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AIMS: Develop a species-specific multiplex PCR to correctly identify Edwardsiella species in routine diagnostic for fish bacterial diseases. METHODS AND RESULTS: The genomes of 62 Edwardsiella spp. isolates available from the National Center for Biotechnology Information (NCBI) database were subjected to taxonomic and pan-genomic analyses to identify unique regions that could be exploited by species-specific PCR. The designed primers were tested against isolated Edwardsiella spp. strains, revealing errors in commercial biochemical tests for bacterial classification regarding Edwardsiella species. CONCLUSION: Some of the genomes of Edwardsiella spp. in the NCBI platform were incorrectly classified, which can lead to errors in some research. A functional mPCR was developed to differentiate between phenotypically and genetically ambiguous Edwardsiella, with which, we detected the presence of Edwardsiella anguillarum affecting fish in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that the misclassification of Edwardsiella spp in Brazil concealed the presence of E. anguillarum in South America. Also, this review of the taxonomic classification of the Edwardsiella genus is a contribution to the field to help researchers with their sequencing and identification of genomes, showing some misclassifications in online databases that must be corrected, as well as developing an easy assay to characterize Edwardsiella species in an end-point mPCR.
Assuntos
Edwardsiella , Infecções por Enterobacteriaceae , Doenças dos Peixes , Animais , Brasil , Edwardsiella/genética , Edwardsiella tarda/genética , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/diagnóstico , Doenças dos Peixes/microbiologia , Peixes/microbiologia , Reação em Cadeia da Polimerase Multiplex/métodosRESUMO
Objective: To conduct an epidemiologic review, analyzing treatment, evolution, and survival of soft tissue sarcomas. Methods: Retrospective study based on medical records of patient with STS treated by the Orthopedic Oncology Group at the Santa Casa de São Paulo, from 2006 to 2019. Data from 121 patients were analyzed according to age, sex, histological type, tumor location, treatment, previous surgery in a non-specialized service, local recurrences, lung metastases, and survival analysis. Results: The most frequent location was the thigh. Patients who underwent surgery with a non-specialized group had higher rates of local recurrence and those with pulmonary metastasis had a lower survival rate. Conclusion: STS can occur at any age and the prevalence of the histological type depends on the patients' age group. Level of Evidence II, Prognostic Study.
Objetivo: Conduzir uma avaliação epidemiológica analisando tratamento, evolução e sobrevida dos sarcomas de partes moles (SPMs). Métodos: Estudo retrospectivo de prontuários de pacientes com SPM tratados pelo Grupo de Oncologia Ortopédica da Santa Casa de Misericórdia de São Paulo, no período de 2006 a 2019. Foram analisados os dados de 121 pacientes referentes a idade, sexo, tipo histológico, localização do tumor, tratamento, cirurgia prévia em serviço não especializado, presença de recidivas, metástases pulmonares e análise de sobrevida. Resultados: A localização mais frequente foi a coxa. Verificou-se que pacientes que realizaram cirurgia com grupo não especializado tiveram maiores índices de recidiva local, e aqueles com metástase pulmonar tiveram menor sobrevida. Conclusão: Os SPMs podem ocorrer em qualquer idade, e a prevalência do tipo histológico depende da faixa etária dos pacientes. Nível de Evidência II, Estudo Prognóstico.
