Uma perspectiva histórica e prospecções da pesquisa biomédica em doenças parasitárias

We all hope that biotechnology will answer some social and economical unavoidable requirements of the modern life. It is necessary to improve agriculture production, food abundance and health quality in a sustainable development. It is indeed a hard task to keep the progress on taking into account the rational use of genetic resources and the conservation of biodiversity. In this context, a historical perspective and prospects of the biomedical research on parasitic diseases is described in a view of three generations of investigators. This work begins with a picture of the scientific progress on biomedical research and human health over the last centuries. This black-and-white picture is painted by dissecting current advancements of molecular biology and modern genetics, which are outlined at the meaning of prospecting achievements in health science for this new millenium.Espera-se que os grandes avanços da pesquisa de genomas alcancem saltos visíveis para a sobrevivência da humanidade dentro do contexto de sustentabilidade global. Vemos atualmente a necessidade imediata de incrementar a produção de alimentos e melhorar a qualidade de vida e saúde humana, mantendo a biodiversidade preservada. A biotecnologia é a utilização racional das ferramentas geradas pela biologia molecular e genética moderna que, dado ao grande avanço dessas áreas, poderá em breve responder essas novas questões. Este trabalho, escrito por três gerações de pesquisadores, inicia com uma perspectiva histórica da pesquisa biomédica em doenças parasitárias nos últimos séculos em nosso país. Em seguida, descrevemos os mais recentes avanços da biologia molecular e genética genômica. Ressaltamos a importância dessas novas conquistas dentro de uma prospecção da pesquisa biomédica deste século e seus possíveis impactos na saúde humana

Predominance of KPC-3 in a Survey for Carbapenemase-Producing Enterobacteriaceae in Portugal

Free PMC Article: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4432220/Among the 2,105 Enterobacteriaceae tested in a survey done in Portugal, 165 were nonsusceptible to carbapenems, from which 35 (26 Klebsiella pneumoniae, 3 Escherichia coli, 2 Enterobacter aerogenes, and 3 Enterobacter cloacae isolates and 1 Klebsiella oxytoca isolate) were confirmed to be carbapenemase producers by the presence of 30 Tn4401d-blaKPC-3, 4 intI3-blaGES-5, and one intI1-blaVIM-2 gene, alone or in combination with other bla genes. The dissemination of blaKPC-3 gene carried by an IncF plasmid suggests lateral gene transfer as a major mechanism of dissemination.V. Manageiro was supported by grant SFRH/BPD/77486/2011 from the Fundação para a Ciência e a Tecnologia, Lisbon, Portugal. J. Almeida was supported by grant BRJ-SUB/01/2012 from the National Institute of Health Dr. Ricardo Jorge, Lisbon, Portugal. R. A. Bonomo was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under awards R01 AI100560 and R01 AI063517, and by the Cleveland Department of Veterans Affairs, the Veterans Affairs Merit Review Program, and the Geriatric Research Education and Clinical Center VISN 10. We thank the Fundação para a Ciência e a Tecnologia (FCT) for project grant PEst-OE/AGR/UI0211/2011-2014, Strategic Project UI211- 2011-2014

Molecular survey of carbapenem resistant Enterobacteriaceae isolates from Portuguese Hospitals: co-production of carbapenemase KPC-3 and the efflux pump OqxAB

