Genomic Characterization of Aeromonas veronii Provides Insights into Taxonomic Assignment and Reveals Widespread Virulence and Resistance Genes throughout the World.

Aeromonas veronii is a Gram-negative bacterial species that causes disease in fish and is nowadays increasingly recurrent in enteric infections of humans. This study was performed to characterize newly sequenced isolates by comparing them with complete genomes deposited at the NCBI (National Center for Biotechnology Information). Nine isolates from fish, environments, and humans from the São Francisco Valley (Petrolina, Pernambuco, Brazil) were sequenced and compared with complete genomes available in public databases to gain insight into taxonomic assignment and to better understand virulence and resistance profiles of this species within the One Health context. One local genome and four NCBI genomes were misidentified as A. veronii. A total of 239 virulence genes were identified in the local genomes, with most encoding adhesion, motility, and secretion systems. In total, 60 genes involved with resistance to 22 classes of antibiotics were identified in the genomes, including mcr-7 and cphA. The results suggest that the use of methods such as ANI is essential to avoid misclassification of the genomes. The virulence content of A. veronii from local isolates is similar to those complete genomes deposited at the NCBI. Genes encoding colistin resistance are widespread in the species, requiring greater attention for surveillance systems.

Genome Sequence of Pseudomonas sp. Strain LAP_36, A Rhizosphere Bacterium Isolated from King George Island, Antarctica.

Pseudomonas sp. strain LAP_36 was isolated from rhizosphere soil from Deschampsia antarctica on King George Island, South Shetland Islands, Antarctica. Here, we report on its draft genome sequence, which consists of 8,794,771 bp with 60.0% GC content and 8,011 protein-coding genes.

Antimicrobial activity of polypyrrole nanoparticles and aqueous extract of Moringa oleifera against Staphylococcus spp. carriers of multi-drug efflux system genes isolated from dairy farms.

Our objectives were to identify genes of the multi-drug efflux system and to evaluate the antimicrobial activities of polypyrrole nanoparticles (PPy-NPs) and aqueous extract of Moringa oleifera against Staphylococcus spp. isolated from dairy farms in Northeast Brazil. Initially, 162 Staphylococcus spp. isolates were subjected to in vitro antimicrobial sensitivity tests. Of these, 35 presented antimicrobial multi-drug resistance phenotypes. These 35 isolates were then referred for the detection of norA, norB, norC, msrA, mgrA, tet-38, and lmrS genes, all of which feature in multi-drug efflux systems. In the isolates carrying the genes, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs and Moringa oleifera aqueous extract were determined. In the molecular analysis of the 35 isolates norA, norC, tet-38, and msrA genes were detected and for the other genes norB, lmrS and mgrA there was no amplification. Antimicrobial activity was verified of PPy-NPs and aqueous extract of Moringa oleifera in Staphylococcus spp. carrying multi-drug efflux system genes. We concluded that there are multi-drug efflux system genes present in the Staphylococcus spp. from the agricultural environment in Northeast Brazil, and that aqueous extract of Moringa oleifera and PPy-NPs show bactericidal activity against these isolates.

Antimicrobial resistance profile of non-aureus Staphylococci isolates from buffalo, goat and sheep mastitis in the Northeast region of Brazil.

The study described in this Research Communication investigated the genotypic and phenotypic profiles of resistance to beta-lactams and other antimicrobials in non-aureus Staphylococci (NAS) isolated from buffalo, goat and sheep mastitis in the Northeast region of Brazil. A total of 190 isolates were analyzed and 42.3, 43.9 and 23.6% of them were positive for blaZ gene in buffalo, goat and sheep, respectively. Regarding the animal groups, in goats, amoxicillin was the antimicrobial with highest resistance index (72.7%), followed by penicillin G in buffaloes (51.9%) and ampicillin in sheep (43.1%). With regard to multiple antimicrobial resistance, 30.8% of NAS isolates from buffalo milk samples, 25.8% from goats and 25.0% from sheep presented multidrug-resistance. In the minimum inhibitory concentration (MIC) technique, amoxicillin MIC50 and MIC90 were 64 and 128 µg/ml, respectively, among isolates of the three animal species. In conclusion, high rates of resistance to beta-lactams are presented among NAS isolated from mastitis cases in buffaloes, goats and sheep in Northeast region of Brazil. These results provide an alert to animal and human health researchers, suggesting that the frequency of NAS needs to be reduced because they carry resistance genes which might increase the existing levels of antimicrobial resistance.

