Bacteriemia por Enterobacter cancerogenus en un paciente con trauma pélvico / Enterobacter cancerogenus bacteremia in a patient with pelvic trauma

Resumen Existen escasas comunicaciones de infecciones por Enterobacter cancerogenus. Aunque ha sido aislado en heridas contaminadas asociadas a traumas, también ha sido reportado como agente etiológico en una amplia variedad de otras infecciones, siendo infrecuente la presencia de bacteriemia. Presentamos el primer caso reportado en Chile de una bacteriemia causada por este microorganismo, en un varón de 28 años con una fractura de pelvis estable debido a un accidente de tránsito de alta energía. Tuvo una buena respuesta clínica al tratamiento con ertapenem.

Abstract There are few cases reports of Enterobacter cancerogenus infections. Although it has been isolated in contaminated wounds associated with trauma, it has also been reported as an etiological agent in a wide variety of other infections, with the presence of bacteremia being infrequent. We present the first case reported in Chile of a bacteremia caused by this agent, in a 28-year-old patient with a stable pelvic fracture due a high-energy traffic accident. He had a good clinical response to treatment with ertapenem.

The importance of environments and surfaces hygienization in pandemic times (Covid-19): a microbiological analysis in places for physical activity practice

The objective of this study was to analyse microbiological organisms in different locations and regions for physical activity in the city of João Pessoa, Brazil. Samples were collected on various objects used, such as: mattresses, drinking fountains, gloves, cell phones and others. The samples were collected in João Pessoa-PB, following the Standard Operating Procedure-SOP/ Microbiology of a specialized laboratory. The collection took place in the five macro-regions: North, South, East, West and Center. Foreach region samples were collected in one public place (square), a private one (gym) and one school (public or private), totaling fifteen collected sites and 450 samples. The following microorganisms were studied in all analyzed surfaces: Bacillus sp, Escherichia Coli, Klebsiella sppor Enterobacter sppand Coag. Neg. Staphylococcus.All regions had a high contamination level by some microorganism. The highest rates were found in the western, central and northern regions -96, 94 and 93% respectively. The Coag. Neg.Staphylococcus presented the highest and lowest incidence rates in the South and East regions, with 43.33 and 6.67%, respectively, as well as Klebsiella sppor Enterobacter spp, which presented high levels. It is concluded that there is a microorganisms' contamination in the most varied places and regions where physical activity practices are developed, with a predominance of Coag. Neg.Staphylococcusand Klebsiella sppor Enterobacter spp. These results lead to a warning about the hygiene importance in places for physical activity practice, especially in pandemic times (COVID-19), since almost all the evaluated surfaces were contaminated.

Calidad microbiológica del aire en seis áreas de la microestación biológica- Zoocriadero de la Universidad Nacional Agraria de la Selva ­ Tingo María ­ Perú / Microbiological quality of air in six areas of the biological micro station - Zoocriadero of the Universidad Nacional Agraria de la Selva - Tingo Maria ­ Peru

El presente trabajo de investigación tiene como objetivo evaluar la calidad microbiológica del aire en seis áreas de la Microestación Biológica-Zoocriadero de la Universidad Nacional Agraria de la Selva-Tingo María-Perú; procediéndose de la siguiente manera: Se seleccionaron seis áreas de muestreo (A1: Boletería, A2: Caseta gallito de las rocas, A3: Zona de las tortugas, A4: Caseta del otorongo, A5: Caseta de los monos y A6: Cocina), de los cuales para la determinación de los parámetros físicos (PAS, temperatura, humedad relativa, precipitación y rosa de viento) se obtuvo información de la estación meteorológica de la UNAS y de lecturas directas en campo, para la determinación de las PAS se realizó por el método pasivo obteniéndose la mayor concentración de PAS en el área 1 "Boletería" (12.61 t/km2/mes, 11.08 t/km2/mes y 8.4 t/km2/mes) y la más baja en el área 5 "caseta de los monos" (2.80 t/km2/mes, 2.80 t/km2/mes y 3.2 t/km2/mes); para el análisis microbiológico se realizó por el método de borboteo en líquidos, identificándose 27 géneros entre bacterias y hongos, siendo las bacterias más frecuentes Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae y algunas bacterias patógenas para el hombre como Streptococcus sp., Clostridium perfringens, Bacillus anthracis. Los hongos de mayor frecuencia son Penicillium sp., Aspergillius sp., Geotrichum sp., algunos hongos patógenos identificados: Microsporum sp., Epirophyton sp. Por otro lado, para la percepción del público como trabajadores se aplicó encuestas, donde indicaron que la calidad microbiológica del aire en la Microestación Biológica-Zoocriadero es regular(AU)

The objective of this research paper is to assess the microbiological quality of the air in six areas of the Biologial Micro-station and animal breeding farm of the Peruvian Univeristy "Universidad Nacional Agraria de la Selva in Tingo María, Perú", proceeding as follows: Six sampling areas were selected (A1: Ticket Office, A2: Cock-of-the-Rock hut, A3: Turtle area, A4: Otorongo hut, A5: Monkey hut, A6: Kitchen), of wich for the determination of the phisical parameters (SBP, temperature, relative humidity, precipitation and wind rose) information was obtained from the UNAS meteorological station and direct readings in the field. For the determination of SBP was performed by the passive method obtaining the high concentration of SBP in area 1 "Ticket Office" (12.61 t/km2/month, 11.08 t/km2/month and 8.4 t/km2/month) and the lowest in area 5 "monkey house" (2.80 t/km2/month, 2.80 t/km2/month and 3.2 t/km2/month); microbiological analysis was performed by the liquid bubbling method, identifying 27 genera incluiding bacteria and fungi, with the most frequent bacteria being Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae and some bacteria pathogenic for humans such as Streptococcus sp., Clostridium perfringens and Bacillus anthracis. The most frequent fungi are Penicillium sp., Aspergillius sp. and Geotrichum sp.; some pathogenic fungi identified: Microsporum sp., Epirophyton sp. On the other hand,regarding the perception of the public and workers, surveys were applied, wich indicated that the microbiologial quality of the air in the Biological Micro-station and animal breeding farm is regular(AU)

