephalus sanguineus complex are known as the brown dog ticks. This complex groups at least 12 species of ticks that are distributed worldwide. On the American continents, R. sanguineus sensu stricto (s.s.), is distributed in temperate areas, while Rhipicephalus sanguineus sensu lato (s.l.), also called "tropical lineage" is distributed in tropical regions. Previous analyses of brown dog ticks from Mexico have identified the so-called tropical lineage and the country generally has a climate more favorable for these ticks (> 20o C in average). In addition, some pathogens thought to be transmitted by this lineage (such as Ehrlichia canis, and Rickettsia rickettsii) are prevalent in Mexico. Herein we aim to contribute to the study of brown dog ticks by providing morphological identification and molecular analysis of mt 12S rDNA and 16S rDNA sequences from ticks collected from 12 states in Mexico. Our results indicate that the tropical lineage of R. sanguineus s.l., recently redescribed as R. linnaei is widely distributed in Mexico.

Diversity of Culturable Bacteria from Endemic Medicinal Plants of the Highlands of the Province of Parinacota, Chile.

Endemic medicinal plants that grow at altitudes in northern Chile have been traditionally used for therapeutic applications by Aymara doctors. Several studies have analyzed the biological properties of these plants for therapeutic purposes. The aim was to characterize at molecular and biochemical levels the bacteria that live in the rhizosphere and roots from endemic medicinal plants that grow between 3681-5104 m.a.s.l. in the province of Parinacota. Thirty-nine bacteria were isolated from nine medicinal plants under our laboratory conditions. These bacteria were characterized by Gram stain, hydrolase production, plant-growth promotion, anti-fungal and antibacterial activities, and 16S rDNA sequencing. A phylogenetic study revealed the presence of three major phyla, Actinomycetota (46.2%), Bacillota (43.6%), and Pseudomonadota (10.3%). The rhizobacteria strains associated with the Aymara medicinal plant exhibited several interesting biological activities, such as hydrolytic enzymes, plant-growth-promoting traits, and antibacterial and antifungal properties, indicating their potential for developing new bio-based products for agricultural or clinical applications. These results are promising and highlight the need to point toward the search for explanations of the bio-molecular basis of the therapeutic effects of medicinal plants.

Vachellia farnesiana Pods or a Polyphenolic Extract Derived from Them Exert Immunomodulatory, Metabolic, Renoprotective, and Prebiotic Effects in Mice Fed a High-Fat Diet.

Obesity causes systemic inflammation, hepatic and renal damage, as well as gut microbiota dysbiosis. Alternative vegetable sources rich in polyphenols are known to prevent or delay the progression of metabolic abnormalities during obesity. Vachellia farnesiana (VF) is a potent source of polyphenols with antioxidant and anti-inflammatory activities with potential anti-obesity effects. We performed an in vivo preventive or an interventional experimental study in mice and in vitro experiments with different cell types. In the preventive study, male C57BL/6 mice were fed with a Control diet, a high-fat diet, or a high-fat diet containing either 0.1% methyl gallate, 10% powdered VFP, or 0.5%, 1%, or 2% of a polyphenolic extract (PE) derived from VFP (Vachellia farnesiana pods) for 14 weeks. In the intervention study, two groups of mice were fed for 14 weeks with a high-fat diet and then one switched to a high-fat diet with 10% powdered VFP for ten additional weeks. In the in vitro studies, we evaluated the effect of a VFPE (Vachellia farnesiana polyphenolic extract) on glucose-stimulated insulin secretion in INS-1E cells or of naringenin or methyl gallate on mitochondrial activity in primary hepatocytes and C2C12 myotubes. VFP or a VFPE increased whole-body energy expenditure and mitochondrial activity in skeletal muscle; prevented insulin resistance, hepatic steatosis, and kidney damage; exerted immunomodulatory effects; and reshaped fecal gut microbiota composition in mice fed a high-fat diet. VFPE decreased insulin secretion in INS-1E cells, and its isolated compounds naringenin and methyl gallate increased mitochondrial activity in primary hepatocytes and C2C12 myotubes. In conclusion VFP or a VFPE prevented systemic inflammation, insulin resistance, and hepatic and renal damage in mice fed a high-fat diet associated with increased energy expenditure, improved mitochondrial function, and reduction in insulin secretion.

Genetic variation and cryptic lineage among the sergestid shrimp Acetes americanus (Decapoda).

The taxonomic status of the sergestid shrimp, Acetes americanus, has been questioned for several decades. No specific study has been performed thus far to resolve the incongruences. This species has a wide geographical range in the western Atlantic and is represented by two formally accepted subspecies: Acetes americanus carolinae, distributed in North America, and Acetes americanus americanus, present in South America. However, there are regions where the coexistence of both subspecies has been reported, such as Central America. This study aimed to genetically compare specimens of A. a. americanus collected in South America with A. a. carolinae sampled in North America to check for possible differences and the existence of more than one subspecies of A. americanus on the Brazilian coast. Based on the sequences of two informative markers, the cytochrome oxidase I region (COI) and 16S rRNA, phylogenetic reconstruction demonstrated well-defined clades with high support values, reinforcing the idea that A. a. americanus is genetically different from A. a. carolinae. Our hypothesis was corroborated as the specimens collected in Brazil were divided into two distinct lineages: the first composed of A. a. americanus sensu stricto (Brazil 1) and the second by Acetes americanus (Brazil 2). The three groups evidenced in the haplotype network were the same as those observed in the phylogenetic tree. The morphometric character (height/length of the thelycum) was effective in distinguishing A. a. Brazil 1 from A. a. carolinae. However, more detailed and conclusive studies comprising other characteristics to propose and describe a possible new entity are necessary. To the best of our knowledge, for the first time, the results of this study provide some insights into the taxonomic status of the sergestid shrimp A. americanus in the western Atlantic.

Metaprofiling of the Bacterial Community in Colonized Compost Extracts by Agaricus subrufescens.

It is well-known that bacteria and fungi play important roles in the relationships between mycelium growth and the formation of fruiting bodies. The sun mushroom, Agaricus subrufescens, was discovered in Brazil ca. 1960 and it has become known worldwide due to its medicinal and nutritional properties. This work evaluated the bacterial community present in mushroom-colonized compost extract (MCCE) prepared from cultivation of A. subrufescens, its dynamics with two different soaking times and the influence of the application of those extracts on the casing layer of a new compost block for A. subrufescens cultivation. MCCEs were prepared through initial submersion of the colonized compost for 1 h or 24 h in water followed by application on casing under semi-controlled conditions. Full-length 16S rRNA genes of 1 h and 24 h soaked MCCE were amplified and sequenced using nanopore technology. Proteobacteria, followed by Firmicutes and Planctomycetes, were found to be the most abundant phyla in both the 1 h and 24 h soaked MCCE. A total of 275 different bacterial species were classified from 1 h soaked MCCE samples and 166 species from 24 h soaked MCCE, indicating a decrease in the bacterial diversity with longer soaking time during the preparation of MCCE. The application of 24 h soaked MCCE provided increases of 25% in biological efficiency, 16% in precociousness, 53% in the number of mushrooms and 40% in mushroom weight compared to control. Further investigation is required to determine strategies to enhance the yield and quality of the agronomic traits in commercial mushroom cultivation.

A reappraisal of microbiome dysbiosis during experimental periodontitis.

Periodontitis is a chronic inflammatory disease associated with the presence of dysbiotic microbial communities. Several studies interrogating periodontitis pathogenesis have utilized the murine ligature-induced periodontitis (LIP) model and have further examined the ligature-associated microbiome relying on 16S rRNA-based sequencing techniques. However, it is often very challenging to compare microbial profiles across studies due to important differences in bioinformatic processing and databases used for taxonomic assignment. Thus, our study aim was to reanalyze microbiome sequencing datasets from studies utilizing the LIP model through a standardized bioinformatic analysis pipeline, generating a comprehensive overview of microbial dysbiosis during experimental periodontitis.We conducted a reanalysis of 16S rDNA gene sequencing datasets from nine published studies utilizing the LIP model. Reads were grouped according to the hypervariable region of the 16S rDNA gene amplified (V1-V3 and V4), preprocessed, binned into operational taxonomic units and classified utilizing relevant databases. Alpha- and beta-diversity analyses were conducted, along with relative abundance profiling of microbial communities. Our findings revealed similar microbial richness and diversity across studies and determined shifts in microbial community structure determined by periodontitis induction and study of origin. Clear variations in the relative abundance of bacterial taxa were observed starting on day 5 after ligation and onward, consistent with a distinct microbial composition during health and experimental periodontitis. We also uncovered differentially represented bacterial taxa across studies, dominating periodontal health and LIP-associated communities. Collectively, this reanalysis provides a unified overview of microbial dysbiosis during the LIP model, providing new insights that aim to inform further studies dedicated to unraveling oral host-microbial interactions.

An outlook on fluorescent in situ hybridization coupled to flow cytometry as a versatile technique to evaluate the effects of foods and dietary interventions on gut microbiota.

The increasing interest in the effects of the gut microbiota on host health has stimulated the investigation of the composition of this microbial community and the factors affecting these microorganisms. This review discusses the recent advances and progress applications in the use of the fluorescent in situ hybridization (FISH) coupled to flow cytometry (FC) technique (FISH-FC) in studies evaluating the gut microbiota published in the last 10 years, with particular emphasis on the effects of foods and dietary interventions. These studies have shown that FISH-FC technique is capable of detecting and quantifying several groups of bacteria found as part of the gut microbiota. FISH-FC can be considered an effective, versatile, and rapid technique to evaluate alterations in gut microbiota composition caused by different foods as assessed in studies in vitro, in vivo, and in clinical trials. Some specific probes have been most used to represent the general gut microbiota, such as those specific to Lactobacillus spp./Enterococcus spp., Bacteroidaceae/Prevotellaceae, Clostridium histolyticum, and Bifidobacterium spp. FISH-FC technique could have an important opportunity for application in studies with next-generation probiotics belonging to the gut microbiota. Optimizations of FISH-FC protocols could allow more discoveries about the gut microbiota, including the development of new probes targeting microorganisms still not explored, the analysis of individual portions of the intestine, and the proposition of novel quantitative approaches.

Bacterial diversity analysis of freshwater sources for human use in rural areas of the tropical Andean region of Colombia / Análisis de la diversidad bacteriana en aguas superficiales usadas para consumo humano en zonas rurales de la región andina colombiana

Abstract Potable water supply and sanitization in rural areas in developing countries are still inadequate. The main risk associated with unsafe drinking water is the infection with pathogenic microorganisms. Objective: In this study, we investigate the bacterial diversity and the potentially pathogenic bacteria in water samples from diffe rent points of distribution in three rural villages from the Andean region of Colombia. Methods: Illumina libraries for water samples were prepared and sequenced using 300 bp paired-end MiSeq protocol, the bioinformatic analyses were performed with Mothur pipeline and the phyloseq package in Rstudio. Results: The mi crobial community composition showed statistically significant differences according to the village and the sample origin. Alpha, Beta, and Gammaproteobacteria were the dominant class detected in all water samples. The most relevant pathogenic genera detected in the surface were Legionella, Mycobacterium, Yersinia, Burkholderia, and Rickettsia. In the tap water samples, potential pathogens like Streptococcus, Staphylococcus, Corynebacterium, Nocardia, and Escherichia/Shige lla were detected.

Resumen El suministro y potabilización del agua de consumo humano en las zonas rurales de los países en vías de desarrollo sigue siendo limitado. El principal riesgo asociado con el uso de agua no potable es la infección con microorganismos patógenos. Objetivo: En este estudio se investigó la diversidad bacteriana y la presencia de bacterias potencialmente patógenas en muestras de agua de diferentes puntos de distribución en tres asentamientos rurales de la región andina de Colombia. Métodos: Se prepararon y secuenciaron bibliotecas de amplicones (rDNA 16S) para muestras de agua utilizando la plataforma Illumina MiSeq con lecturas pareadas de 300 bases. Los análisis bioinformáticos se realizaron con el programa Mothur y el paquete estadístico Phyloseq en Rstudio. Resultados: La composición de la comunidad microbiana mostró diferencias estadísticamente significativas según el sitio y el origen de la muestra. Alfa, Beta y Gammaproteobac terias fueron las clase dominantes detectadas en todas las muestras de agua. Los géneros patógenos más relevantes detectados fueron Legionella, Mycobacterium, Yersinia, Burkholderia y Rickettsia. En las muestras de agua del grifo se detectaron patógenos potenciales como Streptococcus, Staphylococcus, Corynebacterium, Nocardia y Escherichia /Shigella.

Anaerobic biofilm enriched with an ammonia tolerant methanogenic consortium to improve wastewater treatment in the fishing industry.

The digestion efficiency of liquid industrial wastes increases when using bioreactors colonized by microbial biofilms. High concentrations of proteins derived from the fish processing industry lead to the production of ammonia, which inhibits methane production. Two bioreactors were constructed to compare methanogenic activity: one enriched with mMPA (methylaminotrofic methane production archaea) consortia (control bioreactor), and the second with NH3 tolerant consortia (treatment bioreactor). Ammonia tolerant activity was assessed by applying an ammonia shock (755 mg NH3/L). Methane production, consumption of total organic carbon (TOC) and the taxonomic composition of bacteria and archaea was evaluated using 16S rDNA in the acclimatization, ammonia shock, and recovery phases.The ammonia shock significantly affected both methane production and the consumption of TOC in the control reactor (p < 0.05) and taxonomical composition of the microbial consortia (OTU). These values remained constant in the treatment reactor. The analysis of biofilm composition showed a predominance of Methanosarcinaceae (Methanomethylovorans sp., and probably two different species of Methanosarcina sp.) in bioreactors. These results demonstrate that using acclimated biofilms enriched with ammonia tolerant methanogens control the inhibitory effect of ammonia on methanogenesis.

