Padronização de uma PCR multiplex para a detecção de fraude em leite bubalino e caprino por adição de leite bovino / Standardization of a multiplex PCR for detection of fraud in buffalo and goat milk by addition of bovine milk

O consumo de leite de espécies como bubalino e caprino tem se popularizado por representarem uma alternativa para indivíduos que possuem restrições alimentares relacionadas ao leite bovino e em virtude das propriedades nutricionais desses alimentos. No entanto, fatores como a baixa produção e a sazonalidade predispõem a adulterações destes alimentos, principalmente pela adição de leite bovino, visando maior rendimento e lucratividade. Assim, o objetivo do estudo foi padronizar um método de PCR multiplex para autenticação de leites bubalino e caprino. Para isso, amostras de leite exclusivamente de cada espécie foram utilizados para a padronização da técnica. Em seguida, foi realizada a fraude pela adição de leite bovino ao caprino e ao bubalino, em proporções de 0,1% até 100%. A técnica foi eficaz, precisa, rápida e prática para a detecção do DNA de bovino, bubalino e caprino, separadamente e em conjunto. Na fraude experimental, o limite de detecção da técnica ocorreu a partir do menor percentual testado (0,1%) tanto no leite caprino quanto no bubalino. Dessa forma, a PCR multiplex testada mostrou ser uma importante ferramenta para a autenticação de leite, pendendo ser utilizada para fins de fiscalização por órgãos competentes.

Milk consumption of species such as buffalo and goat has become popular due to the nutritional properties of these foods and because they represent an alternative for individuals who have dietary restrictions related to bovine milk. However, factors such as low production and seasonality predispose to adulteration, mainly by the addition of bovine milk, aiming at higher yield and profitability. Thus, the aim of the present study was to standard a multiplex PCR method for buffalo and goat milks authentication. For this, the milks exclusively of each species were used to standardize the technique. Subsequently, fraud was performed by the addition of bovine milk to goat and buffalo in proportions from 0.1% to 100%. The technique was effective and accurate for detecting bovine, buffalo and goat DNA separately and together quickly and practically. In experimental fraud, the detection limit of the technique occurred from the lowest percentage tested (0.1%) in both goat and buffalo milk. Thus, the multiplex PCR tested proved to be an important tool for milk authentication, pending to be used for supervision by competent agencies.

Potentially pathogenic Staphylococcus aureus and Listeria spp. in Brazilian unpasteurized cheese production / Staphylococcus aureus e Listeria spp. potencialmente patogênicos na produção de queijos não pasteurizados

This study aimed to identify potentially pathogenic microorganisms (Listeria innocua, L. seeligeri, L. ivanovii, L. monocytogenes, Staphylococcus aureus, and several virulence genes) in unpasteurized cheese production in the northeastern region of the state of São Paulo, Brazil. Listeria species were detected in 68 (64.14%) out of 106 samples of bovine feces, swabs from milkers' and cheese handlers' hands, milking buckets, raw milk, whey, water, cheese processing surface,s and utensils. All the samples collected at one farm were contaminated with Listeria spp. L. innocua, L. seeligeri, L. ivanovii, or L. monocytogenes were not detected in the samples collected in this study. A set of 391 Staphylococcus spp. isolates were obtained in these samples, from which 60 (15.31%) were identified as S. aureus using PCR (Polymerase Chain Reaction). S. aureus carrying virulence genes (eta, hlg, seg, seh, sei) were detected in milk, in swabs from cheese handler's hands, whey, milk, sieves, buckets, and cheese. The hlg gene (encodes gamma hemolysin) was detected in all the S. aureus isolates. These findings show that poor hygienic practice is associated with a higher risk of pathogenic bacteria in milk or cheese, providing useful information for public health authorities to increase food safety surveillance and prevent the dissemination of pathogens.

O objetivo desse estudo foi identificar microrganismos potencialmente patogênicos (Listeria innocua, L. seeligeri, L. ivanovii, L. monocytogenes, Staphylococcus aureus e diversos genes de virulência) na produção de queijos de leite cru na região noroeste do Estado de São Paulo, Brasil. Listeria foram detectadas em 68 (64,14%) das 106 amostras obtidas de fezes bovinas, suabes das mãos de ordenhadores e queijeiros, baldes, leite cru, soro, água, superfícies e utensílios da produção de queijos. Todas as amostras coletadas em uma fazenda estavam contaminadas com Listeria spp. L. innocua, L. seeligeri, L. ivanovii, e L. monocytogenes não foram detectadas nas amostras coletadas nesse estudo. Um conjunto de 391 isolados de Staphylococcus spp. foram obtidos das amostras, e desses 60 (15,31%) foram identificados como S. aureus pela PCR (Polymerase Chain Reaction). S. aureus contendo genes de virulência (eta, hlg, seg, seh, sei) foram detectados em leite, mãos dos ordenhadores, soro, utensílios e queijos. O gene hlg (gama-hemolisina)foi detectado em todos os isolados de S. aureus.Esses resultados demonstram que práticas inadequadas de higiene estão associadas com um maior risco da presença de bactérias patogênicas no leite e queijos crus, fornecendo informações para as autoridades de saúde pública para incrementarem a vigilância e prevenirem a disseminação de patógenos.

Ocorrência de perigos físicos em alimentos / Occurrence of physical hazards in food

A ocorrência de matérias estranhas com potencial perigo físico em alimentos é um grande desafio para a indústria alimentícia, e a análise microscópica é provavelmente o instrumento mais útil para detectá-las e identificá-las. Considerando a escassez de dados sobre os perigos físicos em produtos alimentícios no Brasil, o objetivo deste estudo foi descrever essa ocorrência nos produtos analisados no Núcleo de Morfologia e Microscopia do Centro de Alimentos do Laboratório Central do Instituto Adolfo Lutz (IAL), em São Paulo, no período de 2008 a 2020. Os resultados revelaram que, das 7.221 amostras de alimentos analisadas, 89 (1,2%) estavam em desacordo com a legislação em vigor, por conterem perigos físicos. De acordo com a categoria de alimentos, a ocorrência foi maior para bebidas (43%), seguida de cereais, farinha e farelo (22%). Quanto ao tipo de matéria estranha, os plásticos (duros e flexíveis) foram os mais frequentes (48%), seguidos dos metais (15%) entre as partículas perigosas detectadas nas amostras. A detecção e identificação de perigos físicos, por meio de análises microscópicas, contribuem para a segurança e qualidade dos produtos alimentícios oferecidos à população (AU).

The occurrence of physical hazards in food is a great challenge for the food industry, and microscopic analysis is probably the most useful instrument to detect and identify them. Considering the scarcity of data on physical hazards in food products in Brazil, the aim of this study was to describe this occurrence in the products analyzed at the Nucleus of Morphology and Microscopy of the Food Center of the Adolf Lutz Institute's Central Laboratory (IAL) in São Paulo, from 2008 to 2020. Results revealed that of the 7,221 analyzed food samples, 89 (1.2%) did not comply with the legislation due to the presence of physical hazards. According to the food category, the occurrence was higher in beverages (43%), followed by cereals, yeasts, flour and bran (22%). Regarding the type of foreign matter, among the dangerous particles detected, the most frequently found were plastics (48%) (hard and flexible), followed by metals (15%). The detection and identification of physical hazards by microscopic analysis contributes to the safety and quality of food products offered to the public. (AU)

Vigilancia epidemiológica de brotes de enfermedades trasmitidas por alimentos en la provincia de Santiago de Cuba / Epidemiologic surveillance of diseases outbreaks transmitted by foods in Santiago de Cuba province

Introducción: Las enfermedades trasmitidas por alimentos se producen por la ingestión de alimentos y/o bebidas contaminados. Objetivo: Identificar los agentes causales que influyen en la aparición de estas enfermedades. Métodos: Se realizó un estudio observacional, descriptivo y transversal del total de brotes de enfermedades trasmitidas por alimentos en la provincia de Santiago de Cuba, desde enero de 2018 hasta diciembre de 2019. Las variables analizadas fueron: número de brotes, muestras de alimentos y muestras de heces fecales para coprocultivo, entre otras. Resultados: En todos los brotes se aisló un solo tipo de agente bacteriano, con predominio de las bacterias gramnegativas, donde la Salmonella fue el microorganismo más identificado. Los principales grupos de alimentos relacionados con la aparición de dichos brotes resultaron ser la carne y sus derivados, así como la ensalada fría. Conclusiones: La vigilancia epidemiológica de brotes de enfermedades transmitidas por alimentos, permitió identificar el verdadero agente causal en los brotes de ETA, al demostrarse identidad entre los aislamientos bacterianos obtenidos de muestras de alimentos y heces fecales.

Introduction: Diseases transmitted by foods take place due to the ingestion of foods and/or contaminated drinks. Objective: To identify the causal agents that influence in the emergence of these diseases. Methods: An observational, descriptive and cross-sectional study of all the diseases outbreaks transmitted by foods was carried out in Santiago de Cuba, from January, 2018 to December, 2019. The analyzed variables were: number of outbreaks, samples of foods and samples of stools for coproculture, among others. Results: In all the outbreaks a single type of bacterial agent was isolated, with prevalence of the gram-negative bacterias, where Salmonella was the most identified microorganism. The main groups of foods related to the emergence of these outbreaks were meat and its derived products, as well as cold salad. Conclusions: The epidemiologic surveillance of diseases outbreaks transmitted by foods, allowed to identify the true causal agent in the FTD outbreaks, when the identity between the bacterial isolations obtained from samples of foods and stools was demonstrated.

Aflatoxins and fungal community distribution during harvesting and processing of Platycladi Semen / 中国中药杂志

This study aimed to analyze aflatoxins content and fungal community distribution in the harvesting and processing of Platycladi Semen, and explore the key link that affects aflatoxins contamination. The related Platycladi Semen samples of different maturity periods(cone non-rupture period, early rupture, and complete rupture period) and different processing periods(before drying, during 2-d drying, during 7-d drying, before and after seed scale removal, before and after peeling, 1 d after color sorting, and 7 d after color sorting) were collected for identifying the fungal community composition on sample surface by ITS amplicon sequencing. Then the content of aflatoxins B_1, B_2, G_1 and G_2 was determined by HPLC-MS/MS. The results showed that during the harvesting of Platycladi Semen from cone non-rupture to complete rupture, aflatoxins were only detected in the seed scale and seed coat, with aflatoxin G_2 in the seed scale and aflatoxin B_1 in the seed coat. During the drying, with the prolongation of drying time, aflatoxins B_1 and G_2 were detected simultaneously in the seed scale, aflatoxin B_1 in the seed coat, and low-content aflatoxin B_1 in the seed kernel. During subsequent processing, the aflatoxin content in seed kernel during subsequent processing was slighted increased. As demonstrated by fungal detection, Aspergillus flavus was not present during the harvesting of Platycladi Semen, but present during the drying and processing. Its content in the seed coat during the drying process was relatively higher. In short, Platycladi Semen should be harvested as soon as possible after it becomes fully mature. Drying process is the key link of preventing aflatoxin contamination. It is advised to build a sunlight room or adopt similar settings, standardize the operations in other processes, and keep the surrounding environment clean to minimize aflatoxin contamination.

Methods for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigation on its contamination / 中国中药杂志

This study aimed to establish a method for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigate its contamination with mycotoxins, so as to provide technical guidance for monitoring the quality of Chinese medicinal materials and medication safety. The sample was extracted with 80% acetonitrile in an oscillator for 1 h, purified using the modified QuEChERS purifying agent(0.1 g PSA + 0.3 g C_(18) + 0.3 g MgSO_4), and separated on a Waters HSS T3 chromatographic column(2.1 mm×100 mm, 1.8 μm). The gradient elution was carried out with 0.1% formic acid in water and acetonitrile, followed by the scanning in the multi-reaction monitoring(MRM) mode and the analysis of mycotoxin contamination in 26 Pseudostellariae Radix samples. The recovery rates of the established method were within the range of 82.17%-113.6%, with the RSD values less than 7% and the limits of quantification(LOQ) being 0.019-0.976 μg·kg~(-1). The detection rate of 14 mycotoxins in 26 batches of medicinal materials was 53.85%. The detection rate of sterigmatocystin(ST) was the highest, followed by those of zearalenone(ZEN), aflatoxin G_2(AFG_2), fumonisin B_1(FB_1), HT-2 toxin, and nivalenol(NIV). Their respective detection rates were 38.46%, 26.92%, 23.08%, 11.54%, 11.54%, and 7.69%, with the pollution ranges being 1.48-69.65, 0.11-31.05, 0.11-0.66, 0.28-0.83, 20.86-42.56, and 0.46-1.84 μg·kg~(-1), respectively. The established method for the detection of 14 mycotoxins is accurate, fast and reliable. The research results have very important practical significance for guiding the monitoring and prevention and control of exogenous fungal contamination of Chinese medicinal materials.

Occurrence of potentially highly pathogenic free-living amoebae in ready made salads from restaurants in Jatai, Goias, Brazil

Free-living amoebae (FLA) are part of a group of protozoa found worldwide and in the most diverse environments. They resist various temperatures and disinfection methods, and are a risk to human health. Pathogenic strains grow at high temperatures and under hyperosmolarity conditions. Some FLA genera are mainly related to primary amoebic meningoencephalitis (PAM), skin ulcerations, corneal lesions, kidney and lung infections and keratitis. Therefore, studies that assess the pathogenic potential of FLA are public health issues of great concern. We aimed to evaluate the pathogenic potential of FLA isolated in salads from restaurants in vitro, using osmotolerance and thermotolerance tests. Forty-five isolates were used from ready-made salads purchased in restaurants in Jatai, Goias. Twelve isolates subjected to the osmotolerance test (26.6%) showed growth in 0.5 M mannitol, 18 (40.0%) in 1.0 M mannitol and 16 (35.5%) in 1.5 M mannitol, 13 (28.8%) isolates did not show growth. Four isolates that underwent the thermotolerance test (8.9%) showed growth at 25°C, 8 (17.8%) showed growth at 30°C, 3 (6.7%) showed growth at 37°C and 30 (66.7%) did not show growth. With the indices obtained in the present study, we concluded that 15.6% of the isolates were osmotolerant and thermotolerant. Our findings highlight a public health problem once these FLA are associated with harboring or being harbored by microorganisms responsible for diseases such as diarrhea and meningitis. Measures are required to improve food hygiene and so avoid FLA-related health problems.

Infestación de Musca domestica L. (Díptera, Muscidae) en faenadoras avícolas / Infestation of Musca domestica L. (Diptera, Muscidae) in poultry slaughterhouses

Musca domestica (Diptera, Muscidae) es la mosca más común en todo el mundo. Más de 100 patógenos pueden causar enfermedades en humanos y animales por este tipo de insectos. Estos patógenos incluyen: diarrea infantil, ántrax, cólera, oftalmía, disentería bacilar, fiebre tifoidea y tuberculosis. Además, las moscas domésticas transmiten muchos de los huevos de helmintos como Enterobius vermicularis, Strongyloides stercoralis, Trichuris trichiura, Toxocara canis, y especies de Dipylidium, Diphyllobothriam, Hymenolepis, Taenia y Trichomonas. También puede transmitir quistes de protozoos y trofozoítos como E. histolytica y Giardia lamblia. Se determinó el índice de infestación por Musca domestica L. (Díptera, Muscidae) en una faenadora avícola ubicada en el Perú. Para ello se evaluó no sólo el tratamiento aplicado: protocolo de cola entomológica (T1), monitoreo de insfectación de mosca (T2) o comincación de lámparas LED UV más tabla de pegamento adhesivo (T3), sino también las diferentes zonas de faenadora: zonas sucias de descarga y estabulación (ZDE), preparación (ZP), zonas adicionales de sacrificio sanitario (ZSS), preparación de residuos orgánicos (ZPRO), conservación de los residuos orgánicos (ZCRO) y de conservación de sangre (ZCS). Los resultados demoestraron que, independientemente de la zona de operación, el tratamiento más efectivo para determinar la infestación por la mosca doméstica fue el combinado de lámpara LED junto con tablas de pegamento (T3), seguido del tratamiento de protocolo de cola entomológica (T1) y el tratamiento por Monitoreo de infestación de moscas (T2) independientemente de los días de recolección. Es importante seguir las diferentes normativas a fin de controlar y eliminar la presencia de moscas (y de otros insectos voladores) para evitar la contaminación y, por ende, enfermedades(AU)

Musca domestica (Diptera, Muscidae) is the most common fly in the world. More than 100 pathogens can cause diseases in humans and animals by these types of insects. These pathogens include: infant diarrhea, anthrax, cholera, ophthalmia, bacillary dysentery, typhoid fever, and tuberculosis. In addition, houseflies transmit many of the eggs of helminths such as Enterobius vermicularis, Strongyloides stercoralis, Trichuris trichiura, Toxocara canis, and Dipylidium, Diphyllobothriam, Hymenolepis, Taenia, and Trichomonas species. It can also transmit cysts of protozoa and trophozoites such as E. histolytica and Giardia lamblia. The infestation index by Musca domestica L. (Diptera, Muscidae) was determined in a poultry slaughterhouse located in Peru. For this, not only the applied treatment was evaluated: entomological glue protocol (T1), fly infestation monitoring (T2) or combination of UV LED lamps plus adhesive glue table (T3), but also the different slaughter areas: unloading and holding areas (ZDE), preparation (ZP), additional stamping-out areas (ZSS), preparation of organic waste (ZPRO), conservation of organic waste (ZCRO) and blood conservation (ZCS). The results showed that, regardless of the area of operation, the most effective treatment to determine housefly infestation was the LED lamp combined with glue boards (T3), followed by the entomological glue protocol treatment (T1 ) and the treatment by Monitoring of fly infestation (T2) regardless of the days of collection. It is important to follow the different regulations in order to control and eliminate the presence of flies (and other flying insects) to avoid contamination and, therefore, diseases(AU)

Parasitological analysis of vegetables sold in supermarkets and free markets in the city of Taguatinga, Federal District, Brazil

Parasitosis are the most common diseases in the world, they are responsible for relevant negative consequences in the individuals´ health. The consumption of vegetables in Brazil and in the world is common due to their high nutritional value and they are recommended to be included in the population´s diet. Contaminations in vegetables are most often caused by irrigation water which is contaminated by feces, other forms of contamination are through organic manure containing fecal waste and the contact of vegetables with animals where they are grown. This research evaluated the contamination in vegetables with the purpose of warning the community about hygiene and the correct approach of food handling in order to prevent parasitic infections, contributing to people´s health. Samples were collected from supermarkets and farmers´ markets, where we obtained a total of 30 samples that were analyzed using Hoffman, centrifugation and slide reading´s techniques. In all samples, parasites were found. In six samples non-pathogenic amoebas were found, however this finding means that there was contamination from human or animal feces. And, in the other 24 samples, parasitic structures that cause damage to health were found. This work showed a high level of contamination of the vegetables that are consumed daily by the population and the diseases caused by food which are a concern for the public health. Contaminated vegetables are dangerous considering the transport of parasites that may generate human infections, therefore the importance to create public policies in order to avoid parasitic infections.

Evaluation of vegetables grown in dry mountainous regions for soil transmitted helminths contamination / Avaliação de vegetais para contaminação por helmintos transmitida pelo solo em quatro distritos da divisão de Malakand: noroeste do Paquistão

Abstract Infection caused by geo-helminth parasites are called geohelminthiasis are one of the global health problems. Vegetables eaten raw is the principal source of transmission of geo-helminth parasites. Pakistani people believe that eating raw vegetables are a significant source to get important vitamins and minerals. Based on the high incidence of pathogenic parasites and cultivating different vegetable types in the study areas, we conducted this study to evaluate the geo-helminth contamination of raw vegetables in northwest Khyber Pakhtunkhwa, Pakistan. This is a descriptive study comprised, 1942 samples of 25 various types of vegetables. The samples were examined in physiological saline solution using sedimentation and centrifugation methods. The findings were analyzed by Graph-Pad version 5. P value less than 0.05 (95% CI) was considered significant. Results showed that 16.5% (n=322) of all vegetables were contaminated with one or more type of geo-helminth parasites. Garlic was the highest (35%) and cauliflower the lowest (4%) contaminated samples respectively. Ascaris lumbricoides was the most common geo-helminth found followed by hook worm species while Trichuris trichura was the least in all the vegetable samples. Leafy vegetables were highly contaminated 25.3% than vegetables with root parts 21.2% and fruity 9.09%. More than half of the contaminated vegetables were contaminated with single species of geo-helminth (P<0.05) while less than half with multiple types of geo-helminth contamination. Ninety two vegetables samples were contaminated with 2 species of parasites (P<0.05) and 45 with 3 (P>0.05) species of geo-helminth parasites. Education level of vendors and means of display were not significantly associated while types of vegetable used were significantly associated with the prevalence of parasites. The findings of this study provide evidence that consumption of raw vegetable has a high risk of acquiring geo-helminth infections. The authors believe that preventing the human to enter to the vegetable farmland for defecation, avoiding the irrigation of agricultural fields via night soil, and educating the people on proper washing and cooking of vegetables may be useful in reducing parasitic infections.

Resumo As infecções causadas por parasitas geo-helmínticos são chamados de geohelmintíases e são um dos problemas de saúde globais. Os vegetais comidos crus são a principal fonte de transmissão dos parasitas geo-helmínticos. O povo paquistanês acredita que comer vegetais crus é uma fonte significativa para obter vitaminas e minerais importantes. Com base na alta incidência de parasitas patogênicos e no cultivo de diferentes tipos de vegetais nas áreas de estudo, conduzimos este estudo para avaliar a contaminação por geo-helmintos de vegetais crus no noroeste de Khyber Pakhtunkhwa, Paquistão. Trata-se de um estudo descritivo composto por 1942 amostras de 25 tipos diversos de vegetais. As amostras foram examinadas em solução salina fisiológica utilizando métodos de sedimentação e centrifugação. Os achados foram analisados ​​pelo Graph-Pad versão 5. O valor de P menor que 0,05 (IC 95%) foi considerado significativo. Os resultados mostraram que 16,5% (n = 322) de todas as hortaliças estavam contaminadas com um ou mais tipos de parasitas geo-helmínticos. O alho foi a amostra mais contaminada (35%) e a couve-flor a menos (4%), respectivamente. Ascaris lumbricoides foi o geo-helmíntico mais comum encontrado, seguido por espécies de verme-anzol, enquanto Trichuris trichura foi o menos encontrado em todas as amostras de vegetais. Os vegetais folhosos foram altamente contaminados 25,3% do que os vegetais com partes de raiz 21,2% e frutados 9,09%. Mais da metade dos vegetais contaminados estavam contaminados com uma única espécie de geo-helmintos (P <0,05), enquanto menos da metade com vários tipos de contaminação com geo-helmintos. Noventa e duas amostras de vegetais estavam contaminadas com 2 espécies de parasitas (P <0,05) e 45 com 3 (P> 0,05) espécies de parasitas geo-helmínticos. O nível de escolaridade dos vendedores e os meios de exibição não foram significativamente associados, enquanto os tipos de vegetais usados ​​foram significativamente associados à prevalência de parasitas. Os resultados deste estudo fornecem evidências de que o consumo de vegetais crus tem um alto risco de adquirir infecções por geo-helmintos. Os autores acreditam que impedir que o ser humano entre na terra de hortaliças para defecação, evitar a irrigação de campos agrícolas via solo noturno e educar as pessoas sobre a lavagem e cozimento adequados de vegetais pode ser útil na redução de infecções parasitárias.

Detection of enteropathogens and research of pesticide residues in Lactuca sativa from traditional and agroecological fairs / Detecção de enteropatógenos e pesquisa de resíduos de agrotóxicos em Lactuca sativa de feiras tradicionais e agroecológicas

Abstract Agroecological production represents a new reality in vegetable farming. Thus, the study aimed to search for the presence of pesticides in agroecological samples and microbiological and parasitological contamination in Lactuca sativa L. of agroecological and conventional fairs in Alagoas. Thirty-two samples were collected, a hygienic-sanitary checklist was performed, along with coliform, Salmonella spp., Staphylococcus aureus research and Gas Chromatography with Mass Spectrometer (GC-MS) for pesticide research; using two methodologies for parasitological studies. Thirty-two samples were analyzed, with a sanitary adequacy level at 45% and 38% for fairs (agroecological and conventional) respectively, with three samples (9.4%) in total, being adequate to the microbiological pattern according to RDC no. 12/2001. Among the 23 samples which were tested positive for parasites, 69.5% originated from samples extracted at conventional fairs, and 53% of every positive samples presented Entamoeba coli as the main contaminant. All together, no traces of deltamethrin were detected, but in two of the samples a peak retention for diphenoconazole was detected. In conclusion, the fairs represent an adequate environment for human pathogens and indicators. Therefore greater hygiene training of vegetable traders and consumers in the state of Alagoas is required, especially in conventional fairs, which presented more critical microbiological and parasitological indexes comparing to fairs with agroecological products; however, the presence of difenoconazole in two samples of agricultural fairs suggests accidental contamination or indiscriminate use of this chemical in the production of Lactuca sativa, however, products with agroecological origin represents a better choice for the consumers.

Resumo A produção agroecológica representa uma nova realidade no cultivo das hortaliças. Assim, o estudo teve como objetivo verificar a presença de agrotóxicos em amostras agroecológicas e analisar a contaminação microbiológico e parasitológico em Lactuca sativa L. de feiras agroecológicas e convencionais de Alagoas. Foram coletadas 32 amostras, realizando checklist higiênico-sanitário, análises de coliformes, Salmonella spp., Staphylococcus aureus e Cromatografia Gasosa com Espectrômetro de Massas (GC-MS) para pesquisa de agrotóxicos; utilizando-se duas metodologias para estudos parasitológicos. Foram analisadas 32 amostras, com nível de adequação sanitária de 45% e 38% para feiras (agroecológicas e convencionais) respectivamente, sendo três amostras no total (9,4%) adequadas a padrão microbiológico segundo RDC n° 12//2001; Dentre as 23 amostras positivas para parasitos, 69,5% teve origem em amostras de feiras convencionais, e 53% de todas as amostras positivas apresentaram Entamoeba coli como principal contaminante, juntamente, não foi detectado traços de deltametrina, porem foi visualizado em duas amostras, pico de retenção para padrão de difenoconazol. Concluindo-se que as feiras representam um ambiente adequado para patógenos humanos e indicadores, necessitando de uma maior higienização e treinamento dos comerciantes e consumidores de hortaliças do estado de Alagoas, principalmente em feiras convencionais, que apresentaram índices microbiológicos e parasitológicos mais críticos em relação a feiras com produtos agroecológicos; mas, a presença de difenoconazole em duas amostras de feiras agroecológicas sugerem contaminação acidental ou uso indiscriminado deste químico em produção da Lactuca sativa L., todavia os produtos de origem agroecológica representam uma melhor escolha para o consumidor.

Parasitic contamination of fresh vegetables sold in open markets: a public health threat / Contaminação parasítica de vegetais frescos vendidos em mercados abertos: uma ameaça para a saúde pública

Abstract Vegetables eating raw are a leading source of transmission of infective forms of pathogenic internal parasites among human beings. This research was conducted from April to October, 2017 to assess the parasitic contamination of vegetables sold at main vegetable markets in districts Lower Dir and Peshawar, Pakistan. Eight hundred specimens of different vegetables were purchased and soaked in physiological saline solution, shaken with a mechanical shaker for 20 minutes and processed by sedimentation concentration method. Results revealed that only 19.7% (n=158/800) of the vegetables were found to be contaminated with single or multiple parasite species. Ascaris lumbricoides (the large round worm) 12.3% (n=99/800) was the most commonly detected pathogen and Taenia saginata (the beef tapeworm) 1.62% (n=13/800) was the least frequently detected one. Interestingly, significant p value (p>0.05 at 95%CI) between the number of examined and contaminated for all the variables studied including education status of the vendors, markets location, type of vegetables, means of display, washed before display, washing source of water and market type. The findings of this study evidenced that consumption of raw vegetables possesses great risk of getting parasitic infections in Lower Dir and Peshawar districts, Pakistan. Instructing the sellers and the public about parasitic disease transfer and their hygiene can reduce the infection rate of parasites of human origin.

Resumo Os vegetais crus são a principal fonte de transmissão de formas infecciosas de parasitas internos patogênicos entre os seres humanos. Esta pesquisa foi realizada de abril a outubro de 2017 para avaliar a contaminação parasitária de vegetais vendidos nos principais mercados de vegetais nos distritos de Lower Dir e Peshawar, Paquistão. Oitocentos exemplares de diferentes vegetais foram adquiridos e embebidos em solução salina fisiológica, agitados em agitador mecânico por 20 minutos e processados ​​pelo método de concentração por sedimentação. Os resultados revelaram que apenas 19,7% (n = 158/800) das hortaliças estavam contaminadas com uma ou várias espécies de parasitas. Ascaris lumbricoides (o verme redondo grande) 12,3% (n = 99/800) foi o patógeno mais comumente detectado e Taenia saginata (a tênia da carne) 1,62% (n = 13/800) foi o menos freqüentemente detectado. Curiosamente, valor de p significativo (p> 0,05 em IC de 95%) entre o número de examinados e contaminados para todas as variáveis ​​estudadas incluindo escolaridade dos vendedores, localização dos mercados, tipo de vegetais, meios de exposição, lavado antes da exposição, fonte de lavagem de água e tipo de mercado. Os resultados deste estudo evidenciaram que o consumo de vegetais crus possui grande risco de contrair infecções parasitárias nos distritos de Lower Dir e Peshawar, no Paquistão. Instruir os vendedores e o público sobre a transferência de doenças parasitárias e sua higiene pode reduzir a taxa de infecção de parasitas de origem humana.

Application and progress in high-throughput sequencing technology for meat adulteration detection / 生物工程学报

Adulteration in meat products is a widespread issue that could lead to serious threats to public health and religious violations. Technology that offers rapid, sensitive, accurate and reliable detection of meat species is the key to an effectual monitoring and control against meat adulteration. In recent years, high-throughput sequencing-based DNA metabarcoding technology has developed rapidly. With the characteristics of being high-throughput, highly precise and high-speed, this technology can simultaneously identify multiple species in complex samples, thus offering pronounced advantages in the surveillance of adulteration in meat and meat products. Starting with an introduction of the major developments in the high-throughput sequencing technology in the past two decades, this review provides an overview of the technical characteristics and research methods of DNA metabarcoding, summarizes the application of DNA metabarcoding technology in meat adulteration detection over the last few years, discusses the challenges of using DNA metabarcoding technology in the detection of meat adulteration, and provides future prospects on the development of this technology.

Dairy goat production in the semi-arid region: productive and reproductive analysis, and the influence of the adoption of hygienic practices on milk quality / Produção de cabras leiteiras no semiárido: análise produtiva e reprodutiva, bem como influência da adoção de práticas higiênicas na qualidade do leite

The aim of this study was to evaluate the productive and reproductive performance of dairy goat genotypes, as well as the influence of hygienic practices by farmers on the quality of goat milk. Productive and reproductive data of 81 Anglo Nubian and Alpine crossbred goats, as well as data on total milk production of goats over 100 days of lactation and partial production up to 56 days of lactation. Also, in natura milk samples from 160 properties in the region were evaluated. At the time of collection, a questionnaire was applied to diagnose goat milk production systems. Data from productive and reproductive performance were evaluated by Tukey test and descriptive analysis using the SPSS program. The herd presented a variation of the total milk production in lactation (TMP) from 267.40kg to 468.55kg, with lactation length ranging from 157 to 247 days, and average daily production between 1.43 and 1.89kg/day. Fertility rates were satisfactory, with the lowest rate being 76% and the highest 92%, with an average of 85.24% considering the six seasons of birth. The means of the gestation periods varied between 144 and 152 days. Regarding the sanitary characterization, 73% of farmers performed a cleaning of the room before and after milking. However, 94.8% of farmers did not eliminate the first jets of milk and only 29.2% used the screened mug test to identify clinical mastitis. Only 41% of farmers performed pre and post-dipping and 30.2% applied the iodine solution. Only 8.3% of farmers used disposable paper towels. However, 92% of producers still used fabric towel. It was also observed that 99% of the properties stored milk in buckets or cans without refrigeration. In the microbiological analysis, a small amount of milk samples (5.6%) was contaminated with Staphylococcus aureus, however the total coliform count was high. Regarding the somatic cell count, it was found that 86% of the properties presented values above one million cells per mL of sample. The study demonstrated the prevalence of several factors that contribute to the vulnerability of milk contamination in various stages of production such as milking and processing. Thus, the guidance and awareness of those responsible is extremely important to improve goat milk quality in the semi-arid region of Paraíba.(AU)

O objetivo do presente estudo foi avaliar o desempenho produtivo e reprodutivo de genótipos caprinos leiteiros, assim como a influência da adoção das práticas higiênicas pelos produtores sobre a qualidade do leite caprino. A primeira parte da pesquisa foi realizada na Estação Experimental Pendência-PB. Foram analisados dados produtivos e reprodutivos de 81 cabras Anglo-Nubiana e alpinas mestiças, bem como dados de produção total de leite das cabras acima de 100 dias de lactação e produção parcial até 56 dias de lactação. Também foram avaliadas amostras de leite in natura de 160 propriedades da região. No momento da coleta, foi aplicado um questionário de diagnóstico dos sistemas de produção de leite caprino. Os dados de desempenho produtivo e reprodutivo foram avaliados pelo teste de Tukey e pela análise descritiva utilizando o programa SPSS. O rebanho apresentou uma variação da produção de leite total na lactação desde 267,40kg a 468,55kg, com durações de lactação variando de 157 e 247 dias, e produção média diária entre 1,43 e 1,89kg/dia. As taxas de fertilidade foram satisfatórias, sendo a menor taxa de 76% e a maior de 92%, com uma média de 85,24% considerando as seis estações de parição. As médias dos períodos de gestação variaram entre 144 e 152 dias. Quanto à caracterização da sanidade, 73% dos produtores realizam limpeza da sala antes e após a ordenha. No entanto, 94,8% dos produtores não eliminam os primeiros jatos de leite e somente 29,2% utilizam o teste da caneca telada para identificação de mastite clínica. Apenas 41% dos produtores realizam pré-dipping e pós-dipping e 30,2% aplicam a solução de iodo. Somente 8,3% dos produtores usam toalhas descartáveis e 92% ainda utilizam toalha de tecido. Observou-se, ainda, que 99% das propriedades armazenam o leite ordenhado em baldes ou latões, sem refrigeração. Pequenas quantidades de amostras (5,6%) estavam contaminadas por Staphylococcus aureus, porém a contagem de coliformes totais teve valor elevado. Em relação à contagem de células somáticas, verificou-se que 86% das propriedades apresentaram CCS acima de um milhão de células por mL, devendo-se ajustar corretamente os manejos alimentar, produtivo e reprodutivo. As épocas de parição, lactação e o genótipo influenciaram de forma direta os índices produtivos e reprodutivos dos animais. Obsevou-se a prevalência de vários fatores que contribuem para a vulnerabilidade de contaminação do leite em diversas etapas de produção, tais como ordenha e processamento. Portanto, a orientação adequada e a conscientização dos responsáveis são de extrema importância para melhorar a qualidade do leite de cabra na região semiárida da Paraíba.(AU)

Evaluation of physic contamiants and contamination with coliforms, molds and yeasts of honey from the Northern Brazil / Avaliação de contaminantes físicos e contaminação com coliformes, bolores e leveduras de mel do Norte do Brasil

The Mercosur Technical Regulation for honey determines microbiological parameters for total coliforms and molds and/ yeasts, and absence of physical contaminants, of any nature. The purpose of this study was to verify the biological and physical contamin of Apis mellifera ligustica and Melipona fasciculata, commercialized in the northeastern region of the State of Pará, Brazil. The fourteen samples from informal trade of the municipalities of Bragança, Capanema, Nova Timboteua, São João de Pirabas, Salinas and Tracauateua were analyzed for the Most Probable Number of colifroms at 35ºC, Fungus and Yeast counts, besides the detection of dirt by Macro and Microscopy techniques. The results obtained from the microbiological analyzes were compared to the standard required by MERCOSUR, for coliforms. 57.14% of the samples were non-standard and ranged from <3.0 NMP/g to 20 NMP/g. The molds and/ yeast counts ranged from 1x10¹ CFU/g to 8.6x10² CFU/g, with atotal of 21.42% of the samples being in disagreement with the legislation. From the macroscopic analysis, it was observed that, although 50% of the samples did not present any apparent dirt, microscopically it was possible to observe dirt in 100% of the samples of honey analyzed, which is out of the standard required by the Brazilian legislation. This shows the low sanitary quality of honey commercialized in this region of the state of Pará.

O Regulamento Técnico do Mercosul para mel, determina parâmetros microbiológicos para coliformes totais e fungos e/leveduras, e ausência de contaminantes físicos, de qualquer natureza. A proposta do presente estudo foi verificar a contaminação biológica e física em méis de abelha das espécies Apis mellifera ligustica e Melipona fasciculata, comercializados na região Nordeste do Estado do Pará, Brasil. As amostras provenientes do comercio informal dos de municípios de, Bragança, Capanema, Nova Timboteua, São João de Pirabas, Salinas e Tracauateua, foram analisadas quanto a determinação do Número Mais Provável de coliformes a 35ºC, Contagem de Fungos e/leveduras, além da detecção de sujidades pelas técnicas de Macro e Microscopia. Os resultados obtidos a partir das análises microbiológicas foram comparadas ao padrão exigido pelo MERCOSUL, para coliformes, 57,14% das amostras encontravam-se fora do padrão variaram de <3,0 NMP/g a 20NMP/g. Já a contagem de fungos e/leveduras variou de 1x10¹ UFC/g a 8,6x10² UFC/g, no total de 21,42 % das amostras apresentaram-se em desacordo com a legislação. A partir da análise macroscópica observou-se que, embora 50% das amostras não apresentassem sujidades aparentes, porém microscopicamente foi possível observar a presença sujidades em 100% das amostras de méis analisadas encontrando-se fora do padrão exigido pela legislação Brasileira. Demostrando a baixa qualidade sanitária do mel comercializado nessa região do estado do Pará.

Estimation of chronic dietary intake of pesticide residues / Estimativa de ingestão crônica de resíduos de agrotóxicos por meio da dieta

ABSTRACT OBJECTIVE To estimate the maximum theoretical daily intake of pesticides potentially consumed, chronically, by the Brazilian population. METHOD By using data from the food consumption section of the 2008-2009 Household Budget Survey to characterize the population diet, a database was built to group the foods based on the NOVA classification. Considering the maximum residue limit values of each pesticide authorized in the country until 2016, the limits of all consumed foods were added and multiplied by the amount consumed, resulting in the maximum theoretical intake index, which was compared with the acceptable daily intake. RESULTS The results show that, of the 283 pesticides considered in the database, 71 (25%) compounds had estimates of zero intake, 144 compounds (50.8%) reached acceptable daily intake values and 68 compounds (24%) showed median intake that exceeded the acceptable daily value. The pesticide intake estimation according to the different regions of the country showed a variation in the amount of compounds that exceeded the acceptable daily intake (48 to 69 substances) due to the different consumption patterns. The categories of products that most exceeded the limits were the insecticides, herbicides and fungicides. CONCLUSION The application of this methodology is valid for the first step in risk assessment, but the resulting values may be different from the actual exposure since they do not include other factors, such as the combined use of pesticides or unauthorized products. The importance of developing research on specific national food consumption data in a systematic way is emphasized, which generates data and analyses that allow a detailed risk assessment.

RESUMO OBJETIVO Estimar a ingestão diária máxima teórica dos agrotóxicos potencialmente consumidos, de forma crônica, pela população brasileira. MÉTODO Utilizando os dados do bloco de consumo alimentar da Pesquisa de Orçamentos Familiares de 2008-2009 para caracterização da dieta da população, foi construído um banco de dados agrupando os alimentos com base na classificação NOVA. Considerando os valores de limite máximo de resíduos de cada agrotóxico autorizado no país até o ano de 2016, foram somados os limites de todos os alimentos consumidos, multiplicados pela quantidade consumida, gerando o índice de ingestão teórica máxima, que foi comparado com a ingestão diária aceitável. RESULTADOS Os resultados mostram que dos 283 agrotóxicos considerados no banco de dados, 71 compostos tiveram estimativas de ingestão zero (25%), 144 compostos (50,8%) atingiram aos valores de ingestão diária aceitável e 68 compostos (24%) apresentaram mediana de ingestão que excedeu o valor diário aceitável. Quando realizada a estimativa de ingestão de agrotóxicos discriminando as distintas regiões do país, houve variação (entre 48 e 69 substâncias) na quantidade de compostos que excederam a ingestão diária aceitável devido aos diferentes padrões de consumo da população. As categorias dos produtos que mais excederam as estimativas são inseticidas, herbicidas e fungicidas. CONCLUSÃO A aplicação dessa metodologia é válida para o primeiro passo na avaliação de risco, porém os valores resultantes podem ser diferentes da exposição real por não englobar outros fatores, como o uso combinado de agrotóxicos ou de produtos de uso não autorizado. É ressaltada a importância do desenvolvimento de pesquisas de dados específicos de consumo de alimentos nacionais de forma sistemática, gerando dados e análises que viabilizem uma avaliação pormenorizada sobre riscos.