Objectives: Although there are important studies regarding the different carbapenemase (CARB)-producing Gram-negative bacteria, little is known concerning their molecular epidemiology in Portugal. The main aim of this study was to characterize, by phenotype and molecular typing methods, CARB-producing Enterobacteriaceae isolates recovered from Portuguese health care institutions, and evaluate its impact on treatment strategy. Methods: This study included 2105 clinical isolates, collected between April/2006 and February/2013, in different Portuguese healthcare institutions. Screening of antimicrobial susceptibility was performed by disc diffusion method. Clinical isolates with resistance or with decreased susceptibility to ertapenem were considered presumptively CARB-producers; in these isolates, PCR and sequencing were applied to detect and identify CARB-encoding genes, as well as other bla and plasmid-mediated quinolone resistance (PMQRs) genes. MICs of CARB-producing isolates were tested by microdilution (EUCAST breakpoints). The plasmids obtained from clinical isolates were characterized by PCR-based replicon typing (PBRT). Clonal relatedness of K. pneumoniae isolates was investigated by multilocus sequence typing (MLST), using the protocol developed by the Institute Pasteur (www.pasteur.fr/mlst/Kpneumoniae.html). Results: Among the 2105 isolates tested, 165 (7.8%) were putative CARB-producers and were selected for further analysis. Thirty-five (21.2%) of the 165 positive isolates were confirmed to be CARB-producers, of which the majority were collected from the urine (54.3%) of elderly (≥65 years old) male patients (54.3%), and admitted at the emergency room/ambulatory (22.9%) or internal medicine (17.1%) wards. All were multidrug-resistant, with nonsusceptibility to at least one carbapenem, and with consistent susceptibility only to colistin. In those isolates was detected the following beta-lactamases: 30 KPC-3 (22 K. pneumoniae, 3 Escherichia coli, 2 Enterobacter aerogenes and 3 Enterobacter clocae), 4 GES-5 (K. pneumoniae) and one VIM-2 (Klebsiella oxytoca). CARB-encoding genes were present alone or in combination with other bla genes, such as blaSHV-12, blaSHV-14, blaSHV-26, blaSHV-36, blaCTX-M-15, and the blaSHV-164. PMQR-encoding genes were also detected, namely qnrA, qnrB, aac(6’)-Ib-cr and the recently identified oqxAB. All blaKPC-3 genes were located on a Tn3-based transposon, Tn4401, while blaGES-5 and blaVIM-2 genes were associated with class 3 and 1 integrons, respectively. In our study, the majority of the blaCARB-harbouring plasmids were nonconjugative, having been typed as IncFrepB by PBRT. Clonal relatedness of the 26 K. pneumoniae isolates, obtained by MLST, showed that they were from distinct STs, namely ST14, ST15, ST34, ST59, ST147, ST416, ST698, and from the two novels ST: ST960 and, among all, the predominant ST1138 (corresponding to KPC-3 plus SHV-36 producers). Conclusion: In conclusion, this study provides new data regarding the molecular epidemiology of CARB-producing Enterobacteriaceae, which appears to be widespread in Portugal. Dissemination of blaCARB seems to be due to carriage of similar CARB-harbouring plasmids within genetically diverse clinical strains. Overall, our results emphasize the need of a concerted action to manage carbapenem use

Impact of antihelminthic treatment on infection by Ascaris lumbricoides, Trichuris trichiura and hookworms in Covas, a rural community of Pernambuco, Brazil

This work aims to evaluate the impact of drug treatment on infection by Ascaris lumbricoides (Al), Trichuris trichiura (Tt) and hookworms (Hook) in a rural community from the sugar-cane zone of Pernambuco, Brazil. Four parasitological surveys were carried out from March 2001 to March 2002. Individual diagnosis was based on eight slides (four by the Kato-Katz method and four by the Hoffman method) per survey. Infected subjects were assigned to two groups for treatment with either albendazole (n = 62) or mebendazole (n = 57). Prevalence of infection fell significantly (p < 0.05) one month after treatment: Al (from 47.7% to 6.6%); Tt (from 45.7% to 31.8%) and Hook (from 47.7% to 24.5%). One year after treatment, infections by Tt and Hook remained significantly below pre-control levels. A substantial decrease in single-infection cases and multiple infections was found. Egg-negative rate was significant for Al (94.0%), Hook (68.3%) but not for Tt (45.5%), and did not differ significantly between subjects treated with mebendazole or albendazole. Egg counts fell significantly in the individuals remaining positive for Tt. It is recommended that antihelminthic treatment should be selective and given at yearly intervals preferably with albendazole, due to its cost-effectiveness.Este trabalho avalia o impacto do tratamento antihelmíntico sobre a infecção por A. lumbricoides (Al), T. trichiura (Tt) e ancilostomídeos (Anc) na população de Covas, Pernambuco. Durante março/2001 e março/2002, quatro inquéritos parasitológicos foram realizados. Em cada um foram coletados dois exames de fezes por morador. O diagnóstico foi feito pelos métodos de Kato-Katz e Hoffmann. Em abril/01, os indivíduos positivos foram separados em dois grupos para tratamento com albendazol (n = 62) ou mebendazol (n = 57). As proporções de indivíduos positivos reduziram-se significativamente um mês pós-tratamento: Al (de 47,7% para 6,6%), Tt (de 45,7% para 31,8%) e Anc (de 47,7% para 24,5%); permanecendo abaixo do nível inicial um ano pós-tratamento. Os casos de monoinfecção, exceto os por Tt, e infecções múltiplas reduziram-se após o tratamento. A negativação foi significativa para Al (94,0%), Anc (68,3%), mas não para Tt (45,5%) e foi maior para Al. A negativação não diferiu significativamente entre os tratados com mebendazol ou albendazol. A intensidade da infecção reduziu-se significativamente nos indivíduos que permaneceram positivos para Tt. Recomenda-se que o tratamento seja seletivo, administrado anualmente, de preferência com albendazole, devido ao seu custo-benefício