Eosinophilic cystitis, a rare cause of persistent haematuria in a diabetic dog / Cistite eosinofílica, uma causa rara de hematúria persistente em um cão diabético

Eosinophilic cystitis is a rare inflammatory disorder characterized by eosinophilic infiltration of entire layers of the bladder wall. The condition has been described in adults, children, and dogs. However, there are no consensus guidelines for the treatment of eosinophilic cystitis. Although human and veterinary literature reviews show some effectiveness in management with corticosteroids, antihistamines, and antibiotics, a variety of serious and frequent side effects are associated with steroid therapy. As a result, steroids are relatively contraindicated for patients with diabetes mellitus and Cushing's syndrome. A five-year-old neutered male chow-chow with controlled diabetes was referred with an 18-month history of malodorous urine, gross haematuria, and dysuria that were nonresponsive to antibiotics. The findings on general examination were unremarkable except for abdominal suprapubic discomfort. The complete blood count and biochemical profile (such as urea and creatinine) were normal except for mild peripheral eosinophilia. Although ultrasonography, bladder contrast radiography, and urine cytology findings indicated malignancy, with the presence of atypical urothelial cells, histopathology confirmed eosinophilic cystitis. Management with cyclosporine was adequate with complete remission of haematuria. This case report presents the first reported successful use of cyclosporine for the treatment of eosinophilic cystitis in a dog with diabetes.(AU)

A cistite eosinofílica é uma doença inflamatória rara caracterizada por infiltração eosinofílica de todas as camadas da parede da bexiga. Essa enfermidade já foi descrita em adultos, crianças e cães. No entanto, não há um consenso de diretrizes sobre o seu tratamento. Mesmo que as literaturas humana e veterinária mostrem alguma eficácia no manejo com corticosteroides, anti-histamínicos e antibióticos, uma variedade de efeitos colaterais graves e frequentes está associada à terapia com esteroides. Dessa forma, o uso de esteroides é relativamente contraindicado para pacientes com diabetes mellitus e síndrome de Cushing, por exemplo. Um chow-chow, macho, castrado, de cinco anos e diabético estável foi encaminhado para atendimento com histórico de urina fétida, hematúria macroscópica e disúria não responsiva a antibióticos há 18 meses. A avaliação dos parâmetros físicos estava dentro dos padrões, exceto por desconforto abdominal suprapúbico à palpação. O hemograma e o perfil bioquímico (como a ureia e a creatinina) estavam dentro da normalidade para a espécie, exceto por eosinofilia periférica leve. Embora a ultrassonografia, a radiografia contrastada da bexiga e os achados da urinálise indicassem malignidade, com a presença de células uroteliais atípicas, a histopatologia confirmou o diagnóstico definitivo de cistite eosinofílica. O manejo com ciclosporina foi satisfatório, com ausência completa da hematúria. Este relato de caso apresenta o primeiro uso documentado de ciclosporina para o tratamento de cistite eosinofílica com sucesso em um cão com diabetes.(AU)

Chronic osteomyelitis in canine penile bone: case report / Osteomielite crônica em osso peniano canino: relato de caso

A 10-year-old male mixed-breed dog was admitted for recurrent signs of urinary tract infection (UTI). Urinary bladder ultrasonography revealed decreased thickness of its wall with floating hyperopic particles within its lumen. Ultrasonography revealed a structure invading the dorsal wall of the penile urethral lumen, located in a segment distal to the bladder. Radiographies showed bone resorption with proliferation at the caudal aspect of the penile bone, stricture of the final aspect of the penile urethra, and no radiopaque images compatible with a urethrolith. Computed tomography showed bone proliferation causing stricture of the urethral lumen at two different sites. Presumptive diagnosis of penile neoplasia was considered more likely and the dog underwent penectomy along with orchiectomy and scrotal urethrostomy. Enterobacter spp. was cultured from the urine sample and antibiotic sensitivity tests revealed that the bacterium was susceptible to amikacin, imipenem, and meropenem. Histopathology revealed severe suppurative urethritis, bone resorption, and hyperostosis, suggestive of osteomyelitis of the penile bone. Neoplastic cells were not observed at any part of the examined tissue. The findings in the present case suggest that osteomyelitis of the penile bone should be included in differential diagnosis for partial and complete urethral obstruction in dogs with recurrent UTI.(AU)

Um cão mestiço, com 10 anos, foi admitido por sinais recorrentes de infecção do trato urinário (ITU). A ultrassonografia da bexiga urinária revelou diminuição da espessura de sua parede com partículas flutuantes dentro de seu lúmen. A ultrassonografia demonstrou estrutura invadindo a parede dorsal do lúmen da uretra peniana, localizada em segmento distal à bexiga. Radiografias evidenciaram reabsorção óssea com proliferação no aspecto caudal do osso peniano, estenose do aspecto final da uretra peniana e ausência de imagens radiopacas compatíveis com uretrólito. Pela tomografia computadorizada, observou-se proliferação óssea causando estreitamento da luz uretral em dois locais diferentes. Diagnóstico presuntivo de neoplasia peniana foi considerado mais provável e o cão foi submetido à penectomia, juntamente com orquiectomia e uretrostomia escrotal. Enterobacter spp. foi cultivada da amostra de urina e testes de sensibilidade revelaram susceptibilidade ao amicacina, imipenem e ao meropenem. A histopatologia revelou uretrite supurativa grave, reabsorção óssea e hiperostose compatível com osteomielite do osso peniano. Células neoplásicas não foram observadas em nenhuma parte do tecido examinado. Os achados do presente caso sugerem que a osteomielite do osso peniano deve ser incluída no diagnóstico diferencial de obstrução uretral parcial e completa em cães com ITU recorrente.(AU)

Reutilização do detergente enzimático: avaliação do impacto da contaminação microbiana da solução na efetividade da limpeza de aparelhos endoscópicos gastrointestinais / Enzymatic detergent reuse: evaluation of the impact of the solution's microbial contamination on the cleaning effectiveness of gastrointestinal endoscopes

No Brasil, é recomendado que durante a limpeza dos Produtos para Saúde (PPS) o detergente utilizado possua ação enzimática. Embora a Resolução da Diretoria Colegiada nº 55 de 14 de novembro de 2012 da Agência Nacional de Vigilância Sanitária desaconselhe a reutilização desta solução de limpeza, sabe-se que na prática clínica elas são reaproveitadas por diversas vezes para imersão de PPS, como os aparelhos endoscópicos, o que pode comprometer a efetividade da ação do detergente enzimático e com isso a segurança no processamento do PPS. Esta pesquisa objetivou avaliar a carga microbiana presente na solução de detergente enzimático durante sua reutilização na limpeza manual de aparelhos endoscópicos gastrointestinais. Tratou-se de um estudo transversal realizado em um serviço de endoscopia digestiva de um hospital universitário de Belo Horizonte e no Laboratório de Microbiologia Oral e Anaeróbios do ICB/UFMG. A amostra foi composta por 57 aparelhos endoscópicos e 76 alíquotas de solução de detergente enzimáticos coletadas de diversos reusos de 19 diferentes soluções. O material coletado foi agitado em vórtex, acrescido a Caldo Letheem Modificado e submetido a filtração em membrana Millipore® 0,45µm. A membrana foi depositada em Tryptic Soy Ágar para crescimento microbiano. A identificação presuntiva dos micro-organismos foi realizada manualmente considerando-se aspectos morfotintoriais e reações bioquímico/fisiológicas. As variáveis foram descritas utilizando frequências, porcentagens e medidas de tendência central. O projeto foi aprovado pelo Comitê de Ética e Pesquisa da Universidade Federal de Minas Gerais (CAAE ­ 67493417.1.0000.5149). As médias das cargas microbianas na solução de detergente enzimático variaram de 19,9 UFC/mL após primeiro uso, 51,1 UFC/mL após terceiro uso e 67,1UFC/mL após o quinto reuso. Nos canais de ar/água e biópsia houve aumento de micro-organismos Gram negativos ao longo das reutilizações do detergente. Foram recuperados, Enterobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas spp., Staphyloccocus aureus, Staphyloccocus coagulase negativa. Pseudomonas spp. foi o micro-organismo mais identificado em todas as alíquotas coletadas. Verificou-se a importância da escovação do canal de biópsia para correta remoção de micro-organismos. Conclui-se que a reutilização das soluções de detergente enzimático contribuiu para contaminação dos aparelhos endoscópicos com micro-organismos potencialmente patogênicos. Faz-se necessário a reavaliação de protocolos institucionais, no sentido de que seja cumprida a orientação da Anvisa por meio da RDC nº 55 de 14 de novembro de 2012 de que os detergentes enzimáticos não devem ser reutilizados sob perda da eficiência do produto. As características físico químicas dos detergentes enzimáticos devem ser respeitadas pelos serviços de saúde conforme parâmetros estabelecidos pelos fabricantes.(AU)

In Brazil, it is recommended that during the cleaning of Health Products the detergent used has enzymatic action. Although Collegiate Board Resolution No. 55 of November 14, 2012 of the National Agency of Sanitary Surveillance advises against the reuse of this cleaning solution, it is known that in clinical practice they are reused several times for immersion of health products, such a gastrointestinal endoscope, which may compromise the effectiveness of the enzymatic detergent action and thus the safety in the processing. This research aimed to evaluate the microbial load present in the enzymatic detergent solution during its reuse in the manual cleaning of endoscopic gastrointestinal devices. This was a cross-sectional study performed at a gastrointestinal endoscopy service at a university hospital in Belo Horizonte and at the Oral Microbiology and Anaerobic Laboratory of ICB/UFMG. The sample consisted of 57 endoscopes and 76 aliquots of enzymatic detergent solution collected from several replicates of 19 different solutions. The collected material was vortexed, added to Modified Letheem Broth and subjected to Millipore® 0.45 µm membrane filtration. The membrane was deposited in Tryptic Soy Ágar for microbial growth. The identification of the microorganisms was performed manually considering morphotintorial aspects and biochemical/physiological reactions. The variables were described using frequencies, percentages and measures of central tendency. The project was approved by the Ethics and Research Committee of the Federal University of Minas Gerais (CAAE - 67493417.1.0000.5149). The mean values of the microbial loads in the enzymatic detergent solution varied from 19.9 UFC/mL after first use, 51.1 UFC/mL after third use and 67.1 UFC/mL after the fifth reuse. In the air/water and biopsy channels there was an increase of Gram negative microorganisms along the reuse of the detergent. Enterobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas spp., Staphylococcus aureus, Coagulase-negative Staphyloccocus were recovered. Pseudomonas spp. was the most identified microorganism in all aliquots collected. It was verified the importance of brushing the biopsy channel for correct removal of microorganisms. It was concluded that the reuse of enzyme detergent solutions contributed to the contamination of the endoscopes with potentially pathogenic microorganisms. It is necessary to re-evaluate institutional protocols, in order to comply with Anvisa's guidance through RDC nº. 55 of November 14, 2012 that enzymatic detergents should not be reused under loss of product efficiency. The physical characteristics of the enzymatic detergents must be observed by the health services according to the parameters established by the manufacturers.(AU)

Isolation, identification, and biocontrol of antagonistic bacterium against Botrytis cinerea after tomato harvest

ABSTRACT Tomato is one of the most important vegetables in the world. Decay after harvest is a major issue in the development of tomato industry. Currently, the most effective method for controlling decay after harvest is storage of tomato at low temperature combined with usage of chemical bactericide; however, long-term usage of chemical bactericide not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after tomato harvest has great practical significance. In this study, antagonistic bacterium B-6-1 strain was isolated from the surface of tomato and identified as Enterobacter cowanii based on morphological characteristics and physiological and biochemical features combined with sequence analysis of 16SrDNA and ropB gene and construction of dendrogram. Effects of different concentrations of antagonistic bacterium E. cowanii suspension on antifungal activity after tomato harvest were analyzed by mycelium growth rate method. Results revealed that antifungal activity was also enhanced with increasing concentrations of antagonistic bacterium; inhibitory rates of 1 × 105 colony-forming units (cfu)/mL antagonistic bacterial solution on Fusarium verticillioides, Alternaria tenuissima, and Botrytis cinerea were 46.31%, 67.48%, and 75.67%, respectively. By using in vivo inoculation method, it was further confirmed that antagonistic bacterium could effectively inhibit the occurrence of B. cinerae after tomato harvest, biocontrol effect of 1 × 109 cfu/mL zymotic fluid reached up to 95.24%, and antagonistic bacterium E. cowanii has biocontrol potential against B. cinerea after harvest of fruits and vegetables.

Antimicrobial activity evaluation and comparison of methods of susceptibility for Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacter spp. isolates

Abstract The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.

Identificación bacteriana en superficies de resina acrílica / Bacterial identification on acrylic resin surfaces

Introducción: el material usado para fabricar aparatos protésicos parciales o totales es el polimetilmetacrilato, el cual forma una superficie sólida que se encuentra en íntimo contacto con la mucosa bucal del paciente, esta superfi cie puede presentar defectos (poros, grietas e irregularidades), que se producen al momento de su elaboracióny varían según la técnica de procesado, actuando como reservorios que contribuyen a la adherencia y proliferación de microorganismos,dentro de los cuales, el más frecuentemente aislado en pacientesportadores de prótesis es Candida albicans. Objetivo: Identifi car lasbacterias presentes en la superfi cie de una resina acrílica para base dedentadura (ProBase Hot®, Ivoclar, Vivadent). Material y métodos: Se seleccionaron 10 pacientes de ambos sexos, entre 25 y 30 años de edad que acudían a la clínica de prótesis. Las impresiones de alginato de los pacientes se utilizaron para crear modelos de yeso, que luegoconfeccionaron paladares de acrílico termocurado que los pacientes llevaban por un periodo de 24 horas. Una muestra del acrílico se tomóposteriormente para fi nes de identifi cación bacteriológica. El análisisestadístico consistió en estadística descriptiva con la distribución defrecuencia y porcentajes, realizando tablas de contingencia y respuestamúltiple (Programa IBM SPSS STATISTICS 21.0). Resultados: Labacteria identifi cada mayor número de veces fue Klebsiella pneumoniae,mientras que de las aisladas en menor frecuencia correspondió tanto a Escherichia coli como a Enterobacter cloacae, Pseudomonas aeruginosa en tres oportunidades, seguido de Enterococcus faecalis, Streptococcus alfa hemolítico y Streptococcus hyicus solo un par de veces. Conclusiones: La resina acrílica usada en este estudio diopositivo a diferentes especies bacterianas y las más frecuentementeaisladas pertenecen a la familia de las enterobacterias.

Introduction: the material most commonly used to make full orpartial prosthetic dental appliances is polymethyl methacrylate, whichprovides a solid surface that is in close contact with the patient’s oralmucosa. During the production of the dental prosthetic, defects suchas holes, cracks, and other irregularities can appear on this surface(depending on the method used), which act as reservoirs that stimulatethe proliferation of microorganisms and make it easier for these toadhere to the surface. The most frequently isolated microorganism indenture wearers is Candida albicans. Objective: To identify the bacteriapresent on the acrylic resin surface of dentures bases (ProBase Hot®,Ivoclar, Vivadent). Material and methods: 10 subjects of both sexesaged between 25 and 30 years were selected from among patientsattending a prosthodontics clinic. Alginate impressions of the patientswere used to create plaster molds, which were then used to constructheat-cured acrylic resin palatal plates that the patients wore for aperiod of 24 hours. A sample of the acrylic was subsequently takenfor bacteriological identifi cation purposes. Statistical analysis wasperformed based on a descriptive analysis of frequency distributionand percentages, and crossover and multiple response tables created(IBM SPSS STATISTICS 21.0 software). Results: The most frequentlyidentifi ed bacterium in this study was Klebsiella pneumoniae, whilethe least frequently isolated were Escherichia coli and Enterobactercloacae (once each), Pseudomonas aeruginosa (in three cases),and Enterococcus faecalis, Streptococcus alpha hemolytic andStreptococcus hyicus (two each). Conclusions: The acrylic resin usedin this study tested positive for various bacterial species, the mostfrequently isolated of these belonging to the family Enterobacteriaceae.

Prevalência de enterobactérias produtoras de Klebsiella pneumoniae carbapenemase em culturas de vigilância epidemiológica em unidade de terapia intensiva de um hospital de ensino de Minas Gerais / Prevalence of enterobacteria producers of Klebsiella pneumoniae carbapenemase in epidemiological surveillance cultures in intensive care unit of a teaching hospital in Minas Gerais, Brazil

O crescente aumento de bactérias multirresistentes no ambiente hospitalar e a falta de opções terapêuticas a curto e médio prazo tem se tornado um grande desafio para o controle das infecções relacionadas à assistência à saúde. As infecções por enterobactérias produtoras de Klebsiella pneumoniae carbapenemase (KPC), vem se destacando como a de maior risco para os pacientes debilitados que internam nas Unidades de Terapia Intensiva (UTIs). O objetivo do trabalho foi identificar a prevalência de KPC em culturas de vigilância epidemiológica de amostras de swab retal de pacientes internados nas UTI´s adulto, neonatal e pediátrica de um Hospital de Ensino de Minas Gerais, no período de janeiro a julho de 2014. Realizou-se um estudo transversal descritivo retrospectivo onde os dados foram analisados a partir dos registros dos livros do laboratório de microbiologia do hospital. Este estudo foi aprovado pelo Comitê de Ética e Pesquisa número do parecer 948.342. Foram analisadas 422 amostras de swab retal, sendo que 367 (86,9%) eram provenientes das UTIs adulto e 55 (13%) da UTI neonatal e pediátrica. Foram positivas para KPC 31 (7,3%) das quais 21 eram da UTI adulto e 10 da UTI neonatal e pediátrica. Das 31 culturas positivas para KPC uma (3%) foi em Escherichia coli, quatro (13%) em Enterobacter sp e 26 (84%) em Klebsiella pneumoniae. A detecção laboratorial de enterobactérias produtoras de KPC exprime a importância das culturas de vigilância epidemiológica na rotina como medida de prevenção e controle da disseminação desses microrganismos multirresistentes, principalmente nas UTIs.

The increasing number of multidrug-resistant bacteria in the hospital environment and the lack of treatment options in the short and medium term have become a major challenge for the control of infections related to health care. Infections caused by Enterobacteriaceae producing Klebsiella pneumoniae carbapenemase (KPC) have emerged as the highest risk for debilitated patients hospitalized in Intensive Care Units (ICUs). The aim of the study was to identify the prevalence of KPC in epidemiological surveillance cultures of samples of rectal swab of patients admitted to the adult, neonatal and pediatric ICUs, of a teaching hospital in Minas Gerais, Brazil in the period from January to July 2014. We conducted a retrospective descriptive cross-sectional study in which data were analyzed from the records of the hospital's microbiology lab books. This study was approved by the Research Ethics Committee, protocol number 948 342. 422 samples of rectal swab were analyzed, of which 367 (86.9%) were from the adult ICU's and 55 (13%) of neonatal and pediatric ICU. Thirty-one samples (7.3%) were positive for KPC, 21 in adult ICU and 10 in neonatal and pediatric ICU. Of the 31 positive cultures for KPC, one (3%) was for Escherichia coli, four (13%) for Enterobacter sp. and 26 (84%) for Klebsiella pneumoniae. Laboratory detection of KPC-producing Enterobacteriaceae expresses the importance of routine epidemiological surveillance cultures as a preventive and control measure for the spread of these multiresistant microorganisms, especially in ICUs.

Estudo genotípico e fenotípico de bacilos Gram-negativos produtores de carbapenemase do tipo New Delhi metalo-ß-lactamase / Genotypic and fenotypic study of Gram-negative bacilli producers carbapenemase type New Delhi metallo--ß-lactamase

Os carbapenêmicos são os antimicrobianos mais amplamente utilizados no tratamento empírico de infecções graves por bacilos Gram-negativos. A pressão seletiva gerada pelo uso desses antimicrobianos ao longo das últimas três décadas contribuiu para a disseminação de enterobactérias e Gram-negativos não fermentadores produtores de carbapenemases, particularmente as do tipo KPC e NDM. Os genes que codificam essas enzimas usualmente estão localizados em plasmídeos e/ou transpósons. A hipótese atualmente mais aceita é que o gene blaNDM-1 seja uma quimera criada em Acinetobacter baumannii. A NDM-1 foi descrita em paciente proveniente da Índia e subsequentemente evidenciou-se sua ampla disseminação nesse país. A epidemiologia que tem sido observada nos casos detectados na Europa e Estados Unidos tem sido viagem à Índia, ou seja, sem casos autóctones. No Brasil, os primeiros casos foram identificados no Rio Grande do Sul, e a seguir no Rio de Janeiro e em São Paulo. Diferentemente dos casos da Europa e América do Norte, os casos do Brasil não tem relação epidemiológica com a Índia. O sequenciamento integral dos plasmídeos e cromossomos albergando o gene blaNDM permitirá entender como ocorre a disseminação desse mecanismo de resistência no Brasil. Para isso, foi avaliado o perfil de susceptibilidade dos isolados, bem como a capacidade conjugativa e clonalidade. Das vinte e oito amostras utilizadas neste trabalho, treze delas pertencem à espécie Enterobacter hormaechei, uma à espécie Citrobacter freundii, sete à espécie Escherichia coli, quatro à Klebsiella pneumoniae e três ao gênero Acinetobacter spp. Os primeiros isolados incluídos neste estudo (Escherichia coli e Enterobacter hormaechei produzindo NDM-1) foram isolados em agosto de 2013, de uma mesma amostra de swab retal de um paciente do Rio de Janeiro que nunca viajou para o exterior. O sequenciamento completo do DNA plasmidial utilizando a plataforma Illumina e a anotação de ambos os plasmídeos albergando o gene blaNDM-1 revelou que estes pertencem a grupos de incompatibilidade diferentes, IncFIIK (E. hormaechei) e IncX3 (E. coli), e abrigam um novo transpóson composto designado Tn3000. A comparação da sequência nucleotídica do Tn3000 com aquelas disponíveis no GenBank evidencia que a mesma estrutura está presente em plasmídeos de isolados da cidade de Porto Alegre e também em diferentes continentes. As espécies de Acinetobacter (A. radioresistens, A. ursingii e A. guillouiae) isoladas em São Paulo e Porto Alegre, possuem o gene blaNDM-1 albergados em um mesmo plasmídeo não tipável de 41.087 pb. A avaliação da clonalidade dos isolados de Enterobacter hormaechei "subsp. oharae" mostrou dois perfis diferentes através da técnica de PFGE, sendo que todos os microrganismos foram isolados de um surto no mesmo hospital no Rio de Janeiro. Isolados de Klebsiella pneumoniae de uma mesma paciente internada em hospital em Salvador, de sítios distintos - swab retal, hemocultura e urina, em ordem cronológica - obtiveram o mesmo perfil clonal pela técnica de PFGE. O mesmo ocorreu com três isolados de Escherichia coli, de um mesmo paciente do Rio de Janeiro, em amostras de swab retal. Os achados deste estudo evidenciam que no Brasil, Nepal, Marrocos e Índia há uma disseminação do gene blaNDM-1 mediada por um novo elemento móvel designado Tn3000 em enterobactérias. A detecção de um mesmo plasmídeo em diferentes espécies de Acinetobacter evidencia que neste gênero bacteriano, no Brasil, a disseminação do gene blaNDM-1 ocorre por conjugação.

Carbapenems are the antimicrobials most widely used in the empirical treatment of severe infections caused by Gram-negative bacilli. The selective pressure generated by the use of these antibiotics over the last three decades has contributed to the spread of enterobacteria and Gram-negative non-fermenting producing carbapenemases, mainly KPC and NDM. Genes encoding these enzymes are usually located in plasmids and/or transposons. Currently the most accepted hypothesis is that the blaNDM-1 gene is a chimera created in Acinetobacter baumannii. The NDM-1 was described in a patient from India and subsequently was reported to be broadly disseminate in this country. The epidemiology that has been observed in cases detected in Europe and United States is traveling to India, but no autochthonous cases. In Brazil, the first cases were identified in Rio Grande do Sul, and then in Rio de Janeiro and São Paulo. Differently from the cases described in Europe and North America, the cases from Brazil have no epidemiological link with India. The complete sequencing of plasmids and chromosomes harboring blaNDM gene will understanding how the dissemination of this resistance mechanism in Brazil occurs. In this work we will be evaluate the susceptibility profile of the isolates, and their conjugal capacity and clonality. Of the twenty-eight samples used in this study, thirteen of them belong to the species Enterobacter hormaechei, one to Citrobacter freundii, seven to Escherichia coli, four to Klebsiella pneumoniae and three to the genus Acinetobacter sp. The first two isolates included in this study (Escherichia coli and Enterobacter hormaechei) were isolated in August 2013, from the same rectal swab sample from a patient from Rio de Janeiro that never traveled abroad. Complete sequencing of plasmid DNA using Illumina platform and annotation of both plasmids harboring the blaNDM-1 gene revealed that they belong to different incompatibility groups, IncFIIK (E. hormaechei) and IncX3 (E. coli), and are harbor to a new transposon designated Tn3000. The comparison of the Tn3000 nucleotide sequence with those available at GenBank shows that the same structure is present in plasmids from other Porto Alegre and also in different continents. The Acinetobacter species (A. radioresistens, A. ursingii and A. guillouiae) isolated in São Paulo and Porto Alegre, have the blaNDM-1 gene harbored in a single non-typing plasmid of 41,087 bp. The evaluation of clonal relationship of Enterobacter hormaechei "subsp. oharae" showed two different profiles by PFGE technique; of note all microorganisms were isolated from an outbreak in the same hospital in Rio de Janeiro. Isolates of Klebsiella pneumoniae from a single patient hospitalized in Salvador, from different anatomical sites - rectal swab, blood culture and urine, in chronological order - obtained the same clonal profile by the PFGE technique. The same occurred with three Escherichia coli isolates, from the same patient from Rio de Janeiro, in swab rectal strains. Our findings suggest that in Brazil, Nepal, Morocco and India there is a spread of blaNDM-1 gene mediated by Tn3000 in enterobacteria. The detection of a same plasmid in different species of Acinetobacter shows that in this bacterial genus, in Brazil, the dissemination of the blaNDM-1 gene occurs by conjugation.

Molecular identification of multidrug resistant Enterobacter hormaechei in Venezuela / Identificación molecular de Enterobacter hormaechei multidrogoresistente en Venezuela

Besides the importance of Enterobacter cloacae species complex as a nosocomial pathogen, little is known about the frequency of each species/genotype. Here, we describe a strain of E. hormaechei subsp. hormaechei isolated from a bronchial secretion of a patient, in the Intensive Care Unit at the General Hospital of Cumaná, Venezuela, who died due to complications of his infection. The molecular identification was done by sequencing the 16S rRNA gene and comparing it to sequences from the GenBank. This strain showed resistance to multiple families of antibiotics (MDR), and the genes blaKPC and blaVIM were detected by PCR. This is the first time E. hormaechei has been identified in Venezuela.

A pesar de la importancia de las especies del complejo Enterobacter cloacae como patógeno nosocomial, poco se conoce sobre la frecuencia de cada especie/genotipo. Aquí se describe una cepa de E. hormaechei subsp. hormaechei aislada de una secreción bronquial de un paciente internado en la Unidad de Cuidados Intensivos del Hospital General de Cumaná, Venezuela, quien murió producto de complicaciones de su infección. La identificación molecular fue hecha por secuenciación del gen ARNr 16S y porcomparación con las secuencias del GenBank. Esta cepa mostró resistencia a múltiples familias de antibióticos (MDR) y se detectaron los genes blaKPCyblaVIMpor PCR. Este es el primer reporte de E. hormaechei en Venezuela.

