Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 34
Filtrar
Más filtros










Intervalo de año de publicación
1.
Animals (Basel) ; 13(16)2023 Aug 13.
Artículo en Inglés | MEDLINE | ID: mdl-37627402

RESUMEN

Brachyspira hyodysenteriae and Lawsonia intracellularis coinfection has been observed in the diagnostic routine; however, no studies have evaluated their interaction. This study aimed to characterize lesions and possible synergisms in experimentally infected pigs. Four groups of piglets, coinfection (CO), B. hyodysenteriae (BRA), L. intracellularis (LAW), and negative control (NEG), were used. Clinical signals were evaluated, and fecal samples were collected for qPCR. At 21 days post infection (dpi), all animals were euthanized. Gross lesions, bacterial isolation, histopathology, immunohistochemistry, and fecal microbiome analyses were performed. Diarrhea started at 12 dpi, affecting 11/12 pigs in the CO group and 5/11 pigs in the BRA group. Histopathological lesions were significantly more severe in the CO than the other groups. B. hyodysenteriae was isolated from 11/12 pigs in CO and 5/11 BRA groups. Pigs started shedding L. intracellularis at 3 dpi, and all inoculated pigs tested positive on day 21. A total of 10/12 CO and 7/11 BRA animals tested positive for B. hyodysenteriae by qPCR. A relatively low abundance of microbiota was observed in the CO group. Clinical signs and macroscopic and microscopic lesions were significantly more severe in the CO group compared to the other groups. The presence of L. intracellularis in the CO group increased the severity of swine dysentery.

2.
J Am Soc Mass Spectrom ; 33(11): 2055-2062, 2022 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-36239433

RESUMEN

Biochemical, serological, and molecular methods have been developed for the laboratory diagnosis of diseases caused by C. pseudotuberculosis (CP), but the identification of the pathogen and biovars differentiation may be time-consuming, expensive, and confusing compared with other bacteria. This study aimed to evaluate MALDI Biotyper and Overall Genome Relatedness Index (OGRI) analysis to optimize the identification and differentiation of biovars of C. pseudotuberculosis. Out of 230 strains isolated from several hosts and countries, 202 (87.8%) were precisely classified using MALDI Biotyper and the BioNumerics platform. The classification accuracies for the Ovis and Equi biovars were 80 (88.75%) and 82 (92.68%), respectively. When analyzing a sampling of these strains by Average Nucleotide Identity based on BLAST and TETRA analyses using genomic sequence data, it was possible to differentiate 100% of the strains in Equi and Ovis. Our data show that MALDI Biotyper and OGRI analysis help identify C. pseudotuberculosis at the species and biovar levels.


Asunto(s)
Corynebacterium pseudotuberculosis , Ovinos , Animales , Corynebacterium pseudotuberculosis/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
3.
Data Brief ; 34: 106746, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33532526

RESUMEN

The genus Klebsiella comprises species that cause nosocomial and community-acquired infections. A dataset was created to compile the sequence type (ST) and capsule type (K-locus) information predicted for 172 worldwide isolates of Klebsiella spp. whose complete genomes could be retrieved from the GenBank (NCBI) repository. The dataset also includes information related to one multidrug-resistant strain (B31) isolated from a patient who was admitted to an intensive care unit in the Northeast region of Brazil. This strain was phenotypically characterized and submitted to whole-genome sequencing and comparative genomics analysis as we recently reported [1]. The dataset also compiles information on Pathogenicity Islands (PIs), Resistance Islands (RIs) and Miscellaneous Islands (MIS) present in the genome of strain B31. The information provided here may support outbreak prevention policies and future epidemiological studies involving Klebsiella spp.

4.
Gene ; 772: 145386, 2021 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-33373662

RESUMEN

The emergence of community acquired infections increases the public health concern on K. pneumoniae and closely related bacteria among which antimicrobial resistance spreads. We report a multidrug-resistant K. pneumoniae isolate, B31, of a patient infected in the community and admitted to an intensive care unit in Northeast Brazil. Antimicrobial susceptibility and genome information were thoroughly investigated to characterize B31 in front of 172 sequenced strains of different countries. Assigned to the Sequence Type 15, which is globally spread, B31 presented extended spectrum beta-lactamase, tigecycline and ciprofloxacin resistance. Genome sequencing revealed most resistance genes being carried by plasmids with high dissemination potential. The absence of main virulence factors, like yersiniabactin and colibactin, apparently suggests a mild pathogenic strain which, on the contrary, persisted and caused severe infection in a previously healthy patient. The present study contributes to unveil the unclear genomic scenario of virulent and multidrug-resistant K. pneumoniae in Brazil.


