[Microbiological and parasitological risk analysis of novel food sources obtained with the use of insects].

The problem of food safety being an important component of the country's food security, provides not only for continuous improvement of the methodology of hygienic standardization, but also for the formation of requirements for novel food, in particular, those obtained from non-traditional sources. The accumulated practical and theoretical competence in the food hygiene area, as well as knowledge of current trends of the food base broadening, allow us to analyze the risks associated with novel food obtained of insects. The purpose of the research was to analyze the microbiological and parasitological risk of novel food sources obtained with the use of insects, suggest the effective risk management measures. Material and methods. The analytical part of the work included literature search, collection of information and statistical materials published in domestic and foreign scientific editions. The search was carried out using the Google Academy retrieval system and electronic databases (PubMed, MEDLINE, EMBASE, Scopus, Web of Science, eLIBRARY), mainly in the last 25 years, using the keywords: Hermetia illucens, Tenebrio molitor, Acheta domesticus, insects, parasite, nematode, pathogen, cysts. Results. Based on the published materials' analysis, a systematization of microbiological and parasitological factors potentially capable of colonizing edible insects has been carried out. There were identified representatives of 24 groups of pathogenic and 18 opportunistic microorganisms and helminths related to microbial and parasitic pathogens, the spread of which is significantly influenced by inappropriate conditions of feeding and keeping insects. As there are currently no veterinary requirements for insect breeding and farming conditions, contamination of end products with infectious and parasitic pathogens can vary over a very wide range. Conclusion. The use of native insect biomass carries certain risks associated with its microbial and parasitic contamination, and the development of measures to prevent them requires significant resources. The possibility of deep processing of such raw materials can be considered as one of the solutions to mitigate these risks. For use in the food industry, insects should be subjected to processing similar to that currently used for soybean seeds, which includes separation of protein (entomoprotein), fat and chitin fractions, each of which would have an independent use. Thus, at present, insects should be considered as a source of novel food ingredients, first of all, complete protein of animal origin.

Prevalence and isolation of Toxoplasma gondii in goats slaughtered for human consumption in the semi-arid of northeastern Brazil.

The aim of this study was to determine the seroprevalence of anti-Toxoplasma gondii antibodies and factors associated with infection in goats, and to isolate protozoan strains in tissue samples from seropositive goats that were destined for human consumption in the state of Paraíba, northeastern Brazil. Serum samples from 229 slaughtered goats were tested using the indirect fluorescence antibody test (IFAT), with a cutoff point of 1:64. Epidemiological questionnaires were applied to the producers, to acquire information about the sanitary management used in their herds. Tissue samples from the animals were collected during slaughter, in order to perform bioassays in mice. The seroprevalence found was 21.39% (49/229), with antibody titers ranging from 1:64 to 1:32,768. The municipalities of origin, Patos (OR: 3.047; CI: 1.384-6.706) and Sousa (OR: 3.355; CI: 1.536-7.327), were considered to be factors associated with infection by T. gondii. Thirty-eight bioassays were performed in mice, using tissues from seropositive goats, with an isolation rate of 50% (19/38). There was no correlation between isolation rate and antibody titers. Only one mouse died, at 30 days post-infection, which demonstrated that the strains isolated had low virulence towards mice. It was concluded that there is high seroprevalence in goats in northeastern Brazil, as well as a high percentage of viable tissue cysts in slaughtered animals destined for human consumption. These results demonstrate that there is an imminent one health problem relating to toxoplasmosis, especially in the most populous municipalities in the study (Patos and Sousa), which were identified as factors associated with T. gondii infection in goats.

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

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.

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.

Prospect of microbial food borne diseases in Pakistan: a review / Perspectiva de doenças microbianas transmitidas por alimentos no Paquistão: uma revisão

Abstract Nowadays food borne illness is most common in people due to their epidemic nature. These diseases affect the human digestive system through bacteria, viruses and parasites. The agents of illness are transmitted in our body through various types of food items, water and uncooked. Pathogens show drastic changes in immunosuppressant people. This review gives general insights to harmful microbial life. Pakistan is a developed country and because of its improper food management, a lot of gastrointestinal problems are noted in many patients. Bacteria are most common agents to spread diarrhoea, villi infection, constipation and dysenteric disease in human and induce the rejection of organ transplant. Enhancement of their lifestyle, properly cooked food should be used and to overcome the outbreak of the diseases.

