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Pesqui. vet. bras ; 39(10): 807-815, Oct. 2019. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1056902


The most consumed cheese in Brazil, Minas Frescal cheese (MFC) is highly susceptible to microbial contamination and clandestine production and commercialization can pose a risk to consumer health. The storage of this fresh product under refrigeration, although more appropriate, may favor the growth of spoilage psychrotrophic bacteria. The objective of this study was to quantify and compare Pseudomonas spp. and other psychrotrophic bacteria in inspected and non-inspected MFC samples, evaluate their lipolytic and proteolytic activities and their metalloprotease production potentials. Twenty MFC samples were evaluated: 10 inspected and 10 non-inspected. Counts of psychrotrophic bacteria and Pseudomonas spp., evaluation of the proteolytic and lipolytic potential of the isolates, and identification of potential producers of alkaline metalloprotease (AprX) were assessed. The mean total psychrotrophic counts were 1.07 (±2.18) × 109CFU/g in the inspected samples and 4.5 (±5.86) × 108CFU/g in the non-inspected, with no significant difference (p=0.37). The average score of Pseudomonas spp. was 6.86 (±18.6) × 105 and 2.08 (±3.65) × 106 CFU/g for the inspected and non-inspected MFC samples, respectively, with no significant difference (p=0.1). Pseudomonas spp. represented 0.06% and 0.004% of psychrotrophic bacteria found in inspected and non-inspected MFC samples, respectively. Collectively, 694 psychrotrophic strains and 47Pseudomonas spp. were isolated, of which 59.9% and 68.1% were simultaneously proteolytic and lipolytic, respectively. Of the 470 psychrotrophs isolated from inspected and 224 from non-inspected cheese samples, 5.74% and 2.23% contained aprX, respectively, while 100 and 86.96% of the Pseudomonas spp. isolates in inspected and non-inspected cheese samples contained the gene. The production potential of AprX did not, however, determine the proteolytic activity on plates of these isolates under the conditions evaluated in this study. Of total, 65.63% of the psychrotrophs that contained aprX gene were confirmed as Pseudomonas spp., using genus-specific PCR. Phylogenetic analysis of the 16S rRNA gene of the other psychrotrophs that were potential producers of AprX identified them as Serratia spp. (n=7), Raoultella ornithinolytica (n=1), and Acinetobacter schindleri (n=1) in the inspected samples and Psychrobacter sanguinis (n=1) and Leuconostoc mesenteroides (n=1) in the non-inspected samples. The production conditions of Brazilian MFC of these samples, while meeting the legal determinations, are not sufficient to control Pseudomonas and other spoilage-related psychrotrophs. Thus, stricter hygienic measures are required during the formal production of this type of cheese.(AU)

O mais consumido no Brasil, o queijo Minas Frescal (QMF) é altamente suscetível à contaminação microbiana e a produção e comercialização clandestina podem representar um risco para a saúde do consumidor. O armazenamento deste produto fresco sob refrigeração, embora mais apropriado, pode favorecer a multiplicação de bactérias psicrotróficas deteriorantes. O objetivo deste estudo foi quantificar e comparar Pseudomonas spp. e outras bactérias psicrotróficas em amostras de QMF inspecionadas e não inspecionadas, avaliar o potencial lipolítico, proteolítico e de produção de metaloprotease alcalina. Vinte amostras de QMF foram avaliadas: 10 inspecionadas e 10 não inspecionadas. Foram avaliadas as contagens de bactérias psicrotróficas e Pseudomonas spp., o potencial proteolítico e lipolítico dos isolados e a identificação de potenciais produtores de metaloprotease alcalina (AprX). A média total das contagens de bactérias psicrotróficas foi de 1,07 (±2,18) × 109UFC/g nas amostras inspecionadas e 4,5 (±5,86) × 108UFC/g nas não inspecionadas, sem diferença significativa (p=0,37). A média de Pseudomonasspp. foi de 6,86 (±18,6) × 105 e 2,08 (±3,65) × 106UFC/g para as amostras QMF inspecionadas e não-inspecionadas, respectivamente, sem diferença significativa (p=0,1). Pseudomonas spp. representaram 0,06% e 0,004% de bactérias psicrotróficas encontradas em amostras QMF inspecionadas e não-inspecionadas, respectivamente. Das amostras inspecionadas e não inspecionadas, foram isoladas 694 colônias psicrotróficas e 47 Pseudomonasspp., dos quais 59,9% e 68,1% foram simultaneamente proteolíticos e lipolíticos, respectivamente. Dos 470 isolados de psicrotróficos das amostras de queijo inspecionados e dos 224 isolados das não inspecionadas, 5,74% e 2,23% continham o gene aprX, respectivamente, enquanto 100 e 86,96% das Pseudomonasspp. isoladas em amostras de queijo inspecionadas e não inspecionadas continham o potencial de expressão de AprX. Esse potencial, no entanto, não determinou a atividade proteolítica em placas desses isolados nas condições avaliadas neste estudo. Do total, 65,63% dos psicrotróficos que continham o gene aprX foram confirmados como Pseudomonasspp., utilizando PCR gênero-específico. A análise filogenética do gene 16S rRNA dos outros psicrotróficos que foram produtores potenciais de AprX os identificou como Serratia spp. (n=7), Raoultella ornithinolytica (n=1) e Acinetobacter schindleri (n=1) nas amostras inspecionadas e Psychrobacter sanguinis (n=1) e Leuconostoc mesenteroides (n=1) nas amostras não inspecionadas. As condições de produção do QMF dessas amostras, atendendo às determinações legais, não são suficientes para controlar a Pseudomonas e outros psicrotróficos relacionados à deterioração. Assim, medidas higiênicas mais rígidas são necessárias durante a produção formal deste tipo de queijo.(AU)