In vitro algicidal effect of polypyrrole on Prototheca species isolates from bovine mastitisAlgicidal activity of polypyrrole on Prototheca spp.

Algae of the genus Prototheca are microorganisms involved in the occurrence of diseases in humans and animals. In bovine species, Prototheca spp. cause environmental mastitis, productive losses in dairy herds, mainly leading to the discard of infected cows. Currently, there are no effective anti-Prototheca spp. drugs to combat this infection. Thus, the search for an efficacious therapy for Prototheca spp. infections have become essential. Highly soluble polypyrrole (Ppy) is a molecule with known antimicrobial activity. This study aimed to characterize Prototheca spp. isolates from bovine mastitis as well as to evaluate the susceptibility profile and to verify the morphological alterations on Prototheca spp. isolates treated with Ppy. In this research, 36 Brazilian isolates of Prototheca spp. were characterized by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) assay for the mitochondrial cytB gene. Additionally, Ppy algicidal activity against these isolates of Prototheca spp. was assessed by minimal microbicidal concentration method in microplates. Further, scanning electron microscopy (SEM) was performed in order to verify the morphological alterations on Prototheca spp. isolates in response to Ppy. The isolates were characterized as belonging to Prototheca zopfii genotype 2 (35/36) and Prototheca blaschkeae (1/36). Ppy had an algicidal effect on all isolates tested at concentrations ranging from 15.625 µg ml-1 to 62.5 µg ml-1. SEM showed changes on planktonic and sessile P. zopfii, including a decrease of the number of cells with the presence of an amorphous substance involving the cells. The algicidal activity of Ppy suggests the therapeutic potential of this molecule in the prevention and treatment of Prototheca spp. in bovine mastitis.

Biofilm formation by Prototheca zopfii isolated from clinical and subclinical bovine mastitis in distinct growth conditions under different dyes / Formação de biofilme por Prototheca zopfii isolados de mastite bovina clínica e subclínica em distintas condições de crescimento sob diferentes corantes

ABSTRACT: Prototheca spp. have been reported as an emergent environmental mastitis pathogen in several countries. Biofilm formation is a significant factor associated with different degrees of virulence developed by many microorganisms, including Prototheca spp. The present study aimed to compare two growth conditions and two staining dyes to determine which combination was more appropriate to evaluate qualitatively and quantitatively the production of biofilm by P. zopfii. Biofilm formation was evaluated in polystyrene microplates under static and dynamic growth conditions and staining with crystal violet or cotton blue dye. All P. zopfii isolates from cows with mastitis were classified as biofilm-producers in all growth conditions and staining. The cotton blue dye proved to be more appropriate method to classify the intensity of P. zopfii biofilm production.

RESUMO: Prototheca spp. tem sido relatado como um patógeno ambiental causador de mastite bovina em vários países. A formação de biofilme é um fator associado a diferentes graus de virulência desenvolvidos por muitos microrganismos, incluindo Prototheca zopfii. O presente estudo teve como objetivo comparar duas condições de crescimento e dois corantes para determinar a combinação mais adequada para avaliar qualitativa e quantitativamente a produção de biofilme por P. zopfii. A formação de biofilme foi avaliada em microplacas de poliestireno sob condições estáticas e dinâmicas de crescimento e coloração com cristal violeta ou azul de algodão. Todos os isolados de P. zopfii de vacas com mastite foram caracterizados como produtores de biofilme, independentemente das condições de crescimento e coloração. O corante azul de algodão demonstrou ser o método mais adequado para classificar a intensidade de produção de biofilme de P. zopfii.

