BackgroundThe role of Mycoplasma hominis and M. genitalium in urogenital tract infections remains unknown. Furthermore these mollicutes present a complex relationship with the host immune response. The role of inflammatory cytokines in infections also makes them good candidates to investigate bacterial vaginosis and mycoplasma genital infections. Therefore, the aim of this study was to detect the above-mentioned mollicutes by quantitative Polymerase Chain Reaction (qPCR) methodologies in vaginal swabs and dosage of cytokines.MethodsVaginal swabs and peripheral blood were collected from 302 women, including healthy individuals. The molecular findings were correlated with some individual behavioral variables, clinical and demographic characteristics, presence of other important microorganisms in vaginal swabs, and levels of interleukin (IL)-1β and IL-6.ResultsM. hominis and M. genitalium were detected in 31.8% and 28.1% of samples, respectively. The qPCR results were associated with clinical signs and symptoms of the infections studied. The frequency of Trichomonas vaginalis, Gardnerella vaginalis, Neisseria gonorrhoeae and Chlamydia trachomatis was 3.0%, 21.5%, 42.4%, and 1.7% respectively. Increased levels of IL-1β were associated with the presence of M. hominis and signs and/or symptoms of the genital infection of women studied.ConclusionIL-1β production was associated with the detection of M. hominis by qPCR. The sexual behavior of women studied was associated with the detection of mycoplasma and other agents of genital infections.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-015-0792-4) contains supplementary material, which is available to authorized users.

BackgroundBacterial pathogens have many strategies for infecting and persisting in host cells. Adhesion, invasion and intracellular life are important features in the biology of mollicutes. The intracellular location of Ureaplasma diversum may trigger disturbances in the host cell. This includes activation or inhibition of pro and anti-apoptotic factors, which facilitate the development of host damage. The aim of the present study was to associate U. diversum infection in HEp-2 cells and apoptosis induction. Cells were infected for 72hs with four U. diversum clinical isolates and an ATCC strain. The U. diversum invasion was analyzed by Confocal Laser Scanning Microscopy and gentamicin invasion assay. The apoptosis was evaluated using pro-apoptotic and anti-apoptotic gene expression, and FITC Annexin V/Dead Cell Apoptosis Kit.ResultsThe number of internalized ureaplasma in HEp-2 cells increased significantly throughout the infection. The flow cytometry analysis with fluorochromes to detect membrane depolarization and gene expression for caspase 2, 3 and 9 increased in infected cells after 24 hours. However, after 72 hours a considerable decrease of apoptotic cells was observed.ConclusionsThe data suggests that apoptosis may be initially induced by some isolates in association with HEp-2 cells, but over time, there was no evidence of apoptosis in the presence of ureaplasma and HEp-2 cells. The initial increase and then decrease in apoptosis could be related to bacterial pathogen-associated molecular pattern (PAMPS). Moreover, the isolates of U. diversum presented differences in the studied parameters for apoptosis. It was also observed that the amount of microorganisms was not proportional to the induction of apoptosis in HEp-2 cells.

Species of Staphylococcus are common in hospital infection (HI). Methicillin resistant S. aureus(MRSA) has also become a serious problem in Brazilian HI. The aim of this study was to characterize the pathogenicity of methicillin-resistant S. aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) isolated in public hospitals. The clinical isolates were obtained from intensive care unit. The MRSA and MSSA strains were genotyped by PCR for detection genes related to virulence factors. Moreover, the strains were tested for biofilm formation and cytokine induction in macrophages. Three strains of MRSA (9.68%) expressed the Sea gene, one (3.23%) Seb, 17 (54.84%) Spa and seven (22.58%) had PVL. Two MSSA strains (2.98%) expressed the Sea gene, three (4.48%) Seb, 18 (26.87%) Spa and 11 (16.42%) showed positive results for the PVL gene. There was no expression of Sec and CflA between MRSA and MSSA strains. Among MRSA and MSSA isolates, none statistical differences were observed in biofilm production. The analysis of cytokine induction in the inflammatory response of J774 macrophages by MRSA and MSSA isolates did not show statistical difference. Understanding the mechanisms of pathogenesis of S. aureus could provide important clues for both preventing and treating infection caused by these organisms.

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.

This study investigates the biofilm formation, presence and distribution of virulence genes and the capacity to induce an inflammatory response in strains of Staphylococcus aureus isolated from milk samples in Bahia, Brazil. A total of 132 samples of raw milk were collected from four dairy farms (designated A to D) located in southwestern Bahia, in the municipality of Vitória da Conquista, from October/2009 to September/2010. After processing of the samples, 94 (71.2%) S. aureus isolates were obtained. These strains were subjected to the antibiogram method MIC (Minimum Inhibitory Concentration). As for the pathogenicity, tests were performed in vitro biofilm formation induced by glucose. Moreover, 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) and analysis of cytokine induction in the inflammatory response of J774 macrophages by exocellular lipoteichoic acid. No isolates were resistant to oxacillin and vancomycin. In biofilm production, 5.31% (5/94) isolates did not produce biofilm, 5.31% (5/94) of the samples were poor producers, 15.96% (15/94) strains were moderate producers, 18.09% (17/94) were producers and 55.32% (55/94) of isolates were strong biofilm producers. One (1.06%) isolate expressed the seb gene, one (1.06%) sec, 18 (19.2%) cflA and 44 (46.8%) had spa. There was no expression of sea and pvl between isolates analyzed. The analysis of cytokine induction in the inflammatory response did not show statistical difference in the levels of IL-6, TNF-α and IL-10 induction. However, there was statistical difference in IL-1 induction between isolates from different farms. Thus, it

In recent years, certain Lactobacillus sp. have emerged in health care as an alternative therapy for various diseases. Based on this, this study is aimed at evaluating in vitro the potential probiotics of five lactobacilli strains isolated from pulp of cupuaçu fruit fermentation against Gardnerella vaginalis and Neisseria gonorrhoeae. Our lactobacilli strains were classified as safe for use in humans, and they were tolerant to heat and pH. Our strains were biofilm producers, while hydrophobicity and autoaggregation varied from 13% to 86% and 13% to 25%, respectively. The coaggregation of lactobacilli used in this study with G. vaginalis and N. gonorrhoeae ranged from 15% to 36% and 32% to 52%, respectively. Antimicrobial activity was present in all tested Lactobacillus strains against both pathogens, and the growth of pathogens in coculture was reduced by the presence of our lactobacilli. Also, all tested lactobacilli reduced the pH of the culture, even in incubation with pathogens after 24 hours. The cell-free culture supernatants (CFCS) of all five lactobacilli demonstrated activity against the two pathogens with a halo presence and CFCS characterization assay together with gas chromatography revealed that lactic acid was the most abundant organic acid in the samples (50% to 62%). Our results demonstrated that the organic acid production profile is strain-specific. This study revealed that cupuaçu is a promising source of microorganisms with probiotic properties against genital pathogens. We demonstrated by in vitro tests that our Lactobacillus strains have probiotic properties. However, the absence of in vivo tests is a limitation of our work due to the need to evaluate the interaction of our lactobacilli with pathogens in the vaginal mucosa. We believe that these findings may be useful in developing a product containing our lactobacilli and their supernatants in order to support with vaginal health.

Significance and impact of the study: Our data suggest that Lactobacillus plantarum Lp62 could be used as a potential treatment against genital infections, especially bacterial vaginosis caused by Gardnerella vaginalis. We hypothesize that this strain has significant therapeutic potential as it topically helps restore vaginal microbiota.