Resumo Hoje em dia, as doenças transmitidas por alimentos são mais comuns em pessoas devido à sua natureza epidêmica. Essas doenças afetam o sistema digestivo humano por meio de bactérias, vírus e parasitas. Os agentes das doenças são transmitidos em nosso corpo por meio de diversos tipos de alimentos, água e crus. Os patógenos mostram mudanças drásticas em pessoas imunossupressoras. Esta revisão fornece uma visão geral da vida microbiana prejudicial. O Paquistão é um país desenvolvido e, devido ao seu manejo alimentar inadequado, muitos problemas gastrointestinais são observados em muitos pacientes. As bactérias são os agentes mais comuns para espalhar diarreia, infecção de vilosidades, obstipação e doença disentérica em humanos e induzem a rejeição de transplantes de órgãos. Melhoria de seu estilo de vida, alimentos devidamente cozidos devem ser utilizados e para superar o aparecimento de doenças.

Quantification of viable protozoan parasites on leafy greens using molecular methods.

Protozoan contamination in produce is of growing importance due to their capacity to cause illnesses in consumers of fresh leafy greens. Viability assays are essential to accurately estimate health risk caused by viable parasites that contaminate food. We evaluated the efficacy of reverse transcription quantitative PCR (RT-qPCR), propidium monoazide coupled with (q)PCR, and viability staining using propidium iodide through systematic laboratory spiking experiments for selective detection of viable Cryptosporidium parvum, Giardia enterica, and Toxoplasma gondii. In the presence of only viable protozoa, the RT-qPCR assays could accurately detect two to nine (oo)cysts/g spinach (in 10 g processed). When different proportions of viable and inactivated parasite were spiked, mRNA concentrations correlated with increasing proportions of viable (oo)cysts, although low levels of false-positive mRNA signals were detectable in the presence of high amounts of inactivated protozoa. Our study demonstrated that among the methods tested, RT-qPCR performed more effectively to discriminate viable from inactivated C. parvum, G. enterica and T. gondii on spinach. This application of viability methods on leafy greens can be adopted by the produce industry and regulatory agencies charged with protection of human public health to screen leafy greens for the presence of viable protozoan pathogen contamination.

Efficacy of different hermetic bag storage technologies against insect pests and aflatoxin incidence in stored maize grain.

The performance of six grain storage technologies for the control of insect pests in maize was evaluated over a 36-week (9-month) storage period. The six technologies used were: two ZeroFly® hermetic bag brands (laminated and non-laminated); Purdue Improved Crop Storage (PICS) bag; non-hermetic ZeroFly® bag; woven polypropylene (PP) bag containing maize grain treated with Actellic Gold® Dust (pirimiphos-methyl 1.6% + thiamethoxam 0.3%) and woven PP bag containing untreated grain. Each bag was filled with 50 kg maize grain and four replicates of each were set up. With the exception of the non-hermetic ZeroFly® bag, 50 live adults of the larger grain borer Prostephanus truncatus and of the maize weevil Sitophilus zeamais, were introduced into all the bags. Insects were not introduced into the non-hermetic ZeroFly® bag to assess its effectiveness in repelling infestation from outside. Parameters recorded were gas composition (oxygen and carbon dioxide) levels inside the bags; weight of flour generated by insect feeding activities; grain moisture level; live adult insect counts; grain damage and weight loss; grain germination rate and aflatoxin level. At termination, the plastic liners of the hermetic bags were examined for perforations. Results show that oxygen depletion and carbon dioxide evolution were faster in ZeroFly® hermetic compared to PICS bags. Throughout the 36-week storage trial, grain damage remained below 4% and weight loss below 3% in all the treatments except in the untreated PP bags in which it increased to 81.1 and 25.5%, respectively. The hermetic PICS, ZeroFly® and Actellic Gold dust-treated PP bags maintained grain germination at 60%, which was lower than the initial 90%, while in untreated control, it reduced to 4.7%. The mean aflatoxin levels fluctuated between 0.39 and 3.56 parts per billion (ppb) during 24 weeks of storage in all the technologies tested, which is below the acceptable maximum level of 10 ppb in maize. Based on the evaluation results, it can be concluded that hermetic PICS and ZeroFly® bags and woven PP bag with Actellic Gold dust-treated grain effectively protected stored maize grain from insect attack and weight losses. Appropriate strategies and mechanisms for the effective and efficient adoption of hermetic storage bag technology at scale would contribute towards global food security.

The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats.

The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Foodborne transmission of Toxoplasma gondii infection in the last decade. An overview.

Toxoplasmosis is a zoonotic disease of global distribution and importance. It is caused by the protozoan parasite Toxoplasma gondii, the only species in the Toxoplasma genus. This parasite can infect most warm-blooded animals, including humans and livestock. Main routes of transmission are by ingestion of tissue cysts in raw or undercooked meat of infected animals, ingestion of raw vegetables or water contaminated with T. gondii oocysts from cat feces, and transplacental. Around one-third of human beings are chronically infected with T. gondii. Most infections appear to be asymptomatic in immunocompetent persons, but toxoplasmosis can be fatal to the fetus and immunocompromised adults. Water and foodborne outbreaks have been caused by this parasite worldwide, but few are well documented. Importantly, T. gondii is a parasite of high importance in animal health, causing reproductive failure, particularly in small ruminants, and clinical toxoplasmosis in many species. This overview discusses the knowledge of T. gondii infections in the last decade focusing on the foodborne transmission of this parasite.

The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats / O óleo de alho, Allium sativum L. (Amaryllidaceae), como potencial protetor contra Anisakis spp. Infecção tipo II (L3) (Nematoda) em ratos Wistar

Abstract The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Resumo O consumo de peixe inadequadamente tratado termicamente representa um risco para a saúde pública, com a possibilidade da propagação de larvas de Anisakis. O presente estudo demonstrou as alterações fisiológicas e histopatológicas acompanhadas de inoculação oral de extractos brutos de Anisakis tipo II (L3) frescos e termicamente tratados em ratos. Os vermes foram isolados de um peixe marinho, examinados e identificados por microscopia de luz e eletrônica de varredura. O estudo foi conduzido em 6 grupos de ratos: controle (I), óleo de alho (GO) inoculado (II), L3 fresco inoculado (III), L3 tratado termicamente inoculado (IV), L3 fresco + GO inoculado (V), e um grupo L3 + GO tratado termicamente inoculado (VI). Observou-se que ratos inoculados com L3 fresco e tratados termicamente mostraram funções hepáticas e renais anormais, associadas à destruição da sua arquitetura normal. GO produziu um efeito protector em grupos de ratos inoculados com extractos L3 + GO através da melhoria das funções do fígado e dos rins, o que foi confirmado pela estrutura normal marcada da sua histologia. A cozedura de peixes infectados com L3 induziu alterações mais graves do que os peixes não cozidos. Recomenda-se a administração de alho antes e depois do consumo de peixe, para evitar o efeito perigoso dos anisakids, mesmo que sejam cozidos.

Fatal Aberrant Parasite Migration of Echinuria uncinata in Two African Pygmy Geese (Nettapus auritus).

Two adult African pygmy geese (Nettapus auritus) were found dead with minimal clinical disease signs. Necropsy revealed aberrant migration of the nematode parasite Echinuria uncinata, as confirmed morphologically and through DNA sequencing. This common waterfowl parasite typically lives in the proventriculus, burying headfirst into the mucosa and laying eggs into the gastrointestinal lumen. In these geese, the parasites tunneled through the gastrointestinal tract wall to invade the coelomic cavity; from which, a substantial quantity of eggs found their way into the coelomic space and into the air sacs and lungs. This potential parasite migration should be monitored for in Anseriformes species that present with similar disease conditions, and the use of Daphnia species, the intermediate host, as a waterfowl feed source is not recommended.

Reduction of Reinfection Rates with Opisthorchis viverrini through a Three-Year Management Program in Endemic Areas of Northeastern Thailand.

To clarify the reinfection profile associated with risk factors of opisthorchiasis, we conducted an epidemiological study on the chemotherapeutic effects on reinfection with O. viverrini in the endemic areas of Northeastern Thailand for 3 years. A total of 3,674 fecal samples were collected from participants in villages of 5 provinces. They were examined microscopically using a modified technique of formalin ethyl-acetate concentration. Egg-positive residents were reexamined year (2018) by year (2019) after treatment with a single dose (40 mg/kg) of praziquantel. Health education was provided to the participants yearly. The egg-positive rate of O. viverrini was 14.3%, and was highest (22.2%) in the 20-30 year-old group in 2017. The egg positive rate was 15.3% in dogs and 11.4% cats. Human reinfection rate was 15.5% and 6.3% in next 2 years, and was highest (23.2%) among the fishermen. Relative risk factors of reinfection were significantly higher for males, over 40-year-old age, or working as fishermen or farmers, and eating uncooked fish within the preceding year. A significant difference resulting from a health education program was observed in the third year. Therefore, health education and sustainable surveillance for opisthorchiasis should be maintained to decrease the risk of reinfection.

