{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T13:38:44Z","timestamp":1762522724859,"version":"build-2065373602"},"reference-count":70,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,15]],"date-time":"2022-07-15T00:00:00Z","timestamp":1657843200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"Horizon 2020 project SafeConsume","doi-asserted-by":"publisher","award":["727580"],"award-info":[{"award-number":["727580"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Applied Sciences"],"abstract":"<jats:p>The monitoring of the microbial quality of fresh products in the industrial environment has mainly focused on bacterial indicators. Protozoa, such as Giardia duodenalis, Cryptosporidium spp., Toxoplasma gondii, and Cyclospora cayetanensis, are routinely excluded from detection and surveillance systems, despite guidelines and regulations that support the need for tracking and monitoring these pathogens in fresh food products. Previous studies performed by our laboratory, within the scope of the SafeConsume project, clearly indicated that consumption of fresh produce may be a source of T. gondii, thus posing a risk for the contraction of toxoplasmosis for susceptible consumers. Therefore, preliminary work was performed in order to assess the microbiological quality of vegetables, highlighting not only bacteria (Escherichia. coli, Listeria monocytogenes, and Salmonella spp.), but also the zoonotic protozoa G. duodenalis and Cryptosporidium spp. Although all samples were found to be acceptable based on bacteriological parameters, cysts of G. duodenalis and oocysts of Cryptosporidium spp. were observed in vegetables. Moreover, it was possible to genetically characterize G. duodenalis positive samples as assemblage A, a genotype that poses risks to human health. Although these are preliminary results, they highlight the need to include protozoa in the microbiological criteria for foodstuffs, as required by EU Law No. 1441\/2007, and to improve inactivation and removal procedures of (oo)cysts in fresh produce and water.<\/jats:p>","DOI":"10.3390\/app12147145","type":"journal-article","created":{"date-parts":[[2022,7,17]],"date-time":"2022-07-17T21:00:28Z","timestamp":1658091628000},"page":"7145","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Protozoa as the \u201cUnderdogs\u201d for Microbiological Quality Evaluation of Fresh Vegetables"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1492-4506","authenticated-orcid":false,"given":"Cl\u00e1udia S.","family":"Marques","sequence":"first","affiliation":[{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for the Study in Animal Science (CECA), University of Porto, Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for Parasite Biology and Immunology, Department of Infectious Diseases, National Health Institute Dr. Ricardo Jorge (INSARJ), Rua de Alexandre Herculano, 321, 4000-055 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3891-3924","authenticated-orcid":false,"given":"Susana","family":"Sousa","sequence":"additional","affiliation":[{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for the Study in Animal Science (CECA), University of Porto, Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for Parasite Biology and Immunology, Department of Infectious Diseases, National Health Institute Dr. Ricardo Jorge (INSARJ), Rua de Alexandre Herculano, 321, 4000-055 Porto, Portugal"}]},{"given":"Ant\u00f3nio","family":"Castro","sequence":"additional","affiliation":[{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for the Study in Animal Science (CECA), University of Porto, Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for Parasite Biology and Immunology, Department of Infectious Diseases, National Health Institute Dr. Ricardo Jorge (INSARJ), Rua de Alexandre Herculano, 321, 4000-055 Porto, Portugal"}]},{"given":"V\u00e2nia","family":"Ferreira","sequence":"additional","affiliation":[{"name":"CBQF\u2014Centro de Biotecnologia e Qu\u00edmica Fina\u2014Laborat\u00f3rio Associado, Escola Superior de Biotecnologia, Universidade Cat\u00f3lica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6296-5137","authenticated-orcid":false,"given":"Paula","family":"Teixeira","sequence":"additional","affiliation":[{"name":"CBQF\u2014Centro de Biotecnologia e Qu\u00edmica Fina\u2014Laborat\u00f3rio Associado, Escola Superior de Biotecnologia, Universidade Cat\u00f3lica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, Portugal"}]},{"given":"Jos\u00e9 M. Correia","family":"da Costa","sequence":"additional","affiliation":[{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for the Study in Animal Science (CECA), University of Porto, Pra\u00e7a do Coronel Pacheco 42, 4050-083 Porto, Portugal"},{"name":"Centre for Parasite Biology and Immunology, Department of Infectious Diseases, National Health Institute Dr. Ricardo Jorge (INSARJ), Rua de Alexandre Herculano, 321, 4000-055 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,15]]},"reference":[{"key":"ref_1","unstructured":"(2020, February 25). WHO Fact Sheet N_237, Food Safety and Foodborne Illness. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/food-safety."},{"key":"ref_2","first-page":"464","article-title":"Diseases due to unhealthy environments: An updated estimate of the global burden of disease attributable to environmental determinants of health","volume":"39","author":"Wolf","year":"2016","journal-title":"J. Public Health"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.ijfoodmicro.2015.05.026","article-title":"Microbiological survey of raw and ready-to-eat leafy green vegetables marketed in Italy","volume":"210","author":"Losio","year":"2015","journal-title":"Int. J. Food Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Macieira, A., Barbosa, J., and Teixeira, P. (2021). Food Safety in Local Farming of Fruits and Vegetables. Int. J. Environ. Res. Public Health, 18.","DOI":"10.3390\/ijerph18189733"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Gajadhar, A.A. (2015). Transmission dynamics of foodborne parasites on fresh produce. Foodborne Parasites in the Food Supply Web, Woodhead Publishing Series in Food Science, Technology and Nutrition, Woodhead Publishing. Chapter 13.","DOI":"10.1016\/B978-1-78242-332-4.00001-1"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"McKerr, C., Adak, G.K., Nichols, G., Gorton, R., Chalmers, R.M., Kafatos, G., Cosford, P., Charlett, A., Reacher, M., and Pollock, K.G. (2015). An Outbreak of Cryptosporidium parvum across England & Scotland Associated with Consumption of Fresh Pre-Cut Salad Leaves, May 2012. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0125955"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1588","DOI":"10.1093\/cid\/cis255","article-title":"Outbreak of Shiga Toxin-Producing Escherichia coli O104:H4 Associated With Organic Fenugreek Sprouts, France, June 2011","volume":"54","author":"King","year":"2012","journal-title":"Clin. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"108849","DOI":"10.1016\/j.ijfoodmicro.2020.108849","article-title":"Occurrence of Listeria and Escherichia coli in frozen fruit and vegetables collected from retail and catering premises in England 2018\u20132019","volume":"334","author":"Willis","year":"2020","journal-title":"Int. J. Food Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ijpara.2017.09.004","article-title":"Foodborne cryptosporidiosis","volume":"48","author":"Ryan","year":"2018","journal-title":"Int. J. Parasitol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.3201\/eid2512.181565","article-title":"Patterns of Transmission and Sources of Infection in Outbreaks of Human Toxoplasmosis","volume":"25","author":"Caldart","year":"2019","journal-title":"Emerg. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e00142","DOI":"10.1016\/j.fawpar.2021.e00142","article-title":"Use of metagenomic microbial source tracking to investigate the source of a foodborne outbreak of cryptosporidiosis","volume":"26","author":"Ahlinder","year":"2021","journal-title":"Food Waterborne Parasitol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Hadjilouka, A., and Tsaltas, D. (2020). Cyclospora Cayetanensis\u2014Major Outbreaks from Ready to Eat Fresh Fruits and Vegetables. Foods, 9.","DOI":"10.3390\/foods9111703"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1900241","DOI":"10.2807\/1560-7917.ES.2019.24.43.1900241","article-title":"Substantial prevalence of enteroparasites Cryptosporidium spp., Giardia duodenalis and Blastocystis sp. in asymptomatic schoolchildren in Madrid, Spain, November 2017 to June 2018","volume":"24","author":"Reh","year":"2019","journal-title":"Eurosurveillance"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"107900","DOI":"10.1016\/j.exppara.2020.107900","article-title":"Why we need a European focus on foodborne parasites","volume":"214","author":"Robertson","year":"2020","journal-title":"Exp. Parasitol."},{"key":"ref_15","unstructured":"Food Standards Australia New Zealand (FSANZ) (2022, May 20). Compendium of Microbiological Criteria for Food, Available online: https:\/\/www.foodstandards.gov.au\/publications\/Documents\/Compedium%20of%20Microbiological%20Criteria\/Compendium_revised-jan-2018.pdf."},{"key":"ref_16","unstructured":"NSW Food Authority (2022, May 20). Microbiological Quality of Fresh Cut Vegetables\u2014A Survey to Determine the Safety of Fresh Cut Leafy Salad Vegetables Sold in NSW, Available online: https:\/\/www.foodauthority.nsw.gov.au\/sites\/default\/files\/_Documents\/scienceandtechnical\/microbiological_quality_fresh_cut_vegetables.pdf."