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Towards a standardised methodology of Trichinella larval counting for research purposes

Published online by Cambridge University Press:  21 November 2025

D.S. Ashour Open the ORCID record for D.S. Ashour [Opens in a new window] ,
F.M. Elgazzar ,
A.A. Othman  and
H.S. Zoghroban
D.S. Ashour
Affiliation:
Medical Parasitology Department, Faculty of Medicine, Tanta University, Tanta, Egypt
F.M. Elgazzar
Affiliation:
Forensic Medicine and Clinical Toxicology, Faculty of Medicine, Tanta University, Tanta, Egypt
A.A. Othman
Affiliation:
Medical Parasitology Department, Faculty of Medicine, Tanta University, Tanta, Egypt
H.S. Zoghroban*
Affiliation:
Medical Parasitology Department, Faculty of Medicine, Tanta University, Tanta, Egypt
*
Corresponding author: H.S. Zoghroban; Emails: hager.zoghroban@med.tanta.edu.eg; dr.hager.radwan@gmail.com
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Abstract

Trichinella spiralis (T. spiralis) is a zoonotic parasitic worm that has a significant impact on both public health and the livestock industry. It is widely used in experimental studies because of several unique features of its life cycle that can be completed in a single host, including distinct intestinal and muscular phases of infection, and it can be easily maintained in laboratory animals. These characteristics make T. spiralis a valuable model for evaluating the efficacy of new drugs and vaccines against parasitic infections. The current work aims to evaluate the procedures used for Trichinella larval counting in experimental studies by comparing the magnetic stirring digestion method (MSDM) with individual muscle digestion and compression techniques under standardised, controlled conditions to assess their effectiveness and applicability. Fifteen male Swiss albino mice were orally infected with T. spiralis and sacrificed on day 35 post-infection. They were divided into three groups: group 1—MSDM for total larval count; group 2—individual digestion of masseter, diaphragm, gastrocnemius, and tongue muscles; and group 3—slide compression method of the same muscles. MSDM yielded the highest larval counts, with individual muscle digestion and compression methods generally producing significantly lower results, except tongue digestion, while correlation and concordance analyses identified gastrocnemius compression as an alternative despite overall poor agreement with the reference method. Therefore, MSDM is the most accurate method for larval counting in experimental studies, compared to individual muscle digestion or compression methods.

Keywords

Trichinella spiralislarval countmuscle digestionmagnetic stirring digestion techniquecompression methods

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Type
Research Paper
Information
Journal of Helminthology , Volume 99 , 2025 , e123
Copyright
© The Author(s), 2025. Published by Cambridge University Press