Asunto(s)
Infecciones Comunitarias Adquiridas/microbiología , Farmacorresistencia Bacteriana Múltiple , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/clasificación , Secuenciación Completa del Genoma/métodos , Adulto , Ciprofloxacina/farmacología , Femenino , Genoma Bacteriano , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Klebsiella pneumoniae/genética , Tipificación de Secuencias Multilocus , Plásmidos/genética , Tigeciclina/farmacología
5.
Exp Parasitol ; 218: 107964, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32822697

RESUMEN

Protozoan parasites of the genus Leishmania are causative agents of leishmaniasis, a wide range of diseases affecting 12 million people worldwide. The species L. infantum and L. amazonensis are etiologic agents of visceral and cutaneous leishmaniasis, respectively. Most proteome analyses of Leishmania have been carried out on whole-cell extracts, but such an approach tends to underrepresent membrane-associated proteins due to their high hydrophobicity and low solubility. Considering the relevance of this category of proteins in virulence, invasiveness and the host-parasite interface, this study applied label-free proteomics to assess the plasma membrane sub-proteome of L. infantum and L. amazonensis. The number of proteins identified in L. infantum and L. amazonensis promastigotes was 1168 and 1455, respectively. After rigorous data processing and mining, 157 proteins were classified as putative plasma membrane-associated proteins, of which 56 proteins were detected in both species, six proteins were detected only in L. infantum and 39 proteins were exclusive to L. amazonensis. The quantitative analysis revealed that two proteins were more abundant in L. infantum, including the glucose transporter 2, and five proteins were more abundant in L. amazonensis. The identified proteins associated with distinct processes and functions. In this regard, proteins of L. infantum were linked to metabolic processes whereas L. amazonensis proteins were involved in signal transduction. Moreover, transmembrane transport was a significant process among the group of proteins detected in both species and members of the superfamily of ABC transporters were highly represented. Interestingly, some proteins of this family were solely detected in L. amazonensis, such as ABCA9. GP63, a well-known virulence factor, was the only GPI-anchored protein identified in the membrane preparations of both species. Finally, we found several proteins with uncharacterized functions, including differentially abundant ones, highlighting a gap in the study of Leishmania proteins. Proteins characterization could provide a better biological understanding of these parasites and deliver new possibilities regarding the discovery of therapeutic targets, drug resistance and vaccine candidates.


Asunto(s)
Leishmania infantum/química , Leishmania mexicana/química , Proteínas de la Membrana/análisis , Proteómica/métodos , Proteínas Protozoarias/análisis , Animales , Membrana Celular/química , Cromatografía Liquida , Biología Computacional , Cricetinae , Transportador de Glucosa de Tipo 2/análisis , Interacciones Huésped-Parásitos , Leishmania infantum/metabolismo , Leishmania infantum/patogenicidad , Leishmania infantum/ultraestructura , Leishmania mexicana/ultraestructura , Macrófagos Peritoneales/parasitología , Espectrometría de Masas , Mesocricetus , Metaloendopeptidasas/análisis , Ratones , Ratones Endogámicos BALB C , Transducción de Señal , Espectrometría de Masas en Tándem , Virulencia
6.
Front Microbiol ; 11: 549254, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33584554

RESUMEN

The rapid emergence of multidrug-resistant (MDR) bacteria is a global health problem. Mobile genetic elements like conjugative plasmids, transposons, and integrons are the major players in spreading resistance genes in uropathogenic Escherichia coli (UPEC) pathotype. The E. coli BH100 strain was isolated from the urinary tract of a Brazilian woman in 1974. This strain presents two plasmids carrying MDR cassettes, pBH100, and pAp, with conjugative and mobilization properties, respectively. However, its transposable elements have not been characterized. In this study, we attempted to unravel the factors involved in the mobilization of virulence and drug-resistance genes by assessing genomic rearrangements in four BH100 sub-strains (BH100 MG2014, BH100 MG2017, BH100L MG2017, and BH100N MG2017). Therefore, the complete genomes of the BH100 sub-strains were achieved through Next Generation Sequencing and submitted to comparative genomic analyses. Our data shows recombination events between the two plasmids in the sub-strain BH100 MG2017 and between pBH100 and the chromosome in BH100L MG2017. In both cases, IS3 and IS21 elements were detected upstream of Tn21 family transposons associated with MDR genes at the recombined region. These results integrated with Genomic island analysis suggest pBH100 might be involved in the spreading of drug resistance through the formation of resistance islands. Regarding pathogenicity, our results reveal that BH100 strain is closely related to UPEC strains and contains many IS3 and IS21-transposase-enriched genomic islands associated with virulence. This study concludes that those IS elements are vital for the evolution and adaptation of BH100 strain.

7.
Dis Aquat Organ ; 134(2): 107-111, 2019 May 02.
Artículo en Inglés | MEDLINE | ID: mdl-31043578

RESUMEN

This report describes a severe outbreak of the gill fluke Centrocestus formosanus in farm-raised platies Xiphophorus maculatus in Brazil, with mortality rate approaching 95%. Typical clinical signs of infection were observed, with microscopic examinations of fresh gills revealing multiple cysts containing a once-folded metacercaria with an X-shaped excretory bladder. The 18S subunit of the metacercariae (BR1) was amplified by PCR, sequenced and analyzed by BLAST. Subsequent phylogenetic analyses revealed that the BR1 isolate was closely related to C. formosanus from Thailand. This is the first report of C. formosanus in ornamental fish in Brazil. Our observations suggest that platies are highly sensitive to this digenetic parasite. Controlling population densities of the parasite's intermediate host, the snail Melanoides tuberculata, would help to reduce outbreaks.


Asunto(s)
Ciprinodontiformes , Enfermedades de los Peces , Heterophyidae , Trematodos , Infecciones por Trematodos , Animales , Brasil , Brotes de Enfermedades , Granjas , Filogenia , Tailandia , Infecciones por Trematodos/veterinaria
8.
Emerg Infect Dis ; 25(4): 776-779, 2019 04.
Artículo en Inglés | MEDLINE | ID: mdl-30882311

RESUMEN

In 2016 and 2017, we characterized outbreaks caused by Streptococcus agalactiae serotype III sequence type (ST) 283 in Nile tilapia farms in Brazil. Whole-genome multilocus sequence typing clustered the fish isolates together with the zoonotic ST283 and other STs related to cases in humans, frogs, dogs, cattle, and dolphins.


Asunto(s)
Animales Domésticos , Enfermedades de los Peces/epidemiología , Enfermedades de los Peces/microbiología , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae/clasificación , Streptococcus agalactiae/genética , Animales , Brasil/epidemiología , Genoma Bacteriano , Genómica/métodos , Humanos , Tipificación de Secuencias Multilocus , Filogenia , Vigilancia en Salud Pública , Streptococcus agalactiae/aislamiento & purificación
9.
Prev Vet Med ; 162: 131-135, 2019 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-30621892

RESUMEN

Infection with Streptococcus agalactiae causes mortality and major economic losses in Nile tilapia (Oreochromis niloticus) farming worldwide. In Brazil, serotype strains Ia, Ib and III have been isolated in streptococcosis outbreaks, but serotype Ib is the most prevalent. Vaccination is considered an effective method to prevent economically-important diseases in aquaculture and has been associated with decreased use of antibiotics and improvements in fish survival. We developed a flexible partial-budget model to undertake an economic appraisal of vaccination against Streptococcus agalactiae in Nile tilapia farmed in net cages in large reservoirs. The model considers the benefits and costs that are likely to be associated with vaccination at the farm-level, in one production cycle. We built three epidemiological scenarios of cumulative mortality attributable to S. agalactiae (5%, 10%, and 20%, per production cycle) in a non-vaccinated farm. For each scenario, we applied a stochastic model to simulate the net return of vaccination, given a combination of values of "vaccine efficacy", "gain in feed conversion ratio", "feed price", "fish market price ", and "cost of vaccine dose". In the 20% cumulative mortality scenario, the net return would break-even (benefits ≥ costs) in at least 97.9% of interactions. Should cumulative mortality be lower than 10%, the profitability of vaccination would be more dependent on better feed conversion ratio. The inputs "feed price" and "cost of vaccine" had minor effects on the output, in all pre-vaccination mortality scenarios. Although our simulations are based on conservative values and consider uncertainty about the modeled parameters, we conclude that vaccination against S. agalactiae is likely to be profitable in Nile tilapia farms, under similar production conditions.


Asunto(s)
Enfermedades de los Peces/prevención & control , Infecciones Estreptocócicas/veterinaria , Vacunas Estreptocócicas/economía , Streptococcus agalactiae/inmunología , Tilapia/microbiología , Animales , Acuicultura/economía , Brasil , Análisis Costo-Beneficio , Enfermedades de los Peces/economía , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Modelos Económicos , Infecciones Estreptocócicas/economía , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/prevención & control , Vacunas Estreptocócicas/uso terapéutico
10.
Front Microbiol ; 9: 2639, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30450092

RESUMEN

Streptococcus agalactiae is one of the most important pathogens associated with streptococcosis outbreaks in Nile tilapia farms worldwide. High water temperature (above 27°C) has been described as a predisposing factor for the disease in fish. At low temperatures (below 25°C), fish mortalities are not usually observed in farms. Temperature variation can modulate the expression of genes and proteins involved in metabolism, adaptation, and bacterial pathogenicity, thus increasing or decreasing the ability to infect the host. This study aimed to evaluate the transcriptome and proteome of a fish-pathogenic S. agalactiae strain SA53 subjected to in vitro growth at different temperatures using a microarray and label-free shotgun LC-HDMSE approach. Biological triplicates of isolates were cultured in BHIT broth at 22 or 32°C for RNA and protein isolation and submitted for transcriptomic and proteomic analyses. In total, 1,730 transcripts were identified in SA53, with 107 genes being differentially expressed between the temperatures evaluated. A higher number of genes related to metabolism, mainly from the phosphotransferase system (PTS) and ATP-binding cassette (ABC) transport system, were upregulated at 32°C. In the proteome analysis, 1,046 proteins were identified in SA53, of which 81 were differentially regulated between 22 and 32°C. Proteins involved in defense mechanisms, lipid transport and metabolism, and nucleotide transport and metabolism were upregulated at 32°C. A higher number of interactions were observed in proteins involved in nucleotide transport and metabolism. We observed a low correlation between the transcriptome and proteome datasets. Our study indicates that the transcriptome and proteome of a fish-adapted S. agalactiae strain are modulated by temperature, particularly showing differential expression of genes/proteins involved in metabolism, virulence factors, and adaptation.

11.
Stand Genomic Sci ; 13: 21, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30338024

RESUMEN

Corynebacterium pseudotuberculosis is a pathogenic bacterium which has been rapidly spreading all over the world, causing economic losses in the agricultural sector and sporadically infecting humans. Six C. pseudotuberculosis strains were isolated from goats, sheep, and horses with distinct abscess locations. For the first time, Mexican genomes of this bacterium were sequenced and studied in silico. All strains were sequenced using Ion Personal Genome Machine sequencer, assembled using Newbler and SPAdes software. The automatic genome annotation was done using the software RAST and in-house scripts for transference, followed by manual curation using Artemis software and BLAST against NCBI and UniProt databases. The six genomes are publicly available in NCBI database. The analysis of nucleotide sequence similarity and the generated phylogenetic tree led to the observation that the Mexican strains are more similar between strains from the same host, but the genetic structure is probably more influenced by transportation of animals between farms than host preference. Also, a putative drug target was predicted and in silico analysis of 46 strains showed two gene clusters capable of differentiating the biovars equi and ovis: Restriction Modification system and CRISPR-Cas cluster.

