Characterization of Enterococcus spp. isolated from a fish farming environment in southern Brazil / Caracterização de Enterococcus spp. isolado de um ambiente de piscicultura no sul do Brasil

Abstract The aim of present study is to characterize the resistance and virulence profile of enterococci isolated from aquaculture excavated ponds and masonry tanks (6 samples) in southern Brazil. Samples were cultured in selective medium, 10 colonies were randomly selected from each sample, which were identified by MALDI-TOF and tested against 13 antimicrobials. The presence of resistance (tetL, tetM, tetS, ermB and msrC) and virulence (ace, esp, agg, cylA and gelE) genes were determined by PCR. A total of 79 enterococci were identified, and Entecococcus faecalis (44.3%) and E. casseliflavus (36.7%) were the most prevalent species isolated. Sixty-five strains (82.3%) were resistant to at least one of the antimicrobials tested, whereas 27 (34.2%) strains were multiresistant. The overall percentages of antimicrobial resistant isolates were: 58.2% to rifampicin, 40.5% to fluoroquinolones, 36.7% to erythromycin and 30.4% to tetracycline. The tetL and tetM genes were found in 57.7% of the tetracycline-resistant strains; and msrC in 31.01% of erythromycin-resistant strains. The most frequently detected virulence factors were ace and gelE genes. Although limited to a single farm, these data suggest that aquaculture may be a reservoir of resistant and virulent enterococci. This study is the first step towards enhancing our understandingof distribution, resistance and virulence profile in enterococci isolated from fish farming environments in the south Brazil.

Resumo O objetivo do estudo apresentado é caracterizar o perfil de resistência e virulência de enterococos isolados de viveiros escavados e tanques de alvenaria (6 amostras) de uma pisicultura no Sul do Brasil. As amostras foram cultivadas em meio seletivo, 10 colônias foram selecionadas aleatoriamente de cada amostra, que foram identificadas por MALDI-TOF e testadas contra 13 antimicrobianos. A presença de genes de resistência (tetL, tetM, tetS, ermB e msrC) e virulência (ace, esp, agg, cylA e gelE) foi determinada por PCR. Foram identificados 79 enterococos, sendo Entecococcus faecalis (44,3%) e E. casseliflavus (36,7%) as espécies mais frequentes isoladas. Sessenta e cinco cepas (82,3%) eram resistentes a pelo menos um dos antimicrobianos testados, enquanto 27 (34,2%) eram multirresistentes. As porcentagens gerais de isolados resistentes a antimicrobianos foram: 58,2% para rifampicina, 40,5% para fluoroquinolonas, 36,7% para eritromicina e 30,4% para tetraciclina. Os genes tetL e tetM foram encontrados em 57,7% das cepas resistentes à tetraciclina; e msrC em 31,01% das cepas resistentes à eritromicina. Os fatores de virulência mais comumente detectados foram ace e gelE. Embora limitados a uma única fazenda, esses dados indicam que a aquicultura pode ser uma fonte de enterococos resistentes e virulentos. Este estudo é o primeiro passo para melhorar nosso entendimento da distribuição, resistência e perfil de virulência em enterococos isolados de ambientes de piscicultura no sul do Brasil.

Evaluation of antitumor potential of cashew gum extracted from Anacardium occidentale Linn.

This study aims to determine the antitumor potential of cashew gum in vitro and in vivo. The cashew gum (CG) structure is similar to already showed in literature. The cytotoxicity effect of CG was performed by MTT assay, and B16-F10 melanoma model was used to evaluate antitumor effect. The tumor inhibition was calculated based on tumor weight. Hematological, histopathological, FTIR, oxidative stress and Western Blot analysis were performed to elucidate the mechanism of inhibition and toxic effects. As results, CG did not demonstrate cytotoxicity in vitro, however showed a significant tumor inhibition in vivo, with about 36.9 to 43% of reduction in tumor mass, with no toxicity to organs. Animals treated with CG did not show toxicity in normal tissues, FTIR spectrum and oxidative stress analysis of the tumor tissue indicated that CG cause tumor inhibition with the presence of apoptosis morphotype cells, without alterations in the levels of antioxidants components. In addition, it was observed that CG reduced the expression of γH2AX without changing the expression of caspase-3. With this, we can suggest that this polymer can assist in the anticancer activity and/or decrease the side effects of standard drugs used in treatment of cancer.