Desafios e avanços no controle de resíduos de agrotóxicos no Brasil: 15 anos do Programa de Análise de Resíduos de Agrotóxicos em Alimentos / Challenges and strides in the control of pesticide residues in Brazil: 15 years of the Program for Analysis of Pesticide Residues in Food Products / Desafíos y avances en el control de residuos de pesticidas en Brasil: 15 años del Programa de Análisis de Residuos de Pesticidas en Alimentos

Resumo: No Brasil, desde 2001, o monitoramento da presença de agrotóxicos nos alimentos ocorre por meio do Programa de Análise de Resíduos de Agrotóxicos em Alimentos (PARA). O objetivo foi analisar os desafios e avanços no controle de resíduos de agrotóxicos nos alimentos, no Brasil, com base nos resultados do PARA, entre os anos 2001 e 2015. Foi realizada pesquisa descritivo-analítica documental, que utilizou dados provenientes dos relatórios do PARA, disponíveis no sítio eletrônico da Agência Nacional de Vigilância Sanitária. Entre os principais desafios apresentados pelo PARA estão a grande variabilidade no número de amostras e ingredientes ativos analisados, bem como no formato dos relatórios apresentados. Existe a necessidade do aprofundamento das discussões a respeito dos critérios usados para a classificação e autorização dos ingredientes ativos no Brasil, bem como da transparência destas informações para a sociedade. No total, foram analisados 25 tipos de alimentos e mais de 30 mil amostras ao longo da vigência do PARA, porém, o máximo de ingredientes ativos analisados foi 228. A presença de ingredientes ativos não autorizados para a cultura e não autorizados no país foi a principal causa de irregularidades diagnosticadas nos alimentos. Embora ainda existam diversos desafios apresentados no PARA, ele contribui para disparar a reflexão sobre o tema junto à sociedade. Haja vista a atual conjuntura do governo, de maior flexibilização na liberação de ingredientes ativos no país, é necessário que o PARA seja fortalecido.

Abstract: In Brazil, since 2001, pesticides in food products have been monitored by the Program for Analysis of Pesticide Residues in Food Products (PARA, in Portuguese). The study aimed to analyze the challenges and strides in the control of pesticide residues in food products in Brazil based on the program's results from 2001 to 2015. A descriptive-analytical document review was conducted that used data from reports by the PARA, available at the website of the Brazilian Health Regulatory Agency. The program's main challenges feature the wide variation in the number of samples and active ingredients analyzed, as well as the reports' format. There is a need for more in-depth discussions of the criteria used for classification and authorization of active ingredients in Brazil, as well as the information's transparency to society. A total of 25 types of foods were analyzed, with more than 30,000 samples, since the PARA has been in force, but the maximum number of active ingredients analyzed was 228. The presence of active ingredients unauthorized for crops and unauthorized in the country was the principal cause of irregularities diagnosed in the foods. Although the program still faces various challenges, it helps spawn debate on the issue in society. Given the current government context of greater flexibility in the authorization of active ingredients in Brazil, it is necessary to strengthen the PARA.

Resumen: En Brasil, desde 2001, la supervisión de la presencia de pesticidas en los alimentos se produce mediante el Programa de Análisis de Residuos de Pesticidas en Alimentos (PARA). El objetivo fue analizar los desafíos y avances en el control de residuos de pesticidas en los alimentos, en Brasil, a partir de los resultados del PARA, entre los años 2001 y 2015. Investigación descriptivo-analítica documental, que utilizó datos provenientes de los informes del PARA, disponibles en la página web de la Agencia Nacional de Vigilancia Sanitaria. Entre los principales desafíos presentados por el PARA, están la gran variabilidad en el número de muestras e ingredientes activos analizados, así como en el formato de los informes presentados. Existe la necesidad de profundizar en las discusiones, respecto a los criterios utilizados para la clasificación y autorización de los ingredientes activos en Brasil, así como la transparencia de esta información para la sociedad. En total, se analizaron 25 tipos de alimentos y más de 30 mil muestras, a lo largo de la vigencia del PARA, no obstante, el máximo de ingredientes activos analizados fue 228. La presencia de ingredientes activos no autorizados para el cultivo y no autorizados en el país fue la principal causa de irregularidades diagnosticadas en los alimentos. A pesar de que todavía existan diversos desafíos presentados en el PARA, contribuye a activar la reflexión sobre este tema junto a la sociedad. Ante la actual coyuntura de gobierno, con una mayor flexibilización en la autorización de ingredientes activos en el país, es necesario que el PARA sea fortalecido.

Hygienic-sanitary quality and antimicrobial sensitivity profile of Escherichia coli in milk and cheese sold illegally in municipalities of northern Mato Grosso, Brazil

Products such as milk and cheese produced by hand and sold by small producers in open markets and at home are a reality in Brazil, despite legal prohibitions. In many cases, this leads to the production of food without hygienic conditions, which may constitute an important source of transmission of foodborne diseases and a danger to public health. This study proposes to examine the hygienic-sanitary quality of milk and cheese sold illegally in municipalities of northern Mato Grosso, Brazil, to undertake a phenotypical investigation of the presence of resistance of isolated colonies to antimicrobials and to detect the production of ß-lactamase enzymes: extended-spectrum ß-lactamase (ESBL), AmpC ß-lactamases (AmpC) and carbapenemases. The 25 milk and 37 cheese samples analyzed were subjected to the most probable number (MPN) test, isolation on eosin-methylene blue agar (EMB) agar and Escherichia coli identification by biochemical tests and disk diffusion test. Results showed that 76% of the milk samples and 67.57% of the cheese samples had thermotolerant coliform counts above the value allowed by the legislation. The milk and cheese isolates showed 15.79 and 5.88% resistance, respectively, to at least one of the tested antimicrobials. No ß-lactamase enzyme production was observed in the isolates.