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Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for causing Coronavirus Disease-2019 (COVID-19), a heterogeneous clinical condition that manifests varying symptom severity according to the demographic profile of the studied population. While many studies have focused on the spread of COVID-19 in large urban centers in Brazil, few have evaluated medium or small cities in the Northeast region. The aims of this study were: (i) to identify risk factors for mortality from SARS-CoV-2 infection, (ii) to evaluate the gene expression patterns of key immune response pathways using nasopharyngeal swabs of COVID-19 patients, and (iii) to identify the circulating SARS-CoV-2 variants in the residents of a medium-sized city in Northeast Brazil. A total of 783 patients infected with SARS-CoV-2 between May 2020 and August 2021 were included in this study. Clinical-epidemiological data from patients who died and those who survived were compared. Patients were also retrospectively divided into three groups based on disease severity: asymptomatic, mild, and moderate/severe. Samples were added to a qPCR array for analyses of 84 genes involved with immune response pathways and sequenced using the Oxford Nanopore MinION technology. Having pre-existing comorbidity; being male; having cardiovascular disease, diabetes, and/or chronic obstructive pulmonary disease; and PCR cycle threshold (Ct) values under 22 were identified as risk factors for mortality. Analysis of the expression profiles of inflammatory pathway genes showed that the greater the infection severity, the greater the activation of inflammatory pathways, triggering the cytokine storm and downregulating anti-inflammatory pathways. Viral genome analysis revealed the circulation of multiple lineages, such as B.1, B.1.1.28, Alpha, and Gamma, suggesting that multiple introduction events had occurred over time. This study's findings help identify the specific strains and increase our understanding of the true state of local health. In addition, our data demonstrate that epidemiological and genomic surveillance together can help formulate public health strategies to guide governmental actions.
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COVID-19 , SARS-CoV-2 , Humanos , Masculino , Femenino , SARS-CoV-2/genética , COVID-19/epidemiología , Estudios Retrospectivos , Brasil/epidemiologíaRESUMEN
The present study evaluated the gut microbiota profiles of 40 women and correlated them with their nutritional, inflammatory, and hormonal profiles. Stool and blood samples were collected, and anthropometric measurements were obtained from 20 women diagnosed with obesity ("case" group) and 20 women with weight in the normal range ("control" group). Bacteria belonging to two phyla, Firmicutes and Bacteroidetes, one class, Mollicutes, and four genera were evaluated by real-time polymerase chain reaction. Levels of 18 inflammatory cytokines were measured using the Luminex assay, and ghrelin and leptin levels were measured using enzymatic immunoadsorption assay. Mollicutes proportion differed significantly between the case and control groups, and a significant positive association was detected between the presence of Mollicutes and obesity. Statistically significant differences were observed between the proportions of Firmicutes and Bacteroidetes in the two groups, with a higher proportion of Firmicutes/Bacteroidetes ratio among the gut microbiota of women in the case group compared to those of the control group. Higher counts of Escherichia coli and Clostridium spp. were observed in the control group than in the case group, whereas higher counts of Lactobacillus spp. and Bacteroides spp. were detected in the case group than in the control group. There was a positive correlation between interleukin-6 (IL-6) and interferon-γ (IFN-γ) levels and the anthropometric variables and a negative correlation between IL-10 and these variables. Leptin and ghrelin concentrations differed significantly between the two groups and showed positive and negative correlation with obesity predictors, respectively. Therefore, gut microbiota was associated with obesity in women from this study group. Moreover, this microbiota was associated with inflammatory profiles and alterations in ghrelin and leptin levels.
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Leptina , Microbiota , Bacteroidetes , Heces/microbiología , Femenino , Ghrelina , Humanos , Obesidad/microbiologíaRESUMEN
Brazil has a herd of over 9 million goats, and the northeast of Brazil is home to over 93% of this herd. Caprine mycoplasmosis are widely disseminated worldwide, being highly contagious with high rates of morbidity and mortality, causing considerable economic loss to goat herders. In addition, there has been a lack of research using molecular testing to monitor the health and detect Mollicutes in this herd in Brazil. Therefore, the aim of this study is to associate animal management with the presence of the caprine origin Mollicutes in goats, in the southwest region of the state of Bahia, Brazil. A cross-sectional study was conducted on twelve farms, and statistical analyses were performed to identify associations between the presence of Mollicutes and the management of goats. Molecular testing identified Mollicutes class, Mycoplasma agalactiae (Ma) and M. conjunctivae (Mc), in the samples analyzed. Statistical associations were observed between animals from intensive livestock facilities and the presence of Mollicutes in nasal samples and dairy ranch animals and the presence of Mollicutes in ocular samples and animals from extensive ranching sites and positive results of Mollicutes in genital samples. We conclude that mycoplasmas are present in goat herds in the southwestern region of Bahia, which supports the need for more focused studies of mycoplasmas throughout the country. Our research also demonstrated the presence of two important opportunistic bacteria, Mc and Ma, and, to the best of our knowledge, this is the first time that M. conjunctivae was detected in Brazilian goats by molecular testing.
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Enfermedades de las Cabras , Infecciones por Mycoplasma , Mycoplasma agalactiae , Mycoplasma conjunctivae , Animales , Brasil/epidemiología , Estudios Transversales , Enfermedades de las Cabras/diagnóstico , Enfermedades de las Cabras/epidemiología , Cabras , Infecciones por Mycoplasma/epidemiología , Infecciones por Mycoplasma/veterinaria , Reacción en Cadena de la PolimerasaRESUMEN
BACKGROUND: Ureaplasma diversum is a pathogen found in the genital tract of cattle and associated with genital disorders such as infertility, placentitis, abortion, birth of weak calves, low sperm motility, seminal vesiculitis and epididymitis. There are few studies evaluating the genetic diversity of U. diversum strains and their influence on the immune response in cattle. Therefore, to better understand genetic relationships of the pathogenicity of U. diversum, a multilocus sequence typing (MLST) scheme was performed to characterize the ATCC 49782 strain and another 40 isolates recovered from different Brazilian states. RESULTS: Primers were designed for housekeeping genes ftsH, polC, rpL22, rpoB, valS and ureA and for virulence genes, phospholipase D (pld), triacylglycerol lipase (tgl), hemolysin (hlyA), MIB-MIP system (mib,mip), MBA (mba), VsA (VsA) and ribose transporter (tABC). PCRs were performed and the targeted gene products were purified and sequenced. Sequence types (STs), and clonal complexes (CCs) were assigned and the phylogenetic relationship was also evaluated. Thus, a total of 19 STs and 4 CCs were studied. Following the molecular analysis, six isolates of U. diversum were selected, inoculated into bovine monocyte/macrophage culture and evaluated for gene expression of the cytokines TNF-α, IL-1, IL-6, IL-10 and IL-17. Differences were detected in the induction of cytokines, especially between isolates 198 and BA78, promoted inflammatory and anti-inflammatory profiles, respectively, and they also differed in virulence factors. CONCLUSION: It was observed that intra-species variability between isolates of U. diversum can induce variations of virulent determinants and, consequently, modulate the expression of the triggered immune response.
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Infecciones por Ureaplasma/veterinaria , Ureaplasma/genética , Ureaplasma/inmunología , Animales , Brasil , Bovinos , Enfermedades de los Bovinos/microbiología , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Femenino , Masculino , Tipificación de Secuencias Multilocus/veterinaria , Filogenia , Ureaplasma/clasificación , Ureaplasma/patogenicidad , Infecciones por Ureaplasma/inmunología , Virulencia/genéticaRESUMEN
BACKGROUND: Staphylococcus aureus is a human pathogen of clinical importance related to a variety of infections. AIM: The objective of this study was to analyze the molecular and epidemiological characteristics of S. aureus obtained from healthcare professionals (HCP) of a hospital in southwestern Bahia, Brazil. METHODS: Samples were collected from hands, nasal cavity, and laboratory coats of 80 HCP. The bacterial isolates recovered from 240 samples were identified as S. aureus, and then analyzed for their antimicrobial resistance profile, genotypic characterization, and pathogenicity. FINDINGS: 178 isolates were identified as S. aureus, being mostly isolated from the nasal cavity. Thirty isolates (16.8%) were characterized as MRSA. The virulence gene frequency varied according to isolate source. All virulence genes were identified in at least one hand isolate. Isolates from laboratory coats did not show seb and pvl. Isolates from the nasal cavity did not exhibit pvl. The SCCmec type I was identified in 56.7% of MRSA isolates. Among MRSA isolates, 14 PFGE pulsotypes were characterized, with profile A being predominant (nine isolates). Clonal complexes CC5, CC45, and CC398 were found. MRSA isolates induced cytokine gene expression in macrophages, with IL-10 and IL-17 being expressed more often. CONCLUSION: We found a high colonization rate for S. aureus among HCP. Moreover, we observed that MRSA strains presented different virulence factors and could induce cytokine gene expression, indicating an urgent need to control colonization rates of HCP by MRSA isolates in order to protect hospital patients and the general public.
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PURPOSE: Comparative genomic analysis of strains may help us to better understand the wide diversity of their genetic profiles. The aim of this study was to analyse the genomic features of the resistome and virulome of Brazilian first methicillin-resistant Staphylococcus aureus (MRSA) isolates and their relationship to other Brazilian and international MRSA strains. METHODOLOGY: The whole genomes of three MRSA strains previously isolated in Vitória da Conquista were sequenced, assembled, annotated and compared with other MRSA genomes. A phylogenetic tree was constructed and the pan-genome and accessory and core genomes were constructed. The resistomes and virulomes of all strains were identified.Results/Key findings. Phylogenetic analysis of all 49 strains indicated different clones showing high similarity. The pan-genome of the analysed strains consisted of 4484 genes, with 31 % comprising the gene portion of the core genome, 47 % comprising the accessory genome and 22 % being singletons. Most strains showed at least one gene related to virulence factors associated with immune system evasion, followed by enterotoxins. The strains showed multiresistance, with the most recurrent genes conferring resistance to beta-lactams, fluoroquinolones, aminoglycosides and macrolides. CONCLUSIONS: Our comparative genomic analysis showed that there is no pattern of virulence gene distribution among the clones analysed in the different regions. The Brazilian strains showed similarity with clones from several continents.
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Genoma Bacteriano , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/virología , Filogenia , Factores de Virulencia/genética , Antibacterianos/farmacología , Brasil , Enterotoxinas/genética , Fluoroquinolonas/farmacología , Genómica , Humanos , Macrólidos/farmacologíaRESUMEN
Objectives:Ureaplasma diversum is a pathogen of cows that may cause intense inflammatory responses in the reproductive tract and interfere with bovine reproduction. The aims of this study were to evaluate the immune response of bovine blastocysts and macrophages to U. diversum infection and to evaluate the invasion capacity of this microorganism in bovine blastocysts. Methods: Viable and heat-inactivated U. diversum strains ATCC 49782 and CI-GOTA and their extracted membrane lipoproteins were inoculated in macrophages in the presence or absence of signaling blockers of Toll-Like Receptor (TLR) 4, TLR2/4, and Nuclear Factor KB (NF-κB). In addition, the same viable U. diversum strains were used to infect bovine blastocysts. RNA was extracted from infected and lipoprotein-exposed macrophages and infected blastocysts and assayed by qPCR to evaluate the expression of Interleukin 1 beta (IL-1ß), Tumor Necrosis Factor Alpha (TNF-α), TLR2 and TLR4 genes. U. diversum internalization in blastocysts was followed by confocal microscopy. Results: Both Ureaplasma strains and different concentrations of extracted lipoproteins induced a higher gene expression of IL-1ß, TNF-α, TLR2, and TLR4 in macrophages (p < 0.05) when compared to non-infected cells. The used blockers inhibited the expression of IL-1ß and TNF-α in all treatments. Moreover, U. diversum was able to internalize within blastocysts and induce a higher gene expression of IL-1b and TNF- α when compared to non-infected blastocysts (p < 0.05). Conclusion: The obtained results strongly suggest that U. diversum and its lipoproteins interact with TLR4 in a signaling pathway acting via NF-kB signaling to stimulate the inflammatory response. This is the first study to evaluate the in vitro immunological response of macrophages and bovine blastocysts against U. diversum. These results may contribute to a better understanding of the immunomodulatory activity and pathogenicity of this infectious agent.
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OBJECTIVE: To detect Mollicutes in women with endometriosis and healthy peritoneal tissues and evaluate the participation of these bacteria in the immune response during endometriosis. DESIGN: Cross-sectional study. SETTING: University hospitals. PATIENT (S): Women with endometriosis (n = 73) and without endometriosis (n = 31). INTERVENTION(S): Endocervical swabs, peritoneal fluid, and biopsied lesions of endometriosis of women with endometriosis (study group) and healthy peritoneal tissues (control group) were collected during surgery. Clinical characteristics were registered before surgery. MAIN OUTCOME MEASURE(S): We determined the infectious agents with the use of quantitative polymerase chain reaction (PCR). The cytokine secretion profile was determined with the use of Luminex. The expression of immune response related genes was determined with the use of a PCR array kit. RESULT(S): All target microorganisms were detected at least once in the swab samples analyzed. It was possible to observe higher diversity of microorganisms in the samples of swab and peritoneal fluid in the study group compared with the control. Ureaplasma parvum was associated with the severity of the symptom dyspareunia. Mycoplasma genitalium was associated with higher production of interferon-γ and interleukin-1ß. Genes of inflammatory response activation and antigen presentation were up-regulated in biopsied tissue of women with endometriosis. In women with endometriosis, peritoneal fluid cells showed a down-regulation of genes associated with the inflammatory response. This down-regulation profile was higher in presence of M. genitalium. CONCLUSION(S): Mycoplasma genitalium may play a key role in the immune tolerance process and, especially, the aggravation of this profile. More studies are needed to understand this immune tolerance profile of bacterial infections.
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Endometriosis/inmunología , Endometriosis/microbiología , Tolerancia Inmunológica , Infecciones por Mycoplasma/inmunología , Infecciones por Mycoplasma/microbiología , Mycoplasma genitalium/inmunología , Adolescente , Adulto , Líquido Ascítico/inmunología , Líquido Ascítico/microbiología , Estudios de Casos y Controles , Cuello del Útero/inmunología , Cuello del Útero/microbiología , Estudios Transversales , Endometriosis/genética , Endometriosis/metabolismo , Femenino , Regulación de la Expresión Génica , Hospitales Universitarios , Interacciones Huésped-Patógeno , Humanos , Tolerancia Inmunológica/genética , Mediadores de Inflamación/metabolismo , Interferón gamma/genética , Interferón gamma/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Persona de Mediana Edad , Infecciones por Mycoplasma/genética , Infecciones por Mycoplasma/metabolismo , Mycoplasma genitalium/patogenicidad , Adulto JovenRESUMEN
Abstract Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) have increasingly been reported in healthy communities. This study aimed to assess the rate of S. aureus in general and MRSA in particular from nasal secretion of children in daycare centers in Vitória da Conquista, Brazil. The isolates were identified based on morphology, biochemical tests and by PCR. Detection of virulence genes, biofilm production, and susceptibility test by disk diffusion agar were performed. MRSA isolates were characterized by spa, SCCmec, and multilocus sequence typing (MLST). S. aureus were recovered from 70 (47.3%) of 148 children. Among the 11 MRSA strains (15.7%), two SCCmec types (IV and V) were detected. MLST identified four STs related to three clonal complexes (CC): 5, 45, and 398. Four spa types were found circulating in this setting. Resistance of S. aureus isolates to ampicillin, erythromycin, ciprofloxacin, clindamycin, and tetracycline was 80%, 32.8%, 7.1%, 7.1% and 4.3%, respectively. One isolate presented intermediate resistance to vancomycin detected by Etest methodology. All strains were biofilm producers. The virulence genes seb, sec, spa, and pvl were detected in some isolates. This study revealed a high rate of children carrying MRSA among healthy attendees in daycare centers in Vitória da Conquista, Brazil.
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Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Infecciones Estafilocócicas/microbiología , Guarderías Infantiles , Nariz/microbiología , Infecciones Comunitarias Adquiridas/microbiología , Biopelículas/crecimiento & desarrollo , Factores de Virulencia , Pruebas Antimicrobianas de Difusión por Disco , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Genotipo , Antibacterianos/farmacologíaRESUMEN
Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) have increasingly been reported in healthy communities. This study aimed to assess the rate of S. aureus in general and MRSA in particular from nasal secretion of children in daycare centers in Vitória da Conquista, Brazil. The isolates were identified based on morphology, biochemical tests and by PCR. Detection of virulence genes, biofilm production, and susceptibility test by disk diffusion agar were performed. MRSA isolates were characterized by spa, SCCmec, and multilocus sequence typing (MLST). S. aureus were recovered from 70 (47.3%) of 148 children. Among the 11 MRSA strains (15.7%), two SCCmec types (IV and V) were detected. MLST identified four STs related to three clonal complexes (CC): 5, 45, and 398. Four spa types were found circulating in this setting. Resistance of S. aureus isolates to ampicillin, erythromycin, ciprofloxacin, clindamycin, and tetracycline was 80%, 32.8%, 7.1%, 7.1% and 4.3%, respectively. One isolate presented intermediate resistance to vancomycin detected by Etest methodology. All strains were biofilm producers. The virulence genes seb, sec, spa, and pvl were detected in some isolates. This study revealed a high rate of children carrying MRSA among healthy attendees in daycare centers in Vitória da Conquista, Brazil.
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Guarderías Infantiles , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Nariz/microbiología , Infecciones Estafilocócicas/microbiología , Antibacterianos/farmacología , Biopelículas/crecimiento & desarrollo , Niño , Preescolar , Infecciones Comunitarias Adquiridas/microbiología , Pruebas Antimicrobianas de Difusión por Disco , Femenino , Genotipo , Humanos , Lactante , Masculino , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Factores de VirulenciaRESUMEN
Whole genome sequencing and analyses of Ureaplasma diversum ATCC 49782 was undertaken as a step towards understanding U. diversum biology and pathogenicity. The complete genome showed 973,501 bp in a single circular chromosome, with 28.2% of G+C content. A total of 782 coding DNA sequences (CDSs), and 6 rRNA and 32 tRNA genes were predicted and annotated. The metabolic pathways are identical to other human ureaplasmas, including the production of ATP via hydrolysis of the urea. Genes related to pathogenicity, such as urease, phospholipase, hemolysin, and a Mycoplasma Ig binding protein (MIB)-Mycoplasma Ig protease (MIP) system were identified. More interestingly, a large number of genes (n = 40) encoding surface molecules were annotated in the genome (lipoproteins, multiple-banded antigen like protein, membrane nuclease lipoprotein and variable surface antigens lipoprotein). In addition, a gene encoding glycosyltransferase was also found. This enzyme has been associated with the production of capsule in mycoplasmas and ureaplasma. We then sought to detect the presence of a capsule in this organism. A polysaccharide capsule from 11 to 17 nm of U. diversum was observed trough electron microscopy and using specific dyes. This structure contained arabinose, xylose, mannose, galactose and glucose. In order to understand the inflammatory response against these surface molecules, we evaluated the response of murine macrophages J774 against viable and non-viable U. diversum. As with viable bacteria, non-viable bacteria were capable of promoting a significant inflammatory response by activation of Toll like receptor 2 (TLR2), indicating that surface molecules are important for the activation of inflammatory response. Furthermore, a cascade of genes related to the inflammasome pathway of macrophages was also up-regulated during infection with viable organisms when compared to non-infected cells. In conclusion, U. diversum has a typical ureaplasma genome and metabolism, and its surface molecules, including the identified capsular material, represent major components of the organism immunopathogenesis.
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Genoma Bacteriano/genética , Interacciones Huésped-Patógeno/genética , Infecciones por Ureaplasma/genética , Ureaplasma/genética , Composición de Base/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Inflamasomas/genética , Lipoproteínas/genética , Redes y Vías Metabólicas/genética , Anotación de Secuencia Molecular , Mycoplasma/genética , Mycoplasma/patogenicidad , Fosfolipasas/genética , Receptores Toll-Like/genética , Ureaplasma/patogenicidad , Infecciones por Ureaplasma/microbiología , Infecciones por Ureaplasma/patología , Ureasa/genéticaRESUMEN
Here, we report the complete genome sequence of Ureaplasma diversum strain ATCC 49782. This species is of bovine origin, having an association with reproductive disorders in cattle, including placentitis, fetal alveolitis, abortion, and birth of weak calves. It has a small circular chromosome of 975,425 bp.
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Currently, hospital infection is a serious public health problem, and several factors may influence the occurrence of these infections, including the presence of insects, which are carriers of multidrug-resistant bacterial species. The aim of this study was to isolate staphylococci carried by insects in two public hospitals of Vitoria da Conquista, Bahia and to identify the resistance profile, pathogenicity and efficacy of disinfection of the premises. A total of 91 insects were collected in 21 strategic points of these hospitals, and 32 isolated strains ofStaphylococcus aureus were isolated. Based on antibiogram and Minimum Inhibitory Concentration results, 95% of these strains were susceptible to oxacillin. These strains were also evaluated for the presence of resistance genes encoding resistance to oxacillin/methicillin by polymerase chain reaction, but the sample was negative for this gene. Pathogenicity tests were performed in vitro biofilm formation induced by glucose, where it was found that eight (27.58%) strains were classified as biofilm producers and 21 (72.4%) as stronger producers. In addition, we performed PCR for their virulence genes: Sea (enterotoxin A), SEB (B), Sec (C), PVL (Panton-Valentine Leukocidin), ClfA (clumping factor A) and Spa (protein A). Of these, Sea, Spa PVL were positive in 7 (21.8%), 2 (6.3%) and 1 (3.1%) samples, respectively. The analysis of cytokine induction in the inflammatory response of J774 macrophages by isolates from the two hospitals did not show statistical difference at the levels of IL-6, TNF-α, IL-1 and IL-10 production. In addition, we verified the antimicrobial activity of disinfecting agents on these strains, quaternary ammonium, 0.5% sodium hypochlorite, 1% sodium hypochlorite, 2% sodium hypochlorite, 2% glutaraldehyde, Lysoform®, 70% alcohol solution of chlorhexidine digluconate, 2% peracetic acid, and 100% vinegar. Resistance was seen in only for the following two disinfectants: 70% alcohol in 31 (96.8%) samples tested and vinegar in 30 (93.8%) samples. The study demonstrated the presence of resistant and pathogenic organisms conveyed by insects, thus suggesting improvement in efforts to control these vectors.
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Animales , Antibacterianos/farmacología , Desinfección/métodos , Insectos/microbiología , Staphylococcus aureus , Brasil , Biopelículas/crecimiento & desarrollo , ADN Bacteriano/genética , Genotipo , Hospitales Públicos , Insectos/clasificación , Pruebas de Sensibilidad Microbiana , Resistencia a la Meticilina/genética , Reacción en Cadena de la Polimerasa , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/aislamiento & purificación , Staphylococcus aureus/patogenicidad , Staphylococcus aureus/fisiología , Factores de Virulencia/genéticaRESUMEN
Currently, hospital infection is a serious public health problem, and several factors may influence the occurrence of these infections, including the presence of insects, which are carriers of multidrug-resistant bacterial species. The aim of this study was to isolate staphylococci carried by insects in two public hospitals of Vitoria da Conquista, Bahia and to identify the resistance profile, pathogenicity and efficacy of disinfection of the premises. A total of 91 insects were collected in 21 strategic points of these hospitals, and 32 isolated strains of Staphylococcus aureus were isolated. Based on antibiogram and Minimum Inhibitory Concentration results, 95% of these strains were susceptible to oxacillin. These strains were also evaluated for the presence of resistance genes encoding resistance to oxacillin/methicillin by polymerase chain reaction, but the sample was negative for this gene. Pathogenicity tests were performed in vitro biofilm formation induced by glucose, where it was found that eight (27.58%) strains were classified as biofilm producers and 21 (72.4%) as stronger producers. In addition, we performed PCR for their virulence genes: Sea (enterotoxin A), SEB (B), Sec (C), PVL (Panton-Valentine Leukocidin), ClfA (clumping factor A) and Spa (protein A). Of these, Sea, Spa PVL were positive in 7 (21.8%), 2 (6.3%) and 1 (3.1%) samples, respectively. The analysis of cytokine induction in the inflammatory response of J774 macrophages by isolates from the two hospitals did not show statistical difference at the levels of IL-6, TNF-α, IL-1 and IL-10 production. In addition, we verified the antimicrobial activity of disinfecting agents on these strains, quaternary ammonium, 0.5% sodium hypochlorite, 1% sodium hypochlorite, 2% sodium hypochlorite, 2% glutaraldehyde, Lysoform(®), 70% alcohol solution of chlorhexidine digluconate, 2% peracetic acid, and 100% vinegar. Resistance was seen in only for the following two disinfectants: 70% alcohol in 31 (96.8%) samples tested and vinegar in 30 (93.8%) samples. The study demonstrated the presence of resistant and pathogenic organisms conveyed by insects, thus suggesting improvement in efforts to control these vectors.
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Antibacterianos/farmacología , Desinfección/métodos , Insectos/microbiología , Staphylococcus aureus , Animales , Biopelículas/crecimiento & desarrollo , Brasil , ADN Bacteriano/genética , Genotipo , Hospitales Públicos , Insectos/clasificación , Resistencia a la Meticilina/genética , Pruebas de Sensibilidad Microbiana , Reacción en Cadena de la Polimerasa , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/aislamiento & purificación , Staphylococcus aureus/patogenicidad , Staphylococcus aureus/fisiología , Factores de Virulencia/genéticaRESUMEN
Ureaplasma diversum in veterinary studies is an undesirable microbe, which may cause infection in bulls and may result in seminal vesiculitis, balanopostitis, and alterations in spermatozoids, whereas in cows, it may cause placentitis, fetal alveolitis, abortion, and birth of weak calves. U. diversum is released through organic secretions, especially semen, preputial and vaginal mucus, conjunctival secretion, and milk. The aim of the present study was to develop a TaqMan probe, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of U. diversum from genital swabs of bovines. Primers and probes specific to U. diversum 16S rRNA gene were designed. The specificity, detection limit, intra- and inter-assay variability of qPCR to detect this ureaplasma was compared with the results of the conventional PCR assay (cPCR). Swabs of vaginal mucus from 169 cows were tested. The qPCR assay detected as few as 10 copies of U. diversum and was 100-fold more sensitive than the cPCR. No cross-reactivity with other Mollicutes or eubacteria was observed. U. diversum was detected in 79 swabs (46.42%) by qPCR, while using cPCR it was detected in 42 (25%) samples. The difference in cPCR and qPCR ureaplasma detection between healthy and sick animals was not statistically significant. But the U. diversum load in samples from animals with genital disorders was higher than in healthy animals. The qPCR assay developed herein is highly sensitive and specific for the detection and quantification of U. diversum in vaginal bovine samples.
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Enfermedades de los Bovinos/diagnóstico , Reacción en Cadena de la Polimerasa/veterinaria , Infecciones por Ureaplasma/veterinaria , Ureaplasma/genética , Animales , Bovinos , Femenino , ARN Ribosómico 16S/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Infecciones por Ureaplasma/diagnóstico , Vagina/microbiologíaRESUMEN
The aim of the present study was to isolate S. aureus strains resistant to antibiotics, characterize the genotype profiles of resistance staphylococci, and evaluate the efficacy of antiseptic agents and disinfectants used in two public hospitals of Vitoria da Conquista, Bahia, Brazil. Clinical samples were obtained from ICU environments and equipment surfaces in two public hospitals in Vitoria da Conquista. Broth cultures were plated onto mannitol salt agar, and antimicrobial susceptibility testing was performed by the broth microdilution method according to CLSI. MRSA strains were submitted to PCR for detecting the mecA gene. PCR products were purified and sequenced for SCCmec type identification. Moreover, the strains were tested for efficacy of different disinfectant solutions. S. aureus were isolated from 31 and 67 sites in each hospital, respectively. Among the isolates from hospital 1, 07 (22.6%) were resistant to oxacillin while 28 (41.8%) were resistant in hospital 2. Thirty-one were positive for the mecA gene. All isolates showed SCCmec type III genotype characteristics of the Brazilian epidemic clone. In disinfectant tests, sodium hypochlorite (0.5, 1.0 and 2.0%), 2% chlorhexidine gluconate, quaternary ammonium, peracetic acid and formaldehyde were effective against the isolates tested. The strains showed higher resistance to vinegar (4% acetic acid), alcohol and glutaraldehyde. The findings of this study should assist in reducing the occurrence of nosocomial infections and therefore the morbidity, mortality and socio-economic burden caused by prolonged hospitalization.