In vitro antitumor effect of a lignan isolated from Combretum fruticosum, trachelogenin, in HCT-116 human colon cancer cells.

The use of natural products in therapeutics has been growing over the years. Lignans are compounds with large pharmaceutical use, which has aroused interest in the search for new drugs to treat diseases. The present study evaluated the cytotoxicity of (-)-trachelogenin, a dibenzylbutyrolactone type lignan isolated from Combretum fruticosum, against several tumor and non-tumor cell lines using the MTT assay and its possible mechanism of action. (-)-Trachelogenin showed IC50 values ranging of 0.8-32.4µM in SF-295 and HL-60 cell lines, respectively and IC50 values >64µM in non-tumor cell lines. (-)-trachelogenin persistently induced autophagic cell death, with cytoplasmic vacuolization and formation of autophagosomes mediated by increasing LC3 activation and altering the expression levels of Beclin-1.

Characterization of Enterococcus spp. isolated from a fish farming environment in southern Brazil

Abstract The aim of present study is to characterize the resistance and virulence profile of enterococci isolated from aquaculture excavated ponds and masonry tanks (6 samples) in southern Brazil. Samples were cultured in selective medium, 10 colonies were randomly selected from each sample, which were identified by MALDI-TOF and tested against 13 antimicrobials. The presence of resistance (tetL, tetM, tetS, ermB and msrC) and virulence (ace, esp, agg, cylA and gelE) genes were determined by PCR. A total of 79 enterococci were identified, and Entecococcus faecalis (44.3%) and E. casseliflavus (36.7%) were the most prevalent species isolated. Sixty-five strains (82.3%) were resistant to at least one of the antimicrobials tested, whereas 27 (34.2%) strains were multiresistant. The overall percentages of antimicrobial resistant isolates were: 58.2% to rifampicin, 40.5% to fluoroquinolones, 36.7% to erythromycin and 30.4% to tetracycline. The tetL and tetM genes were found in 57.7% of the tetracycline-resistant strains; and msrC in 31.01% of erythromycin-resistant strains. The most frequently detected virulence factors were ace and gelE genes. Although limited to a single farm, these data suggest that aquaculture may be a reservoir of resistant and virulent enterococci. This study is the first step towards enhancing our understandingof distribution, resistance and virulence profile in enterococci isolated from fish farming environments in the south Brazil.

Resumo O objetivo do estudo apresentado é caracterizar o perfil de resistência e virulência de enterococos isolados de viveiros escavados e tanques de alvenaria (6 amostras) de uma pisicultura no Sul do Brasil. As amostras foram cultivadas em meio seletivo, 10 colônias foram selecionadas aleatoriamente de cada amostra, que foram identificadas por MALDI-TOF e testadas contra 13 antimicrobianos. A presença de genes de resistência (tetL, tetM, tetS, ermB e msrC) e virulência (ace, esp, agg, cylA e gelE) foi determinada por PCR. Foram identificados 79 enterococos, sendo Entecococcus faecalis (44,3%) e E. casseliflavus (36,7%) as espécies mais frequentes isoladas. Sessenta e cinco cepas (82,3%) eram resistentes a pelo menos um dos antimicrobianos testados, enquanto 27 (34,2%) eram multirresistentes. As porcentagens gerais de isolados resistentes a antimicrobianos foram: 58,2% para rifampicina, 40,5% para fluoroquinolonas, 36,7% para eritromicina e 30,4% para tetraciclina. Os genes tetL e tetM foram encontrados em 57,7% das cepas resistentes à tetraciclina; e msrC em 31,01% das cepas resistentes à eritromicina. Os fatores de virulência mais comumente detectados foram ace e gelE. Embora limitados a uma única fazenda, esses dados indicam que a aquicultura pode ser uma fonte de enterococos resistentes e virulentos. Este estudo é o primeiro passo para melhorar nosso entendimento da distribuição, resistência e perfil de virulência em enterococos isolados de ambientes de piscicultura no sul do Brasil.

