RESUMO

Aeromonas are Gram-negative rods widely distributed in the environment. They can cause severe infections in fish related to financial losses in the fish industry, and are considered opportunistic pathogens of humans causing infections ranging from diarrhea to septicemia. The objective of this study was to determine in silico the contribution of genomic islands to A. hydrophila. The complete genomes of 17 A. hydrophila isolates, which were separated into two phylogenetic groups, were analyzed using a genomic island (GI) predictor. The number of predicted GIs and their characteristics varied among strains. Strains from group 1, which contains mainly fish pathogens, generally have a higher number of predicted GIs, and with larger size, than strains from group 2 constituted by strains recovered from distinct sources. Only a few predicted GIs were shared among them and contained mostly genes from the core genome. Features related to virulence, metabolism, and resistance were found in the predicted GIs, but strains varied in relation to their gene content. In strains from group 1, O Ag biosynthesis clusters OX1 and OX6 were identified, while strains from group 2 each had unique clusters. Metabolic pathways for myo-inositol, L-fucose, sialic acid, and a cluster encoding QueDEC, tgtA5, and proteins related to DNA metabolism were identified in strains of group 1, which share a high number of predicted GIs. No distinctive features of group 2 strains were identified in their predicted GIs, which are more diverse and possibly better represent GIs in this species. However, some strains have several resistance attributes encoded by their predicted GIs. Several predicted GIs encode hypothetical proteins and phage proteins whose functions have not been identified but may contribute to Aeromonas fitness. In summary, features with functions identified on predicted GIs may confer advantages to host colonization and competitiveness in the environment.

RESUMO

Muitos métodos têm sido empregados na produção de peptídeos bioativos para a promoção da saúde. O objetivo deste estudo foi produzir caseinofosfopeptídeos por hidrólise tríptica do caseinato de sódio usando diferentes temperaturas (37 e 50 °C) e tempos de reação (2 e 4 h), caracterizá-los e analisar sua influência nas atividades citotóxica, antimicrobiana e antioxidante. Caseinofosfopeptídeos foram caracterizados através da composição centesimal, eletroforese em gel de poliacrilamida, Espectrometria de Massas e Cromatografia Líquida de Alta Eficiência. Toxicidade para leucócitos humanos, atividade antimicrobiana utilizando o teste de microdiluição em caldo e determinação da capacidade antioxidante pelo método de espécies reativas ao ácido tiobarbitúrico foram os ensaios biológicos realizados. Os resultados mostraram que as quatro frações peptídicas obtidas apresentaram-se com baixo peso molecular e elevados teores proteico e mineral; quanto ao perfil aminoacídico, apresentaram elevadas e diferenciadas quantidades de ácido glutâmico e serina, que pouco variaram de acordo com o processo de obtenção; não se mostraram tóxicos para leucócitos humanos; demonstraram atividade antimicrobiana para Escherichia coli e Salmonella Enteritidis e elevada capacidade antioxidante. Os resultados físico-químicos das frações de caseinofosfopeptídeos demonstraram elevada composição nutricional em termos de proteína e, principalmente, cálcio. O conjunto de dados indicou que alterações no tempo e na temperatura de reação para a obtenção dos hidrolisados não interferem nas suas qualidades biológicas, mostrando serem seguros para a promoção da saúde e para a aplicação em situações especiais, que envolvem pacientes desnutridos, imunossuprimidos, com comprometimento ósseo ou gastrintestinal decorrentes de inflamações e infecções.

Several methods have been employed for the production of bioactive peptides for health promotion. The aim of this study was to produce and characterize Casein phosphopeptides obtained by sodium caseinate tryptic hydrolysis under different temperatures (37 and 50 °C) and reaction times (2 and 4 h), and evaluate their biological capabilities. They have been characterized by assessing their centesimal composition, by polyacrylamide gel electrophoresis, Mass Spectrometry, and High-Performance Liquid Chromatography. The biological activities tested included toxicity for human leukocytes, antimicrobial assay using the microdilution test, and determination of the antioxidant capacity by the thiobarbituric acid reactive species method. The results showed that the four fractions obtained were of low molecular weight with high protein and mineral contents; their amino acid profile showed high and differentiated amounts of glutamic acid and serine independent of the methodological procedures. The results also showed no toxicity for human peripheral leukocytes, demonstrated antimicrobial activity against Escherichia coli and Salmonella Enteritidis as well as high antioxidant capacity. The results of the physicochemical Casein phosphopeptides' fractions showed high nutritional composition in terms of protein and, particularly, calcium. The biological assays indicated that time and temperature changes in the process for obtaining casein hydrolysates have not interfered with their biological qualities. In addition, they have proven safe in promoting health in special conditions involving malnourished and/or, immunocompromised patients or those with bone and/or gastrointestinal impairment due to inflammations and infections.

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RESUMO

BACKGROUND: Diarrheagenic Escherichia coli (DEC) strains are important causes of diarrhea. However, they cannot be distinguished from E. coli of the intestinal microbiota by conventional microbiological tests. METHODS: This work presents a two-system multiplex PCR for detection of DEC. Primers for 16S rRNA gene were added as internal amplification control to validate negative reactions. The multiplex-PCR system 1 contains primers for detection of Shiga toxin producing E. coli (STEC; stx1, stx2), enteropathogenic E. coli (EPEC; eae, bfpA), atypical enteropathogenic E. coli (aEPEc; eae), enteroinvasive E. coli (ETEC; lt, st), enteroinvasive E. coli (EIEC; ial), and the internal amplification control 16S rRNA. The system 2 contains primers for EIEC (ipaH), enteroaggregative E. coli (CVD432), diffusely adherent E. coli (daaE), and 16S rRNA. The protocol was tested with E. coli reference strains, and also with cultures of fecal specimens of people with diarrhea and healthy controls. RESULTS: The protocol correctly identified the DEC reference strains. No DEC marker was amplified for negative controls; these results were validated by the amplification of a fragment of the 16S rRNA gene. The frequency of DEC was 7.6% for both patients and healthy controls; two Shigella sonnei strains were detected in the group with diarrhea. The identity of the amplicons was confirmed by DNA sequencing. CONCLUSION: The protocol is specific for DEC Shigella and is suitable for clinical laboratories.

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Seven fungi were isolated from 50 samples of cosmetic powders. Morphological analyses and ribosomal DNA Internal Transcribed Spacers sequencing were performed which allowed the discrimination of the isolated fungi as Aspergillus fumigatus, Penicillium sp., and Cladosporium sp. which could have, among their species, potentially pathogenic microorganisms.

RESUMO

Aeromonas spp. were identified in five (2,7%) of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil.

Assuntos

RESUMO

Aeromonas spp. were identified in five (2,7%) of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil.