Introduction and objective:
Official food control laboratories ensure food safety using reliable, validated methods. Council Regulations (EC) No. 853/2004, 854/2004 and 882/2004 of the European Parliament established hygiene rules the production of food of animal origin, together with requirements for official controls. This leads to detailed requirements for Trichinella control set out in Commission Implementing Regulation (EU) 2015/1375 of 10 August 2015. These regulations require the laboratory to participate in proficiency testing (PT) to confirm their competence and improve the quality of testing, and require the PT Organizer to use methods for the preparation and preservation of parasite larvae in order to evaluate and improve detection. Traditional methods of preparing such larvae expose them to rapid degradation, making it necessary to simultaneously isolate the larvae and place them in meatballs to ensure quality.

Material and methods:
We developed a technique for preserving of Trichinella spp. for quality control such as PT sample preparation. The procedure protects larvae against toxic oxygen activity and bacterial destruction via a gelatin barrier. To estimate the viability of larvae preserved by this method, gelatin capsules with 10 larvae of T. spiralis in each were stored (4–8 °C) during 45 days of an experiment. Samples were tested at 2 day intervals (3 samples each day of testing).

In total, 75 samples were tested. Larvae remained alive up to 3 weeks. The number of living larvae diminished after 27 days through day 43, after which no living larvae were observed.

The gelatin medium procedure facilitated easy, high-throughput sample preparation and supported 100% recovery for 3 weeks. The method allows fast, efficient and accurate PT sample preparation.

Shimoni Z, Froom P. Uncertainties in diagnosis, treatment and prevention of trichinellosis. Exp Rev Anti-infective Therapy. 2015;13(10):1279–1288.
Mohib O, Clevenbergh P, Truyens C, Morissens M, Castro Rodriguez J. Trichinella spiralis-associated myocarditis mimicking acute myocardial infarction. Acta Clin Belg. 2022;77(1): 147–152.
Rostami A, Gamble HR, Dupouy-Camet J, Khazan H, Bruschi F. Meat sources of infection for outbreaks of human trichinellosis. Food Microbiol. 2017;64:65–71. doi:
Ilic N, Vasilev S, Gruden-Movsesijan A, Gnjatovic M, Sofronic-Milosavljevic L, Mitic I. Long lasting immunity in trichinellosis – insight from a small study group. J Helminthol. 2022; 96: E35.
Lupu MA, Pavel R, Lazureanu VE, Popovici ED, Olariu TR. Trichinellosis in hospitalized patients from Western Romania: A retrospective study. Exp Ther Med. 2021; 22: 895.
Despotović D, Nenadović K, Sladojević Ž, Dimitrijević S, Ilić T. Epidemiology and risk factors of trichinellosis in Bosnia and Herzegovina from 1961 to 2021. Parasitol Res. 2023;122(2):635–643.
Commission U. Commission Implementing Regulation (EU) 2015/1375 of 10 August 2015 laying down specific rules on official controls for Trichinella in meat (codification). 2015. doi:
Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. 2004.
Regulation (EU) 2017/625 of the European Parliament and of the Council of 15 March 2017 on official controls and other official activities performed to ensure the application of food and feed law, rules on animal health and welfare, plant health and plant protection products, amending Regulations (EC) No 999/2001, (EC) No 396/2005, (EC) No 1069/2009, (EC) No 1107/2009, (EU) No 1151/2012, (EU) No 652/2014, (EU) 2016/429 and (EU) 2016/2031 of the European Parliament and of the Council, Council Regulations (EC) No 1/2005 and (EC) No 1099/2009 and Council Directives 98/58/EC, 1999/74/EC, 2007/43/EC, 2008/119/EC and 2008/120/EC, and repealing Regulations (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council, Council Directives 89/608/EEC, 89/662/EEC, 90/425/EEC, 91/496/EEC, 96/23/EC, 96/93/EC and 97/78/EC and Council Decision 92/438/EEC (Official Controls Regulation)Text with EEA relevance. 2007.
Mayer-Scholl A, Pozio E, Gayda J, Thaben N, Bahn P, Nöckler K. Magnetic stirrer method for the detection of Trichinella larvae in muscle samples. JoVE. 2017;(121).
ISO/IEC 17043:2023 Conformity assessment — General requirements for the competence of proficiency testing providers. 2023.
van der Giessen J, Franssen F, Fonville M, et al. How safe is the meat inspection based on artificial digestion of pooled samples for Trichinella in pork? A scenario from wildlife to a human patient in a non-endemic region of Europe. Vet Parasitol. 2013;194(2):110–112. doi:
Marucci G, Tonanzi D, Cherchi S, et al. Proficiency testing to detect Trichinella larvae in meat in the European Union. Vet Parasitol. 2016;231:145–149. doi:
Rossi P, Marucci G, Lalle M, Casulli A, Possenti A, Pozio E. Proficiency testing carried out by the European Union Reference Laboratory for Parasites. Accreditation Qual Assur. 2015;20(4):311–317. doi:10.1007/s00769-015-1142-3.
Gajadhar AA, Noeckler K, Boireau P, Rossi P, Scandrett B, Gamble HR. International Commission on Trichinellosis: Recommendations for quality assurance in digestion testing programs for Trichinella. Food and Waterborne Parasitol. 2019;16:e00059. doi:
Forbes LB, Rajic A, Gajadhar AA. Proficiency Samples for Quality Assurance in Trichinella Digestion Tests. J Food Prot. 1998;61(10):1396–1399. doi:
Winter M, Pasqualetti M, Farina F, et al.Trichinellosis surveillance in wildlife in northeastern argentine patagonia. Vet Parasitol Reg Stud. 2018;11:32–35.
Vallée I, Macé P, Forbes L, et al. Use of Proficiency Samples To Assess Diagnostic Laboratories in France Performing a Trichinella Digestion Assay. J Food Prot. 2007;70(7):1685–1690. doi:–0....
Różycki M, Korpysa-Dzirba W, Bełcik A, et al. Validation of the Magnetic Stirrer Method for the Detection of Trichinella Larvae in Muscle Samples Based on Proficiency Tests Results. Foods. 2022;11(4).
Różycki M, Korpysa-Dzirba W, Bełcik A, et al. Results of Proficiency Testing for Trichinella in Poland, 2015–2019. J Clin Med. 2021;10(22).
ISO 18743:2015 Microbiology of the food chain — Detection of Trichinella larvae in meat by artificial digestion method. 2015.
Zarlenga DS, Chute MB, Martin A, Kapel CMO. A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella. Int J Parasitol. 1999;29(11):1859–1867. doi:–....
Noeckler K, Pozio E, van der Giessen J, Hill DE, Gamble HR. International Commission on Trichinellosis: Recommendations on post-harvest control of Trichinella in food animals. Food Waterborne Parasitol. 2019;14:e00041. doi: