Biofilm formation in drug-resistant Acinetobacter baumannii and Acinetobacter nosocomialis isolates obtained from a university hospital in Pelotas, RS, Brazil.

This study aimed to investigate the antibiotic resistance and biofilm formation of Acinetobacter calcoaceticus-A. baumannii (ACB) complex isolates recovered from a university hospital in Pelotas, RS, Brazil. The species were confirmed using gyrB multiplex and blaOXA-51-like genes PCR. The presence of the bfmRS virulence gene was evaluated by the PCR, and the isolates were classified based on their biofilm-forming ability on polystyrene (PO) and glass surfaces (TM). Out of 50 ACB complex isolates evaluated, 41 were identified as A. baumannii and nine as A. nosocomialis. The bfmRS gene was detected in 97.6% (40/41) of A. baumannii and 33.3% (3/9) of A. nosocomialis species. Forty-nine isolates exhibited a multidrug-resistant (MDR) profile, while one A. nosocomialis isolate presented an extensively drug-resistant (XDR) profile. All isolates were able of forming biofilms on PO surfaces and 98% (49/50) on TM surfaces. A significant correlation was observed between biofilm production on PO and TM surfaces (P < 0.05). However, no correlation was found between biofilms forming and the presence of the bfmRS gene or displaying a certain antibiotic resistance profile. In conclusion, A. baumannii and A. nosocomialis are frequent species causing nosocomial infections in a hospital in Pelotas, RS, Brazil, and both are capable of forming biofilms.

Origanum vulgare essential oil: antibacterial activities and synergistic effect with polymyxin B against multidrug-resistant Acinetobacter baumannii.

Antimicrobial resistance is increasing around the world and the search for effective treatment options, such as new antibiotics and combination therapy is urgently needed. The present study evaluates oregano essential oil (OEO) antibacterial activities against reference and multidrug-resistant clinical isolates of Acinetobacter baumannii (Ab-MDR). Additionally, the combination of the OEO and polymyxin B was evaluated against Ab-MDR. Ten clinical isolates were characterized at the species level through multiplex polymerase chain reaction (PCR) for the gyrB and blaOXA-51-like genes. The isolates were resistant to at least four different classes of antimicrobial agents, namely, aminoglycosides, cephems, carbapenems, and fluoroquinolones. All isolates were metallo-ß-lactamase (MßL) and carbapenemase producers. The major component of OEO was found to be carvacrol (71.0%) followed by ß-caryophyllene (4.0%), γ-terpinene (4.5%), p-cymene (3,5%), and thymol (3.0%). OEO showed antibacterial effect against all Ab-MDR tested, with minimum inhibitory concentrations (MIC) ranging from 1.75 to 3.50 mg mL-1. Flow cytometry demonstrated that the OEO causes destabilization and rupture of the bacterial cell membrane resulting in apoptosis of A. baumannii cells (p < 0.05). Synergic interaction between OEO and polymyxin B (FICI: 0.18 to 0.37) was observed, using a checkerboard assay. When combined, OEO presented until 16-fold reduction of the polymyxin B MIC. The results presented here indicate that the OEO used alone or in combination with polymyxin B in the treatment of Ab-MDR infections is promising. To the best of our knowledge, this is the first report of OEO and polymyxin B association against Ab-MDR clinical isolates.

Molecular Characterization of Carbapenem-Resistant Acinetobacter baumannii Associated with Nosocomial Infection in the Pelotas, RS, Brazil.

Acinetobacter calcoaceticus-Acinetobacter baumannii complex (ACB) comprises some opportunistic pathogens associated with infectious outbreaks in hospital settings. A. baumannii is the most relevant species owing to its capacity to develop resistance to the different classes of antimicrobials. The aim of this study was to identify the species, establish the genetic patterns, resistance and biofilm profiles in ACB isolates associated with nosocomial infection in a hospital of Pelotas, Rio Grande do Sul, Brazil. Twenty-two clinical isolates were characterized at the species level through multiplex polymerase chain reaction (PCR) for the gyrB and blaOXA51-like genes, and the genetic relationship was determined through pulsed-field gel electrophoresis (PFGE). Their antibiotic resistance profiles and carbapenemases synthesis were evaluated following CLSI guidelines. PCR was carried out to evaluate the presence of carbapenemases genes and the isolates were classified for their biofilm-forming ability. All isolates obtained in the study were identified as A. baumannii and 72.7% of the isolates were classified as strong biofilm formers. In the class carbapenems, 95.4% and 77.3% of the isolates were resistant to meropenem and imipenem, respectively. The blaVIM gene was identified in 90.9% of isolates and carbapenemases synthesis were confirmed in 95.4% of the isolates. Fourteen genetic patterns were confirmed through PFGE analyses. The isolates collected within a time gap of 2 years demonstrated a genetic relationship, and the same clone was identified in different departments in the hospital. To the best of our knowledge, this is the first report of identification and characterization of A. baumannii nosocomial isolates in Pelotas, RS, Brazil.