Pseudomonas/isolation & purification , Cheese/microbiology , Quality Control
Braz. j. microbiol ; 49(4): 695-702, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974298


ABSTRACT Antarctica harbors a great diversity of microorganisms, including bacteria, archaea, microalgae and yeasts. The Pseudomonas genus is one of the most diverse and successful bacterial groups described to date, but only eight species isolated from Antarctica have been characterized. Here, we present three potentially novel species isolated on King George Island. The most abundant isolates from four different environments, were genotypically and phenotypically characterized. Multilocus sequence analysis and 16S rRNA gene analysis of a sequence concatenate for six genes (16S, aroE, glnS, gyrB, ileS and rpoD), determined one of the isolates to be a new Pseudomonas mandelii strain, while the other three are good candidates for new Pseudomonas species. Additionally, genotype analyses showed the three candidates to be part of a new subgroup within the Pseudomonas fluorescens complex, together with the Antarctic species Pseudomonas antarctica and Pseudomonas extremaustralis. We propose terming this new subgroup P. antarctica. Likewise, phenotypic analyses using API 20 NE and BIOLOG® corroborated the genotyping results, confirming that all presented isolates form part of the P. fluorescens complex. Pseudomonas genus research on the Antarctic continent is in its infancy. To understand these microorganisms' role in this extreme environment, the characterization and description of new species is vital.

Phylogeny , Pseudomonas/isolation & purification , Pseudomonas/classification , Phenotype , Pseudomonas/genetics , Soil Microbiology , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , RNA, Ribosomal, 16S/genetics , Multilocus Sequence Typing , Islands , Genotype , Antarctic Regions
Braz. j. microbiol ; 49(1): 54-59, Jan.-Mar. 2018. tab
Article in English | LILACS | ID: biblio-889195


ABSTRACT Plant Growth Promoting Rhizobacteria (PGPR) have different mechanisms of action in the development of plants, such as growth promotion, production of phytohormones and antibiotic substances and changes in root exudates. These help to control plant diseases. In order to evaluate the potential of microorganisms in the control of Meloidogyne javanica and Ditylenchus spp., five rhizobacteria isolated from rhizosphere of garlic cultivated in the Curitibanos (SC) region were tested. Hatching chambers were set on Petri dishes, in which were added 10 mL of bacterial suspension and 1 mL of M. javanica eggs suspension, at the rate of 4500, on the filter paper of each chamber. The same procedure was performed with 300 juvenile Ditylenchus spp. The experimental design was completely randomized, with four replications. The evaluations were performed every 72 h for nine days. The antagonized population of nematodes was determined in Peters counting chamber, determining the percentage hatching (for M. javanica) and motility (for Ditylenchus spp). Isolates CBSAL02 and CBSAL05 significantly reduced the hatching of M. javanica eggs (74% and 54.77%, respectively) and the motility of Ditylenchus spp. (55.19% and 53.53%, respectively) in vitro. Isolates were identified as belonging to the genera Pseudomonas (CBSAL05) and Bacillus (CBSAL02).

Animals , Bacillus/physiology , Plant Diseases/prevention & control , Pseudomonas/physiology , Tylenchoidea/microbiology , Bacillus/genetics , Bacillus/isolation & purification , Pest Control, Biological , Plant Diseases/parasitology , Pseudomonas/genetics , Pseudomonas/isolation & purification , Tylenchoidea/physiology
Hig. aliment ; 32(276/277): 108-112, fev. 27, 2018.
Article in Portuguese | LILACS | ID: biblio-884014