Toxoplasma gondii seroprevalence in beef cattle raised in Italy: a multicenter study.

Toxoplasmosis represents an important public health issue, with the consumption of raw or undercooked meat being a major way of human infection. The role of beef in the transmission of the parasite to humans is questioned due to lower quantity of tissue cysts compared with other meat-producing species. However, the habit of consuming raw beef is regionally diffused, and the risk posed by Toxoplasma gondii infection in cattle should not be overlooked. Therefore, to update information on T. gondii in cattle reared in Italy, a multicentric seroepidemiological survey was designed and implemented in four Northern regions (Liguria, Lombardy, Piedmont, and Trentino Alto Adige) and Sardinia. Overall, a convenience sampling was performed, collecting 1444 serum samples from 57 beef cattle herds. Thirteen beef breeds were sampled, besides cross-breed; bovines age varied from 3 months to over 12 years. Sera were tested with a commercial ELISA for the detection of anti-T. gondii antibodies. Individual and herd data were analyzed by binary logistic regression analysis. A T. gondii seroprevalence of 10.2% was recorded, with differences among regions and values ranging from 5.3% in Liguria to 18.6% in the Piedmont region (p value = 0.0001). Both young and adult animals and males and females tested positive, without any significant difference (age and gender: p value > 0.05). Lower seroprevalence values were recorded in cattle born in Italy (8.7%) if compared with animals imported from abroad (13.4%) (p value = 0.046). The spread of T. gondii in beef cattle destined to Italian consumers is confirmed, suggesting the need of continuous monitoring of the infection.

Anisakiasis and Anisakis: An underdiagnosed emerging disease and its main etiological agents.

Anisakiasis or anisakiosis is a human parasitic infection caused by the third-stage larvae (L3) of nematodes of the genus Anisakis, although the term is also used in medical literature for the much less frequent (<3% of cases) infection by L3 of other genera of anisakids, particularly Pseudoterranova. These parasites have a marine lifecycle. Humans are infected by the L3 through ingesting of fish and squid, the intermediate/paratenic hosts. The live larvae generally penetrate the wall of the stomach or intestine causing, among other symptoms, intense pain or allergic symptoms. These are emerging, cosmopolite illnesses. Diagnosis and treatment is usually by endoscopy and extraction and identification of the larvae. Allergic forms are usually diagnosed by prick-test and/or allergen-specific IgE detection and treated with a suitable anti-allergy treatment. The patient is also warned against further consumption of marine fish or squid, as these may be infected with Anisakis. The most common method of prevention is thermal treatment of the entire fish or squid prior to consumption (>60 °C, >1 min or - 20 °C, >24 h). Useful measures for the control of anisakiasis would be to establish a national register of cases, to initiate educational campaigns for the general public and consciousness-raising and training campaigns for health professionals. These would be complemented by control measures for the relevant sectors of the economy: fish operators, fish farming, fishermen, fishmongers, fish industry and catering facilities. Possible genetic predisposition for allergy to Anisakis and the possible relationship between anisakiasis and cancer would also require further investigation.

The socio-economic burden of cystic echinococcosis in Morocco: A combination of estimation method.

Cystic echinococcosis (CE) is a major zoonosis in Morocco despite the launch of a national control programme in 2005. As its economic consequences have not been studied yet in Morocco, this study estimated CE impact in terms of monetary losses, disability-adjusted life years (DALY), and DALY for zoonotic diseases (zDALY) in the entire country and in specific regions for the 2011 to 2014 period. The direct monetary losses were related to organ seizure from infected animal in slaughterhouses, and to healthcare expenses as well as lost wages for infected humans. Animal production losses concerned milk yield, fertility, carcass weight, and wool production. Losses due to human infection were also composed of disability and productivity losses at work. Monte Carlo simulations were used to estimate monetary losses and zDALY values. Nationwide, the estimated DALY was 0.5 years per 100,000 persons per year, and the zDALY was 55 years per 100,000 persons per year. Total yearly losses were estimated at 73 million USD (54-92 million USD). However, losses differed significantly among regions. Most of the economic losses consisted of unperceived consequences, i.e. decreased animal production and reduced productivity of asymptomatic individuals. Future studies should determine the socioeconomic and epidemiological factors underlying the differences in economic losses among regions to develop better adapted control programmes.