Murine Experimental Root Canal Infection: Cytokine Expression in Response to F. nucleatum and E. faecalis

Abstract The aim of this study was to evaluate the gene expression of proinflammatory (RANKL, TNF-a and IFN-g) and regulatory (TGF-b and IL-10) cytokines as reaction to experimental infection by mono or bi-association of Fusobacterium nucleatum (ATCC 10953) and Enterococcus faecalis (ATCC 19433). F. nucleatum and E. faecalis, either in mono- or bi-association were inoculated into the root canal system (RCS) of Balb/c mice. Animals were sacrificed at 10 and 20 days after infection and periapical tissues surrounding the root were collected. The mRNA expression of the cytokines RANKL, TNF-a, IFN- g, TGF-b and IL-10 was assessed using real-time PCR. The Kruskal-Wallis test was used for statistical analysis. F. nucleatum mono-infection induced high expression of RANKL and TNF-a, while its modulation was due to IL-10. High expression of IFN-g at day 20 was up-regulated by E. faecalis and RANKL; TNF-a was up-regulated by an independent mechanism via IL-10 and TGF-b. Bi-association (F. nucleatum and E. faecalis) stimulated high expression of RANKL, TNF-a and IFN-g, which seemed to be modulated by TGF-b 20 days later. The gene expression of proinflammatory cytokines was more prominent in the earlier periods of the experimental periapical infection, which concomitantly decreased in the later period. This expression may be regulated by IL-10 and TGF-b in an infection-specific condition

Resumo O objetivo deste trabalho foi avaliar a expressão gênica de citocinas pró-inflamatórias (RANKL, TNF-a e IFN-g) e regulatórias (TGF-b e IL-10) em resposta à infecção experimental por Fusobacterium nucleatum (ATCC 10953) e Enterococcus faecalis (ATCC 19433) como mono-infecção ou em bi-associação. F. nucleatum e E. faecalis foram inoculados no sistema de canais radiculares de camundongos Balb/c, tanto isoladas como em bi-associação. Os animais foram sacrificados em 10 e 20 dias após a infecção, e os tecidos periapicais foram coletados. As expressões do mRNA das citocinas RANKL, TNF-a, IFN-g, TGF-b e IL-10 foram analisadas por meio do real-time PCR. O teste de Kruskal-Wallis foi utilizado para análise estatística. A mono-infecção com F. nucleatum induziu alta expressão de RANKL e TNF-a, enquanto sua modulação ocorreu devido à IL-10. A alta expressão de IFN-g no dia 20 foi regulada positivamente por E. faecalis e RANKL; TNF-a foi regulada positivamente por um mecanismo independente via IL-10 e TGF-b. A bi-associação (F. nucleatum e E. faecalis) estimulou uma alta expressão de RANKL, TNF-a e IFN-g, que parece ser modulada por TGF-b após 20 dias. A expressão gênica de citocinas pró-inflamatórias foi mais proeminente nos estágios iniciais da infecção periapical experimental, com concomitante redução no período tardio. Este fenômeno pode ser regulado por IL-10 e TGF-b em uma condição de infecção específica.

Antimicrobial Effect and Surface Tension of Some Chelating Solutions with Added Surfactants

Abstract This study assessed the antimicrobial efficacy and surface tension of established irrigating solutions with a new experimental chelating solution in infected dentin tubes. Twenty-five specimens were randomly assigned to each of the irrigating solutions. Twenty specimens were used as negative and positive controls. After 21 days of contamination with E. faecalis, the irrigating solutions MTAD, QMiX and Tetraclean NA were delivered into each infected root canal. The solutions were removed and dentin samples were withdrawn from the root canals with sterile low-speed round burs with increasing ISO diameters. The dentin powder samples obtained with each bur were immediately collected in separate test tubes containing 3 mL of BHI broth. After that, 100 μL from each test tube was cultured on blood agar. The grown colonies were counted and recorded as colony-forming units (CFU). The surface tension of the irrigants was measured using a Cahn DCA-322 Dynamic Contact Angle Analyzer. A Kruskal Wallis nonparametric ANOVA and a Friedman test were used (p<0.05). Tetraclean NA showed lower surface tension and CFU values than MTAD and QMiX. Better antibacterial action and low surface tension were observed for Tetraclean NA, probably due to the improved penetration into the root canal and dentinal tubes

Resumo Este estudo avaliou a eficácia antimicrobiana e tensão superficial de soluções irrigadoras e uma nova solução quelante em tubos de dentina infectada. Vinte e cinco espécimes foram aleatoriamente distribuídos conforme as soluções irrigantes. Decorrifdos 21 dias de contaminação com E. faecalis, a soluções de irrigação MTAD, QMiX e Tetraclean NA foram distribuídas em cada canal radicular infectado. As soluções foram removidas e as amostras de dentina foram retiradas dos canais radiculares com brocas esféricas de baixa velocidade com diâmetros ISO sucessivamente maiores. As amostras do pó de dentina obtidas com cada broca foram imediatamente colocadas em tubos de ensaio separados contendo 3 mL de caldo BHI. A seguir, 100 μL de cada amostra do tubo de teste foi cultivada em agar de sangue. As colônias crescidas foram contadas e registadas como unidades formadoras de colônias (UFC). A tensão superficial das soluções irrigantes foi medida utilizando o método de Wilhelmy. A análise não paramétrica de Kruskal-Wallis e o teste de Friedman foram utilizados (p<0,05). Tetraclean NA apresentou menor tensão de superfície e menores valores de UFC do que MTAD e QMiX. A melhor ação antibacteriana e baixa tensão superficial foram observadas para Tetraclean NA, provavelmente devido à melhor penetração no canal radicular e túbulos dentinários.