[{"text": "Este trabalho objetivou avaliar a\r\ninfluência da cloração da água utilizada\r\nna higienização de tanques de\r\nexpansão na contagem de Escherichia\r\ncoli e Pseudomonas sp. do leite\r\ncru refrigerado. Amostras de leite cru\r\ne de água de18 tanques de expansão\r\nforam avaliadas por um ano, sendo\r\nque em 9 tanques não havia sistema\r\nde cloração da água implantado no\r\ndecorrer de 12 meses e nos outros\r\n9, por 6 meses não houve cloração\r\nda água e nos 6 meses seguintes o\r\nsistema foi implantado. Para determinação\r\nde E. coli no leite cru e na\r\nágua utilizou-se a técnica do Número\r\nMais Provável (NMP) e para\r\na contagem de Pseudomonas sp.\r\nutilizou-se Ágar Para Isolamento de\r\nPseudomonas (PIA). Constataram-\r\n-se valores médios de 1,5NMP/mL e\r\n1,6NMP/mLde E. coli no leite e na\r\nágua, respectivamente, nas amostras\r\nprovenientes dos 9 tanques em que a\r\nágua utilizada não foiclorada por 12\r\nmeses. As amostras de leite e água\r\nprocedentes dos 9 tanques que receberam\r\nágua clorada durante a limpeza\r\npor 6 meses apresentaram em média\r\n1,8NMP/mL e < 1,1 NMP/mL de\r\nE.coli, respectivamente. A contagem\r\nmédia de Pseudomonas sp. nas amostras\r\nde água procedentes dos 9 tanques\r\nem que a água utilizada no processo\r\nde limpeza não recebeu cloração por\r\n6 meses e que posteriormente passou\r\na ser clorada foi 1,1x103UFC/mL e\r\n1,2x102UFC/mL, respectivamente.\r\nPor outro lado,amédia das contagens\r\nde Pseudomonas sp. foi de 9,8x104\r\nUFC/mL e 5,1x105 UFC/mL nas\r\namostras de leite procedentes dos tanques\r\nem que a água utilizada no processo\r\nde limpeza não recebeu cloração\r\npor 6 meses e que posteriormente\r\npassou a ser clorada, respectivamente,\r\no que indica que esta bactéria acessa o\r\nleite cru a partir de diferentes fontes\r\nde contaminação, além da água. Assim,\r\na cloração foi eficiente apenas\r\nna redução da contagem de E. coli e\r\nPseudomonas sp. na água.(AU)", "_i": "pt"}]

Humans , Water Samples , Chlorine/analysis , Water Disinfection/methods , Milk/microbiology , Food Storage , Pseudomonas/isolation & purification , Dairying , Escherichia coli/isolation & purification , Water Contamination Control
Belo Horizonte; s.n; 2018. 92 p. tab, graf, ilus.
Thesis in Portuguese | LILACS, BDENF | ID: biblio-963973


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)

Humans , Infection Control/methods , Endoscopes/microbiology , Detergents/isolation & purification , Detergents/standards , Pseudomonas/isolation & purification , Staphylococcus aureus/isolation & purification , Containment of Biohazards , Academic Dissertation , Enterobacter/isolation & purification , Enzymes , Escherichia coli/isolation & purification , Klebsiella/isolation & purification
Pesqui. vet. bras ; 37(9): 941-948, Sept. 2017. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-895520


This study focused on isolating Pseudomonas spp. during milking process in ten dairy farms with manual and mechanical milking systems during dry and rainy seasons, and evaluating DNA homology and patterns of distribution between isolates, in order to identify main sources of milk contamination by Pseudomonas spp. A total of 167 isolates of Pseudomonas spp. were obtained from water, milkers' hands, cows' teats, teat cups, cooling tanks and raw milk. Bacteria of Pseudomonas spp. genus were isolated from 85 and 82 sampling points in dairy farms with manual and mechanical milking system, respectively. A significant difference (p=0.02) on Pseudomonas spp. isolation was observed among samples of surface of cows' teats before and after pre-dipping, but no significant difference (p>0.05) was observed among milking systems or seasons. The possibility of the same Pseudomonas spp. patterns are distributed in different farms and seasons using Amplified Fragment Length Polymorphism (AFLP) technique was demonstrated. Milkers' hands, surface of cows' teats, teat cups and cooling tanks were associated with raw milk contamination with Pseudomonas spp. on farms with manual and mechanical milking system, showing that regardless of the type of milking system and season, proper hygiene procedures of equipment, utensils and workers' hands are essential to avoid contamination of the milk and, therefore, improve milk quality.(AU)

Este estudo se propôs a isolar Pseudomonas spp. durante o processo de ordenha em dez fazendas com sistemas manuais e mecanizados, durante as estações seca e chuvosa, além de avaliar a homologia do DNA e seus padrões de distribuição entre os isolados, a fim de se determinar as principais fontes de contaminação do leite. Cento e sessenta e sete isolados de Pseudomonas spp. foram obtidos a partir de amostras de água, mãos de ordenhadores, tetos, teteiras, tanques de resfriamento e leite cru armazenado, sendo 85 e 82 pontos de amostragem em fazendas com sistemas de ordenha manual e mecânico, respectivamente. Diferença estatisticamente significativa foi encontrada entre os isolados observados entre a superfície dos tetos antes e após o pré-dipping (p=0,02), mas nenhuma diferença foi encontrada entre sistemas de ordenha ou estações (p>0,05). A possibilidade do mesmo padrão de Pseudomonas spp. estar distribuído em diferentes fazendas ou estações foi avaliada pela técnica de Polimorfismo do Tamanho de Fragmento Amplificado (AFLP). As mãos de ordenhadores, superfície dos tetos das vacas, teteiras e tanques de resfriamento foram associados com a contaminação do leite cru, demonstrando que independente do tipo de ordenha e estação, a higiene adequada de equipamentos, utensílios e mãos dos ordenhadores é essencial para evitar contaminação do leite, e consequentemente aumentar sua qualidade.(AU)

Humans , Animals , Cattle , Pseudomonas/isolation & purification , Stabilization Ponds/analysis , Milk/microbiology , Food Contamination , Amplified Fragment Length Polymorphism Analysis/veterinary , Farms
Braz. dent. j ; 28(4): 423-427, July-Aug. 2017. tab
Article in English | LILACS | ID: biblio-888675


Abstract The aim of this study was to isolate Enterobacteria and Pseudomonas from the oral cavity of hospitalized newborns (NB) and determine their prevalence and the sensitivity profile to most commonly used antibiotics for this age group. Samples from the oral cavity of NB from 24 to 48 h age were collected using swabs. The samples were inoculated on MacConkey agar, incubated and the colonies counted and identified. For each strain, the minimum inhibitory concentration (MIC) was determined using agar dilution test. Tests for enterobacteria producing extended spectrumβ-lactamases (ESBL) were performed using agar diffusion. Descriptive statistics was used for data analysis. Two of the isolated strains were submitted to the susceptibility test in biofilm. Of the collected samples, 8% presented Enterobacteria (mean of 6,141 CFU/mL) and no Pseudomona species was isolated. Positive samples were from NB in accommodation set or in the NB nursery. Enterobacter was the most prevalent genus and some strains were resistant to ampicillin, gentamicin and cephalothin. No ESBL strain was detected. Microorganisms in biofilms were resistant to all antibiotics, with concentrations four times higher than MIC. The presence of enterobacteria in the oral cavity of newborns, especially some strains resistant to normally used antibiotics, warns to the need for care to avoid the early colonization of this niche and the occurrence of a possible hospital infection in this age group.

Resumo O objetivo foi isolar enterobactérias e Pseudomonas da cavidade oral de recém-nascidos hospitalizados (RN) e determinar a prevalência e o perfil de sensibilidade aos antibióticos mais comumente utilizados para este grupo etário. Foram coletadas amostras da cavidade oral de NB com idade de 24-48 horas, usando swab. As amostras foram inoculadas em ágar MacConkey, incubadas e, as colônias contadas e identificadas. Para cada cepa, a concentração inibitória mínima (CIM) foi determinada utilizando teste de ágar diluição. Testes para enterobactérias produtoras de b-lactamases de espectro estendido (ESBL) foram realizados utilizando difusão em ágar. Estatística descritiva foi utilizada para análise dos resultados. Duas das cepas isoladas foram submetidas ao teste de susceptibilidade em biofilme. Das amostras coletadas, 8% apresentaram enterobactérias (média de 6,141 UFC / ml) e nenhuma espécie de Pseudomonas foi isolada. As amostras positivas foram de RN de alojamento conjunto ou RN de berçário. Enterobacter foi o gênero mais prevalente e algumas cepas foram resistentes à ampicilina gentamicina e cefalotina. Não foi detectada cepa ESBL. Micro-organismos em biofilme foram resistentes a todos os antibióticos, em concentrações quatro vezes superiores ao MIC. A presença de enterobactérias em cavidade oral de recém-nascidos, especialmente algumas cepas resistentes aos antibióticos normalmente utilizados, alerta para a necessidade de cuidados, evitando a colonização precoce deste nicho e a ocorrência de possível infecção nosocomial neste grupo etário.

Humans , Infant, Newborn , Anti-Bacterial Agents/pharmacology , Enterobacteriaceae/drug effects , Mouth/microbiology , Pseudomonas/drug effects , Biofilms , Colony Count, Microbial , Drug Resistance, Bacterial , Enterobacteriaceae/isolation & purification , Microbial Sensitivity Tests , Pseudomonas/isolation & purification
Braz. j. microbiol ; 48(2): 187-188, April.-June 2017.
Article in English | LILACS | ID: biblio-839392


Abstract Pseudomonas taiwanensis strain SJ9 is a caprolactam degrader, isolated from industrial wastewater in South Korea and considered to have the potential for caprolactam bioremediation. The genome of this strain is approximately 6.2 Mb (G + C content, 61.75%) with 6,010 protein-coding sequences (CDS), of which 46% are assigned to recognized functional genes. This draft genome of strain SJ9 will provide insights into the genetic basis of its caprolactam-degradation ability.

