Inactivation of Toxoplasma gondii in dry sausage and processed pork, and quantification of the pathogen in pig tissues prior to production

IF 2.9 Q2 PARASITOLOGY
Filip Dámek , Bastien Fremaux , Dominique Aubert , Sandra Thoumire , Maxime Delsart , Jean-Luc Martin , Sandra Vuillermet , Marieke Opsteegh , Pikka Jokelainen , Delphine Le Roux , Pascal Boireau , Isabelle Villena , Radu Blaga
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引用次数: 1

Abstract

Toxoplasma gondii is an important zoonotic foodborne parasite. Meat of infected animals appears to be a major source of infection in Europe. Pork is the most consumed meat in France, with dry sausages well represented. The risk of transmission via consumption of processed pork products is largely unknown, mainly since processing will affect viability but may not entirely inactivate all T. gondii parasites.

We investigated the presence and concentration of T. gondii DNA in the shoulder, breast, ham, and heart of pigs orally inoculated with 1000 oocysts (n = 3) or tissue cysts (n = 3) and naturally infected pigs (n = 2), by means of magnetic capture qPCR (MC-qPCR). Muscle tissues of experimentally infected pigs were further used to evaluate the impact of manufacturing processes of dry sausages, including different concentrations of nitrates (0, 60, 120, 200 ppm), nitrites (0, 60, 120 ppm), and NaCl (0, 20, 26 g/kg), ripening (2 days at 16–24 °C) and drying (up to 30 days at 13 °C), by a combination of mouse bioassay, qPCR and MC-qPCR.

DNA of T. gondii was detected in all eight pigs, including in 41.7% (10/24) of muscle samples (shoulder, breast and ham) and 87.5% (7/8) of hearts by MC-qPCR. The number of parasites per gram of tissue was estimated to be the lowest in the hams (arithmetic mean (M) = 1, standard deviation (SD) = 2) and the highest in the hearts (M = 147, SD = 233). However, the T. gondii burden estimates varied on the individual animal level, the tissue tested and the parasitic stage used for the experimental infection (oocysts or tissue cysts). Of dry sausages and processed pork, 94.4% (51/54) were positive for T. gondii by MC-qPCR or qPCR, with the mean T. gondii burden estimate equivalent to 31 parasites per gram (SD = 93). Only the untreated processed pork sample collected on the day of production was positive by mouse bioassay.

The results suggest an uneven distribution of T. gondii in the tissues examined, and possibly an absence or a concentration below the detection limit in some of them. Moreover, the processing of dry sausages and processed pork with NaCl, nitrates, and nitrites has an impact on the viability of T. gondii from the first day of production. Results are valuable input for future risk assessments aiming to estimate the relative contribution of different sources of T. gondii human infections.

Abstract Image

干香肠和加工猪肉中弓形虫的灭活以及生产前猪组织中病原体的定量
弓形虫是一种重要的人畜共患食源性寄生虫。受感染动物的肉似乎是欧洲的主要感染源。猪肉是法国消费量最大的肉类,干香肠很有代表性。通过食用加工猪肉产品传播的风险在很大程度上是未知的,主要是因为加工会影响生存能力,但可能不会完全灭活所有弓形虫寄生虫。我们通过磁性捕获qPCR(MC-qPCR)研究了经口接种1000个卵囊(n=3)或组织囊肿(n=3)的猪和自然感染的猪(n=2)的肩部、乳房、火腿和心脏中弓形虫DNA的存在和浓度。实验感染猪的肌肉组织进一步用于评估干香肠生产过程的影响,包括不同浓度的硝酸盐(0,60,120,200 ppm)、亚硝酸盐(0,60120 ppm)和NaCl(0,20,26 g/kg)、成熟(16-24°C下2天)和干燥(13°C下最多30天),通过小鼠生物测定相结合,qPCR和MC-qPCR.DNA在8头猪中均检测到弓形虫DNA,包括41.7%(10/24)的肌肉(肩、胸和火腿)和87.5%(7/8)的心脏。据估计,每克组织中的寄生虫数量在火腿中最低(算术平均值(M)=1,标准差(SD)=2),在心脏中最高(M=147,SD=233)。然而,弓形虫的负担估计因个体动物水平、测试的组织和实验感染所用的寄生虫阶段(卵囊或组织囊肿)而异。在干香肠和加工猪肉中,通过MC-qPCR或qPCR,94.4%(51/54)的弓形虫呈阳性,平均弓形虫负荷估计相当于每克31个寄生虫(SD=93)。只有在生产当天收集的未经处理的加工猪肉样品通过小鼠生物测定呈阳性。结果表明,弓形虫在所检查的组织中分布不均,其中一些组织可能不存在或浓度低于检测限。此外,用NaCl、硝酸盐和亚硝酸盐加工干香肠和加工猪肉从生产的第一天起就对弓形虫的生存能力产生影响。结果对未来的风险评估是有价值的投入,旨在评估不同来源的弓形虫人类感染的相对贡献。
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来源期刊
Food and Waterborne Parasitology
Food and Waterborne Parasitology Immunology and Microbiology-Parasitology
CiteScore
5.10
自引率
4.00%
发文量
38
审稿时长
13 weeks
期刊介绍: Food and Waterborne Parasitology publishes high quality papers containing original research findings, investigative reports, and scientific proceedings on parasites which are transmitted to humans via the consumption of food or water. The relevant parasites include protozoa, nematodes, cestodes and trematodes which are transmitted by food or water and capable of infecting humans. Pertinent food includes products of animal or plant origin which are domestic or wild, and consumed by humans. Animals and plants from both terrestrial and aquatic sources are included, as well as studies related to potable and other types of water which serve to harbor, perpetuate or disseminate food and waterborne parasites. Studies dealing with prevalence, transmission, epidemiology, risk assessment and mitigation, including control measures and test methodologies for parasites in food and water are of particular interest. Evidence of the emergence of such parasites and interactions among domestic animals, wildlife and humans are of interest. The impact of parasites on the health and welfare of humans is viewed as very important and within scope of the journal. Manuscripts with scientifically generated information on associations between food and waterborne parasitic diseases and lifestyle, culture and economies are also welcome. Studies involving animal experiments must meet the International Guiding Principles for Biomedical Research Involving Animals as issued by the Council for International Organizations of Medical Sciences.
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