},{"key":"ref_17","unstructured":"(2022, May 20). COMMISSION REGULATION (EC) No. 1441\/2007 of 5 December 2007 Amending Regulation (EC) No. 2073\/2005 on Micro-Biological Criteria for Foodstuffs. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=CELEX:32007R1441&from=EN."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority (EFSA), and European Centre for Disease Prevention and Control (ECDPC) (2021). The European Union One Health 2019 Zoonoses Report. EFSA J., 19.","DOI":"10.2903\/j.efsa.2021.6406"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"529","DOI":"10.3934\/microbiol.2017.3.529","article-title":"Foodborne pathogens","volume":"3","author":"Bintsis","year":"2017","journal-title":"AIMS Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1016\/j.vetpar.2009.05.026","article-title":"Emerging food-borne parasites","volume":"163","author":"Dorny","year":"2009","journal-title":"Veter. Parasitol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.vetpar.2011.07.017","article-title":"The socioeconomic burden of parasitic zoonoses: Global trends","volume":"182","author":"Torgerson","year":"2011","journal-title":"Veter. Parasitol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1016\/S0020-7519(00)00128-4","article-title":"Emerging parasite zoonoses associated with water and food","volume":"30","author":"Slifko","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-021-04769-4","article-title":"Outbreaks of clinical toxoplasmosis in humans: Five decades of personal experience, perspectives and lessons learned","volume":"14","author":"Dubey","year":"2021","journal-title":"Parasites Vectors"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1128\/CMR.00033-10","article-title":"Zoonotic Potential and Molecular Epidemiology of Giardia Species and Giardiasis","volume":"24","author":"Feng","year":"2011","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.rvsc.2020.09.034","article-title":"Giardia duodenalis in humans and animals\u2014Transmission and disease","volume":"135","author":"Dixon","year":"2020","journal-title":"Res. Veter. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Bouzid, M., Kintz, E., and Hunter, P. (2018). Risk factors for Cryptosporidium infection in low and middle income countries: A systematic review and meta-analysis. PLOS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006553"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.1017\/S0031182021001013","article-title":"Congenital toxoplasmosis in humans: An update of worldwide rate of congenital infections","volume":"148","author":"Dubey","year":"2021","journal-title":"Parasitology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1016\/S0140-6736(04)16412-X","article-title":"Toxoplasmosis","volume":"363","author":"Montoya","year":"2004","journal-title":"Lancet"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.vetpar.2004.09.011","article-title":"Cyclospora cayetanensis, a food- and waterborne coccidian parasite","volume":"126","author":"Mansfield","year":"2004","journal-title":"Veter. Parasitol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Mathison, B.A., and Pritt, B.S. (2021). Cyclosporiasis\u2014Updates on Clinical Presentation, Pathology, Clinical Diagnosis, and Treatment. Microorganisms, 9.","DOI":"10.3390\/microorganisms9091863"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.ijfoodmicro.2004.12.032","article-title":"Foodborne protozoan parasites","volume":"103","author":"Dawson","year":"2005","journal-title":"Int. J. Food Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1051\/parasite\/2018009","article-title":"Assessing viability and infectivity of foodborne and waterborne stages (cysts\/oocysts) of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: A review of methods","volume":"25","author":"Rousseau","year":"2018","journal-title":"Parasite"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1080\/20477724.2018.1514137","article-title":"A review on inactivation methods of Toxoplasma gondii in foods","volume":"112","author":"Jazaeri","year":"2018","journal-title":"Pathog. Glob. Health"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.vetpar.2008.02.004","article-title":"Effects of ozone and ultraviolet radiation treatments on the infectivity of Toxoplasma gondii oocysts","volume":"153","author":"Baffet","year":"2008","journal-title":"Veter. Parasitol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"113595","DOI":"10.1016\/j.ijheh.2020.113595","article-title":"Systematic review and meta-analysis of time-temperature pathogen inactivation","volume":"230","author":"Espinosa","year":"2020","journal-title":"Int. J. Hyg. Environ. Health"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Adeyemo, F.E., Singh, G., Reddy, P., Bux, F., and Stenstr\u00f6m, T.A. (2019). Efficiency of chlorine and UV in the inactivation of Cryptosporidium and Giardia in wastewater. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0216040"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2786","DOI":"10.4315\/0362-028X-69.11.2786","article-title":"Inactivation of Protozoan Parasites in Food, Water, and Environmental Systems","volume":"69","author":"Erickson","year":"2006","journal-title":"J. Food Prot."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Gajadhar, A., Scandrett, W.B., and Forbes, L.B. (2006). Overview of food- and water-borne zoonotic parasites at the farm level. Rev. Sci. Tech., 25.","DOI":"10.20506\/rst.25.2.1679"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.fawpar.2018.04.002","article-title":"Parasite contamination of berries: Risk, occurrence, and approaches for mitigation","volume":"10","author":"Tefera","year":"2018","journal-title":"Food Waterborne Parasitol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2385","DOI":"10.1111\/j.1462-2920.2010.02297.x","article-title":"Fresh fruit and vegetables as vehicles for the transmission of human pathogens","volume":"12","author":"Berger","year":"2010","journal-title":"Environ. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1727","DOI":"10.1111\/1541-4337.12487","article-title":"Microbial Contamination of Fresh Produce: What, Where, and How?","volume":"18","author":"Richards","year":"2019","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1186\/s13071-020-04040-2","article-title":"Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits","volume":"13","author":"Marques","year":"2020","journal-title":"Parasites Vectors"},{"key":"ref_43","unstructured":"United States Environmental Protection Agency (EPA) (2022, May 20). Method 1623.1: Cryptosporidium and Giardia in Water by Filtra-Tion\/IMS\/FA, Available online: https:\/\/nepis.epa.gov\/Exe\/ZyPDF.cgi\/P100J7G4.PDF?Dockey=P100J7G4.PDF."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"43","DOI":"10.3347\/kjp.2010.48.1.43","article-title":"Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal","volume":"48","author":"Almeida","year":"2010","journal-title":"Korean J. Parasitol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-020-64590-4","article-title":"rTgOWP1-f, a specific biomarker for Toxoplasma gondii oocysts","volume":"10","author":"Sousa","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"105","DOI":"10.3347\/kjp.2010.48.2.105","article-title":"Biological and Genetic Characterization of Cryptosporidium spp. and Giardia duodenalis Isolates from Five Hydrographical Basins in Northern Portugal","volume":"48","author":"Almeida","year":"2010","journal-title":"Korean J. Parasitol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.vetpar.2007.03.019","article-title":"Molecular characterization of Cryptosporidium and Giardia isolates from cattle from Portugal","volume":"147","author":"Almeida","year":"2007","journal-title":"Veter. Parasitol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"S177","DOI":"10.1111\/j.1550-7408.2006.00222.x","article-title":"Genotype Analysis of Giardia Isolated from Asymptomatic Children in Northern Portugal","volume":"53","author":"Almeida","year":"2006","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"S26","DOI":"10.1111\/j.1550-7408.2006.00163.x","article-title":"Genetic Characterization of Cryptosporidium Isolates from Humans in Northern Portugal","volume":"53","author":"Almeida","year":"2006","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1128\/AEM.67.3.1097-1101.2001","article-title":"Molecular Characterization of Cryptosporidium Oocysts in Samples of Raw Surface Water and Wastewater","volume":"67","author":"Xiao","year":"2001","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/S0020-7519(02)00068-1","article-title":"Sequence analysis of the \u03b2-giardin gene and development of a polymerase chain reaction\u2013restriction fragment length polymorphism assay to genotype Giardiaduodenalis cysts from human faecal samples","volume":"32","author":"Pozio","year":"2002","journal-title":"Int. J. Parasitol."},{"key":"ref_52","unstructured":"(2013). Microbiology of the Food Chain\u2014Horizontal Method for the Enumeration of Microorganisms\u2014Part 1: Colony Count at 30 \u00b0C by the Pour Plate Technique (Standard No. ISO 4833)."},{"key":"ref_53","unstructured":"(2016). Microbiology of the food chain\u2014Method validation\u2014Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (Standard No. ISO 16140-2:2016)."},{"key":"ref_54","unstructured":"(2017). Microbiology of the Food Chain-Horizontal Method for the Detection and Enumeration of Enterobacteriaceae\u2014Part 2: Colony-Count Technique (Standard No. ISO 21528-2)."},{"key":"ref_55","unstructured":"(2001). Microbiology of Food and Animal Feeding Stuffs\u2014Horizontal Method for the Enumeration of Beta-Glucuronidase-Positive Escherichia coli\u2014Part 2: Colony-Count Technique at 44 \u00b0C Using 5-bromo-4-chloro-3-indolyl beta-D-glucuronide (Standard No. ISO 16649-2)."},{"key":"ref_56","unstructured":"(2017). Microbiology of the Food Chain\u2014Horizontal Method for the Detection and Enumeration of Listeria Monocytogenes and of Listeria spp.\u2014Part 2: Enumeration Method (Standard No. ISO 11290-2)."},{"key":"ref_57","unstructured":"(2008). Microbiology of Food and Animal Feeding Stuffs\u2014Horizontal Method for the Enumeration of Yeasts and Moulds\u2014Part 1: Colony Count Technique in Products with Water Activity Greater Than 0.95 (Standard No. ISO 21527-1)."},{"key":"ref_58","unstructured":"(2017). Microbiology of the Food Chain\u2014Horizontal Method for the Detection and Enumeration of Listeria Monocytogenes and of Listeria spp.\u2014Part 1: Detection Method (Standard No. ISO 11290-1)."},{"key":"ref_59","unstructured":"(2017). Microbiology of the Food Chain\u2014Horizontal Method for the Detection, Enumeration and Serotyping of Salmonella\u2014Part 1: Detection of Salmonella spp (Standard No. ISO 6579-1)."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1017\/S0031182020000086","article-title":"Detection methods and prevalence of transmission stages of Toxoplasma gondii, Giardia duodenalis and Cryptosporidium spp. in fresh vegetables: A review","volume":"147","author":"Berrouch","year":"2020","journal-title":"Parasitology"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1016\/j.pt.2019.07.002","article-title":"Foodborne Parasites in Europe: Present Status and Future Trends","volume":"35","author":"Trevisan","year":"2019","journal-title":"Trends Parasitol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.fm.2017.06.006","article-title":"Detection and prevalence of protozoan parasites in ready-to-eat packaged salads on sale in Italy","volume":"67","author":"Caradonna","year":"2017","journal-title":"Food Microbiol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.ijfoodmicro.2017.05.009","article-title":"Keeping it cool: Survival of Giardia cysts and Cryptosporidium oocysts on lettuce leaves","volume":"255","author":"Utaaker","year":"2017","journal-title":"Int. J. Food Microbiol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"4584","DOI":"10.1111\/1541-4337.12786","article-title":"Practical in-storage interventions to control foodborne pathogens on fresh produce","volume":"20","author":"Sheng","year":"2021","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Ferreira, C., Lopes, F., Costa, R., Komora, N., Ferreira, V., Fernandes, V.C., Delerue-Matos, C., and Teixeira, P. (2020). Microbiological and Chemical Quality of Portuguese Lettuce\u2014Results of a Case Study. Foods, 9.","DOI":"10.3390\/foods9091274"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1111\/jam.14935","article-title":"Microbiological quality and safety of minimally processed parsley (Petroselinum crispum) sold in food markets, southeastern Brazil","volume":"131","author":"Finger","year":"2020","journal-title":"J. Appl. Microbiol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"108855","DOI":"10.1016\/j.ijfoodmicro.2020.108855","article-title":"Microbiological safety of ready-to-eat fresh-cut fruits and vegetables sold on the Canadian retail market","volume":"335","author":"Zhang","year":"2020","journal-title":"Int. J. Food Microbiol."},{"key":"ref_68","unstructured":"Food Safety Authority of Ireland (2020). Guidelines for the Interpretation of Results of Microbiological Testing of Ready-to-Eat Foods Placed on the Market (Revision 4), FSAI."},{"key":"ref_69","unstructured":"Saraiva, M., Correia, C.B., Cunha, I.C., Maia, C., Bonito, C.C., Furtado, R., and Calhau, M.A. (2022, May 19). Interpreta\u00e7\u00e3o dos resultados de ensaios microbiol\u00f3gicos em alimentos prontos para consumo e em superf\u00edcies do ambiente de produ\u00e7\u00e3o e distribui\u00e7\u00e3o alimentar: Valores-guia. Reposit\u00f3rio Cient\u00edfico do Instituto Nacional de Sa\u00fade Dr. Ricardo Jorge 2019. Available online: http:\/\/hdl.handle.net\/10400.18\/5610."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"118206","DOI":"10.1016\/j.watres.2022.118206","article-title":"Ozone disinfection of waterborne pathogens and their surrogates: A critical review","volume":"214","author":"Morrison","year":"2022","journal-title":"Water Res."}],"container-title":["Applied Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3417\/12\/14\/7145\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:51:22Z","timestamp":1760140282000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3417\/12\/14\/7145"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,15]]},"references-count":70,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["app12147145"],"URL":"https:\/\/doi.org\/10.3390\/app12147145","relation":{},"ISSN":["2076-3417"],"issn-type":[{"type":"electronic","value":"2076-3417"}],"subject":[],"published":{"date-parts":[[2022,7,15]]}}}