HLA-G alleles and their impacts on placental HSV-1 infection in women from southern Brazil.

The Human Leukocyte Antigen G (HLA-G) is an immunoregulatory molecule with a critical role in pregnancy success. HLA-G alleles are associated with differential susceptibility to multiple conditions, including gestational problems, infectious diseases, and viral persistence. Of note, both herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) can impair HLA-G expression, interfering with HLA-G-associated immunoregulation. On the other hand, the impacts of HLA-G alleles on susceptibility to Herpesviridae infection is a neglected issue. Therefore, this study evaluated HLA-G allele frequencies and their associations with placental Herpesviridae infection in women from southern Brazil. Placenta samples were collected soon after delivery, and detection of viral DNA of HSV-1, HSV-2 and human cytomegalovirus (HCMV) was performed by polymerase chain reaction (PCR). A fragment of HLA-G (exons 2-4) was amplified by PCR, sequenced, and analyzed to allele determination. One hundred and seventy women had their alleles determined. Overall, 25 HLA-G alleles were found, distributed into 56 different genotypes. The most frequent alleles were G* 01:01:01 and G* 01:01:02, found in 37.9 % and 16.5 % of samples, respectively. Among the 170 women, 89 (52.4 %) tested positive for Herpesviridae DNA in the placenta, 55 (32.3 %) tested negative, 3 (1.8 %) were negative for HSV-1 and HSV-2 (with absent HCMV data), and 23 (13.5 %) were undetermined. The G* 01:01:01 allele was significantly associated with an increased risk of placental HSV-1 infection (p = 0.0151; OR=1.837; IC=1.108-3.045). This study describes new information concerning placental HLA-G alleles in women from southern Brazil and helps explain how genetic background can modify susceptibility to placental infections.

Phenotypic and molecular evaluation of biofilm formation in Klebsiella pneumoniae carbapenemase (KPC) isolates obtained from a hospital of Pelotas, RS, Brazil.

Introduction. A significant cause of mortality in the intensive care unit (ICU) is multidrug-resistant (MDR) Gram-negative bacteria, such as Klebsiella pneumoniae carbapenemase (KPC). Biofilm production is a key factor in KPC colonization and persistence in the host, making the treatment difficult.Gap Statement. The aim of this study was to evaluate the antibiotic resistance, molecular and phenotypic biofilm profiles of 12 KPC isolates associated with nosocomial infection in a hospital in Pelotas, Rio Grande do Sul, Brazil.Methodology. Clinical isolates were obtained from different sources, identified and characterized by antibiotic resistance and carbapenemase synthesis following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Polymerase chain reaction (PCR) was used to evaluate the presence of carbapenemase (blaKPC) and biofilm formation-associated genes (fimA, fimH, rmpA, ecpA, mrkD and wabG). Additionally, phenotypic evaluation of in vitro biofilm formation capacity was evaluated by Congo red agar (CRA) assay and the crystal violet staining method.Results. The 12 isolates evaluated in this study presented the blaKPC gene and were positive for synthesizing carbapenemases in vitro. In the carbapenem class, 83.3 % isolates were resistant and 16.7 % intermediately resistant to imipenem and meropenem. Molecular analyses found that the fimA and wabG genes were detected in 75 % of isolates, while fimH and ecpA were detected in 42 % and mrkD were detected in 8.3 % (1). The CRA assay demonstrated that all isolates were slime producers and 91.7 % (11) of isolates were classified as strong and 8.3 % (1) as moderate biofilm producers by the crystal violet staining method. The optical density (OD540nm) for strong biofilm formers ranged from 0.80±0.05 to 2.47±0.28 and was 0.55±0.12 for moderate biofilm formers.Conclusion. Our study revealed a high level of antibiotic resistance and biofilm formation in KPC isolates obtained from a hospital in Pelotas, RS, Brazil.