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Bacterial extracellular vesicles (EVs) are natural lipidic nanoparticles implicated in intercellular communication. Although EV research focused mainly on pathogens, the interest in probiotic-derived EVs is now rising. One example is Propionibacterium freudenreichii, which produces EVs with anti-inflammatory effects on human epithelial cells. Our previous study with P. freudenreichii showed that EVs purified by size exclusion chromatography (SEC) displayed variations in protein content according to bacterial growth conditions. Considering these content variations, we hypothesized that a comparative proteomic analysis of EVs recovered in different conditions would elucidate whether a representative vesicular proteome existed, possibly providing a robust proteome dataset for further analysis. Therefore, P. freudenreichii was grown in two culture media, and EVs were purified by sucrose density gradient ultracentrifugation (UC). Microscopic and size characterization confirmed EV purification, while shotgun proteomics unveiled that they carried a diverse set of proteins. A comparative analysis of the protein content of UC- and SEC-derived EVs, isolated from cultures either in UF (cow milk ultrafiltrate medium) or YEL (laboratory yeast extract lactate medium), showed that EVs from all these conditions shared 308 proteins. This EV core proteome was notably enriched in proteins related to immunomodulation. Moreover, it showed distinctive features, including highly interacting proteins, compositional biases for some specific amino acids, and other biochemical parameters. Overall, this work broadens the toolset for the purification of P. freudenreichii-derived EVs, identifies a representative vesicular proteome, and enumerates conserved features in vesicular proteins. These results hold the potential for providing candidate biomarkers of purification quality, and insights into the mechanisms of EV biogenesis and cargo sorting.
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Stingless bees are a diverse group with a relevant role in pollinating native species. Its diet is rich in carbohydrates and proteins, by collecting pollen and nectar supplies the development of its offspring. Fermentation of these products is associated with microorganisms in the colony. However, the composition of microorganisms that comprise this microbiome and its fundamental role in colony development is still unclear. To characterize the colonizing microorganisms of larval food in the brood cells of stingless bees Frieseomelitta varia, Melipona quadrifasciata, Melipona scutellaris, and Tetragonisca angustula, we have utilized molecular and culture-based techniques. Bacteria of the phyla Firmicutes, Proteobacteria, Actinobacteria, and fungi of the phyla Ascomycota, Basidiomycota, Mucoromycota, and Mortierellomycota were found. Diversity analysis showed that F. varia had a greater diversity of bacteria in its microbiota, and T. angustula had a greater diversity of fungi. The isolation technique allowed the identification of 189 bacteria and 75 fungi. In summary, this research showed bacteria and fungi associated with the species F. varia, M. quadrifasciata, M. scutellaris, and T. angustula, which may play an essential role in the survival of these organisms. Besides that, a biobank with bacteria and fungus isolates from LF of Brazilian stingless bees was created, which can be used for different studies and the prospection of biotechnology compounds.
Assuntos
Fungos , Leveduras , Abelhas , Animais , Larva , Brasil , Fungos/genética , Bactérias/genéticaRESUMO
The bacterium Corynebacterium pseudotuberculosis is of major veterinary importance because it affects livestock, particularly sheep, goats, and horses, in several countries, including Australia, Brazil, the United States, and Canada, resulting in significant economic losses. In the present study, we describe the complete genome of the Corynebacterium pseudotuberculosis Cp316 strain, biovar equi, isolated from the abscess of a North American horse.
Assuntos
Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Abscesso/microbiologia , Abscesso/veterinária , Animais , California , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , Doenças dos Cavalos/microbiologia , Cavalos , Dados de Sequência MolecularRESUMO
Exiguobacterium antarcticum is a psychotropic bacterium isolated for the first time from microbial mats of Lake Fryxell in Antarctica. Many organisms of the genus Exiguobacterium are extremophiles and have properties of biotechnological interest, e.g., the capacity to adapt to cold, which make this genus a target for discovering new enzymes, such as lipases and proteases, in addition to improving our understanding of the mechanisms of adaptation and survival at low temperatures. This study presents the genome of E. antarcticum B7, isolated from a biofilm sample of Ginger Lake on King George Island, Antarctic peninsula.
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Bacillales/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Regiões Antárticas , Bacillales/isolamento & purificação , Bacillales/fisiologia , Biofilmes/crescimento & desenvolvimento , Água Doce/microbiologia , Ilhas , Lagos , Dados de Sequência MolecularRESUMO
In this work we report the genome of Corynebacterium pseudotuberculosis strain 267, isolated from a llama. This pathogen is of great veterinary and economic importance, as it is the cause of caseous lymphadenitis in several livestock species around the world and causes significant losses due to the high cost of treatment.
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Camelídeos Americanos/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Análise de Sequência de DNA , Animais , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Dados de Sequência MolecularRESUMO
Corynebacterium pseudotuberculosis is a pathogen of great veterinary and economic importance, since it affects livestock, mainly sheep and goats, worldwide, together with reports of its presence in camels in several Arabic, Asiatic, and East and West African countries, as well as Australia. In this article, we report the genome sequence of Corynebacterium pseudotuberculosis strain Cp162, collected from the external neck abscess of a camel in the United Kingdom.
Assuntos
Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Abscesso/microbiologia , Abscesso/veterinária , Animais , Camelus , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , Dados de Sequência Molecular , Reino UnidoRESUMO
In this work, we report the complete genome sequence of a Corynebacterium pseudotuberculosis PAT10 isolate, collected from a lung abscess in an Argentine sheep in Patagonia, whose pathogen also required an investigation of its pathogenesis. Thus, the analysis of the genome sequence offers a means to better understanding of the molecular and genetic basis of virulence of this bacterium.
Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Abscesso Pulmonar/microbiologia , Doenças dos Ovinos/microbiologia , Animais , Argentina , Sequência de Bases , Corynebacterium pseudotuberculosis/isolamento & purificação , Corynebacterium pseudotuberculosis/patogenicidade , Dados de Sequência Molecular , Ovinos , VirulênciaRESUMO
In this work, we report the whole-genome sequence of Corynebacterium pseudotuberculosis bv. equi strain CIP 52.97 (Collection Institut Pasteur), isolated in 1952 from a case of ulcerative lymphangitis in a Kenyan horse, which has evidently caused significant losses to agribusiness. Therefore, obtaining this genome will allow the detection of important targets for postgenomic studies, with the aim of minimizing problems caused by this microorganism.
Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Doenças dos Cavalos/microbiologia , Animais , Sequência de Bases , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Cavalos , Quênia , Dados de Sequência MolecularRESUMO
Campylobacter fetus subsp. venerealis is the etiologic agent of bovine genital campylobacteriosis, a sexually transmitted disease of cattle that is of worldwide importance. The complete sequencing and annotation of the genome of the type strain C. fetus subsp. venerealis NCTC 10354(T) are reported.
Assuntos
Infecções por Campylobacter/veterinária , Campylobacter fetus/genética , Doenças dos Bovinos/microbiologia , Genoma Bacteriano , Animais , Sequência de Bases , Infecções por Campylobacter/microbiologia , Campylobacter fetus/isolamento & purificação , Bovinos , Feminino , Masculino , Dados de Sequência MolecularRESUMO
ABSTRACT Objective: To conduct an epidemiologic review, analyzing treatment, evolution, and survival of soft tissue sarcomas. Methods: Retrospective study based on medical records of patient with STS treated by the Orthopedic Oncology Group at the Santa Casa de São Paulo, from 2006 to 2019. Data from 121 patients were analyzed according to age, sex, histological type, tumor location, treatment, previous surgery in a non-specialized service, local recurrences, lung metastases, and survival analysis. Results: The most frequent location was the thigh. Patients who underwent surgery with a non-specialized group had higher rates of local recurrence and those with pulmonary metastasis had a lower survival rate. Conclusion: STS can occur at any age and the prevalence of the histological type depends on the patients' age group. Level of Evidence II, Prognostic Study.
RESUMO Objetivo: Conduzir uma avaliação epidemiológica analisando tratamento, evolução e sobrevida dos sarcomas de partes moles (SPMs). Métodos: Estudo retrospectivo de prontuários de pacientes com SPM tratados pelo Grupo de Oncologia Ortopédica da Santa Casa de Misericórdia de São Paulo, no período de 2006 a 2019. Foram analisados os dados de 121 pacientes referentes a idade, sexo, tipo histológico, localização do tumor, tratamento, cirurgia prévia em serviço não especializado, presença de recidivas, metástases pulmonares e análise de sobrevida. Resultados: A localização mais frequente foi a coxa. Verificou-se que pacientes que realizaram cirurgia com grupo não especializado tiveram maiores índices de recidiva local, e aqueles com metástase pulmonar tiveram menor sobrevida. Conclusão: Os SPMs podem ocorrer em qualquer idade, e a prevalência do tipo histológico depende da faixa etária dos pacientes. Nível de Evidência II, Estudo Prognóstico.
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Melipona scutellaris is a Brazilian stingless bee species and a highly important native pollinator besides its use in rational rearing for honey production. In this study, we present the whole mitochondrial DNA sequence of M. scutellaris from a haploid male. The mitogenome has a size of 14,862 bp and harbors 13 protein-coding genes (PCGs), 2 rRNA genes and 21 tRNA genes.
Assuntos
Abelhas/genética , Genoma Mitocondrial , Animais , Haploidia , Proteínas de Insetos/genética , Masculino , RNA Ribossômico/genética , RNA de Transferência/genéticaRESUMO
New sequencing platforms have enabled rapid decoding of complete prokaryotic genomes at relatively low cost. The Ion Torrent platform is an example of these technologies, characterized by lower coverage, generating challenges for the genome assembly. One particular problem is the lack of genomes that enable reference-based assembly, such as the one used in the present study, Corynebacterium pseudotuberculosis biovar equi, which causes high economic losses in the US equine industry. The quality treatment strategy incorporated into the assembly pipeline enabled a 16-fold greater use of the sequencing data obtained compared with traditional quality filter approaches. Data preprocessing prior to the de novo assembly enabled the use of known methodologies in the next-generation sequencing data assembly. Moreover, manual curation was proved to be essential for ensuring a quality assembly, which was validated by comparative genomics with other species of the genus Corynebacterium. The present study presents a modus operandi that enables a greater and better use of data obtained from semiconductor sequencing for obtaining the complete genome from a prokaryotic microorganism, C. pseudotuberculosis, which is not a traditional biological model such as Escherichia coli.
Assuntos
Biologia Computacional/métodos , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano/genética , Semicondutores , Análise de Sequência de DNA , Animais , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , DNA Bacteriano/análise , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Desenho de Equipamento , Genômica/métodos , Doenças dos Cavalos/microbiologia , Cavalos , Análise de Sequência de DNA/instrumentação , Análise de Sequência de DNA/métodos , SoftwareRESUMO
Although attempts have been made to unveil protein-protein and host-pathogen interactions based on molecular insights of important biological events and pathogenesis in various organisms, these efforts have not yet been reported in Corynebacterium pseudotuberculosis (Cp), the causative agent of Caseous Lymphadenitis (CLA). In this study, we used computational approaches to develop common conserved intra-species protein-protein interaction (PPI) networks first time for four Cp strains (Cp FRC41, Cp 316, Cp 3/99-5, and Cp P54B96) followed by development of a common conserved inter-species bacterial PPI using conserved proteins in multiple pathogens (Y. pestis, M. tuberculosis, C. diphtheriae, C. ulcerans, E. coli, and all four Cp strains) and E. Coli based experimentally validated PPI data. Furthermore, the interacting proteins in the common conserved inter-species bacterial PPI were used to generate a conserved host-pathogen interaction (HP-PPI) network considering human, goat, sheep, bovine, and horse as hosts. The HP-PPI network was validated, and acetate kinase (Ack) was identified as a novel broad spectrum target. Ceftiofur, penicillin, and two natural compounds derived from Piper betel were predicted to inhibit Ack activity. One of these Piper betel compounds found to inhibit E. coli O157:H7 growth similar to penicillin. The target specificity of these betel compounds, their effects on other studied pathogens, and other in silico results are currently being validated and the results are promising.
