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      Complete genome sequence and characterization of a novel Enterococcus faecium with probiotic potential isolated from the gut of Litopenaeus vannamei

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          Abstract

          Litopenaeus vannamei, the Pacific whiteleg shrimp, is one of the most marketable species in aquaculture worldwide. However, it is susceptible to different infections causing considerable losses in production each year. Consequently, using prebiotics that promotes the proliferation of beneficial bacteria and strengthen the immune system is a current strategy for disease control. In this study, we isolated two strains of E. faecium from the gut of L. vannamei fed with agavin-supplemented diets. These isolates showed antibacterial activity against Vibrio parahaemolyticus, Vibrio harveyi and Vibrio alginolyticus , most likely due to peptidoglycan hydrolase (PGH) activity. Furthermore, we sequenced the genome of one isolate. As a result, we observed three proteins related to the production of bacteriocins, a relevant trait for selecting probiotic strains since they can inhibit the invasion of potential pathogens. Additionally, the genome annotation showed genes related to the production of essential nutrients for the host. It lacked two of the most common factors associated with virulence in Enterococcus pathogenic strains (esp and hyl). Thus, this host-probiotic-derived strain has potential application not only in shrimp health but also in alternative aquatic environments, as it is adapted to coexist within the gut shrimp microbiota, independently of the diet.

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          CARD 2020: antibiotic resistome surveillance with the comprehensive antibiotic resistance database

          Abstract The Comprehensive Antibiotic Resistance Database (CARD; https://card.mcmaster.ca) is a curated resource providing reference DNA and protein sequences, detection models and bioinformatics tools on the molecular basis of bacterial antimicrobial resistance (AMR). CARD focuses on providing high-quality reference data and molecular sequences within a controlled vocabulary, the Antibiotic Resistance Ontology (ARO), designed by the CARD biocuration team to integrate with software development efforts for resistome analysis and prediction, such as CARD’s Resistance Gene Identifier (RGI) software. Since 2017, CARD has expanded through extensive curation of reference sequences, revision of the ontological structure, curation of over 500 new AMR detection models, development of a new classification paradigm and expansion of analytical tools. Most notably, a new Resistomes & Variants module provides analysis and statistical summary of in silico predicted resistance variants from 82 pathogens and over 100 000 genomes. By adding these resistance variants to CARD, we are able to summarize predicted resistance using the information included in CARD, identify trends in AMR mobility and determine previously undescribed and novel resistance variants. Here, we describe updates and recent expansions to CARD and its biocuration process, including new resources for community biocuration of AMR molecular reference data.
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            KAAS: an automatic genome annotation and pathway reconstruction server

            The number of complete and draft genomes is rapidly growing in recent years, and it has become increasingly important to automate the identification of functional properties and biological roles of genes in these genomes. In the KEGG database, genes in complete genomes are annotated with the KEGG orthology (KO) identifiers, or the K numbers, based on the best hit information using Smith–Waterman scores as well as by the manual curation. Each K number represents an ortholog group of genes, and it is directly linked to an object in the KEGG pathway map or the BRITE functional hierarchy. Here, we have developed a web-based server called KAAS (KEGG Automatic Annotation Server: http://www.genome.jp/kegg/kaas/) i.e. an implementation of a rapid method to automatically assign K numbers to genes in the genome, enabling reconstruction of KEGG pathways and BRITE hierarchies. The method is based on sequence similarities, bi-directional best hit information and some heuristics, and has achieved a high degree of accuracy when compared with the manually curated KEGG GENES database.
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              16S ribosomal DNA amplification for phylogenetic study.

              A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.
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                Author and article information

                Journal
                Microb Genom
                Microb Genom
                mgen
                mgen
                Microbial Genomics
                Microbiology Society
                2057-5858
                2023
                8 March 2023
                8 March 2023
                : 9
                : 3
                : mgen000938
                Affiliations
                [ 1] departmentDepartamento de Ingeniería Celular y Biocatálisis , Instituto de Biotecnología, UNAM, Avenida Universidad 2001, Col. Chamilpa, 62420 Cuernavaca , Morelos, Mexico
                [ 2] departmentDepartamento de Microbiología Molecular , Instituto de Biotecnología (IBT), Universidad Nacional Autónoma de México (UNAM) Av. Universidad #2001, Col. Chamilpa , Cuernavaca, Morelos 62210, Mexico
                Author notes

                The whole genome sequence was submitted to the NCBI database (GenBank CP101669, BioSample SAMN28936884, BioProject PRJNA847457).

                *Correspondence: Agustín López Munguía, agustin.lopez@ 123456ibt.unam.mx
                *Correspondence: Adrian Ochoa-Leyva, adrian.ochoa@ 123456ibt.unam.mx
                Author information
                https://orcid.org/0000-0002-8860-0996
                https://orcid.org/0000-0002-1157-8698
                https://orcid.org/0000-0001-6789-705X
                https://orcid.org/0000-0002-4701-2303
                https://orcid.org/0000-0003-4204-4936
                Article
                000938
                10.1099/mgen.0.000938
                10132076
                36884014
                717de51d-69bb-42fb-aeaa-d27da5563f47
                © 2023 The Authors

                This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.

                History
                : 22 July 2022
                : 22 November 2022
                Funding
                Funded by: DGAPA PAPIIT-UNAM
                Award ID: IN215520
                Award Recipient : Ochoa-LeyvaAdrian
                Funded by: CONACYT, Mexico
                Award ID: 2019-263986
                Award Recipient : Ochoa-LeyvaAdrian
                Categories
                Research Articles
                Microbial Communities
                Custom metadata
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                shrimp diet,fructans,agavins,litopenaeus vannamei,enterococcus faecium,peptidoglycan hydrolase

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