Growth inhibitory effects of 3'-nitro-3-phenylamino nor-beta-lapachone against HL-60: a redox-dependent mechanism.

In this study, the cytotoxicity, genotoxicity and early ROS generation of 2,2-dimethyl-(3H)-3-(N-3'-nitrophenylamino)naphtho[1,2-b]furan-4,5-dione (QPhNO(2)) were investigated and compared with those of its precursor, nor-beta-lapachone (nor-beta), with the main goal of proposing a mechanism of antitumor action. The results were correlated with those obtained from electrochemical experiments held in protic (acetate buffer pH 4.5) and aprotic (DMF/TBABF(4)) media in the presence and absence of oxygen and with those from dsDNA biosensors and ssDNA in solution, which provided evidence of a positive interaction with DNA in the case of QPhNO(2). QPhNO(2) caused DNA fragmentation and mitochondrial depolarization and induced apoptosis/necrosis in HL-60 cells. Pre-treatment with N-acetyl-l-cysteine partially abolished the observed effects related to the QPhNO(2) treatment, including those involving apoptosis induction, indicating a partially redox-dependent mechanism. These findings point to the potential use of the combination of pharmacology and electrochemistry in medicinal chemistry.

Field performance of a rapid point-of-care diagnostic test for antenatal syphilis screening in the Amazon region, Brazil.

We evaluated an immunochromatographic point-of-care (POC) syphilis test in 712 pregnant women under field conditions in remote communities of the Amazon region (Brazil), and identified risk factors for syphilis. Women were screened by POC test using whole blood obtained by fingerprick, the fluorescent treponemal antibody absorption (FTA-Abs) test as the gold standard and the Venereal Diseases Research Laboratory (VDRL) test to determine test performance in active syphilis. Multivariate analysis was conducted to identify factors associated with syphilis infection. Among women, 2.2% had syphilis (positive FTA-Abs) and 0.8% active syphilis (FTA-Abs and VDRL positive). In all, 2.2% of samples were positive by the POC test. The sensitivity, specificity, positive and negative predictive values were 62.5% (95% confidence interval [CI]: 38.6-81.5), 99.1% (95% CI: 98.1-99.6), 62.5% (95% CI: 38.6-81.5) and 99.1% (95% CI: 98.1-99.6), respectively. The POC test identified 62.5% (10/16) of syphilis cases, 66.7% (4/6) of active syphilis cases and all high-titre syphilis cases (VDRL > 1:8). Older age was associated with syphilis infection. The rapid test performed moderately well as a screening tool for low-risk populations. This combined with on-site testing and same day treatment could expand antenatal syphilis screening programmes in distant communities characterized by difficult access to antenatal services and infrequent clinical follow-up visits.

Development and reproductive performance of Swiss mice in an enriched environment / Desenvolvimento e performance reprodutiva de camundongos Swiss em ambiente enriquecido

The present study investigated the effects of environment enrichment on the development and reproductive performance of an outbred Swiss strain. Physical enrichment consisted of free access of animals to tubular devices of different shapes and sizes. The development evaluation was done by monitoring growth rate during 39 days, from weaning (21 days) to the onset of sexual maturity (60 days of age). Over five consecutive gestations (105 days), the following variables were monitored: litter size, number of animals born alive; number of pups alive at 12 hours and on days 4, 9, 12, 15 and 18; number of animals weaned per litter; average weight of litters on days 4, 9, 12, 15 and 18. The reproductive performance was then evaluated using the inclusion of these variables in the indices of gestation and birth, viability, lactation, survival success, mating success and production. The results showed that the environmental enrichment did not influence, in a significant way, the evaluated parameters. However, there is no reason to deprive the animals from an enriched environment where they can develop their natural instinctive behaviour and guarantee health and well-being.