Pseudomonas/genetics , Pseudomonas/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/chemistry , Caprolactam/metabolism , Genome, Bacterial , Sequence Analysis, DNA , Pseudomonas/isolation & purification , Base Composition , Water Microbiology , Biotransformation , Open Reading Frames , Molecular Sequence Annotation , Industrial Waste , Korea
Braz. j. infect. dis ; 21(1): 98-101, Jan.-Feb. 2017. tab
Article in English | LILACS | ID: biblio-1039180


Abstract Colistin resistance involving Gram-negative bacilli infections is a challenge for health institutions around of the world. Carbapenem-resistance among these isolates makes colistin the last therapeutic option for this treatment. Colistin resistance among Enterobacteriaceae, Acinetobacter spp., and Pseudomonas spp. was evaluated between 2010 and 2014 years, at Hospital das Clínicas, São Paulo, Brazil. Over five years 1346 (4.0%) colistin resistant Gram-negative bacilli were evaluated. Enterobacteriaceae was the most frequent (86.1%) pathogen isolated, followed by Acinetobacter spp. (7.6%), and Pseudomonas spp. (6.3%). By temporal analysis there was a trend for an increase of colistin resistance among Enterobacteriaceae, but not among non-fermentative isolates. Among 1346 colistin resistant isolates, carbapenem susceptibility was observed in 21.5%. Colistin resistance in our hospital has been alarmingly increased among Klebsiella pneumoniae isolates in both KPC positive and negative, thus becoming a therapeutic problem.

Humans , Pseudomonas/drug effects , Acinetobacter/drug effects , Colistin/pharmacology , Drug Resistance, Bacterial/drug effects , Enterobacteriaceae/drug effects , Anti-Bacterial Agents/pharmacology , Pseudomonas/isolation & purification , Time Factors , Acinetobacter/isolation & purification , Brazil , Microbial Sensitivity Tests , Retrospective Studies , Enterobacteriaceae/isolation & purification , Hospitals, University
IBJ-Iranian Biomedical Journal. 2017; 21 (1): 48-56
in English | IMEMR | ID: emr-185667


Background: Alginate is a linear polysaccharide consisting of guluronate [polyG] and mannuronate [polyM] subunits

Methods: In the initial screening of alginate-degrading bacteria from soil, 10 isolates were able to grow on minimal medium containing alginate. The optimization of cell growth and alginate lyase [algL] production was carried out by the addition of 0.8% alginate and 0.2-0.3 M NaCl to the culture medium. Of 10 isolates, one was selected based on its fast growth rate on minimal 9 medium containing 0.4% sodium alginate. The selected bacterium, identified based on morphological and biochemical characteristics, as well as 16S rDNA sequence data, was confirmed to be an isolate belonging to the genus Bacillus and designated as Bacillus sp. TAG8

Results: The results showed the ability of Bacillus sp. TAG8 in utilizing alginate as a sole carbon source. Bacillus sp. TAG8 growth and algL production were augmented with an increase in sodium alginate concentration and also by the addition of 0.2-0.3 M NaCl. Molecular analysis of TAG8 algL gene showed 99% sequence identity with algL of Pseudomonas aeruginosa PAO1. The algL produced by Bacillus sp. TAG8 cleaved both polyM and polyG blocks in alginate molecule, as well as acetylated alginate residues, confirming the bifunctionality of the isolated lyase

Conclusion: The identification of novel algL genes from microbial communities constitutes a new approach for exploring lyases with specific activity against bacterial alginates and may thus contribute to the eradication of persistent biofilms from clinical samples

Substrate Specificity , Bacterial Proteins , Extracellular Matrix Proteins , Pseudomonas/isolation & purification , Bacillus megaterium/isolation & purification
J. appl. oral sci ; 24(5): 462-471, Sept.-Oct. 2016. tab
Article in English | LILACS, BBO | ID: lil-797971


ABSTRACT Aspiration of oral bacteria leads to cardiac and respiratory infectious diseases and dentures can act as a reservoir for pathogenic microorganisms. Objective: To determine the occurrence and the in vitro antimicrobial susceptibility of enteric rods and pseudomonads from the denture biofilm of 52 subjects at the Center for Dental Specialties of Sobral/ Ceara, Brazil. Material and Methods: Denture biofilm was collected and samples plated on MacConkey agar. The isolated bacterial colonies were identified using the BBL Crystal enteric/non-fermenter system. Antibiotic bacterial susceptibility was assessed by the disc diffusion method of amoxicillin, amoxicillin/clavulanic acid, doxycycline, tetracycline, tobramycin, imipenem, cefotaxime, and ciprofloxacin. The Minimum Inhibitory Concentration (MIC) of cefotaxime, tobramycin, doxycycline, imipenem, and ciprofloxacin was determined for 40 species by E-test. Results: 34 subjects (65.4%) harbored enteric rods in their prostheses. Klebsiella pneumoniae (26.5%), Escherichia coli (23.5%), and Enterobacter aerogenes (23.5%) were the most prevalent species. All organisms were susceptible to ciprofloxacin and most species were resistant to amoxicillin or amoxicillin/clavulanic acid, demonstrating variable sensitivity patterns to other antimicrobials. However, the MIC showed the emergence of strains with reduced sensitivity to ciprofloxacin (MIC90≥3 μg/ mL) and cefotaxime (MIC90≥2 μg/mL). Conclusion: The findings show high prevalence of nosocomial diseases-related bacterial species and low susceptibility to antimicrobial drugs. Therefore, these results imply caution against the indiscriminate use of broad spectrum antibiotics in dental practice.

Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Pseudomonas/isolation & purification , Dental Prosthesis/microbiology , Biofilms/growth & development , Enterobacteriaceae/isolation & purification , Pseudomonas/drug effects , Reference Values , Time Factors , Microbial Sensitivity Tests , Sex Distribution , Age Distribution , Statistics, Nonparametric , Drug Resistance, Multiple, Bacterial , Enterobacteriaceae/drug effects , Anti-Bacterial Agents/pharmacology
Braz. j. microbiol ; 46(4): 1037-1044, Oct.-Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769646


In a previous study, three bacterial strains isolated from tropical hydrocarbon-contaminated soils and phylogenetically identified as Achromobacter sp. strain SL1, Pseudomonas sp. strain SL4 and Microbacterium esteraromaticum strain SL6 displayed angular dioxygenation and mineralization of carbazole in batch cultures. In this study, the ability of these isolates to survive and enhance carbazole degradation in soil were tested in field-moist microcosms. Strain SL4 had the highest survival rate (1.8 x 107 cfu/g) after 30 days of incubation in sterilized soil, while there was a decrease in population density in native (unsterilized) soil when compared with the initial population. Gas chromatographic analysis after 30 days of incubation showed that in sterilized soil amended with carbazole (100 mg/kg), 66.96, 82.15 and 68.54% were degraded by strains SL1, SL4 and SL6, respectively, with rates of degradation of 0.093, 0.114 and 0.095 mg kg−1 h−1. The combination of the three isolates as inoculum in sterilized soil degraded 87.13% carbazole at a rate of 0.121 mg kg−1 h−1. In native soil amended with carbazole (100 mg/kg), 91.64, 87.29 and 89.13% were degraded by strains SL1, SL4 and SL6 after 30 days of incubation, with rates of degradation of 0.127, 0.121 and 0.124 mg kg−1 h−1, respectively. This study successfully established the survivability (> 106 cfu/g detected after 30 days) and carbazole-degrading ability of these bacterial strains in soil, and highlights the potential of these isolates as seed for the bioremediation of carbazole-impacted environments.

Achromobacter/chemistry , Achromobacter/genetics , Achromobacter/isolation & purification , Achromobacter/metabolism , Actinobacteria/chemistry , Actinobacteria/genetics , Actinobacteria/isolation & purification , Actinobacteria/metabolism , Biodegradation, Environmental/chemistry , Biodegradation, Environmental/genetics , Biodegradation, Environmental/isolation & purification , Biodegradation, Environmental/metabolism , Carbazoles/chemistry , Carbazoles/genetics , Carbazoles/isolation & purification , Carbazoles/metabolism , Phylogeny/chemistry , Phylogeny/genetics , Phylogeny/isolation & purification , Phylogeny/metabolism , Pseudomonas/chemistry , Pseudomonas/genetics , Pseudomonas/isolation & purification , Pseudomonas/metabolism , Soil Microbiology/chemistry , Soil Microbiology/genetics , Soil Microbiology/isolation & purification , Soil Microbiology/metabolism , Soil Pollutants/chemistry , Soil Pollutants/genetics , Soil Pollutants/isolation & purification , Soil Pollutants/metabolism
Braz. j. microbiol ; 45(2): 621-625, Apr.-June 2014. ilus, tab
Article in English | LILACS | ID: lil-723126


Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR.