Naso-oropharyngeal microbiome from breast cancer patients diagnosed with COVID-19.

Due to immunosuppressive cancer therapies, cancer patients diagnosed with COVID-19 have a higher chance of developing severe symptoms and present a higher mortality rate in comparison to the general population. Here we show a comparative analysis of the microbiome from naso-oropharyngeal samples of breast cancer patients with respect to SARS-CoV-2 status and identified bacteria associated with symptom severity. Total DNA of naso-oropharyngeal swabs from 74 women with or without breast cancer, positive or negative for SARS-CoV-2 were PCR-amplified for 16S-rDNA V3 and V4 regions and submitted to massive parallel sequencing. Sequencing data were analyzed with QIIME2 and taxonomic identification was performed using the q2-feature-classifier QIIME2 plugin, the Greengenes Database, and amplicon sequence variants (ASV) analysis. A total of 486 different bacteria were identified. No difference was found in taxa diversity between sample groups. Cluster analysis did not group the samples concerning SARS-CoV-2 status, breast cancer diagnosis, or symptom severity. Three taxa (Pseudomonas, Moraxella, and Klebsiella,) showed to be overrepresented in women with breast cancer and positive for SARS-CoV-2 when compared to the other women groups, and five bacterial groups were associated with COVID-19 severity among breast cancer patients: Staphylococcus, Staphylococcus epidermidis, Scardovia, Parasegitibacter luogiensis, and Thermomonas. The presence of Staphylococcus in COVID-19 breast cancer patients may possibly be a consequence of nosocomial infection.

Caracterização da microbiota de queijos artesanais provenientes da Serra da Canastra - MG e da cultura iniciadora natural utilizada em sua produção / Microbiota characterization of artisanal cheeses from Serra da Canastra - MG and their endogenous starter culture

O queijo Canastra possui grande importância na cultura e economia local, é parte do Patrimônio Imaterial do Brasil (IPHAN, 2014) e recebeu o selo de produto com designação de origem em 2012 (INPI, 2016). Sua produção utiliza leite, sal, coalho e uma cultura iniciadora natural, chamada popularmente de pingo. Esse estudo visou a caracterização da microbiota presente no queijo maturado da Serra da Canastra e no pingo utilizado em sua produção utilizando técnicas avançadas de sequenciamento em larga escala para identificação das bactérias e fungos ali presentes. Nossos dados da microbiota bacteriana foram comparados com dados da microbiota de outros queijos brasileiros e do mundo disponíveis na literatura. As principais bactérias encontradas em amostras de pingo pertencem aos gêneros Lactococcus (45.6%), Streptococcus (30.3%), Staphylococcus (5.1%), e em amostras de queijo aos gêneros Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8%), Staphylococcus (13.6%), Leuconostoc (6.3%) e Weissella (6%). Os principais gêneros de fungos encontrados nos queijos foram Debaryomycesa (78.6%), Trichosporona (7.8%). Nosso estudo foi capaz de separar a microbiota dos queijos produzidos na Serra da Canastra de outros queijos na Europa e América do Norte, sendo o pH um possível fator de segregação. Também foi observada uma diferença entre a microbiota do queijo Canastra com outros queijos Brasileiros. Além disso, visualizamos que a distância geográfica entre produtores e a sazonalidade possuem um efeito sobre a microbiota dos pingos e queijos. A partir da análise de todos os microrganismos encontrados na microbiota bacteriana, foram detectados táxons que discriminam produtores por suas aplicações de boas práticas de fabricação e por sua infraestrutura. Observamos proporções menores de um táxon de Kocuria Kristinae nos pingos e um de Streptococcus nos queijos e proporções maiores de um táxon de Staphylococcus nos queijos. Também pudemos observar uma diminuição nas proporções de táxons de Debaryomycesa e aumento na proporção de táxons de Trichosporona na composição fúngica dos queijos, possivelmente devido a transição sazonal do período seco para o chuvoso. Usando técnicas moleculares de sequenciamento em larga escala, demonstramos que há uma diferença na microbiota presente em diferentes áreas da Serra da Canastra, um possível efeito da sazonalidade na composição fúngica e bacteriana. E evidenciamos que táxons de Streptococcus, Staphylococcus e Kocuria estão correlacionados às boas práticas de produção e elucidamos a conexão existente entre a microbiota do pingo e a do queijo. Estes resultados podem influenciar o desenvolvimento de métodos de rastreamento de sub-regiões específicas da Canastra e auxiliar os produtores na produção de queijos de boa qualidade, mantendo as características específicas de sua região