O presente estudo investigou efeitos do enriquecimento ambiental sobre o desenvolvimento e a performance reprodutiva de camundongos outbred da linhagem Swiss. O enriquecimento teve como base o livre acesso dos animais a dispositivos tubulares de tamanhos e formas diferentes. A avaliação do desenvolvimento a partir da taxa de crescimento decorreu por um período de 39 dias, desde o desmame (21 dias) até os 60 dias de vida (maturidade sexual). Durante cinco gestações consecutivas (105 dias), foram registradas as seguintes variáveis: média do tamanho da ninhada, número de animais nascidos vivos; número de filhotes vivos em 12 horas, e nos dias 4, 9, 12, 15 e 18; número de animais desmamados por ninhada; e a média do peso dos filhotes nos dias 4, 9, 12, 15 e 18. A performance reprodutiva foi então avaliada a partir da inclusão dessas variáveis nos índices de gestação e nascimento, viabilidade, lactação, sucesso de sobrevivência, sucesso de acasalamento e produção. Os resultados obtidos mostraram que o enriquecimento ambiental não influenciou significativamente no desenvolvimento e na performance reprodutiva de camundongos da linhagem Swiss para nenhum dos parâmetros avaliados. Contudo, não há motivo para privar os animais de um ambiente enriquecido onde possam desenvolver seu comportamento instintivo natural e, consequentemente, viver com saúde e bem-estar.

Sintomas respiratórios e testes espirográficos dos funcionários de uma pedreira do Rio de Janeiro / Respiratory symptoms and spirometric tests of the quarry workers Rio de Janeiro

Foram investigados os sintomas e antecedentes respiratórios, bem como os resultados das provas espirográficas, de 72 dos 86 (83,7 por cento) funcionários de uma pedreira de porte médio do Rio de Janeiro. Setenta e um (98,6 por cento) eram homens, e a média das idades foi de 36,2 + ou - 9,3 anos (20 a 65 anos). Quarenta e dois (58,3 por cento) tinham ocupaçöes consideradas muito expostas ao pó de pedra e 30 (44,7 por cento) pouco expostas. Quarenta (55,6 por cento) eram fumantes, 20 (27,8 por cento) näo fumantes e 12 (16,7 por cento) ex-fumantes. Foi utilizado um questionário padronizado baseado em modelos internacionais e adaptados a condiçöes brasileiras. Houve elevada freqüência de sintomas, com exceçäo da dispnéia: 47 (65,3 por cento) referiram um ou mais sintomas, dos quais 31,9 por cento referiram tosse, 41,7 por cento expectoraçäo, 9,7 por cento dispnéia e 33,3 por cento chiado torácico. Os sintomas se associaram principalmente à presença de antecedentes de doenças respiratórias, havendo também associaçäo com o tabagismo. Um ou outro desses fatores explicaram os sintomas. Näo houve associaçäo com a maior exposiçäo ao pó de pedra, qua näo explicou os sintomas. As provas espirográficas foram realizadas com o espirógrafo de campânula de 6 litros, observando-se discreta reduçäo das médias da capacidade vital forçada (CVF) (89,9 + ou - 11,2 por cento do teórico), do volume de expiraçäo forçada do 1§ segundo (VEF1) (90,1 + ou - 12,9 por cento) e do fluxo expiratório forçado entre 25 por cento e 75 por cento da CVF (FEF 25-75) (92,9 + ou - 32,7 por cento) em relaçäo às médias esperadas numa amostra populacional näo selecionada. Contudo, as proporçöes de valores subnormais foram comparáveis às de amostras näo selecionadas da literatura nacional e internacional. Näo houve influência estatisticamente significativa da maior exposiçäo ao pó de pedra, antecedentes respiratórios e tabagismo sobre as médias dos resultados espirográficos. Concluímos que, neste grupo de funcionários de uma pedreira, näo se demonstrou associaçäo entre a maior exposiçäo ao pó de pedra e os sintomas respiratórios, ou influência dessa exposiçäo sobre as médias dos resultados espirográficos. Com relaçäo aos antecedentes respiratórios e tabagismo, houve associaçäo com os sintomas mas näo influência sobre as médias dos resultados espirográficos.