Azospirillum brasilense/isolation & purification , Bacillus/isolation & purification , Endophytes/isolation & purification , Hordeum/microbiology , Nitrogen Fixation , Pseudomonas/isolation & purification , Seeds/microbiology , Antibiosis , Azospirillum brasilense/classification , Azospirillum brasilense/genetics , Azospirillum brasilense/metabolism , Bacillus/classification , Bacillus/genetics , Bacillus/metabolism , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Endophytes/classification , Endophytes/genetics , Endophytes/metabolism , Indoleacetic Acids/metabolism , Molecular Typing , Pseudomonas/classification , Pseudomonas/genetics , Pseudomonas/metabolism , Random Amplified Polymorphic DNA Technique , /genetics , Sequence Analysis, DNA
Assiut Medical Journal. 2014; 38 (2): 105-110
in English | IMEMR | ID: emr-160291


Carbapenem-resistant Gram negative bacilli [CRGNB] resulting from carbapenemase enzyme production have been reported to be an important cause of nosocomial infection and are a critical therapeutic problem worldwide. This study was conducted to determine the prevalence of carbapenem-resistant Gram negative bacilli in Asiut University Hospital and relevant risk factors associated with its occurrence. The study was conducted over a period of 12 month from January 2011 to January 2012 on 355 specimen of 134 nosocomialy infected patients, aged two month to 82 years, admitted to the ICUs at Assiut University Hospital. Patient demographics, comorbidities, antibiotics, invasive devices were determined to identify risk factors associated with CRGNB infections. Three hundred and fifty five Gram negative bacilli isolates [47.92%] were isolated from patients. Ninety seven [27.17%] of Gram negative bacilli isolates were carbapenem-resistant. Patients aged above 40 years, male sex, prior antibiotic treatment, the presence of a urinary catheter, mechanical ventilation and central line in ICU were the most prevalent risk factors associated with the accurence of nosocomial CRGNB infections. The prevalence of CRGNB was 27.17% among GNB isolates. Patients aged above 40 years, male sex, prior antibiotic treatment, the presence of a urinary catheter, mechanical ventilation and central line in ICU were the most prevalent risk factors Interventions aimed reducing initial broad spectrum antibiotics are clearly needed to help control the spread in of these difficult to treat infections

Pseudomonas/isolation & purification , Enterobacteriaceae/isolation & purification , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae Infections/epidemiology , Prevalence , Cross Infection/epidemiology , Risk Factors , Hospitals, University
Rev. argent. microbiol ; 45(1): 57-60, mar. 2013.
Article in Spanish | LILACS, BINACIS | ID: biblio-1171766


The present work sought to detect the presence of Pseudomonas spp. at different stages of an effluent treatment plant using the Australian system of stabilization ponds, and to determine the susceptibility of those isolates to different antimicrobials. Thirty-four isolates of Pseudomonas spp. derived from effluent treatment station water samples were collected near the transfer ducts between the ponds in November/2008 and december/2009. Among the Pseudomonas spp. isolates, 47.05

showed susceptibility to all antimicrobials tested, 20.58

were resistant to cefepime, and 24

showed intermediate resistance to streptomycin. No Pseudomonas spp. isolates were found in the final pond, or in post-treatment effluents. The Pseudomonas spp. isolates did not exhibit multiresistance to the antimicrobials tested.

Abattoirs , Pseudomonas/drug effects , Drug Resistance, Microbial , Sus scrofa/microbiology , Animals , Anti-Bacterial Agents/pharmacology , Brazil , Waste Disposal, Fluid/methods , Medical Waste Disposal/methods , Species Specificity , Retrospective Studies , Water Microbiology , Pseudomonas/classification , Pseudomonas/isolation & purification , Industrial Waste , Swine , Disk Diffusion Antimicrobial Tests , Waste Water/microbiology
Braz. j. microbiol ; 44(1): 307-315, 2013. ilus, tab
Article in English | LILACS | ID: lil-676922


Microbiological analysis of overburden samples collected from chromite mining areas of Orissa, India revealed that they are rich in microbial density as well as diversity and dominated by Gramnegative (58%) bacteria. The phenotypically distinguishable bacterial isolates (130) showed wide degree of tolerance to chromium (2-8 mM) when tested in peptone yeast extract glucose agar medium. Isolates (92) tolerating 2 mM chromium exhibited different degrees of Cr+6 reducing activity in chemically defined Vogel Bonner (VB) broth and complex KSC medium. Three potent isolates, two belonging to Arthrobacter spp. and one to Pseudomonas sp. were able to reduce more than 50 and 80% of 2 mM chromium in defined and complex media respectively. Along with Cr+6 (MIC 8.6-17.8 mM), the isolates showed tolerance to Ni+2, Fe+3, Cu+2 and Co+2 but were extremely sensitive to Hg+2 followed by Cd+2, Mn+2 and Zn+2. In addition, they were resistant to antibiotics like penicillin, methicillin, ampicillin, neomycin and polymyxin B. During growth under shake-flask conditions, Arthrobacter SUK 1201 and SUK 1205 showed 100% reduction of 2 mM Cr+6 in KSC medium with simultaneous formation of insoluble precipitates of chromium salts. Both the isolates were also equally capable of completely reducing the Cr+6 present in mine seepage when grown in mine seepage supplemented with VB concentrate

Arthrobacter/isolation & purification , Biodiversity , Carcinogens, Environmental , Environmental Microbiology , Metals/analysis , Garbage , Pseudomonas/isolation & purification , Methods , Minerals , Waste Products
Braz. j. microbiol ; 44(2): 629-637, 2013. ilus, tab
Article in English | LILACS | ID: lil-688594


The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (µg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1-0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05) when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plantgrowth promoting agent.

Endophytes/isolation & purification , Endophytes/metabolism , Pseudomonas/isolation & purification , Pseudomonas/metabolism , Sophora/microbiology , Antibiosis , Anti-Bacterial Agents/pharmacology , Cluster Analysis , Carboxylic Acids/metabolism , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Endophytes/classification , Endophytes/genetics , Fungi/growth & development , Hydrogen-Ion Concentration , Indoleacetic Acids/metabolism , Molecular Sequence Data , Phylogeny , Plant Development , Phosphates/metabolism , Plant Roots/microbiology , Pseudomonas/classification , Pseudomonas/genetics , /genetics , Sequence Analysis, DNA , Siderophores/metabolism , Sodium Chloride/metabolism , Sophora/growth & development , Temperature
Braz. j. microbiol ; 42(4): 1321-1328, Oct.-Dec. 2011. tab
Article in English | LILACS | ID: lil-614591


Antimicrobial therapy may cause changes in the resident oral microbiota, with the increase of opportunistic pathogens. The aim of this study was to compare the prevalence of Candida, Staphylococcus, Pseudomonas and Enterobacteriaceae in the oral cavity of fifty patients undergoing antibiotic therapy for pulmonary tuberculosis and systemically healthy controls. Oral rinsing and subgingival samples were obtained, plated in Sabouraud dextrose agar with chloramphenicol, mannitol agar and MacConkey agar, and incubated for 48 h at 37ºC. Candida spp. and coagulase-positive staphylococci were identified by phenotypic tests, C. dubliniensis, by multiplex PCR, and coagulase-negative staphylococci, Enterobacteriaceae and Pseudomonas spp., by the API systems. The number of Candida spp. was significantly higher in tuberculosis patients, and C. albicans was the most prevalent specie. No significant differences in the prevalence of other microorganisms were observed. In conclusion, the antimicrobial therapy for pulmonary tuberculosis induced significant increase only in the amounts of Candida spp.

Humans , Candida , Diagnostic Techniques and Procedures , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/pathogenicity , Genetic Techniques , Enterobacteriaceae Infections/therapy , Pseudomonas/isolation & purification , Methods , Patients , Polymerase Chain Reaction , Prevalence , Methods
Braz. j. microbiol ; 42(4): 1479-1484, Oct.-Dec. 2011. ilus, tab
Article in English | LILACS | ID: lil-614613


The production of extracellular hydrolases by a psychrotrophic bacterium isolated from refrigerated raw milk, and identified as a Pseudomonas sp. belonging to the Pseudomonas jenssenii group, was studied. This bacterium produced proteolytic and lipolytic enzymes in all media investigated (skim milk, cheese whey, casein broth, and tryptone soy broth). High levels of á-glucosidase were produced in skim milk broth. Hydrolytic enzymes detected in skim milk broth are of particular concern, indicating that these enzymes could be produced by Pseudomonas sp. during the cold storage of raw milk, contributing to the spoilage problem in milk and dairy products.

Cooled Foods , Glucosidases/analysis , Lipase/analysis , Pseudomonas/isolation & purification , Food Samples , Milk
Braz. j. microbiol ; 42(3): 927-936, July-Sept. 2011. ilus, tab
Article in English | LILACS | ID: lil-607521


Psychrophilic bacteria, which grow on lactose as a carbon source, were isolated from Antarctic polar sea water. Among the psychrophilic bacteria isolated, strain KNOUC808 was able to grow on lactose at below 5¨¬C, and showed 0.867 unit of o-nitrophenyl ¥â-D-galactopyranoside(ONPG) hydrolyzing activity at 4¨¬C. The isolate was gram-negative, rod, aerobic, catalase positive and oxidase positive. Optimum growth was done at 20¨¬C, pH 6.8-7.2. The composition of major fatty acids in cell of KNOUC801 was C12:0 (5.48 percent), C12:0 3OH (9.21 percent), C16:0 (41.83 percent), C17:0 ¥ø8 (7.24 percent) and C18:1 ¥ø7 (7.04 percent). All suthese results together suggest that it is affiliated with Pseudoalteromonas genus. The 16S rDNA sequence corroborate the phenotypic tests and the novel strain was designated as Pseudoalteromonas sp. KNOUC808. The optimum temperature and pH for lactose hydrolyzing enzyme was 20¨¬C and 7.8, respectively. The enzyme was stable at 4¨¬C for 7 days, but its activity decreased to about 50 percent of initial activity at 37¨¬C in 7 days.

Lactose/analysis , Lactose/isolation & purification , Pseudomonas/genetics , Pseudomonas/isolation & purification , Methodology