The Canastra cheese has great importance for the local culture and economy, being part of the Intangible Heritage of Brazil (IPHAN, 2014). It has received the protected designation of origin certification in 2012 (INPI, 2016). It's made using milk, salt, rennet and a endogenous starter culture, popularly called as "pingo". This study aimed to characterize the microbiota present in the Serra da Canastra's cheese and the pingo used in its production. In order to conduct this research we used next generation sequencing to identify the bacteria and fungi present there. Our bacterial microbiota dataset was compared with microbiota datasets from other Brazilian and world cheeses available in the literature. The main bacteria found were Lactococcus (45.6%), Streptococcus (30.3%) and Staphylococcus (5.1%) in the endogenous starter samples and Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8 %), Staphylococcus (13.6%), Leuconostoc (6.3%) and Weissella (6%) in cheese samples. The main fungi found in the cheeses were Debaryomycesa (78.6%) and Trichosporona (7.8%). We were able to separate the microbiota from Serra da Canastra cheeses and other cheeses in Europe and North America, being the pH a possible segregation factor. Furthermore, a difference was also observed between the microbiota of Canastra and other Brazilian cheeses. In addition, we observed that the geographical distance between producers and the seasonality could be affecting the pingos and cheeses microbiota. We found bacterial taxa that could discriminate producers by their good manufacturing practices and their local infrastructure. Low levels of good manufacturing practices (GMPs) were assigned to bigger proportions of a Kocuria Kristinae taxon in the pingos and a Staphylococcus taxon in the cheeses. Also, higher levels of GMPs were assigned to smaller proportions of Streptococcus taxons in the cheeses. Furthermore We could observe a decrease of Debaryomycesa and an increase of Trichosporona proportions in the fungal composition of cheeses. This could be due to a climate transition: from the dry season to the rainy season. Using large-scale sampling coupled with molecular sequencing techniques, we observe a connection between pingo and cheeses microbiota. We show that the microbiota of different areas in Serra da Canastra is different, also, there is a possible effect of seasonality on fungal and bacterial composition. Furthermore, we could see that Streptococcus, Staphylococcus and Kocuria taxons are correlated with good practices. These results may influence the development of tracking methods for specific Canastra subregions and assist producers to manufacture good quality cheeses while maintaining the specific characteristics of their region

Metagenomic analysis of MWWTP effluent treated via solar photo-Fenton at neutral pH: Effects upon microbial community, priority pathogens, and antibiotic resistance genes.

The effectiveness of advanced technologies on eliminating antibiotic resistant bacteria (ARB) and resistance genes (ARGs) from wastewaters have been recently investigated. Solar photo-Fenton has been proven effective in combating ARB and ARGs from Municipal Wastewater Treatment Plant effluent (MWWTPE). However, most of these studies have relied solely on cultivable methods to assess ARB removal. This is the first study to investigate the effect of solar photo-Fenton upon ARB and ARGs in MWWTPE by high throughput metagenomic analysis (16S rDNA sequencing and Whole Genome Sequencing). Treatment efficiency upon priority pathogens and resistome profile were also investigated. Solar photo-Fenton (30 mg L-1 of Fe2+ intermittent additions and 50 mg L-1 of H2O2) reached 76-86% removal of main phyla present in MWWTPE. An increase in Proteobacteria abundance was observed after solar photo-Fenton and controls in which H2O2 was present as an oxidant (Fenton, H2O2 only, solar/H2O2). Hence, tolerance mechanisms presented by this group should be further assessed. Solar photo-Fenton achieved complete removal of high priority Staphylococcus and Enterococcus, as well as Klebsiella pneumoniae and Pseudomonas aeruginosa. Substantial reduction of intrinsically multi-drug resistant bacteria was detected. Solar photo-Fenton removed nearly 60% of ARGs associated with sulfonamides, macrolides, and tetracyclines, and complete removal of ARGs related to ß-lactams and fluoroquinolones. These results indicate the potential of using solar-enhanced photo-Fenton to limit the spread of antimicrobial resistance, especially in developing tropical countries.