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Recent studies have shown how intestinal parasites can modulate gut microbiota. This observation is not surprising since the human intestinal lumen, like any other niche, is a battlefield of microbial competition, and Eukaryotes can affect bacterial populations. Intestinal pathogenic protist has been associated with reshaping the microbial community structure; however, the interactions between the colonic bacterial communities and parasites like Blastocystis spp., Entamoeba coli, and Endolimax nana have been poorly studied. In this work, we studied the distal intestinal bacterial microbiota of 49 children attending 7 public daycare centers in Medellin, Colombia, and compared the bacterial microbiota structure in the presence or absence of the protists Blastocystis spp., E. coli, and E. nana. Parasite colonization was associated with an increase in bacterial richness. Moreover, Blastocystis spp. presented a positive relationship with Prevotella, since this bacterium was selectively enriched in children carrying it. Remarkably, the E. coli colonized children showed a microbial profile that was closer to uninfected controls, although some bacterial taxa displayed to be enriched. This is the case for Akkermansia, which showed to be favored in E. coli colonized individuals, while notably reduced in the Blastocystis spp. parasitized group.

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb), leading to pulmonary and extrapulmonary TB, whereby Mtb is disseminated to many other organs and tissues. Dissemination occurs early during the disease, and bacteria can be found first in the lymph nodes adjacent to the lungs and then later in the extrapulmonary organs, including the spleen. The early global gene expression response of human tissue macrophages and intracellular clinical isolates of Mtb has been poorly studied. Using dual RNA-seq, we have explored the mRNA profiles of two closely related clinical strains of the Latin American and Mediterranean (LAM) family of Mtb in infected human splenic macrophages (hSMs). This work shows that these pathogens mediate a distinct host response despite their genetic similarity. Using a genome-scale host–pathogen metabolic reconstruction to analyze the data further, we highlight that the infecting Mtb strain also determines the metabolic response of both the host and pathogen. Thus, macrophage ontogeny and the genetic-derived program of Mtb direct the host–pathogen interaction.

Tuberculosis (TB) is caused by

Spiders (Araneae) are the most abundant terrestrial predators and megadiverse on earth. In recent years, the mitochondrial genome of a great diversity of species has been sequenced, mainly for ecological and commercial purposes. These studies have uncovered the existence of a variety of mitochondrial genome rearrangements. However, there is poor genetic information in several taxonomic families of spiders. We have sequenced the complete genome of Phoneutria depilata (Ctenidae) and, based on this, extract the mitogenomes of other ctenid species from published transcriptomes to perform a comparative study among spider species to determine the relationship between the level of mitochondrial rearrangements and its possible relationship with molecular variability in spiders. Complete mitochondrial genomes of eighteen spiders (including eight Ctenidae species) were obtained by two different methodologies (sequencing and transcriptome extraction). Fifty-eight spider mitochondrial genomes were downloaded from the NCBI database for gene order analysis. After verifying the annotation of each mitochondrial gene, a phylogenetic and a gene order analysis from 76 spider mitochondrial genomes were carried out. Our results show a high rate of annotation error in the published spider mitochondrial genomes, which could lead to errors in phylogenetic inference. Moreover, to provide new mitochondrial genomes in spiders by two different methodologies to obtain them, our analysis identifies six different mitochondrial architectures among all spiders. Translocation or tandem duplication random loss (TDRL) events in tRNA genes were identified to explain the evolution of the spider mitochondrial genome. In addition, our findings provide new insights into spider mitochondrial evolution.

Spiders are the most abundant land predators and megadiverse on earth. In recent years, the mitochondrial genome has been sequenced, mainly for ecological and commercial purposes, reporting some level of rearrangements in this genome. However, there is poor genetic information in several taxonomic families of spiders. The aim of this study was to obtain the sequence of the complete genome of Phoneutria boliviensis and, based on this, extract the mitogenomes of other species of the family Ctenidae from published transcriptomes to perform a comparative study among spider species to determine the relationship between the level of mitochondrial rearrangement and its possible relationship with molecular variability in spiders. Complete mitochondrial genomes of eighteen spiders (including nine Ctenidae species) were obtained by two different methodologies (sequencing and transcriptome extraction). Fifty-eight spider mitochondrial genomes were downloaded from the NCBI database for gene order analysis. After verifying the annotation of each mitochondrial gene, a phylogeny and gene order, analysis from 76 spider mitochondrial genomes was obtained. Our results show a high rate of annotation error in the mitochondrial genomes of spiders published in databases, which could lead to false phylogenetic relationships. Moreover, to provide new mitochondrial genomes in spiders by two different methodologies to obtain them, our analysis identifies six different mitochondrial architectures among all spiders. Translocation or tandem duplication random loss (TDRL) events in tRNA genes were identified to explain the evolution of the spider mitochondrial genome. In addition, our findings provide new insights into spider mitochondrial evolution.