Phytochemical screening and antimicrobial activity testing of crude hydroalcoholic extract from leaves of Sphagneticola trilobata (Asteraceae) / Triagem fitoquímica e atividade antimicrobiana do extrato hidroalcoólico bruto das folhas de Sphagneticola trilobata (Asteraceae)

ABSTRACT: This study aimed to perform phytochemical analysis and to test the antimicrobial activity of the crude hydroalcoholic extract obtained from the leaves of Sphagneticola trilobata. Classes of secondary metabolites present in the extract were identified through phytochemical screening using analytical thin-layer chromatography. Antimicrobial activity was evaluated by testing cultures of Staphylococcus aureus, S. epidermidis, Staphylococcus spp., Escherichia coli, Serratia marcescens, Enterococcus faecalis, Pseudomonas aeruginosa, Salmonella Typhimurium, and Klebsiella pneumoniae isolated from human skin and those of Staphylococcus spp. isolated from dog skin using the broth microdilution method. In the phytochemical screening, classes of anthracenic derivatives and mono-, sesqui-, and diterpenes were identified. Colorimetric analysis showed total phenol and total flavonoid contents of 21.7 ± 0.009 mg of gallic acid equivalents per gram of sample and 0.23 ± 0.005 mg of catechin equivalents per gram of sample, respectively. Microbiological analysis revealed that the hydroalcoholic extract of S. trilobata exhibited antimicrobial activity against cultures of Staphylococcus spp., E. coli, S. marcescens, and E. faecalis isolated from human skin and those of Staphylococcus spp. isolated from dog skin. Thus, crude hydroalcoholic extract of leaves of S. trilobata contained flavonoids and terpenoids as secondary metabolites, which contributed to its antimicrobial activity against skin bacteria isolated from different sources.

RESUMO: Este estudo teve como objetivo realizar a triagem fitoquímica preliminar e testar a atividade antimicrobiana do extrato hidroalcoólico bruto das folhas de Sphagneticola trilobata. A identificação das classes de metabólitos secundários presentes no extrato foi realizada através da cromatografia em camada delgada analítica (CCDA). Para determinar a quantidade de fenóis e flavonoides totais foram utilizados os métodos espectrofotométricos de Folin-Ciocalteu e complexação com AlCl3, respectivamente. Para avaliar a atividade antimicrobiana foram testadas culturas de Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus spp., Escherichia coli, Serratia marcescens, Enterococcus faecalis, Pseudomonas aeruginosa, Salmonella Typhimurium, Klebsiella pneumoniae isoladas de pele humana e culturas de Staphylococcus spp. isoladas de pele de cães pelo método de microdiluição em caldo. Na triagem fitoquímica foi verificada reação positiva para a presença de derivados antracênicos, mono, sesqui e diterpenos. As análises colorimétricas mostraram conteúdos de fenóis totais e flavonoides totais de 21,7 ± 0,009 miligramas de equivalentes de ácido gálico por grama de amostra e 0,23 ± 0,005 miligramas de equivalentes de catequina por grama de amostra, respectivamente. Na análise microbiológica, o extrato hidroalcoólico das folhas de Sphagneticola trilobata apresentou atividade antimicrobiana frente às culturas de Staphylococcus spp., Escherichia coli, Serratia marcescens e Enterococcus faecalis. Todas as culturas de Staphylococcus spp. isoladas de pele de cães foram sensíveis ao extrato. Conclui-se que o extrato hidroalcoólico bruto das folhas de Sphagneticola trilobata possui entre seus metabólitos secundários os flavonoides e terpenoides que contribuíram com a atividade antimicrobiana frente às bactérias isoladas de pele de diferentes origens.

Ocorrência de ovinos portadores da infecção por Campylobacter spp. no estado de Pernambuco / Occurrence of sheep carrier of infection with Campylobacter spp. in the state of Pernambuco, Brazil

Objetivou-se com este estudo determinar a ocorrência e os fatores de risco associados à infecção por Campylobacter spp. em criações de ovinos no estado de Pernambuco, Brasil. Foram coletadas 421 amostras fecais de ovinos procedentes de 20 rebanhos para o isolamento de Campylobacter spp. As espécies Campylobacter fetus subsp. fetus e Campylobacter jejuni foram identificadas por Reação em Cadeia da Polimerase (PCR). Para análise dos fatores de risco foi realizada uma análise univariada e posteriormente regressão logística a partir de questionário com perguntas objetivas sobre o manejo higiênico-sanitário e reprodutivo. A ocorrência para Campylobacter spp. foi de 4,5% (19/421; I.C. 2,8% - 7,1%). Das 19 amostras positivas no cultivo, oito (1,9%; I.C. 0,9% - 3,9%) foram classificadas como C. fetus subsp. fetus e sete (1,7%; I.C. 0,7% - 3,6%) como C. jejuni, com co-infecção em quatro amostras (0,95%). O número de focos identificados foi de 35,0% (7/20) das criações de ovinos que apresentavam pelo menos um animal positivo. Na análise de regressão logística não foi identificada nenhuma das variáveis como fator de risco. Este é o primeiro registro da infecção por Campylobacter spp. em rebanhos ovinos no Nordeste do Brasil, concluindo-se que a infecção ocorre nesses rebanhos. Dessa forma, se faz necessário à implementação de medidas de controle e prevenção, para impedir a propagação do agente entre as criações, evitando prejuízos para ovinocultura e riscos para saúde pública, uma vez que a campilobacteriose é considerada uma zoonose emergente.(AU)

The objective of this study was to determine the occurrence and risk factors associated with Campylobacter spp. infection in sheep in the State of Pernambuco, Brazil. A total of 421 fecal samples were collected from 20 herds for the isolation of Campylobacter spp. The species Campylobacter fetus and Campylobacter jejuni were identified by Polymerase Chain Reaction (PCR). To analyze the risk factors, logistic regression was conducted through a questionnaire about the hygienic-sanitary and reproductive management. The occurrence of Campylobacter spp. was 4.5% (19/421; C.I. 2.8 to 7.1%). Of the 19 positive samples isolated, eight (1.9% CI 0.9 to 3.9%) were classified as C. fetus subsp. fetus and seven (1.7% CI 0.7 to 3.5%) as C. jejuni, with co-infection in four samples (0.95%). The number of identified focuses was 35.0% (7/20) of the sheep herds that had at least one positive animal. The logistic regression analysis did not identify any of the variables as a risk factor. This appears to be the first report of infection with Campylobacter spp. in sheep herds in northeastern Brazil. Thus it is necessary to implement measures for control and prevention avoid damage to sheep production and risk to public health, since campylobacteriosis is considered an emerging zoonosis.(AU)

The presence of plasmids in Aeromonas hydrophila and its relationship with antimicrobial and heavy metal-resistance profiles / Presença de plasmídeos em Aeromonas hydrophila e sua relação com perfis de resistência aos antimicrobianos e metais pesados

ABSTRACT: Aeromonas hydrophila is a common fish pathogen that causes extensive damage to aquaculture. To develop and implement a more adequate strategy to farm fish, it is crucial to understand the bacterial-resistance levels and their transference dynamics. The objective of this study was to analyze the resistance profile of isolated Aeromonas hydrophila to antimicrobial agents and heavy metals and draw a correlation of the observed profiles with the presence of plasmids. Resistance of the isolated bacteria to antimicrobial agents (oxacilin, gentamicin, tetracycline, and nalidixic acid) and heavy metals (cadmium, lead, copper, and manganese) was verified using the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) standards. The Multiple Antibiotic Resistance Index (MAR Index) was calculated. Plasmids were extracted by using a common methodology described elsewhere. Mann-Whitney Test, implemented in the R environment, was used to determine the correlation between resistance and plasmids presence. A high resistance to almost all antimicrobial agents and heavy metals was observed, except to gentamicin and cadmium. The MAR index results showed resistance to all antimicrobial profiles. Of the isolated bacteria, 14 showed the presence of plasmids. However, no correlation was noted between the resistance profile and the plasmid presence for these isolates, indicating that the genes responsible for resistance to microbial agents and heavy metals are present in the cromossomic DNA, which in turn suggested the possibility of gene transfer between the isolated bacteria. The resistance to heavy metals can be linked to heavy utilization of fertilizers along the Sao Francisco River.

RESUMO: Bactérias da espécie Aeromonas hydrophila são patógenos que atacam peixes, causando grandes prejuízos à piscicultura. Entender os perfis de resistência dessa bactéria e a capacidade da mesma em transferir tal resistência é importante para implantação de um manejo adequado na produção de peixes. Os objetivos desse estudo foram analisar a resistência de isolados de Aeromonas hydrophila à antimicrobianos e metais pesados e, correlacionar os perfis encontrados com a presença de plasmídeos. A resistência dos isolados aos antimicrobianos (oxacilina, gentamicina, tetraciclina e ácido nalidíxico) e metais pesados (cádmio, chumbo, cobre e manganês) foi verificada pelas técnicas da Concentração Bactericida Mínima (CBM) e Concentração Inibitória Mínima (CIM). Foi calculado o Índice de Resistência Múltipla aos Antimicrobianos (IRMA). Os plasmídeos foram extraídos por metodologias descritas pela literatura. A relação entre a resistência aos antimicrobianos e metais pesados com a presença de plasmídeos foi determinada pelo teste de Mann-Whitney utilizando o ambiente R. Foi observada alta resistência aos antimicrobianos e metais pesados testados, com exceção à gentamicina e cádmio. No IRMA os isolados apresentaram resistência a todos os perfis de antimicrobianos possíveis. Quatorze isolados apresentaram plasmídeos, mas não foi encontrada relação dos perfis de resistência com a presença destes, o que indica que os genes de resistência a esses compostos estejam presentes no DNA cromossômico. Porém apontam a possibilidade de transferência dos genes de resistência entre os isolados. Estes resultados apontam alta resistência dos isolados e capacidade de transmissão dessa resistência a outras bactérias. A resistência aos metais pesados, pode estar ligada ao uso de fertilizantes nas plantações localizadas próximas as margens do Rio São Francisco.

Soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (teste in situ) / Solutions of Zinc oxide and silver nitrate as an alternative to bovine udder antisepsis

Este trabalho teve como objetivo analisar o potencial das soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (in situ), considerando a escassez de literatura a respeito do uso destes compostos na profilaxia de mastites. Primeiramente foram realizados ensaios de microdiluição e CBM (Concentração Bactericida Mínima) a fim de verificar a atividade antimicrobiana das soluções de óxido de zinco e de nitrato de prata sobre 30 isolados de Staphylococcus spp., obtidos de casos de mastite. Todos os isolados apresentaram sensibilidade às duas soluções testadas, mas a solução de nitrato de prata apresentou menores valores de CBMs (3,05 a 97,65 µg/mL), quando comparada as CBMs da solução de óxido de zinco (97,65 a 6.250 µg/mL). Posteriormente, foram conduzidos os ensaios de antissepsia dos tetos (in situ) através da imersão dos mesmos nas soluções antimicrobianas testadas. Para isso, foram utilizados 40 tetos (n=40) oriundos de vacas abatidas, os quais foram divididos em 4 grupos de 10 tetos, destinados ao teste das solução de óxido de zinco a 3% (30mg/mL), de nitrato e prata 1% (10mg/mL) e seus respectivos controles. Os tetos foram submetidos à contagem de microrganismos mesófilos na superfície dos tetos (UFC/cm2) antes e após a imersão nas soluções testadas. Como resultado, todas as soluções (de óxido de zinco e de nitrato de prata) apresentaram significativa redução de UFC/cm2 até aos 60 minutos após a imersão (M 0' a M 60'). A solução de nitrato de prata apresentou redução de UFC/cm2 significativamente maior, quando comparado ao grupo testado com solução de óxido de zinco. Tais resultados validam o potencial das soluções de óxido de zinco e nitrato de prata para utilização no pré e pós-dipping em vacas leiteiras.(AU)

This study aimed to analyze the zinc oxide and silver nitrate solutions as an alternative antiseptic for cow teats (in situ), considering the lack in the literature about these compounds uses in mastitis prophylaxis. First microdilution tests med and MBC (Minimal Bactericidal Concentration were perfor in order to determine the antimicrobial activity of zinc oxide and silver nitrate solutions over 30 Staphylococcus spp. isolates, obtained from cows with mastitis. All strains tested showed sensitivity to both solutions, but the silver nitrate solution had lower MBC values (3,05 to 97,65ug/ml) compared with zinc oxide solution MBCs (97,65 to 6,250 ug/ml). Subsequently, the antiseptic teat tests were conducted (in situ) by immersing the teats in same antimicrobial solutions tested in the first experiment. Therefore, 40 teats were used (n = 40) originating from slaughtered cows were divided into 4 groups of 10 teats, to test test zinc oxide 3% (30mg/mL) and silver nitrate 1% (10mg/mL) solutions and their respective controls. The teats were submitted to mesophilic count on the teat surface (CFU/cm2) before and after immersion in tested solutions. As a result, all the solutions (zinc oxide and silver nitrate) had a significant reduction in CFU/cm2 until 60 minutes after immersion (M 0' to 60 M'). Silver nitrate solution showed a reduction of CFU/cm2 significantly higher compared to the group treated with zinc oxide solution. These results validate the potential use of zinc oxide and silver nitrate solutions in dairy cows as a pre- and post-dipping antiseptic.(AU)

Indirect diagnostic tests for the detection of subclinical mastitis in dairy goats experimentally infected with Staphylococcus aureus / Testes diagnósticos indiretos na detecção da mastite subclínica em cabras leiteiras infectadas experimentalmente com Staphylococcus aureus

ABSTRACT: The aim of the present study was to assess two diagnostic techniques (California mastitis test (CMT) and the somatic cell count (SCC)) that can diagnose mastitis in dairy goats. Experimental infection was conducted using 20 mammary glands, a strain of Staphylococcus aureus, an infectious dose of 1.2x108CFU mL-1 and a volume of 1mL per mammary gland. The CMT and the SCC were used to detect subclinical mastitis. Bacterial culture (BC) was performed immediately after milk collection and was used as the gold standard. Four experimental time points were established (0, 24, 48 and 72 hours post-inoculation). Analysis of the ROC curve confirmed that the best combination of sensitivity and specificity were obtained with a cutoff point of 405.5, 6030.0 and 729.5x103 cells mL-1, respectively at M1, M2 and M3. Furthermore, considering the drop in sensitivity throughout the experimental time points, the use of serial bacterial cultures are recommended, particularly in herds with a high prevalence of S. aureus.

RESUMO: Neste trabalho, objetivou-se avaliar duas técnicas diagnósticas (California mastitis test (CMT) e a contagem de células somáticas (CCS)) disponíveis para o diagnóstico da mastite em cabras leiteiras. Realizou-se infecção experimental em 20 metades mamárias, utilizando-se cepa de S. aureus, em uma dose infectante de 1,2x108 UFC mL-1 e um volume de 1mL/metade mamária. Para detecção da mastite subclínica, foi utilizado o CMT e a CCS. A cultura bacteriológica (CB) foi empregada como padrão ouro, sendo realizada logo após a coleta do leite. Foram estabelecidos quatro momentos experimentais (0, 24, 48 e 72 horas pós-infecção). A análise da curva de ROC confirmou que a melhor combinação (sensibilidade e especificidade) foi obtida com ponto de corte de 405,5, 6030,0 e 729,5x103 cells mL-1, respectivamente, em M1, M2 and M3. Ademais, levando em consideração a queda da sensibilidade ao longo dos momentos experimentais, é relevante a realização da cultura bacteriológica seriada, principalmente em rebanhos com elevada prevalência de S. aureus.

Genotypic and phenotypic detection of efflux pump in Rhodococcus equi

The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90) of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi.

Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections.

Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

Perfil de suscetibilidade antimicrobiana e presença do gene vapA em Rhodococcus equi de origem humana, ambiental e equina / Susceptibility profile of Brazilian Rhodococcus equi isolates to different antimicrobial classes and the presence of vapA gene

Rhodococcus equi é um micro-organismo intracelular facultativo, agente etiológico da rodococose, uma importante enfermidade que acomete principalmente potros com menos de seis meses de idade, causando a morte geralmente em decorrência de lesões pulmonares. Este agente também tem potencial zoonótico e emergiu como um patógeno oportunista no mundo, acometendo humanos imunocomprometidos, especialmente os transplantados e infectados pelo vírus da imunodeficiência humana (HIV). Entretanto, infecções por R. equi em hospedeiros hígidos tem sido relatadas, principalmente em crianças e idosos. Estudos tem mostrado um nível crescente na resistência de isolados de R. equi em relação aos antimicrobianos comumente utilizados no tratamento de animais e seres humanos infectados por este agente. A virulência deste pode estar associada a fatores como a cápsula de polissacarídeo, fosfolipase C e à enzima colesterol oxidase (fator equi). No entanto, uma proteína localizada em um plasmídeo, designada vapA, é essencial para a sobrevivência e replicação do agente em macrófagos. Com isso, os objetivos deste estudo foram avaliar o perfil de suscetibilidade de isolados de R. equi de diferentes fontes em relação aos antimicrobianos mais comumente utilizados na terapêutica animal e humana, bem como verificar a associação entre a presença do gene vapA e o índice de resistência múltipla aos antimicrobianos (IRMA). Neste estudo, 67 isolados brasileiros de R. qui de diferentes fontes foram analisados: 30 provenientes de amostras clínicas de equinos, sete de humanos e 30 ambientais (seis do solo e 24 de fezes de equinos). Para avaliar o perfil de suscetibilidade dos isolados utilizou-se o método de disco difusão, sendo testadas 16 drogas de diferentes classes de antimicrobianos. As amostras clínicas de equinos apresentaram as maiores taxas de resistência à penicilina (86,7%) e lincomicina (30%). Além disso, foram também resistentes a macrolídeos (azitromicina a 6,7%, eritromicina a 6% e ...

Rhodococcus equi is a facultative intracellular bacterium and etiological agent of rodococosis, an important disease that affects specially foals under six months old and leading the death generally due to pulmonary lesions. R. equi also has zoonotic potential, and it has emerged as an opportunistic pathogen in the world, specially infecting solid organ transplant recipients and immunocompromised human patients, mainly those infected by human immunodeficiency virus (HIV). Additionally, R. equi infections by healthy hosts have been reported principally in children and elderly individuals. Studies have shown an increasing level of resistance of isolates of R. equi against antibiotics commonly used to treat affected animals and humans. The virulence of this agent is associated with a protein located on a plasmid, designated vapA, it is essential for survival and replication of R. equi within macrophages. The present study evaluated the susceptibility profile of R.equi isolates from different sources against the antimicrobials most commonly used to treat animals and humans, as well as the occurrence and association of vapA gene and the antimicrobial multiple resistance index (AMRI). Sixty-seven Brazilian isolates of R. equi from different sources were analyzed: 30 clinical samples of horses, seven of human and 30 of environmental (six from soil and 24 from horse feces). To evaluate the susceptibility profile of R. equi isolates, the Kirby Bauer method was performed by using 16 drugs of 11 distinct antimicrobials classes. Additionally, those samples were also resistant to macrolides (azithromycin 6.7%, erythromycin 6% and clarithromycin 3.3%) as well as rifamycin (13%). All human and environmental samples were sensitive to macrolides and rifamycin. However, environmental isolates demonstrated high levels of resistance to penicillin and chloramphenicol. Similarly, human isolates had high level of resistance to ceftiofur, lincomycin and sulfazotrim. AMRI...

Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections

Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

Low level laser therapy (AlGaInP) applied at 5J/cm2 reduces the proliferation of Staphylococcus aureus MRSA in infected wounds and intact skin of rats.

BACKGROUND: Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. OBJECTIVE: To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. METHODS: An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p <0.05. RESULTS: The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. CONCLUSION: The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation.

