Microbiota composition of the dorsal patch of reproductive male Leptonycteris yerbabuenae.

Bacteria and other types of microbes interact with their hosts in several ways, including metabolic pathways, development, and complex behavioral processes such as mate recognition. During the mating season, adult males of the lesser long-nosed agave pollinator bat Leptonycteris yerbabuenae (Phyllostomidae: Glossophaginae) develop a structure called the dorsal patch, which is located in the interscapular region and may play a role in kin recognition and mate selection. Using high-throughput sequencing of the V4 region of the 16S rRNA gene, we identified a total of 2,847 microbial phylotypes in the dorsal patches of eleven specimens. Twenty-six phylotypes were shared among all the patches, accounting for 30 to 75% of their relative abundance. These shared bacteria are distributed among 13 families, 10 orders, 6 classes and 3 phyla. Two of these common bacterial components of the dorsal patch are Lactococcus and Streptococcus. Some of them-Helcococcus, Aggregatibacter, Enterococcus, and Corynebacteriaceae-include bacteria with pathogenic potential. Half of the shared phylotypes belong to Gallicola, Anaerococcus, Peptoniphilus, Proteus, Staphylococcus, Clostridium, and Peptostreptococcus and specialize in fatty acid production through fermentative processes. This work lays the basis for future symbiotic microbe studies focused on communication and reproduction strategies in wildlife

Metagenomic strategies identify diverse integron-integrase and antibiotic resistance genes in the Antarctic environment

The objective of this study is to identify and analyze integrons and antibiotic resistance genes (ARGs) in samples collected from diverse sites in terrestrial Antarctica. Integrons were studied using two independent methods. One involved the construction and analysis of intI gene amplicon libraries. In addition, we sequenced 17 metagenomes of microbial mats and soil by high-throughput sequencing and analyzed these data using the IntegronFinder program. As expected, the metagenomic analysis allowed for the identification of novel predicted intI integrases and gene cassettes (GCs), which mostly encode unknown functions. However, some intI genes are similar to sequences previously identified by amplicon library analysis in soil samples collected from non-Antarctic sites. ARGs were analyzed in the metagenomes using ABRIcate with CARD database and verified if these genes could be classified as GCs by IntegronFinder. We identified 53 ARGs in 15 metagenomes, but only four were classified as GCs, one in MTG12 metagenome (Continental Antarctica), encoding an aminoglycoside-modifying enzyme (AAC(6´)acetyltransferase) and the other three in CS1 metagenome (Maritime Antarctica). One of these genes encodes a class D β-lactamase (blaOXA-205) and the other two are located in the same contig. One is part of a gene encoding the first 76 amino acids of aminoglycoside adenyltransferase (aadA6), and the other is a qacG2 gene.Fil: Antelo, Verónica. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Giménez, Matías. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Azziz, Gastón. Universidad de la Republica. Facultad de Agricultura; UruguayFil: Valdespino Castillo, Patricia. Lawrence Berkeley National Laboratory; Estados UnidosFil: Falcón, Luisa I.. Universidad Nacional Autónoma de México; MéxicoFil: Ruberto, Lucas Adolfo Mauro. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: MacCormack, Walter P.. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Mazel, Didier. Institut Pasteur de Paris.; FranciaFil: Batista, Silvia. Instituto de Investigaciones Biológicas "Clemente Estable"; Urugua

Comparación de tres métodos moleculares para el análisis de procariontes ambientales en el mar del canal de Yucatán, México

In this study we describe a prokaryotic community from a seawater sample obtained in the Yucatan Channel, using for this purpose three molecular methods: 1) T-RFLPs. 2) Sequencing of amplicons (clone libraries). 3) Metagenome shotgun sequencing; the three are useful for the determination of microbial diversity. We also present a comparison of the scope and limits of each method. The comparison took into account three criteria: the number of taxonomic units detected, the taxonomic assignment accuracy and the cost of the study. The most abundant taxa were Candidatus Portiera OTU 3744 (equivalent to SAR86 clade) and Candidatus Pelagibacter. The results showed that the shotgun sequencing strategy is the most powerful in terms of detected taxonomic units, while the data obtained by T-RFLPs and clone library methods represent only a subsample of the shotgun fragment library. Regarding phylogenetic resolution (taxonomical determination), the more accurate approach is the sequencing of clone libraries. The costs of the three strategies vary considerably, but so does its scope. Therefore, it is important to consider that one, or another methodology, can only specifically answer some ecological and evolutionary questions.En este trabajo describimos la comunidad procarionte de una muestra de agua marina del canal de Yucatán. Para la determinación de la diversidad microbiana se usaron tres métodos moleculares: 1) T-RFLPs. 2) Secuenciación de amplicones (bibliotecas de clones). 3) Secuenciación shotgun de un metagenoma. Como un segundo objetivo, se presenta una comparación de los alcances y los límites de cada uno de estos tres métodos. Para esta comparación, se tomaron en cuenta tres criterios: el número de unidades taxonómicas detectadas, la precisión en la asignación taxonómica y el costo del estudio. Los taxa más abundantes fueron Candidatus Portiera OTU 3744 (equivalente al clado SAR86) y Candidatus Pelagibacter. Los resultados mostraron que la estrategia de secuenciación shotgun de ADN es la más poderosa en términos de unidades taxonómicas detectadas, mientras que los datos que se obtuvieron por T-RFLPs y con la biblioteca de clones, representan sólo una submuestra de la biblioteca de fragmentos generados mediante shotgun. En cuanto a resolución filogenética (determinación taxonómica), la aproximación más precisa fue la secuenciación de bibliotecas de clones. Los costos de las tres estrategias varían considerablemente, pero sus alcances también lo hacen. Por lo tanto, es importante tomar en consideración que algunas preguntas ecológicas y evolutivas sólo pueden ser contestadas específicamente por una u otra metodología