Cryptosporidium parasites are ubiquitous and can infect a broad range of vertebrates and are considered the most frequent protozoa associated with waterborne parasitic outbreaks. The intestine is the target of three of the species most frequently found in humans: C. hominis, C. parvum, and. C. meleagridis. Despite the recent advance in genome sequencing projects for this apicomplexan, a broad genomic comparison including the three species most prevalent in humans have not been published so far. In this work, we downloaded raw NGS data, assembled it under normalized conditions, and compared 23 publicly available genomes of C. hominis, C. parvum, and C. meleagridis. Although few genomes showed highly fragmented assemblies, most of them had less than 500 scaffolds and mean coverage that ranged between 35X and 511X. Synonymous single nucleotide variants were the most common in C. hominis and C. meleagridis, while in C. parvum, they accounted for around 50% of the SNV observed. Furthermore, deleterious nucleotide substitutions common to all three species were more common in genes associated with DNA repair, recombination, and chromosome-associated proteins. Indel events were observed in the 23 studied isolates that spanned up to 500 bases. The highest number of deletions was observed in C. meleagridis, followed by C. hominis, with more than 60 species-specific deletions found in some isolates of these two species. Although several genes with indel events have been partially annotated, most of them remain to encode uncharacterized proteins.

Abundance and diversity of microbial communities in biosolids are variable and poorly studied in the tropics, and it is known that rainfall is one of the events that could affect the phylogenetic and functional microbial structure. In the present study, using NGS technics, we studied the microbial diversity as well as the methanogenesis pathway in one of the largest WWTP in Colombia. Besides, we sampled and analyzed biosolids from rainy season and dry season. Phylogenetic classification showed a predominance of bacteria in both samples and difference in the dominant groups depending on the rainfall season. Whereas Pseudomonas was the dominant bacteria in the dry season, Coprothermobacter was in the rainy season. Archaea abundance was higher in the rainy season (11.5%) doubling dry season proportion. The bioreactor biogas production and total solids content showed similar results between rainy and dry season at the sampling dates. The most abundant Archaea related with methanogenesis was Methanosaeta, which is a methanogenic microorganism that exclusively uses acetate to produce methane. Moreover, annotation of the methanogenic pathway in the metagenome showed abundance in genes encoding Acetyl-CoA synthetases (ACSS), an enzyme that catalyzes acetate activation. Our results suggest that the microbial diversity was stable among the two time points tested, rainy season and dry season; and, although there were changes in the microbial abundance of dominant bacterial species, anaerobic digester performance is not affected.

The public-key operation in multivariate encryption and signature schemes evaluates \begin{document}$ m $\end{document} quadratic polynomials in \begin{document}$ n $\end{document} variables. In this paper we analyze how fast this simple operation can be made. We optimize it for different finite fields on modern architectures. We provide an objective and inherent efficiency measure of our implementations, by comparing their performance with the peak performance of the CPU. In order to provide a fair comparison for different parameter sets, we also analyze the expected security based on the algebraic attack taking into consideration the hybrid approach. We compare the attack's efficiency for different finite fields and establish trends. We detail the role that the field equations play in the attack. We then provide a broad picture of efficiency of MQ-public-key operation against security.

The heterogeneity of the clinical outcome of Mycobacterium tuberculosis (Mtb) infection may be due in part to different strategies used by circulating strains to cause disease. This heterogeneity is one of the main limitations to eradicate tuberculosis disease. In this study, we have compared the transcriptional response of two closely related Colombian clinical isolates (UT127 and UT205) of the LAM family under two axenic media conditions. These clinical isolates are phenotypically different at the level of cell death, cytokine production, growth kinetics upon in vitro infection of human tissue macrophages, and membrane vesicle secretion upon culture in synthetic medium. Using RNA-seq, we have identified different pathways that account for two different strategies to cope with the stressful condition of a carbon-poor media such as Sauton's. We showed that the clinical isolate UT205 focus mainly in the activation of virulence systems such as the ESX-1, synthesis of diacyl-trehalose, polyacyl-trehalose, and sulfolipids, while UT127 concentrates its efforts mainly in the survival mode by the activation of the DNA replication, cell division, and lipid biosynthesis. This is an example of two Mtb isolates that belong to the same family and lineage, and even though they have a very similar genome, its transcriptional regulation showed important differences. This results in summary highlight the necessity to reach a better understanding of the heterogeneity in the behavior of these circulating Mtb strains which may help us to design better treatments and vaccines and to identify new targets for drugs.

Anaerobic digestion is a microbe-driven process widely applied to treat activated sludge from municipal wastewater treatment plants. It is one of the most efficient solutions for sludge reduction along with biogas production. However, the knowledge of the microbial consortium involved in this process is still unknown in full-scale anaerobic digesters from Latin America. This study aimed to elucidate the dynamics of the microbial community of a full-scale anaerobic digester for a year using 16S rDNA amplicon sequencing with the Illumina Miseq platform. The results showed fluctuations in the frequencies of dominant phyla with a decrease of Proteobacteria and Bacteroidetes after a temporary suspension of anaerobic digester. The core community was affiliated with bacterial phyla Firmicutes, Actinobacteria, Proteobacteria, and Chloroflexi. The core community was represented by 154 OTUs that accounted for 74% of all the processed reads. The Anaerolineaceae family, within Chloroflexi phylum, was the most frequently observed taxonomic group in all samples analyzed. Despite the microbial fluctuations, the biogas production was stable over the studied year (average 66% methane production), which might indicate a functional redundancy in the microbial consortium.

CARD9 deficiency is an inborn error of immunity that predisposes otherwise healthy humans to mucocutaneous and invasive fungal infections, mostly caused by Candida, but also by dermatophytes, Aspergillus, and other fungi. Phaeohyphomycosis are an emerging group of fungal infections caused by dematiaceous fungi (phaeohyphomycetes) and are being increasingly identified in patients with CARD9 deficiency. The Corynespora genus belongs to phaeohyphomycetes and only one adult patient with CARD9 deficiency has been reported to suffer from invasive disease caused by C. cassiicola. We identified a Colombian child with an early-onset, deep, and destructive mucocutaneous infection due to C. cassiicola and we searched for mutations in CARD9. We reviewed the medical records and immunological findings in the patient. Microbiologic tests and biopsies were performed. Whole-exome sequencing (WES) was made and Sanger sequencing was used to confirm the CARD9 mutations in the patient and her family. Finally, CARD9 protein expression was evaluated in peripheral blood mononuclear cells (PBMC) by western blotting. The patient was affected by a large, indurated, foul-smelling, and verrucous ulcerated lesion on the left side of the face with extensive necrosis and crusting, due to a C. cassiicola infectious disease. WES led to the identification of compound heterozygous mutations in the patient consisting of the previously reported p.Q289* nonsense (c.865C > T, exon 6) mutation, and a novel deletion (c.23_29del; p.Asp8Alafs10*) leading to a frameshift and a premature stop codon in exon 2. CARD9 protein expression was absent in peripheral blood mononuclear cells from the patient. We describe here compound heterozygous loss-of-expression mutations in CARD9 leading to severe deep and destructive mucocutaneous phaeohyphomycosis due to C. cassiicola in a Colombian child.

Background A prime objective in metagenomics is to classify DNA sequence fragments into taxonomic units. It usually requires several stages: read’s quality control, de novo assembly, contig annotation, gene prediction, etc. These stages need very efficient programs because of the number of reads from the projects. Furthermore, the complexity of metagenomes requires efficient and automatic tools that orchestrate the different stages. Method DATMA is a pipeline for fast metagenomic analysis that orchestrates the following: sequencing quality control, 16S rRNA-identification, reads binning, de novo assembly and evaluation, gene prediction, and taxonomic annotation. Its distributed computing model can use multiple computing resources to reduce the analysis time. Results We used a controlled experiment to show DATMA functionality. Two pre-annotated metagenomes to compare its accuracy and speed against other metagenomic frameworks. Then, with DATMA we recovered a draft genome of a novel Anaerolineaceae from a biosolid metagenome. Conclusions DATMA is a bioinformatics tool that automatically analyzes complex metagenomes. It is faster than similar tools and, in some cases, it can extract genomes that the other tools do not. DATMA is freely available at https://github.com/andvides/DATMA.