Genotipificação de isolados de Staphylococcus epidermidis provenientes de casos de mastite caprina / Genotyping Staphylococcus epidermidis isolated from mastitis in goats

Para verificar a dinâmica da resistência aos antimicrobianos em uma propriedade rural no município de Santa Maria da Boa Vista, PE, foram avaliados 14 isolados de Staphylococcus epidermidis de caprinos com mastite subclínica. O perfil de resistência aos antimicrobianos foi determinado pelo teste de difusão em disco. A genotipificação foi realizada empregando o marcador REP (Repetitive Extragenic Palindromic) - PCR, utilizando o primer RW3A, enquanto os graus de similaridade e o fenograma de agrupamento foram estabelecidos por meio do coeficiente de Sorensen-Dice (SD) do algoritmo UPGMA, programa NTSYS-pc, o qual permitiu a identificação de 4 padrões dos 14 isolados de S. epidermidis, sendo oito no perfil A, quatro no perfil B, um no perfil C e um no perfil D. Para todos os grupos, a resistência à penicilina foi observada, enquanto que, para os grupos A e C, esta foi associada à lincomicina, no grupo B, esta foi associada à tetraciclina.

To verify the dynamics of antimicrobial resistance in a property in Santa Maria da Boa Vista city - Pernambuco were evaluated 14 Staphylococcus epidermidis isolates from goats with subclinical mastitis. The profile of antimicrobial resistance was determined by disk diffusion test. The genotyping was performed using the marker REP (Repetitive Extragenic Palindromic) - PCR, using RW3A primer, where the degrees of similarity and clustering phenogram were established by means of the Sorensen-Dice coefficient (SD) algorithm UPGMA, program NTSYS-pc, which allowed the identification of 4 patterns of 14 S. epidermidis isolates, being eight in the profile A, four in a profile B, one in profile C and one in profile D. For all groups to penicillin resistance was observed, while for groups A and C this was associated with lincomycin, for group B this was associated with tetracycline.

Low level laser therapy (AlGaInP) applied at 5J/cm2 reduces the proliferation of Staphylococcus aureus MRSA in infected wounds and intact skin of rats* / Laser de baixa intensidade (AlGaInP) aplicado com 5J/cm2 reduz a proliferação de Staphilococcus aureus MRSA em feridas infectadas e em pele íntegra de ratos

Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p <0.05. The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation. .

Fundamentos: A terapia a laser é um procedimento de baixo custo, não invasiva e com bom desempenho na cicatrização. As dúvidas existentes quanto a sua ação sobre microrganismos justifica a realização de pesquisas visando investigar os possíveis efeitos em feridas infectadas por bactérias. Avaliar o efeito do laser de baixa intensidade sobre a taxa de contaminação bacteriana em feridas infectadas na pele de ratos. Estudo experimental, utilizando 56 ratos machos Wistar. Os animais foram distribuídos aleatoriamente em oito grupos de sete animais. Nos animais dos grupos lesionados foi realizada uma incisão na região dorsal.Os animais dos grupos infectados foram infectados por Staphylococcus aureus MRSA. os animais dos grupos tratados foram tratados com laser de Diodo vermelho (AlGaInP) 658nm, 5J/cm2 em varredura, durante 3 dias consecutivos. Foi colhida uma amostra antes de inocular as bactérias e outra após o tratamento com laser. Para a análise estatística foram utilizados os testes não paramétricos de Wilcoxon (dados pareados). Considerando como significante p<0,05. Através da análise estatística das medianas, observou-se que os grupos submetidos ao laser apresentavam uma proliferação bacteriana menor. O laser (AlGaInP), com uma dose de 5J/cm2, tanto em feridas quanto em pele íntegra de ratos infectados por Staphilococcus aureus MRSA, se mostrou capaz de reduzir a proliferação bacteriana. .

Qualidade microbiológica de manteiga de garrafa comercializada no município de Petrolina, PE / Microbiological quality of bottled butter commercialized in the city of Petrolina, PE

A manteiga de garrafa é um alimento muito apreciado nos estados do Nordeste do Brasil, onde sua produção ocorre predominantemente de forma artesanal. Nesse trabalho, objetivou-se avaliar a qualidade microbiológica da manteiga de garrafa produzida e comercializada no município de Petrolina, PE.