[Frequency of Human Papillomavirus in the placenta, in the colostrum and in the umbilical cord blood]. / Frequência do Papilomavírus Humano na placenta, no colostro e no sangue do cordão umbilical.

PURPOSE: To determine the frequency of Human Papillomavirus (HPV) in the placenta, in the colostrum and in the umbilical cord blood of parturient women and their newborns assisted at the Clinic of Gynecology and Obstetrics of the University Hospital of Rio Grande (RS), Brazil. METHODS: Biopsies were collected from 150 placentas on the maternal side, 150 on the fetal side, 138 samples of umbilical cord blood and 118 of the colostrum. The placenta biopsies were collected from the central and peripheral portions. DNA was extracted according to the manufacturer's protocol and to a reference found in the literature. HPV was detected by the nested polymerase chain reaction (PCR-Nested) using primers MY09/11 and GP5/GP6. Genotyping was performed by direct sequencing. The participants responded to a self-applied questionnaire with demographic and clinical data, in order to characterize the sample. RESULTS: HPV was detected in 4% (6/150) of cases on the mother's side of the placentas, in 3.3% (5/150) on the fetal side, in 2.2% (3/138) in umbilical cord blood and in 0.84% (1/118) in colostrum samples. The vertical transmission rate was 50%. HPV-6 was the low-risk genotype found (60%) and the high-risk genotypes were HPV-16 and HPV-18 (20% each). CONCLUSIONS: These results suggest that HPV can infect the placenta, the colostrum and the umbilical cord blood.

Frequência do Papilomavírus Humano na placenta, no colostro e no sangue do cordão umbilical / Frequency of Human Papillomavirus in the placenta, in the colostrum and in the umbilical cord blood

OBJETIVO: Determinar a frequência do Papilomavírus Humano (HPV) na placenta, no colostro e no sangue do cordão umbilical de parturientes e seus neonatos atendidos no Ambulatório de Ginecologia e Obstetrícia do Hospital Universitário de Rio Grande (RS), Brasil. MÉTODOS: Foram coletadas biópsias de 150 placentas do lado materno, 150 do lado fetal, 138 amostras do sangue do cordão umbilical e 118 amostras de colostro. As biópsias de placenta foram coletadas da porção central e periférica. O DNA foi extraído segundo protocolo do fabricante e conforme referência encontrada na literatura. O HPV foi detectado pela técnica da reação em cadeia da polimerase aninhada (PCR-Nested) com os primers MY09/11 e GP5/GP6. A genotipagem foi por sequenciamento direto. As participantes responderam a um questionário autoaplicado com dados demográficos e clínicos, a fim de caracterizar a amostra. RESULTADOS: O HPV foi detectado em 4% (6/150) do lado materno das placentas, 3,3% (5/150) do lado fetal; 2,2% (3/138) no sangue do cordão e 0,8% (1/118) no colostro. A taxa de transmissão vertical foi de 50%. O genótipo de baixo risco oncogênico encontrado foi o HPV-6 (60%) e de alto risco, os HPV-16 e HPV-18 (20% cada). CONCLUSÕES: Esses resultados sugerem que o HPV pode infectar a placenta, o colostro e o sangue do cordão umbilical. .

PURPOSE: To determine the frequency of Human Papillomavirus (HPV) in the placenta, in the colostrum and in the umbilical cord blood of parturient women and their newborns assisted at the Clinic of Gynecology and Obstetrics of the University Hospital of Rio Grande (RS), Brazil. METHODS: Biopsies were collected from 150 placentas on the maternal side, 150 on the fetal side, 138 samples of umbilical cord blood and 118 of the colostrum. The placenta biopsies were collected from the central and peripheral portions. DNA was extracted according to the manufacturer's protocol and to a reference found in the literature. HPV was detected by the nested polymerase chain reaction (PCR-Nested) using primers MY09/11 and GP5/GP6. Genotyping was performed by direct sequencing. The participants responded to a self-applied questionnaire with demographic and clinical data, in order to characterize the sample. RESULTS: HPV was detected in 4% (6/150) of cases on the mother's side of the placentas, in 3.3% (5/150) on the fetal side, in 2.2% (3/138) in umbilical cord blood and in 0.84% (1/118) in colostrum samples. The vertical transmission rate was 50%. HPV-6 was the low-risk genotype found (60%) and the high-risk genotypes were HPV-16 and HPV-18 (20% each). CONCLUSIONS: These results suggest that HPV can infect the placenta, the colostrum and the umbilical cord blood. .