Background Hot spring bacteria have unique biological adaptations to survive the extreme conditions of these environments; these bacteria produce thermostable enzymes that can be used in biotechnological and industrial applications. However, sequencing these bacteria is complex, since it is not possible to culture them. As an alternative, genome shotgun sequencing of whole microbial communities can be used. The problem is that the classification of sequences within a metagenomic dataset is very challenging particularly when they include unknown microorganisms since they lack genomic reference. We failed to recover a bacterium genome from a hot spring metagenome using the available software tools, so we develop a new tool that allowed us to recover most of this genome. Results We present a proteobacteria draft genome reconstructed from a Colombian’s Andes hot spring metagenome. The genome seems to be from a new lineage within the family Rhodanobacteraceae of the class Gammaproteobacteria, closely related to the genus Dokdonella. We were able to generate this genome thanks to CLAME. CLAME, from Spanish “CLAsificador MEtagenomico”, is a tool to group reads in bins. We show that most reads from each bin belong to a single chromosome. CLAME is very effective recovering most of the reads belonging to the predominant species within a metagenome. Conclusions We developed a tool that can be used to extract genomes (or parts of them) from a complex metagenome. Electronic supplementary material The online version of this article (10.1186/s12864-018-5191-y) contains supplementary material, which is available to authorized users.

Biallelic loss-of-function (LOF) mutations of the NCF4 gene, encoding the p40phox subunit of the phagocyte NADPH oxidase, have been described in only 1 patient. We report on 24 p40phox-deficient patients from 12 additional families in 8 countries. These patients display 8 different in-frame or out-of-frame mutations of NCF4 that are homozygous in 11 of the families and compound heterozygous in another. When overexpressed in NB4 neutrophil-like cells and EBV-transformed B cells in vitro, the mutant alleles were found to be LOF, with the exception of the p.R58C and c.120_134del alleles, which were hypomorphic. Particle-induced NADPH oxidase activity was severely impaired in the patients' neutrophils, whereas PMA-induced dihydrorhodamine-1,2,3 (DHR) oxidation, which is widely used as a diagnostic test for chronic granulomatous disease (CGD), was normal or mildly impaired in the patients. Moreover, the NADPH oxidase activity of EBV-transformed B cells was also severely impaired, whereas that of mononuclear phagocytes was normal. Finally, the killing of Candida albicans and Aspergillus fumigatus hyphae by neutrophils was conserved in these patients, unlike in patients with CGD. The patients suffer from hyperinflammation and peripheral infections, but they do not have any of the invasive bacterial or fungal infections seen in CGD. Inherited p40phox deficiency underlies a distinctive condition, resembling a mild, atypical form of CGD. ispartof: JOURNAL OF CLINICAL INVESTIGATION vol:128 issue:9 pages:3957-3975 ispartof: location:United States status: published

RESUMEN: Objetivo Caracterizar la microbiota bacteriana presente en los biosólidos generados en una de las plantas de tratamiento de aguas residuales más grande de Colombia. Materiales y Métodos Se utilizó la plataforma de secuenciamiento 454 de la compañía Roche para secuenciar las regiones variables V1-V3 y V6-V9 del marcador molecular 16S rRNA y caracterizar la microbiota. Adicionalmente, se aplicaron estrategias filogenéticas para la identificación de especies bacterianas de importancia. Resultados Nuestros análisis muestran que los Phyla más abundantes son Chloroflexi, Proteobacteria, Bacteroidetes, Actinobacteria y Firmicutes. Los géneros clasificados más abundantes fueron Pseudomonas, Dysgonomonas y Proteiniphilum. Sin embargo, el grupo dominante según la región variable V1-V3 es una Anaerolineaceaeque no se ajusta a las especies descritas para esta familia. Conclusiones En las muestras de biosólido analizadas predominan bacterias ambientales que participan en los procesos de estabilización de la materia orgánica durante los tratamientos biológicos de tipo secundario y la digestión anaerobia. Se detectaron secuencias de especies dentro de la familia Anaerolineaceae, los análisis filogenéticos muestran que probablemente se trata de especies no descritas. En el momento del estudio, se encontró que en el sistema de digestión anaerobia se genera biosolido con una baja carga de bacterias potencialmente patógenas. ABSTRACT: Objective To describe bacterial microbiota in the biosolids generated in one of the lar-gest wastewater treatment plants of Colombia.Materials and Methods Using NGS technology, 16S rRNA Gene Amplicon libraries were amplified and sequenced. The Roche 454 FLX Titanium platform was used, while the V1-V3 and V6-V9 hypervariable regions were amplified and analyzed inde-pendently. Amplicon processing and bacterial classification were performed using the AmpliconNoise pipeline and the RDP Classifier tool.Results The analysis showed that the most dominant Phyla in the biosolids were Chlo-roflexi, Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most domi-nant genera were Pseudomonas, Dysgonomonas and Proteiniphilum; however, the dominant group according in the V1-V3 variable region was Anaerolineaceae, which does not conform to the species described for this family. Pathogenic bacteria such as Salmonella and E. coli/Shigella were not detected in the studied biosolid sample. Conclusions In the biosolids samples analyzed, environmental bacteria involved in organic matter stabilization processes during secondary biological treatments and anaerobic digestion were predominant. One of the dominant species in this sludge is a novel species of the Anaerolineaceae group. At the time of the study, it was found that the anaerobic digester system was able to maintain pathogenic bacteria at very low concentrations. 0007506