Ecoepidemiologia da esquistossomose urbana na ilha de Itamaracá, Estado de Pernambuco

INTRODUCTION: In 1988, 22 autochthonous cases from accidental exposure were registered in Forte Orange beach, Itamaracá Island, Pernambuco, Brazil. All cases occurred in middle-to-upper class individuals who were vacationing in the island. After the cases were identified, the major objective was to find breading sites of schistosomiasis vectors and correlate the biological factors with the environmental conditions. METHODS: The environmental characteristics of the beach before human occupation were obtained from several documents. Also, a one-year malacological survey was conducted with monthly collection of mollusks, and the determination of their infectivity rates. RESULTS/CONCLUSIONS: The malacological survey was able to identify 20 breading sites of Biomphalaria glabrata and 28 capture station were set up. Snails were collected and examined each month throughout a whole year. The results show a seasonal variation in the mollusk population density associated with infection rates and types of breading sites. The importance of this new epidemiological profile of schistosomiasis in the State of Pernambuco relies on the fact that it can be related with the drastic human interference on the environment. Ecological, environmental, and demographic factors as well as the epidemiological characteristics of the disease have social and economic repercussions.INTRODUÇÃO: Em 1988, 22 casos autóctones de esquistossomose foram registrados na Praia do Forte Orange, ilha de Itamaracá, Pernambuco. Todos os casos ocorreram em indivíduos de classe média/alta que veraneavam na ilha. Foi realizado estudo com o objetivo de identificar e caracterizar criadouros/focos de vetores da esquistossomose na localidade, correlacionando os determinantes biológicos da doença com o contexto ambiental da sua ocorrência. MÉTODOS: Foram levantados dados secundários para resgatar as características ambientais da área antes da ocupação humana. O inquérito malacológico teve a duração de um ano com mapeamento da área, coleta mensal e exame dos moluscos. RESULTADOS/CONCLUSÕES: Em 1 km de extensão da praia, foram identificados 20 criadouros e demarcadas 28 estações de coleta. Os resultados mostram a variação mensal da densidade populacional de moluscos e das taxas de infecção, correlacionados com sazonalidade e tipos de criadouros. Destaca-se a importância desse novo perfil epidemiológico da esquistossomose em Pernambuco, relacionando o modo de ocupação daquele espaço com o estabelecimento de sítios de transmissão ativa da esquistossomose

Strongyloides spp e outros parasitos encontrados em alfaces (Lactuca sativa)

