A fraction of human Salmonella infections is associated with direct contact with reptiles, yet the number of reptile-associated Salmonellosis cases are believed to be underestimated. Existing data on Salmonella spp. transmission by reptiles in Portugal is extremely scarce. The aim of the present work was to evaluate the prevalence of Salmonella spp. in pet reptiles (snakes, turtles, and lizards), as well as evaluate the isolates' antimicrobial resistance and virulence profiles, including their ability to form biofilm in the air-liquid interface. Additionally, the antimicrobial effect of chlorhexidine gluconate on the isolates was tested. Salmonella was isolated in 41% of the animals sampled and isolates revealed low levels of antimicrobial resistance. Hemolytic and lypolytic phenotypes were detected in all isolates. The majority (90.63%) of the Salmonella isolates were positive for the formation of pellicle in the air-liquid interface. Results indicate chlorhexidine gluconate is an effective antimicrobial agent, against the isolates in both their planktonic and biofilm forms, demonstrating a bactericidal effect in 84.37% of the Salmonella isolates. This study highlights the possible role of pet reptiles in the transmission of non-typhoidal Salmonella to humans, a serious and increasingly relevant route of exposure in the Salmonella public health framework.

We report a Microsporum audouinii infection in a female juvenile chimpanzee (Pan troglodytes) presenting generalized dermatitis compatible with dermatophytosis. Dermatophyte was identified by macro- and microscopic characterization of skin and scales cultures in Mycosel Agar. The topical treatment applied was effective, having the potential for dermatophytosis treatment in immunocompetent primates.

Salmonella 1,4,[5],12:i:- is presently considered one of the major serovars responsible for human salmonellosis worldwide. Due to its recent emergence, studies assessing the demographic characterization and spatial epidemiology of salmonellosis 1,4,[5],12:i:- at local- or country-level are lacking. In this study, a analysis was conducted over a 10year period, from 2000 to the first quarter of 2011 at the Portuguese National Laboratory in Portugal mainland, with a total of 215 Salmonella 1,4,[5],12:i:- serotyped isolates obtained from human infections by a passive surveillance system. Data regarding source, year and month of sampling, gender, age, district and municipality of the patients were registered. Descriptive statistical analysis and a spatial scan statistic combined with a geographic information system were employed to characterize the epidemiology and identify spatial clusters. Results showed that most districts have reports of Salmonella 1,4,[5],12:i:-, with a higher number of cases at the Portuguese coastland, including districts like Porto (n=60, 27.9%), Lisboa (n=29, 13.5%) and Aveiro (n=28, 13.0%). An increased incidence was observed in the period from 2004 to 2011 and most infections occurred during May and October. Spatial analysis revealed 4 clusters of higher than expected infection rates. Three were located in the north of Portugal, including two at the coastland (Cluster 1 [RR=3.58, p≤0.001] and 4 [RR=10.42 p≤0.230]), and one at the countryside (Cluster 3 [RR=17.76, p≤0.001]). A larger cluster was detected involving the center and south of Portugal (Cluster 2 [RR=4.85, p≤0.001]). The present study was elaborated with data provided by a passive surveillance system, which may originate an underestimation of disease burden. However, this is the first report describing the incidence and the distribution of areas with higher risk of infection in Portugal, revealing that Salmonella 1,4,[5],12:i:- displayed a significant geographic clustering and these areas should be further evaluated to identify risk factors in order to establish prevention programs.

The increase in prevalence of Salmonella 1,4,[5],12:i:-related infections over the last few years has been considered a public health issue in many European countries, especially as this serovar may be associated with tetraresistance to ampicillin, streptomycin, sulfonamides, and tetracyclines (R-type ASSuT). Salmonella 1,4,[5],12:i:- isolates (n = 187) obtained by the Portuguese National Laboratory from different sources, including human clinical cases (n = 170), veterinary (n = 10), environmental (n = 6), and food samples (n = 1), were collected from 15 districts between 2006 and 2011. All isolates were serotyped using the slide agglutination method and results were confirmed by multiplex PCR for the monophasic variant. From the confirmed Salmonella 1,4,[5], 12: i:-, R-type ASSuT isolates were selected by disc diffusion and minimal inhibitory concentration (MIC) determination for further characterization by pulsed-field gel electrophoresis restriction with XbaI, virulence genes determination by PCR, additional antimicrobial resistance profiling by disc diffusion, and epidemiological distribution evaluation. Out of the 187 serotyped isolates, 133 were confirmed as Salmonella 1,4,[5],12: i:-with a R-type ASSuT occurrence of 61.7%. Distribution among Portuguese districts showed a higher percentage of reported cases in coastal areas, in particular, in Porto (24.8%), Setubal (13.5%), and Aveiro (12.8%), probably due to the higher population density. Clonality analysis revealed a high diversity of pulsotypes with the majority of human salmonellosis cases being attributed to sporadic events. All isolates harbored 14 out of the 18 virulence genes evaluated and 87.8% of the isolates showed all the resistance genes frequently associated with the European clone, blaTEM(+)sul2(+)straA-straB(+)tetB(+). This study shows that Salmonella 1,4,[5],12:i:- resistant isolates are widely distributed in Portugal. This may be related to a selective advantage offered by R-type ASSuT profile, the presence of multiple virulent features, including the ability to form biofilms, which along with a high diversity of pulsotypes may be responsible for the dissemination through the country.

Reutilization of effluents from wastewater treatment plants (WWTP) for non-potable applications is increasing due to the reduction of sustainable water resources. These products mostly come from municipal WWTP and also from slaughterhouses effluents. The microbiological certification of these products is mandatory before their discharge into the environment. This study evaluates if the treatment applied in WWTP to municipal waters or to poultry slaughterhouse effluents distributed over the Portuguese continental territory is efficient in reducing the microbiological risk associated with the reutilization of those wastewaters and sludges. Fecal indicators Escherichia coli and enterococci were evaluated in 42 and 24 wastewater samples from 14 municipal WWTP and 8 poultry slaughterhouse treatment plants, respectively, by the conventional culture method and a rapid Fluorescent in situ hybridization (FISH) technique. Bacterial enumeration in inflow water from most WWTP was rather high (generally >105 cells/ml), for both E. coli and Enterococcus spp., and the bacterial quantification by FISH was generally higher than enumeration by the conventional culture method. In both types of treatment plants studied, bacterial load from effluents and sludges was not statistically different from the inflows, indicating that the treatment applied seems to be equally unable to reduce the microbiological load of the effluents. These findings may jeopardize the safe reuse of treated wastewaters in agriculture and the quality of the water environment. Therefore, products like water, sewage sludge, and biosolids originated from the municipal and slaughterhouse WWTP studied should not be reutilized, and effluents treatment should be urgently reviewed.

Fish and shellfish products have been used as food supply since immemorial times. Earliest evidences show that for more than one million years ago, manual capture of gastropods and bivalves bordering the sea shore waterfront have been crucial for survival, development and increasing of human communities. Fish products are a unique source of rich nutrients, being easy to digested, representing a very relevant source of indispensable amino acids, fatty acids, minerals and vitamins for many millions of people worldwide. However, only products obtained from healthy fish and maintained in hygienic environments are suitable for consumption and can present those nutritional advantages. If such products are not handled and or correctly processed, consumer health may be put at risk.

Diseases of fish, caused by biological agents (bacteria, fungi, virus or parasites), are better known in farmed or ornamental fish, since the access to the affected fish coming from the natural environment is less probable. In fact, the fish that are infected by a specific pathogenic agent, natural inhabitants of the marine, estuarine or freshwater ecosystems, are rapidly eliminated from its biotopes by other predatory animals, due to its higher vulnerability and susceptibility to the social interactions.Diffusion of pathogenic bacteria, fungi, virus or parasites in the aquatic environments is more efficient than in the terrestrial ones. Diagnosis and therapeutics of fish disease have specific difficulties and the application of preventive measures is a very complex issue. Some of those diseases have the same epidemiological problems of the infectious diseases of the terrestrial animals: High spread of diffusion, very significant economic losses, restrictions to fish travel or commerce and some (few) have zoonotic impacts.

A recent episode of aflatoxin M-1 (AFM(1)) found in cow milk produced in Acores, convoked to the problem of mycotoxins control in feed for dairy cows. Exposure control of milk-producing animals to mycotoxins, requires a systematic monitoring of feeds and raw materials. The European Union established maximum values for aflatoxin B-1 (AFB(1)) in dairy cattle, ewe and goats feeds (5 mu g/kg) (European Commission, 2011) and recommend guidance levels of zearalenone (ZEA; 500 mu g/kg) in complementary and complete feeding stuffs for dairy cattle, sheep and goats (European Commission, 2006a). Portuguese dairy production is largely (70%) dependent on imported cereals and other grains for feed production, coming from the international market. All feed plants or home-mixers are officially approved, according to Regulation (EC) No 183/2005 (European Commission, 2005), enforcing the HACCP system application in each feed plant. However, the production conditions of feedstuffs, especially those imported from developing countries, are difficult to control, concerning the specific risk of mycotoxins contaminations. These kinds of hazards are mostly introduced in feed materials in the pre-harvest period and must be locally controlled through agricultural 'good practices'. In this chapter, authors review published data from occurrence of AFB1 and ZEA applied, particularly, to feed manufactured in Portugal and intended for dairy cattle, goat and ewe feeding. Attention is also given to raw materials used in dairy feed. Finally, the global impacts for the food chain of the presence of these toxins in dairy feeds are discussed, with relevance to the presence of AFM(1) in dairy foods.

Biofilm-forming ability is well established as an important virulence factor. However, there are no studies available regarding biofilm formation of Salmonella Typhimurium 1,4,[5],12:i:-, the new pandemic serovar in Europe. To address this problem, biofilm expression by Salmonella 1,4,[5],12:i:- was evaluated using 133 isolates from clinical, environmental and animal origins, collected in Portugal from 2006 to 2011. Biofilm detection was performed by phenotypic and genotypic methods, such growth characterization in agar and broth medium, optical density determination by microtiter assays and direct observation by fluorescent in situ hybridization. Biofilm-related genes adrA, csgD and gcpA were detected by PCR. A socio-geographic characterization of strains as biofilm producers was also performed. Results showed that biofilm formation in monophasic Salmonella is widely distributed in Portuguese isolates and could be one of the reasons for its dissemination in this country. Biofilm expression varies between locations, showing that isolates from some regions like Lisboa or Ponta Delgada have an increased ability to persist in the environment due to an enhanced biofilm production. Biofilm formation also varies between risk groups, with a higher prevalence in isolates from salmonellosis infections in women. Therefore, the analysis of the socio-geographic distribution of biofilm-forming bacteria should be considered for the establishment of more adequate regulatory measures or therapeutics regimens, especially important due to the continuous increase of infections caused by antimicrobial resistant microorganisms.

Three hundred and thirty-three isolates representing 40 different serotypes of Salmonella enterica, recovered from environmental and faecal samples of breeder and broiler flocks from 2009 to 2011, were studied. Antimicrobial susceptibility was determined by measuring the minimal inhibitory concentration of 11 antimicrobials using the agar dilution method. Salmonella Havana, S. Enteritidis and S. Mbandaka were the most common serotypes isolated from broiler flocks, while S. Enteritidis was the common isolate from breeder flocks. The frequency of non-wild-type Salmonella isolates (those with decreased susceptibility to the different antimicrobials) varied according to serotype. S. Mbandaka in broilers and S. Enteritidis in both breeders and broilers showed higher frequencies of reduced susceptibility to quinolones, but clinical resistance towards ciprofloxacin was not observed. Reduced susceptibility to sulfamethoxazole, tetracycline, ampicillin and streptomycin were common in Salmonella Typhimurium isolates. Two isolates of S. Havana from broilers were resistant to cefotaxime and phenotypically categorised as extended-spectrum β-lactamase producers. The results presented in this study provide useful data on the antimicrobial susceptibility of different Salmonella serotypes and highlight the high diversity of multi-drug resistance patterns present.

Salmonella Typhimurium 1,4,[5],12:i:- is a major serovar responsible for human salmonellosis whose biofilm-forming ability, influenced by environmental conditions like those found in the gastrointestinal tract, is one of the main contributing factors to its ability to persist in the host and thus one of the main causes of chronic relapsing infections. Most studies to evaluate biofilm formation are performed in microtiter assays using standard media. However, no reports are available on the ability of this serovar to produce biofilm under in vitro simulated gastrointestinal conditions which better correlate with the environment found in the gastrointestinal tract. To address this, a modified biofilm assay simulating intestinal fluid was conceived to assess the biofilm formation of 133 Salmonella Typhimurium 1,4,[5],12:i:- isolates with and without agitation and at three different time points (24 h, 48 h, and 72 h). The results were then compared to the existing microtiter method using conventional biofilm growth medium (Mueller Hinton Broth). Statistical analysis revealed significant differences in the results obtained between the three protocols used. The simulated human intestinal environment impaired biofilm production demonstrating that conditions like pH, agitation or the presence of enzymes can influence biofilm production. Therefore, results from in vitro simulation of in vivo conditions may contribute to unravelling factors relating to biofilm formation and persistence in the context of the human host.

While routine monitoring poultry meat was obtained from breeding hens, dioxins contaminations were detected in Portugal. Levels of 430.9 pg PCDD/F-WHO-TEQ/g1 were found, which are higher than the official limits legally allowed for this matrix (1.75 pg PCDD/F-WHO-TEQ/g). To identify the magnitude of the contaminations, 60 samples were collected from poultry farms and different matrices, namely: feed, water, wood shavings from the litters, muscle of the breeding hens, hatching eggs collected in the positive farm and muscle collected from broilers farms supplied by the positive breeding farm. The comparison of the dioxins congeners profiles showed that there was a coincidence of peaks of higher relative concentrations in the wood shavings, with the peaks of the highest relative concentration in the hatching eggs, especially the relative concentrations of the congeners 1,2,3,4,6,7,8-HpCDD and OCDD, which may be explained by the role of VLDLy in the delivery of triglycerides to the oocyte, where they will be used as the energy source for the developing embryo. The comparison of the dioxins congeners profiles of the breeding hens muscle with the poultry muscle, showed a coincidence of peaks of higher relative concentrations in the congeners 1,2,3,7,8-PeCDD, 1,2,3,6,7,8-HxCDD, 1,2,3,4,6,7,8-HpCDD and OCDD which may indicate a dechlorination pathway “in vivo”. Results allowed concluding that those wood shavings, improperly used as poultry litters, were certainly the source of contamination of the food chain.

For a long time, ripened cheeses have been officially recognized as having human health beneficial properties. However, to date only a few studies have investigated the complex microbiota of S?o Jorge and Parmigiano-Reggiano cheeses in order to identify microorganisms with probiotic potential. Hence, with the purpose of analyzing the probiotic potential of autochthone bacteria from S?o Jorge and Parmigiano-Reggiano cheeses, lactic acid bacteria isolated from both products were isolated and their direct biotic interaction against three bacteria found in the human intestine evaluated. From a total of 225 bacterial autochthone isolates, 67 had synergetic behavior with Bifidobacterium animalis , 34 had no synergetic behavior with Shigella dysenteriae and 33 had antagonistic activity with this human pathogen. Biochemical tests were used to identify these cheese isolates with probiotic potential. Lactobacillus paracasei subsp paracasei Lactobacillus arhamnosus were found in São Jorge cheese and Lactobacillus rhamnosus , Lactobacillus buchneri and Lactobacillus curvatus in the Parmigiano cheese. Overall, initial exploratory studies with São Jorge cheese seem to show a higher number of potential probiotic agents than those found in the Parmigiano-Reggiano cheese. However, in order to better understand the functional potential of this traditional Portuguese cheese, further confirmatory studies should be pursued.

This paper presents 3 years of data (2009–2011) on the occurrence of two mycotoxins, aflatoxin B1 (AFB1) and zearalenone (ZEA), in samples of feedstuff for dairy cows (n = 963), ewes (n = 42), and goats (n = 131) produced in Portugal. AFB1 was found in 15 samples of cow feed (1.6 %), 3 samples of ewe feed (2.3 %) and in 2 samples of goat feed (4.8 %). All but two samples contained AFB1 at levels below the European Union maximum level (5 μg/kg). Nearly half (45 %) of the samples were contaminated with ZEA, but its levels were relatively low, at 5–136.9 μg/kg, well below the European Union guidance value (500 μg/kg).

ABSTRACT Objective: to characterize the epidemiological aspects of women with Chagas disease. Method: this is a descriptive, transversal, study conducted from March 2007 to April 2008, with the application of a form to 32 patients in Campo Grande, Mato Grosso do Sul, Brazil. Data were analyzed through descriptive statistics, under the form of table and figure, using the software Statistical Package for the Social Sciences (SPSS), version 13.0. The significance level was 0.05. The research was approved by the Ethics Committee of Universidade Federal de Mato Grosso do Sul (UFMS), under the Protocol 883/2007. Results: the incidence rate was 0.9 per 1,000 pregnant women; there was a predominance of low education, housewives, and white colored people. At birth, they lived in the countryside of Mato Grosso do Sul or in other towns in endemic states, at wooden houses. Conclusion: there was a correlation between low education and Chagas disease. Changes in the electrocardiogram, echocardiogram, and chest x-ray have a significant value in the initial assessment, even in asymptomatic patients. Descriptors: Chagas Disease; Women’s Health; Epidemiological Surveillance; Trypanosoma Cruzi ; Prenatal Care. RESUMO Objetivo: caracterizar os aspectos epidemiologicos de mulheres com doenca de Chagas. Metodo: trata-se de estudo descritivo, transversal, realizado de marco de 2007 a abril de 2008, com a aplicacao de um formulario a 32 pacientes em Campo Grande-MS. Os dados foram analisados por meio da estatistica descritiva, sob a forma de tabela e figura, com uso do programa Statistical Package for the Social Sciences (SPSS), versao 13.0. O nivel de significância foi de 0,05 . A pesquisa foi aprovada pelo Comite de Etica da Universidade Federal de Mato Grosso do Sul (UFMS), sob o Protocolo n. 883/2007. Resultados: a taxa de incidencia foi de 0,9 para cada 1.000 gestantes; houve predominio de baixa escolaridade, de trabalhadoras do lar e de cor branca. Ao nascimento residiam no interior de Mato Grosso do Sul ou em outras cidades de estados endemicos, em casas de madeira. Conclusao: houve correlacao entre baixa escolaridade e doenca de Chagas. Alteracoes do eletrocardiograma, ecocardiograma e radiografia de torax tem valor significativo na avaliacao inicial, mesmo em pacientes assintomaticas. Descritores: Doenca de Chagas; Saude da Mulher; Vigilância Epidemiologica; Trypanosoma Cruzi ; Cuidado Pre-Natal. Resumen Objetivo: caracterizar los aspectos epidemiologicos de mujeres con enfermedad de Chagas. Metodo: esto es un estudio descriptivo, transversal, realizado de marzo de 2007 a abril de 2008, con aplicacion de un formulario a 32 pacientes en Campo Grande, Mato Grosso do Sul, Brazil. Los datos fueron analizados por medio de la estadistica descriptiva, bajo la forma de tabla y figura, con utilizacion del programa Statistical Package for the Social Sciences (SPSS), version 13.0. El nivel de significacion fue de 0,05. La investigacion fue aprobada por el Comite de Etica de la Universidade Federal de Mato Grosso do Sul (UFMS), bajo el Protocolo 883/2007. Resultados: la tasa de incidencia fue de 0,9 por cada 1.000 mujeres embarazadas; hubo predominio de bajo nivel de educacion, de trabajadoras del hogar y del color blanco. Al nacer residian en el interior de Mato Grosso do Sul o en otras ciudades de estados endemicos, en casas de madera. Conclusion: hubo correlacion entre baja educacion y enfermedad de Chagas. Cambios en el electrocardiograma, ecocardiograma y radiografia de torax tienen valor significativo en la evaluacion inicial, incluso en pacientes asintomaticas. Descriptores: Enfermedad de Chagas; Salud de la Mujer; Vigilancia Epidemiologica; Trypanosoma Cruzi ; Atencion Prenatal. Normal 0 21 false false false PT-BR X-NONE X-NONE st1\:*{behavior:url(#ieooui) } /* Style Definitions */ table.MsoNormalTable {mso-style-name:Tabela normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:Times New Roman; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Times New Roman; mso-bidi-theme-font:minor-bidi;}

A water-vapour retrieval algorithm has been developed that uses satellite observations in the microwave region. It is based on neural-network modelling and includes a dedicated calibration scheme for the satellite observations. The water vapour is retrieved for clear and cloudy scenes, over both ocean and land surfaces. Precipitation cases are excluded. The atmospheric relative humidity profile is retrieved on six atmospheric layers, together with the total column water vapour. By-products are also retrieved by the algorithm, including surface temperature and microwave emissivities over the continents and surface wind speed over the ocean. A first version of a retrieval chain has been produced for the French-Indian Megha-Tropiques mission launched on 12 October 2011. The algorithm has been further developed for the instruments AMSR-E/HSB (resp. AMSU-A/MHS) on board the AQUA (resp. MetOp) platform, in order to test it on existing satellite observations. In this article, the principles of the inversion method are presented and the theoretical retrieval uncertainties are assessed using direct tests on simulated data as well as estimations using the traditional information-content analysis. Results of the retrieval algorithm will be evaluated in a companion article for AQUA and MetOp observations using comparisons with European Centre for Medium-Range Weather Forecasts (ECMWF) analysis and radiosonde measurements. Copyright (c) 2012 Royal Meteorological Society