To describe bacterial microbiota in the biosolids generated in one of the largest wastewater treatment plants of Colombia.Using NGS technology, 16S rRNA Gene Amplicon libraries were amplified and sequenced. The Roche 454 FLX Titanium platform was used, while the V1-V3 and V6-V9 hypervariable regions were amplified and analyzed independently. Amplicon processing and bacterial classification were performed using the AmpliconNoise pipeline and the RDP Classifier tool.The analysis showed that the most dominant Phyla in the biosolids were Chlo-roflexi, Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most dominant genera were Pseudomonas, Dysgonomonas and Proteiniphilum; however, the dominant group according in the V1-V3 variable region was Anaerolineaceae, which does not conform to the species described for this family. Pathogenic bacteria such as Salmonella and E. coli/Shigella were not detected in the studied biosolid sample.In the biosolids samples analyzed, environmental bacteria involved in organic matter stabilization processes during secondary biological treatments and anaerobic digestion were predominant. One of the dominant species in this sludge is a novel species of the Anaerolineaceae group. At the time of the study, it was found that the anaerobic digester system was able to maintain pathogenic bacteria at very low concentrations.Caracterizar la microbiota bacteriana presente en los biosólidos generados en una de las plantas de tratamiento de aguas residuales más grande de Colombia.Se utilizó la plataforma de secuenciamiento 454 de la compañía Roche para secuenciar las regiones variables V1-V3 y V6-V9 del marcador molecular 16S rRNA y caracterizar la microbiota. Adicionalmente, se aplicaron estrategias filogenéticas para la identificación de especies bacterianas de importancia.Nuestros análisis muestran que los Phyla más abundantes son Chloro-flexi, Proteobacteria, Bacteroidetes, Actinobacteria y Firmicutes. Los géneros clasificados más abundantes fueron Pseudomonas, Dysgonomonas y Proteiniphilum. Sin embargo, el grupo dominante según la región variable V1-V3 es una Anaerolineaceae que no se ajusta a las especies descritas para esta familia.En las muestras de biosólido analizadas predominan bacterias ambientales que participan en los procesos de estabilización de la materia orgánica durante los tratamientos biológicos de tipo secundario y la digestión anaerobia. Se detectaron secuencias de especies dentro de la familia Anaerolineaceae, los análisis filogenéticos muestran que probablemente se trata de especies no descritas. En el momento del estudio, se encontró que en el sistema de digestión anaerobia se genera biosolido con una baja carga de bacterias potencialmente patógenas.

Biological sequence comparison is one of the most important tasks in Bioinformatics. Owing to the fast growth of databases that contain biological information, sequence comparison represents an important challenge for high-performance computing, especially when very long sequences are compared, i.e. the complete genome of several organisms. The Smith–Waterman (SW) algorithm is an exact method based on dynamic programming to quantify local similarity between sequences. The inherent large parallelism of the algorithm makes it ideal for architectures supporting multiple dimensions of parallelism (TLP, DLP and ILP). Concurrently, there is a paradigm shift towards chip multiprocessors in computer architecture, which offer a huge amount of potential performance that can only be exploited efficiently if applications are effectively mapped and parallelized. In this work, we analyze how large-scale biology sequence comparison takes advantage of the current and future multicore architectures. Our starting point is the performance analysis of the current multicore IBM Cell B.E. processor; we analyze two different SW implementations on the Cell B.E. Then, using simulation tools, we study the performance scalability when a many-core architecture is used for performing long DNA sequence comparison. We investigate the efficient memory organization that delivers the maximum bandwidth with the minimum cost. Our results show that a heterogeneous architecture can be an efficient alternative to execute challenging bioinformatic workloads. Copyright © 2011 John Wiley & Sons, Ltd.