Assuntos
Interações Hospedeiro-Patógeno , Mapas de Interação de Proteínas , Acetato Quinase/metabolismo , Animais , Antibacterianos/farmacologia , Cefalosporinas/farmacologia , Análise por Conglomerados , Corynebacterium/metabolismo , Corynebacterium diphtheriae/metabolismo , Corynebacterium pseudotuberculosis/metabolismo , Escherichia coli/metabolismo , Escherichia coli O157/metabolismo , Genes Bacterianos , Humanos , Mycobacterium tuberculosis/metabolismo , Penicilinas/farmacologia , Piper/química , Especificidade da Espécie , Yersinia pestis/metabolismoRESUMO
Vibrio cholerae is the causal organism of the cholera epidemic, which is mostly prevalent in developing and underdeveloped countries. However, incidences of cholera in developed countries are also alarming. Because of the emergence of new drug-resistant strains, even though several generic drugs and vaccines have been developed over time, Vibrio infections remain a global health problem that appeals for the development of novel drugs and vaccines against the pathogen. Here, applying comparative proteomic and reverse vaccinology approaches to the exoproteome and secretome of the pathogen, we have identified three candidate targets (ompU, uppP and yajC) for most of the pathogenic Vibrio strains. Two targets (uppP and yajC) are novel to Vibrio, and two targets (uppP and ompU) can be used to develop both drugs and vaccines (dual targets) against broad spectrum Vibrio serotypes. Using our novel computational approach, we have identified three peptide vaccine candidates that have high potential to induce both B- and T-cell-mediated immune responses from our identified two dual targets. These two targets were modeled and subjected to virtual screening against natural compounds derived from Piper betel. Seven compounds were identified first time from Piper betel to be highly effective to render the function of these targets to identify them as emerging potential drugs against Vibrio. Our preliminary validation suggests that these identified peptide vaccines and betel compounds are highly effective against Vibrio cholerae. Currently we are exhaustively validating these targets, candidate peptide vaccines, and betel derived lead compounds against a number of Vibrio species.
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Cólera/tratamento farmacológico , Descoberta de Drogas , Piper betle/química , Vibrio cholerae/efeitos dos fármacos , Cólera/imunologia , Cólera/microbiologia , Epitopos de Linfócito T/imunologia , Humanos , Ligantes , Proteoma , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Vacinas de Subunidades Antigênicas/química , Vacinas de Subunidades Antigênicas/farmacologia , Vibrio cholerae/imunologia , Vibrio cholerae/patogenicidadeRESUMO
Caseous lymphadenitis is a chronic goat and sheep disease caused by Corynebacterium pseudotuberculosis (Cp) that accounts for a huge economic loss worldwide. Proper vaccination or medication is not available because of the lack of understanding of molecular biology of the pathogen. In a recent approach, four Cp (CpFrc41, Cp1002, CpC231, and CpI-19) genomes were sequenced to elucidate the molecular pathology of the bacteria. In this study, using these four genome sequences along with other eight genomes (total 12 genomes) and a novel subtractive genomics approach (first time ever applied to a veterinary pathogen), we identified potential conserved common drug and vaccine targets of these four Cp strains along with other Corybacterium, Mycobacterium and Nocardia (CMN) group of human pathogens (Corynebacterium diphtheriae and Mycobacterium tuberculosis) considering goat, sheep, bovine, horse, and human as the most affected hosts. The minimal genome of Cp1002 was found to consist of 724 genes, and 20 conserved common targets (to all Cp strains as well as CMN group of pathogens) from various metabolic pathways (13 from host-pathogen common and seven from pathogen's unique pathways) are potential targets irrespective of all hosts considered. ubiA from host-pathogen common pathway and an ABC-like transporter from unique pathways may serve dual (drug and vaccine) targets. Two Corynebacterium-specific (mscL and resB) and one broad-spectrum (rpmB) novel targets were also identified. Strain-specific targets are also discussed. Six important targets were subjected to virtual screening, and one compound was found to be potent enough to render two targets (cdc and nrdL). We are currently validating all identified targets and lead compounds.
Assuntos
Antibacterianos/farmacologia , Vacinas Bacterianas/imunologia , Hibridização Genômica Comparativa , Corynebacterium pseudotuberculosis/efeitos dos fármacos , Corynebacterium pseudotuberculosis/imunologia , Animais , Corynebacterium pseudotuberculosis/genética , Desenho de Fármacos , Genoma Bacteriano , Humanos , Mycobacterium/efeitos dos fármacos , Mycobacterium/genética , Mycobacterium/imunologia , Nocardia/efeitos dos fármacos , Nocardia/genética , Nocardia/imunologiaRESUMO
Não encontrado.