The Sorcerer II Global Ocean Sampling Expedition: Northwest Atlantic through Eastern Tropical Pacific

The world's oceans contain a complex mixture of micro-organisms that are for the most part, uncharacterized both genetically and biochemically. We report here a metagenomic study of the marine planktonic microbiota in which surface (mostly marine) water samples were analyzed as part of the Sorcerer II Global Ocean Sampling expedition. These samples, collected across a several-thousand km transect from the North Atlantic through the Panama Canal and ending in the South Pacific yielded an extensive dataset consisting of 7.7 million sequencing reads (6.3 billion bp). Though a few major microbial clades dominate the planktonic marine niche, the dataset contains great diversity with 85% of the assembled sequence and 57% of the unassembled data being unique at a 98% sequence identity cutoff. Using the metadata associated with each sample and sequencing library, we developed new comparative genomic and assembly methods. One comparative genomic method, termed “fragment recruitment,” addressed questions of genome structure, evolution, and taxonomic or phylogenetic diversity, as well as the biochemical diversity of genes and gene families. A second method, termed “extreme assembly,” made possible the assembly and reconstruction of large segments of abundant but clearly nonclonal organisms. Within all abundant populations analyzed, we found extensive intra-ribotype diversity in several forms: (1) extensive sequence variation within orthologous regions throughout a given genome; despite coverage of individual ribotypes approaching 500-fold, most individual sequencing reads are unique; (2) numerous changes in gene content some with direct adaptive implications; and (3) hypervariable genomic islands that are too variable to assemble. The intra-ribotype diversity is organized into genetically isolated populations that have overlapping but independent distributions, implying distinct environmental preference. We present novel methods for measuring the genomic similarity between metagenomic samples and show how they may be grouped into several community types. Specific functional adaptations can be identified both within individual ribotypes and across the entire community, including proteorhodopsin spectral tuning and the presence or absence of the phosphate-binding gene PstS

Time to Switch to Second-line Antiretroviral Therapy in Children With Human Immunodeficiency Virus in Europe and Thailand.

Background: Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand. Methods: Children aged <18 years initiating combination ART (≥2 nucleoside reverse transcriptase inhibitors [NRTIs] plus nonnucleoside reverse transcriptase inhibitor [NNRTI] or boosted protease inhibitor [PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of ≥1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks. Results: Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch. Conclusions: One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch

Metagenome of Acropora palmata coral rubble: Potential metabolic pathways and diversity in the reef ecosystem.

Over the past 30 years, the stony coral Acropora palmata has experienced an excessive loss of individuals showing few signs of recovery throughout the Mexican Caribbean, resulting in long stretches of coral rubble structures. When the coral dies, the skeleton begins to be colonized by algae, sponges, virus, bacteria and other microorganisms, forming a new community. Here we analyze, using a metagenomic approach, the diversity and biogeochemical cycles associated to coral rubble in La Bocana (Puerto Morelos, QRoo, Mexico). This study provides the first broad characterization of coral rubble associated communities and their role in biogeochemical cycling, suggesting a potential view of a world where coral reefs are no longer dominated by corals

Diversity of Diazotrophic Unicellular Cyanobacteria in the Tropical North Atlantic Ocean

We present data on the genetic diversity and phylogenetic affinities of N(2)-fixing unicellular cyanobacteria in the plankton of the tropical North Atlantic Ocean. Our dinitrogenase gene (nifH) sequences grouped together with a group of cyanobacteria from the subtropical North Pacific; another subtropical North Pacific group was only distantly related. Most of the 16S ribosomal DNA sequences from our tropical North Atlantic samples were closely allied with sequences from a symbiont of the diatom Climacodium frauenfeldianum. These findings suggest a complex pattern of evolutionary and ecological divergence among unicellular cyanobacteria within and between ocean basins

N(2) Fixation by Unicellular Bacterioplankton from the Atlantic and Pacific Oceans: Phylogeny and In Situ Rates

N(2)-fixing proteobacteria (α and γ) and unicellular cyanobacteria are common in both the tropical North Atlantic and Pacific oceans. In near-surface waters proteobacterial nifH transcripts were present during both night and day while unicellular cyanobacterial nifH transcripts were present during the nighttime only, suggesting separation of N(2) fixation and photosynthesis by unicellular cyanobacteria. Phylogenetic relationships among unicellular cyanobacteria from both oceans were determined after sequencing of a conserved region of 16S ribosomal DNA (rDNA) of cyanobacteria, and results showed that they clustered together, regardless of the ocean of origin. However, sequencing of nifH transcripts of unicellular cyanobacteria from both oceans showed that they clustered separately. This suggests that unicellular cyanobacteria from the tropical North Atlantic and subtropical North Pacific share a common ancestry (16S rDNA) and that potential unicellular N(2) fixers have diverged (nifH). N(2) fixation rates for unicellular bacterioplankton (including small cyanobacteria) from both oceans were determined in situ according to the acetylene reduction and (15)N(2) protocols. The results showed that rates of fixation by bacterioplankton can be almost as high as those of fixation by the colonial N(2)-fixing marine cyanobacteria Trichodesmium spp. in the tropical North Atlantic but that rates are much lower in the subtropical North Pacific