12.
Front Microbiol ; 9: 1807, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30174657

RESUMEN

Propionibacterium freudenreichii is a beneficial Gram-positive bacterium, traditionally used as a cheese-ripening starter, and currently considered as an emerging probiotic. As an example, the P. freudenreichii CIRM-BIA 129 strain recently revealed promising immunomodulatory properties. Its consumption accordingly exerts healing effects in different animal models of colitis, suggesting a potent role in the context of inflammatory bowel diseases. This anti-inflammatory effect depends on surface layer proteins (SLPs). SLPs may be involved in key functions in probiotics, such as persistence within the gut, adhesion to host cells and mucus, or immunomodulation. Several SLPs coexist in P. freudenreichii CIRM-BIA 129 and mediate immunomodulation and adhesion. A mutant P. freudenreichii CIRM-BIA 129ΔslpB (CB129ΔslpB) strain was shown to exhibit decreased adhesion to intestinal epithelial cells. In the present study, we thoroughly analyzed the impact of this mutation on cellular properties. Firstly, we investigated alterations of surface properties in CB129ΔslpB. Surface extractable proteins, surface charges (ζ-potential) and surface hydrophobicity were affected by the mutation. Whole-cell proteomics, using high definition mass spectrometry, identified 1,288 quantifiable proteins in the wild-type strain, i.e., 53% of the theoretical proteome predicted according to P. freudenreichii CIRM-BIA 129 genome sequence. In the mutant strain, we detected 1,252 proteins, including 1,227 proteins in common with the wild-type strain. Comparative quantitative analysis revealed 97 proteins with significant differences between wild-type and mutant strains. These proteins are involved in various cellular process like signaling, metabolism, and DNA repair and replication. Finally, in silico analysis predicted that slpB gene is not part of an operon, thus not affecting the downstream genes after gene knockout. This study, in accordance with the various roles attributed in the literature to SLPs, revealed a pleiotropic effect of a single slpB mutation, in the probiotic P. freudenreichii. This suggests that SlpB may be at a central node of cellular processes and confirms that both nature and amount of SLPs, which are highly variable within the P. freudenreichii species, determine the probiotic abilities of strains.

13.
Gene ; 677: 349-360, 2018 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-30098432

RESUMEN

Corynebacterium pseudotuberculosis has been widely studied in an effort to understand its biological evolution. Transcriptomics has revealed possible candidates for virulence and pathogenicity factors of strain 1002 (biovar Ovis). Because C. pseudotuberculosis is classified into two biovars, Ovis and Equi, it was interesting to assess the transcriptional profile of biovar Equi strain 258, the causative agent of ulcerative lymphangitis. The genome of this strain was re-sequenced; the reassembly was completed using optical mapping technology, and the sequence was subsequently re-annotated. Two growth conditions that occur during the host infection process were simulated for the transcriptome: the osmotic and acid medium. Genes that may be associated with the microorganism's resilience under unfavorable conditions were identified through RNAseq, including genes present in pathogenicity islands. The RT-qPCR was performed to confirm the results in biological triplicate for each condition for some genes. The results extend our knowledge of the factors associated with the spread and persistence of C. pseudotuberculosis during the infection process and suggest possible avenues for studies related to the development of vaccines, diagnosis, and therapies that might help minimize damage to agribusinesses.


Asunto(s)
Corynebacterium pseudotuberculosis/genética , Estrés Fisiológico/genética , Transcriptoma/genética , Animales , Proteínas Bacterianas/genética , Infecciones por Corynebacterium/microbiología , Perfilación de la Expresión Génica/métodos , Genoma Bacteriano/genética , Ovinos , Virulencia/genética , Factores de Virulencia/genética
14.
Reprod Domest Anim ; 53(5): 1243-1246, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-29888513

RESUMEN

The study aimed to describe the Bubalus bubalis seminal plasma proteome using a label-free shotgun UDMSE approach. A total of 859 nonredundant proteins were identified across five biological replicates with stringent identification. Proteins specifically related to sperm maturation and protection, capacitation, fertilization and metabolic activity were detected in the buffalo seminal fluid. In conclusion, we provide a comprehensive proteomic profile of buffalo seminal plasma, which establishes a foundation for further studies designed to understand regulation of sperm function and discovery of novel biomarkers for fertility. MS data are available in the ProteomeXchange with identifier PXD003728.


Asunto(s)
Búfalos/fisiología , Proteoma/fisiología , Semen/fisiología , Animales , Criopreservación/veterinaria , Masculino , Espectrometría de Masas , Proteómica , Análisis de Semen/veterinaria , Preservación de Semen/veterinaria
15.
Front Genet ; 9: 44, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29487617

RESUMEN

Diphtheria is an acute and highly infectious disease, previously regarded as endemic in nature but vaccine-preventable, is caused by Corynebacterium diphtheriae (Cd). In this work, we used an in silico approach along the 13 complete genome sequences of C. diphtheriae followed by a computational assessment of structural information of the binding sites to characterize the "pocketome druggability." To this end, we first computed the "modelome" (3D structures of a complete genome) of a randomly selected reference strain Cd NCTC13129; that had 13,763 open reading frames (ORFs) and resulted in 1,253 (∼9%) structure models. The amino acid sequences of these modeled structures were compared with the remaining 12 genomes and consequently, 438 conserved protein sequences were obtained. The RCSB-PDB database was consulted to check the template structures for these conserved proteins and as a result, 401 adequate 3D models were obtained. We subsequently predicted the protein pockets for the obtained set of models and kept only the conserved pockets that had highly druggable (HD) values (137 across all strains). Later, an off-target host homology analyses was performed considering the human proteome using NCBI database. Furthermore, the gene essentiality analysis was carried out that gave a final set of 10-conserved targets possessing highly druggable protein pockets. To check the target identification robustness of the pipeline used in this work, we crosschecked the final target list with another in-house target identification approach for C. diphtheriae thereby obtaining three common targets, these were; hisE-phosphoribosyl-ATP pyrophosphatase, glpX-fructose 1,6-bisphosphatase II, and rpsH-30S ribosomal protein S8. Our predicted results suggest that the in silico approach used could potentially aid in experimental polypharmacological target determination in C. diphtheriae and other pathogens, thereby, might complement the existing and new drug-discovery pipelines.

16.
Front Microbiol ; 8: 1937, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29075239

RESUMEN

This review gathers recent information about genomic and transcriptomic studies in the Corynebacterium genus, exploring, for example, prediction of pathogenicity islands and stress response in different pathogenic and non-pathogenic species. In addition, is described several phylogeny studies to Corynebacterium, exploring since the identification of species until biological speciation in one species belonging to the genus Corynebacterium. Important concepts associated with virulence highlighting the role of Pld protein and Tox gene. The adhesion, characteristic of virulence factor, was described using the sortase mechanism that is associated to anchorage to the cell wall. In addition, survival inside the host cell and some diseases, were too addressed for pathogenic corynebacteria, while important biochemical pathways and biotechnological applications retain the focus of this review for non-pathogenic corynebacteria. Concluding, this review broadly explores characteristics in genus Corynebacterium showing to have strong relevance inside the medical, veterinary, and biotechnology field.

17.
Sci Rep ; 7(1): 13538, 2017 10 19.
Artículo en Inglés | MEDLINE | ID: mdl-29051505

RESUMEN

Streptococcus agalactiae is a major pathogen and a hindrance on tilapia farming worldwide. The aims of this work were to analyze the genomic evolution of Brazilian strains of S. agalactiae and to establish spatial and temporal relations between strains isolated from different outbreaks of streptococcosis. A total of 39 strains were obtained from outbreaks and their whole genomes were sequenced and annotated for comparative analysis of multilocus sequence typing, genomic similarity and whole genome multilocus sequence typing (wgMLST). The Brazilian strains presented two sequence types, including a newly described ST, and a non-typeable lineage. The use of wgMLST could differentiate each strain in a single clone and was used to establish temporal and geographical correlations among strains. Bayesian phylogenomic analysis suggests that the studied Brazilian population was co-introduced in the country with their host, approximately 60 years ago. Brazilian strains of S. agalactiae were shown to be heterogeneous in their genome sequences and were distributed in different regions of the country according to their genotype, which allowed the use of wgMLST analysis to track each outbreak event individually.


Asunto(s)
Enfermedades de los Peces/microbiología , Genoma Bacteriano , Streptococcus agalactiae/genética , Animales , Teorema de Bayes , Brasil , ADN/química , ADN/aislamiento & purificación , ADN/metabolismo , Evolución Molecular , Enfermedades de los Peces/patología , Explotaciones Pesqueras , Genotipo , Tipificación de Secuencias Multilocus , Filogenia , Serogrupo , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/patología , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae/clasificación , Streptococcus agalactiae/aislamiento & purificación
18.
Front Microbiol ; 8: 1492, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28848512

RESUMEN

Gram-positive cocci, such as Streptococcus agalactiae, Lactococcus garvieae, Streptococcus iniae, and Streptococcus dysgalactiae subsp. dysgalactiae, are found throughout the world, particularly in outbreaks in farmed fish, and are thus associated with high economic losses, especially in the cultivation of Nile Tilapia. The aim of this study was to evaluate the efficacy of matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) as an alternative for the diagnosis of these pathogens. One hundred and thirty-one isolates from Brazilian outbreaks assisted by the national authority were identified using a MALDI Biotyper from Bruker Daltonics. The results showed an agreement with respect to identification (Kappa = 1) between this technique and 16S ribosomal RNA gene sequencing for S. agalactiae and L. garvieae. However, for S. iniae and S. dysgalactiae subsp. dysgalactiae, perfect agreement was only achieved after the creation of a custom main spectra profile, as well as further comparisons with 16S ribosomal RNA and multilocus sequence analysis. MALDI-TOF MS was shown to be an efficient technology for the identification of these Gram-positive pathogens, yielding a quick and precise diagnosis.

19.
Artículo en Inglés | MEDLINE | ID: mdl-28791255

RESUMEN

Corynebacterium pseudotuberculosis biovar equi is the etiologic agent of ulcerative lymphangitis. To investigate proteins that could be related to the virulence of this pathogen, we combined an experimental passage process using a murine model and high-throughput proteomics with a mass spectrometry, data-independent acquisition (LC-MSE) approach to identify and quantify the proteins released into the supernatants of strain 258_equi. To our knowledge, this approach allowed characterization of the exoproteome of a C. pseudotuberculosis equi strain for the first time. Interestingly, the recovery of this strain from infected mouse spleens induced a change in its virulence potential, and it became more virulent in a second infection challenge. Proteomic screening performed from culture supernatant of the control and recovered conditions revealed 104 proteins that were differentially expressed between the two conditions. In this context, proteomic analysis of the recovered condition detected the induction of proteins involved in bacterial pathogenesis, mainly related to iron uptake. In addition, KEGG enrichment analysis showed that ABC transporters, bacterial secretion systems and protein export pathways were significantly altered in the recovered condition. These findings show that secretion and secreted proteins are key elements in the virulence and adaptation of C. pseudotuberculosis. Collectively, bacterial pathogenesis-related proteins were identified that contribute to the processes of adherence, intracellular growth and evasion of the immune system. Moreover, this study enhances our understanding of the factors that may influence the pathogenesis of C. pseudotuberculosis.


Asunto(s)
Proteínas Bacterianas/metabolismo , Infecciones por Corynebacterium/microbiología , Corynebacterium pseudotuberculosis/aislamiento & purificación , Medios de Cultivo/química , Proteoma/análisis , Animales , Cromatografía Liquida , Corynebacterium pseudotuberculosis/crecimiento & desarrollo , Modelos Animales de Enfermedad , Ensayos Analíticos de Alto Rendimiento , Espectrometría de Masas , Ratones , Proteómica
20.
PLoS One ; 12(4): e0175116, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28384209

RESUMEN

Lactococcus lactis subsp. lactis NCDO 2118 was recently reported to alleviate colitis symptoms via its anti-inflammatory and immunomodulatory activities, which are exerted by exported proteins that are not produced by L. lactis subsp. lactis IL1403. Here, we used in vitro and in silico approaches to characterize the genomic structure, the safety aspects, and the immunomodulatory activity of this strain. Through comparative genomics, we identified genomic islands, phage regions, bile salt and acid stress resistance genes, bacteriocins, adhesion-related and antibiotic resistance genes, and genes encoding proteins that are putatively secreted, expressed in vitro and absent from IL1403. The high degree of similarity between all Lactococcus suggests that the Symbiotic Islands commonly shared by both NCDO 2118 and KF147 may be responsible for their close relationship and their adaptation to plants. The predicted bacteriocins may play an important role against the invasion of competing strains. The genes related to the acid and bile salt stresses may play important roles in gastrointestinal tract survival, whereas the adhesion proteins are important for persistence in the gut, culminating in the competitive exclusion of other bacteria. Finally, the five secreted and expressed proteins may be important targets for studies of new anti-inflammatory and immunomodulatory proteins. Altogether, the analyses performed here highlight the potential use of this strain as a target for the future development of probiotic foods.


Asunto(s)
Genes Bacterianos , Lactococcus lactis/genética , Probióticos , Estrés Fisiológico , Ácidos y Sales Biliares/farmacología , Farmacorresistencia Bacteriana , Técnicas In Vitro , Lactococcus lactis/efectos de los fármacos
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...