Detection of human intestinal protozoan parasites in vegetables and fruits: a review.

Diarrheal diseases caused by intestinal protozoan parasites are a major food-borne public health problem across the world. Vegetables and fruits provide important nutrients and minerals, but are also common sources of some food-borne human pathogenic microorganisms. The contamination of raw vegetables and fruits with human pathogenic parasites are now a global public health threat, despite the health benefits of these foods in non-pharmacological prophylaxes against diseases. A large number of reports have documented the contamination of vegetables or fruits with human pathogenic microorganisms. In this paper, we reviewed the contamination and detection methods of human pathogenic intestinal protozoans that are frequently recovered from raw vegetables and fruits. The protozoan parasites include Cryptosporidium spp., Giardia duodenalis, Cyclospora cayetanensis, Entamoeba spp., Toxoplasma gondii, Balantioides coli, Blastocystis sp., Cystoisospora belli and Enterocytozoon bieneusi. The risk factors involved in the contamination of vegetables and fruits with parasites are also assessed.

Risk factors and prevalence of enteroparasitic diseases in shellfish pickers from a lake area in the Northeast of Brazil / Fatores de risco e prevalência de enteroparasitoses em marisqueiras de uma região lagunar no Nordeste do Brasil

INTRODUCTION: Intestinal parasitosis are a public health problem worldwide. There are several risk factors and a high association with some specific labor activities. OBJECTIVE: The present study assessed the risk factors and prevalence of enteroparasitic diseases in shellfish pickers from one district of Maceió, Alagoas state, Brazil. METHODS: Crosssectional study of 41 female shellfish pickers including parasitological tests in fecal samples and a questionnaire with objective and subjective questions. Sand samples from their working environment were also analyzed. RESULTS: At least one species of parasite was found in 19.51% of the fecal samples. Pathogenic species of Giardia lamblia, Trichuris trichiura, Schistosoma mansoni, Ascaris lumbricoides, Enterobius vermicularis, from the Ancylostomatidae family, and non-pathogenic species of Entamoeba coli were found. Polyparasitism was diagnosed in 37.5% of the positive samples. A total of 57.14% of sand samples contained hookworm larvae. Regarding the risk factor, low educational level was statistically associated to the presence of parasites (p<0.05). CONCLUSION: Greater investment in basic education is needed to increase the knowledge about preventive measures against parasitic diseases and the promotion food-handling courses in order to change existing inadequate habits in the community. Basic sanitation is also essential in preventing environmental contamination.

INTRODUÇÃO: As parasitoses intestinais representam um problema de saúde pública mundial. Estão associados inúmeros fatores de risco, bem como atividades laborais específicas. OBJETIVO: O presente estudo avaliou os fatores de risco e a prevalência de enteroparasitoses em marisqueiras de um bairro da cidade de Maceió, Alagoas, Brasil. MÉTODOS: Procedeu-se um estudo de corte transversal, sendo 41 marisqueiras avaliadas mediante exame parasitológico de fezes e questionário contendo questões discursivas e de múltipla escolha. Foram analisadas também amostras de areia do ambiente de trabalho das mesmas. RESULTADOS: A positividade para pelo menos uma espécie de parasito nas fezes foi de 19,51%. Foram encontradas as espécies patogênicas Giardia lamblia, Trichuris trichiura, Schistosoma mansoni, Ascaris lumbricoides, Enterobius vermicularis e da família Ancylostomatidae, e a espécie não patogênica, Entamoeba coli. Poliparasitismo foi diagnosticado em 37,5% dos exames positivos. Um total de 57,14% das amostras de areia continha larvas de ancilostomatídeos. Em relação aos fatores de risco, a baixa escolaridade foi estatisticamente associada à presença de parasitas (p<0,05). CONCLUSÃO: É necessário maior investimento na educação básica para aprimorar o conhecimento das formas de prevenção das parasitoses e a promoção de cursos de boas práticas de manipulação de alimentos, a fim de modificar hábitos errôneos já incorporados na comunidade. O saneamento básico também é fundamental para evitar contaminação ambiental.

Methods to assess the effect of meat processing on viability of Toxoplasma gondii: towards replacement of mouse bioassay by in vitro testing.