Natural vegetables, mainly lettuce (Lactuca saliva), are important vehicles forthe transmission of infectious parasitic diseases. This study tested the presence ofenteroparasites in commercial sources of lettuce cultivated through conventional,organic and hydroponic methods in the city of Recife, Brazil. A total of 66 samples were collected, of which the following cultivation types were taken: 30 conventional,30 organic, and 6 hydroponic. Parasites were detected by sedimentation through theHoffman, Pons &amp; Janer (HPJ) method. Parasitological analysis detected parasitesin 88.8% and 100% of the lettuce collected, in supermarkets and fairs, respectively;predominantly larvae of Strongyloides spp. Additionally, 4/6 vegetables cultivatedthrough hydroponics and bought at supermarkets tested positive for Strongyloidesspp. Other parasites were observed as well: Ancylostoma spp eggs (8.3%) were foundin samples grown through conventional and organic methods, while Entamoeba sppcysts (2%) were observed in vegetables grown through conventional means andcollected from free standing markets. Data presented show that lettuce are belowthe standards of normal consumption as by the Resolution 12/1978 of the NationalCommission of Normal Dietary Standards, which was established to certifythe absence of contamination, e.g., parasites and larvae, in Brazil’s commercialvegetable industry. The elevated presence of Strongyloides spp larvae demonstratethe lack of hygienic and sanitary conditions in the irrigation of commercializedlettuce. The results of this study also indicate the need for a better monitoring system of those commercial enterprises that sell consumable vegetables, as this could possibly result in better quality and hygiene of these products.Hortaliças in natura são importantes veículos de transmissão de doenças infecto- parasitárias. No presente estudo, avaliou-se a presença de enteroparasitos em alfaces (Lactuca sativa) convencionais, orgânicas e hidropônicas in natura comercializadas na cidade do Recife. Os 66 “pés” de alface amostrados foram coletados entre maio e junho de 2007: 18 unidades em supermercados (06 convencionais, 06 hidropônicas, 06 orgânicas) e 48 unidades nas feiras livres (24 orgânicas e 24 convencionais). As amostras foram processadas pelo método diagnóstico de sedimentação espontânea, ou método de Lutz (Hoffmann, Pons &amp; Janer), com adaptações descritas na metodologia. As unidades de análise foram os supermercados e feiras livres que oferecessem hortaliças convencionais, orgânicas e hidropônicas. Para cada Região Administrativa foram selecionados supermercados e feiras que atendessem a esse critério e para cada bairro foi sorteado um estabelecimento. Ficou evidenciada a presença de parasitos em 88,8% das amostras coletadas em supermercados e em todas as amostras coletadas nas feiras livres. Todas as amostras oriundas de feiras e supermercados estavam contaminadas por larvas Strongyloides spp e 4/6 das hortaliças hidropônicas (supermercados) revelaram-se positivas. Foram observados outros parasitos: ovos de Ancylostoma spp (8,3%) em cultivos convencionais e orgânicos e cistos de Entamoeba spp (2%) nas hortaliças convencionais de feiras livres. A elevada presença de larvas de Strongyloides spp aponta para as péssimas condições higiênico-sanitárias na irrigação das alfaces comercializadas, indicando a necessidade de atuação mais intensa do sistema de vigilância sanitária para a melhoria da qualidade desses produtos

The Geographic Information System applied to study schistosomiasis in Pernambuco

OBJECTIVE: Diagnose risk environments for schistosomiasis in coastal localities of Pernambuco using geoprocessing techniques. METHODS: A coproscopic and malacological survey were carried out in the Forte Orange and Serrambi areas. Environmental variables (temperature, salinity, pH, total dissolved solids and water fecal coliform dosage) were collected from Biomphalaria breeding sites or foci. The spatial analysis was performed using ArcGis 10.1 software, applying the kernel estimator, elevation map, and distance map. RESULTS: In Forte Orange, 4.3% of the population had S. mansoni and were found two B. glabrata and 26 B. straminea breeding sites. The breeding sites had temperatures of 25ºC to 41ºC, pH of 6.9 to 11.1, total dissolved solids between 148 and 661, and salinity of 1,000 d. In Serrambi, 4.4% of the population had S. mansoni and were found seven B. straminea and seven B. glabrata breeding sites. Breeding sites had temperatures of 24ºC to 36ºC, pH of 7.1 to 9.8, total dissolved solids between 116 and 855, and salinity of 1,000 d. The kernel estimator shows the clusters of positive patients and foci of Biomphalaria, and the digital elevation map indicates areas of rainwater concentration. The distance map shows the proximity of the snail foci with schools and health facilities. CONCLUSIONS: Geoprocessing techniques prove to be a competent tool for locating and scaling the risk areas for schistosomiasis, and can subsidize the health services control actions.</jats:p