[Respiratory symptoms and spirometric tests of quarry workers in Rio de Janeiro]. / Sintomas respiratórios e testes espirográficos dos funcionários de uma pedreira do Rio de Janeiro.

Respiratory symptoms and past history of chest disease as well as spirometric tests were investigated in 72 of 86 (83.7%) employees of a middle sized quarry in Rio de Janeiro, Brazil. Seventy one were men (98.6%), and the mean age was 36.2 +/- 9.3 years (20-65). Forty two (58.3%) had occupations considered as highly exposed to dust, and 30 (44.7%) were considered as lightly exposed. Forty (55.6%) were smokers, 20 (27.8%) never had been smokers and 12 (16.7%) were former smokers. Symptoms and past history were investigated with a questionnaire based on international models, and adapted for the Brazilian public. A high prevalence of symptoms (except for dyspnea) was noted: 47 (65.3%) had one or more symptoms-there was cough in 31.9%, expectoration in 41.7% dyspnea in 9.7% and wheezing in 33.3%. The symptoms were found to be associated mostly with a past history of chest disease, and also with smoking, factors which explained, in part, the presence of the symptoms. No association with a higher exposure to dust was found. The exposure to dust did not explain the symptoms. The spirometric tests were performed on a 6 liter bell spirometer. The means of the parameters were lower than one would expect in a non selected population sample--89.9 +/- 11.2% of predicted for the Forced Vital Capacity (FVC), 90.1 +/- 12.9% for the Forced Expiratory Volume in 1 second (FEV1) and 92.9 +/- 32.7% for the Forced Expiratory Flow between 25% and 75% of the FVC (FEF25-75). However, the proportions of abnormal results were comparable to the reported ones from unselected samples. There was no statistically significant influence of higher dust exposure, past history or smoking on the means of the results. It is concluded that, in this sample of quarry employees, no association between respiratory symptoms or spirometric results and dust exposure could be demonstrated. There was an association between the symptoms, but not with the spirometric results, and a past history of chest disease, as well as with smoking.

Enteroparasitos em índios Yanomâmi / Intestinal parasites among Yanomâmi indians

The findings of intestinal helminths and protozoans parasites from the Yanomâmi indians of the Roraima State in Brazil are reported. The fecal samples were collected before these communities started a permanent contact with non-indians. Comments are made on the possible ecological and evolutionary factors responsible for the patterns of parasitism observed.

The finding of eggs of Diphyllobothrium in human coprolites (4,100-1,950 B.C.) from Northern Chile.

Os autores realizaram exame parasitologico de vinte e seis coprolitos encontrados em um sitio arqueologico no norte do Chile, Provincia de Iquique.O material foi colhido em duas unidades de escavacao, I e II (sitio Tiliviche) datados respectivamente de 5.900 a.C. a 4.100 a.C. e 4.110 a.C. a 1.950 a.C. e identificados como de origem humana.Na unidade II foram observados ovos de helmintos diagnosticados como de Diphyllobothrium pacificum. Esse achado nos informa sobre a antiguidade da infeccao bem como sobre os habitos alimentares dessas populacoes, uma vez que a contaminacao se da por ingestao de peixes marinhos. E interessante assinalar que Baer (1969), ao descrever pela primeira vez a infeccao humana por esse cestodeo em populacoes atuais do Peru, sugere a sua presenca nas populacoes pre-colombianas