Diversity and capacity to promote maize growth of bacteria isolated from the Amazon region / Diversidade e capacidade de promoção de crescimento de milho de bactérias isoladas da região Amazônica

Maize plants can establish beneficial associations with plant growth-promoting bacteria. However, few studies have been conducted on the characterization and inoculation of these bacteria in the Amazon region. This study aimed to characterize endophytic bacteria isolated from maize in the Amazon region and to assess their capacity to promote plant growth. Fifty-five bacterial isolates were obtained from maize grown in two types of ecosystems, i.e., a cerrado (savanna) and a forest area. The isolates were characterized by the presence of the nifH gene, their ability to synthesize indole-3-acetic acid (IAA) and solubilize calcium phosphate (CaHPO4), and 16S rRNA partial gene sequencing. Twenty-four bacteria contained the nifH gene, of which seven were isolated from maize plants cultivated in a cerrado area and seventeen from a forest area. Fourteen samples showed the capacity to synthesize IAA and only four solubilized calcium phosphate. The following genera were found among these isolates: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia and Bacillus. In addition, eight isolates with plant growth-promoting capacity were selected for a glasshouse experiment involving the inoculation of two maize genotypes (a hybrid and a variety) grown in pots containing soil. Inoculation promoted the development of the maize plants but no significant interaction between maize cultivar and bacterial inoculation was found. A high diversity of endophytic bacteria is present in the Amazon region and these bacteria have potential to promote the development of maize plants.

Plantas de milho podem estabelecer associações benéficas com bactérias promotoras do crescimento vegetal. No entanto, poucos estudos de caracterização e inoculação com essas bactérias foram realizados na região Amazônica. O objetivo deste estudo foi caracterizar bactérias endofíticas isoladas de milho na região Amazônica e avaliar sua capacidade de promover o crescimento. Cinquenta e cinco isolados bacterianos foram obtidos de plantas de milho cultivadas em diferentes tipos de ecossistemas, cerrado (savana) e área de floresta. Estes foram caracterizados pela presença do gene nifH, capacidade de sintetizar ácido indol-3-acético (AIA), solubilizar fosfato de cálcio (CaHPO4) e sequenciamento parcial do gene 16S rRNA. Vinte e quatro bactérias apresentaram o gene nifH. Destas, sete são oriundas de plantas cultivadas em uma área de cerrado e 17 em área de floresta. Quatorze bactérias demonstraram a capacidade de solubilizar AIA e quatro de solubilizar fosfato de cálcio. Entre estas bactérias, foram encontrados os seguintes gêneros: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia e Bacillus. Oito isolados com capacidade de promoção do crescimento vegetal foram selecionados para experimento em casa de vegetação com a inoculação em dois diferentes genótipos de milho (um híbrido e uma variedade) em vasos com solo. Quanto à promoção de crescimento vegetal, a inoculação promoveu o desenvolvimento de plantas de milho, mas não foi encontrada interação significativa entre a cultivar de milho e a inoculação bacteriana. Na região Amazônica há uma diversidade de bactérias endofíticas e estas possuem potencial para promover o desenvolvimento de plantas de milho.

Isolation and characterization of arsenic resistant bacteria from wastewater

The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 ºC and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.

Optimization of biodegradable plastic production on sugar cane molasses in Enterobacter sp. SEL2

Contaminated environments have a large number of bacteria which can accumulate PHA as their energy reserves. Out of 54 isolated bacterial strains from three groups of contaminated sites 48 were found PHA positive. The sites were grouped on the basis of the type of carbon sources i.e. sugars, fatty acids and much diverse type. Strains MFD5, MFD11, UML3, USL2, SEL2, SEL3, SEL10 and PFW1 produced 69.9 ± 0.29, 75.27 ± 0.45, 65.43 ± 0.1, 72.54 ± 0.27, 76.61 ± 0.28, 61.81 ± 0.05, 71.16 ± 0.09 and 74.92 ± 0.5 percent of PHA to their constant cell weight (CCW) respectively in PHA detection media supplemented with 2% glucose. Molasses, whey, crumbs hydrolysate and palm oil were checked as inexpensive carbon sources. Molasses alone could supply the required nutrients for growth and PHA production. Strain SEL2 produced 47.36 ± 0.45% PHA using 2% molasses at 37 °C and pH 7.0. Upon production optimization the best accumulation (80.95 ± 0.01%) was observed in PHA detection media with 0.2% nitrogen source, 3% molasses, pH 5.0 and 37 °C by the strain SEL2. The overall effect of the presence of increased molasses concentration in the media was positive it increased the accumulation period till 72 h. Enterobacter sp. SEL2 (JF901810) is first time being reported for PHA production.

Effects of different osmolarities on bacterial biofilm formation

Biofilm formation depends on several factors. The influence of different osmolarities on bacterial biofilm formation was studied. Two strains (Enterobacter sp. and Stenotrophomonas sp.) exhibited the most remarkable alterations. Biofilm formation is an important trait and its use has been associated to the protection of organisms against environmental stresses.

Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12

A mesophilic Enterobacter sp. Bn12 producing an alkaline thermostable lipase was isolated from soil in Tehran, Iran. The lipase gene (ELBn12) was identified from a genomic library. Sequence analysis of the DNA fragment revealed an open reading frame of 879 bp encoding a lipase with a molecular mass of 31.3 kDa. The deduced amino acid sequence showed 96% identity with a lipase of Enterobacter sp. Ag1 and the identity of their DNA sequences was 88.9%. ELBn12 belongs to the lipase subfamily I.1 and its catalytic triad consists of Ser82, Asp237 and His259. The lipase was expressed in Escherichia coli (BL21) pLysS and partially purified by anion exchange chromatography. The maximum activity of ELBn12 was obtained at temperature of 60 °C and pH 8.0 towards tricaprylin (C8) and its specific activity was around 2900 U/mg. ELBn12 was stable within a broad pH range from 6.0 to 11.0. The enzyme showed high stability in both polar and nonpolar organic solvents at 50% (v/v). The lipase activity was enhanced in the presence of 10 mM of Ca2+, Mg2+ and K+, while heavy metals (Fe3+ and Zn2+) had strong inhibitory effect. ELBn12 showed high activity in the presence of 1% (w/v) nonionic surfactants, however ionic surfactants inhibited the lipolytic activity. ELBn12 characteristics show that it has a potential to be used in various industrial processes.