Consumption of meat containing viable tissue cysts is considered one of the main sources of human infection with Toxoplasma gondii. In contrast to fresh meat, raw meat products usually undergo processing, including salting and mixing with other additives such as sodium acetate and sodium lactate, which affects the viability of T. gondii. However, the experiments described in the literature are not always performed in line with the current processing methods applied in industry. It was our goal to study the effect of salting and additives according to the recipes used by industrial producers. Mouse or cat bioassay is the 'gold standard' to demonstrate the presence of viable T. gondii. However, it is costly, time consuming and for ethical reasons not preferred for large-scale studies.Therefore, we first aimed to develop an alternative for mouse bioassay that can be used to determine the effect of processing on the viability of T. gondii tissue cysts. The assays studied were (i) a cell culture method to determine the parasite's ability to multiply, and (ii) a propidium monoazide (PMA) dye-based assay to selectively detect DNA from intact parasites. Processing experiments were performed with minced meat incubated for 20 h with low concentrations of NaCl, sodium lactate and sodium acetate. NaCl appeared to be the most effective ingredient with only one or two out of eight mice infected after inoculation with pepsin-digest of portions processed with 1.0, 1.2 and 1.6% NaCl. Results of preliminary experiments with the PMA-based method were inconsistent and did not sufficiently discriminate between live and dead parasites. In contrast, the cell culture method showed promising results, but further optimization is needed before it can replace or reduce the number of mouse bioassays needed. In future, standardised in vitro methods are necessary to allow more extensive testing of product-specific processing methods, thereby providing a better indication of the risk of T. gondii infection for consumers.

Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits.

BACKGROUND: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts. METHODS: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light. RESULTS: Forty percent of the analysed samples (95% CI: 25.5-56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6-179.9 oocysts/g. CONCLUSIONS: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts.

Consumer knowledge and practices to pork safety in two Taenia solium cysticercosis endemic districts in Eastern Cape Province of South Africa.

BACKGROUND: Globally, Taenia solium can cause cysticercosis in humans (including neurocysticercosis) and in pigs through ingestion of eggs and taeniasis in humans through ingestion of raw/undercooked pork contaminated with mature cysts. It is now recognised globally as one of the most prevalent food-borne parasitic diseases. The majority of cases have been reported in developing countries where consumption of food produced under unhygienic conditions is prevalent, exacerbated by lack of food safety education. The aim of this study was to determine the knowledge and practices of consumers towards pork safety in two districts of the Eastern Cape Province of South Africa, where T. solium cysticercosis is endemic in pigs and humans. METHODS: Three-hundred-and-sixty-one (361) participants were conveniently interviewed on consumer knowledge (harmfulness of T. solium cysticercosis, ability to identify cysts, trustworthiness of registered butcheries and legal requirements) and practices (storage of pork and method(s) of cooking pork safely) through a structured questionnaire. Chi-square for association of variables was used to compare differences in the districts. RESULTS: Overall, 73.1% of the study group from both districts agreed that pork forms an important part of their diet. Consumers (54.2%: 189/349) agreed that pork infected with T. solium cysts could be harmful, and 57.3% (188/328) indicated their inability to identify T. solium cysts in pork when slaughtered at home. Although 69.5% (234/352) trusted pork bought from butcheries, only 52.2% (187/358) were aware that butcheries must present a registration certificate in order to operate. This coincides with the fact that very few (< 10%) were aware of the legal requirements in terms of disease control, slaughter and food preparation. Most consumers (88.7%: 268/302) kept pork in the fridge and only 11.3% (34/302) kept it in a freezer (p = 0.02). Although not significantly different between the districts (p = 0.15), consumers in Alfred Nzo (71.4%: 152/213) and OR Tambo (61.2%: 74/12) mostly cooked pork as a stew, followed by braai/barbeque and frying or baking. This was in line with the fact that consumers in Alfred Nzo (79%: 147/186) and OR Tambo (80.8%: 120) preferred well-cooked pork; the main reason for this was the belief that cooking kills germs (43.6%: 121/277) followed by rendering the meat tasty (26.4%: 73/277). CONCLUSIONS: Consumers surveyed in the two districts were somewhat aware that T. solium cysticercosis could be harmful, although some were not able to identify T. solium cysts in pork. They also lacked sufficient knowledge regarding butchery certification and other legal requirements related to disease control, slaughter and food preparation. Practices related to cooking have the potential to promote the transmission of human taeniasis and the fact that most respondents preferred stewed pork could be a positive sign, as the cysts are destroyed during the cooking process. Results from this study are useful for the development of a control and prevention strategy targeted towards consumers, and the creation of awareness of food safety, with special emphasis on T, solium cysticercosis.