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References

Abdel-Hafeez, EH, Kamal, AM, Abdelgelil, NH and Abdel-Fatah, MM (2015) Parasites transmitted to human by ingestion of different types of meat, El-Minia city, El-Minia governorate. Journal of the Egyptian Society of Parasitology 45(3), 671–680.Google ScholarPubMed
Abou Rayia, D, Othman, A, Harras, S, Helal, D, Dawood, L and Soliman, S (2022) Bevacizumab: a new take on therapy of muscle phase of Trichinella spiralis infection. Acta Tropica 230, 106409.10.1016/j.actatropica.2022.106409CrossRefGoogle Scholar
Ashour, DS, Abou Rayia, DM, Saad, AE and El-Bakary, RH (2016) Nitazoxanide anthelmintic activity against the enteral and parenteral phases of trichinellosis in experimentally infected rats. Experimental Parasitology 170, 28–35.10.1016/j.exppara.2016.08.009CrossRefGoogle ScholarPubMed
Ashour, DS, Ibrahim, FM, Elshamy, AM and Zoghroban, HS (2022) Trichinella spiralis-derived extracellular vesicles induce a protective immunity against larval challenge in mice. Pathogens and Disease 80(1), ftac040.10.1093/femspd/ftac040CrossRefGoogle ScholarPubMed
Barlow, A, Roy, K, Hawkins, K, Ankarah, AA and Rosenthal, B (2021) A review of testing and assurance methods for Trichinella surveillance programs. Food and Waterborne Parasitology 24, e00129.10.1016/j.fawpar.2021.e00129CrossRefGoogle ScholarPubMed
Basso, W, Marreros, N, Hofmann, L, Salvisberg, C, Lundström-Stadelmann, B and Frey, CF (2022) Evaluation of the PrioCHECK™ Trichinella AAD kit to detect Trichinella spiralis, T. britovi, and T. pseudospiralis larvae in pork using the automated digestion method Trichomatic-35. Parasitology International 86, 102449.10.1016/j.parint.2021.102449CrossRefGoogle Scholar
Blair, LS (1983) Laboratory techniques. In: Campbell, W.C. (ed.) Trichinella and Trichinosis. New York, Plenum Press (Springer US), 593–617.Google Scholar
Codina, AV, Indelman, P, Hinrichsen, LI and Lamas, MC (2025) Significant improvement in bioavailability and therapeutic efficacy of mebendazole oral nano-systems assessed in a murine model with extreme phenotypes of susceptibility to Trichinella spiralis. Pharmaceutics 17(8), 1069.10.3390/pharmaceutics17081069CrossRefGoogle Scholar
Dunn, IJ and Wright, KA (1985) Cell injury caused by Trichinella spiralis in the mucosal epithelium of B10A mice. The Journal of Parasitology 1, 757–766.10.2307/3281709CrossRefGoogle Scholar
Dupouy-Camet, J and Murrell, KD (eds) (2007) FAO/WHO/OIE Guidelines for the Surveillance, Management, Prevention and Control of Trichinellosis. Paris: World Organisation for Animal Health (OIE).Google Scholar
El-Wakil, ES, Abdelmaksoud, HF, AbouShousha, TS, Ghallab, MMI (2021) Evaluation of Annona muricata (Graviola) leaves activity against experimental trichinellosis: in vitro and in vivo studies. Journal of Helminthology 95, e53, 1–8.10.1017/S0022149X21000481CrossRefGoogle ScholarPubMed
Forbes, LB, Parker, S and Scandrett, WB (2003) Comparison of a modified digestion assay with trichinoscopy for the detection of Trichinella larvae in pork. Journal of Food Protection 66(6), 1043–1046.10.4315/0362-028X-66.6.1043CrossRefGoogle ScholarPubMed
Franssen, F, Johne, A, Van der Giessen, J, Nöckler, K and Mayer-Scholl, A (2019) Test sensitivity of a commercial serine protease digestion kit for the detection of Trichinella spiralis and Trichinella pseudospiralis larvae in pig muscle. Food Microbiology 78, 99–103.10.1016/j.fm.2018.10.012CrossRefGoogle ScholarPubMed
Gajadhar, AA, Noeckler, K, Boireau, P, Rossi, P, Scandrett, B and Gamble, HR (2019) International Commission on Trichinellosis: Recommendations for quality assurance in digestion testing programs for Trichinella. Food and Waterborne Parasitology 16, e00059.10.1016/j.fawpar.2019.e00059CrossRefGoogle ScholarPubMed
Gajadhar, AA, Pozio, E, Gamble, HR, Nöckler, K, Maddox-Hyttel, C, Forbes, LB, Vallee, I, Rossi, P, Marinculić, A and Boireau, P (2009) Trichinella diagnostics and control: mandatory and best practices for ensuring food safety. Veterinary Parasitology 159(3–4), 197–205.10.1016/j.vetpar.2008.10.063CrossRefGoogle ScholarPubMed
Gajadhar, A, Konecsni, K, Scandrett, B and Buholzer, P (2018) Validation of a new commercial serine protease artificial digestion assay for the detection of Trichinella larvae in pork. Food and Waterborne Parasitology 10, 6–13.10.1016/j.fawpar.2018.04.001CrossRefGoogle ScholarPubMed
Gamble, HR, Bessonov, AS, Cuperlovic, K, Gajadhar, AA, Van Knapen, F, Noeckler, K, Schenone, H and Zhu, X (2000) International Commission on Trichinellosis: recommendations on methods for the control of Trichinella in domestic and wild animals intended for human consumption. Veterinary Parasitology 93(3–4), 393–408.10.1016/S0304-4017(00)00354-XCrossRefGoogle ScholarPubMed
Kapel, CM, Webster, P and Gamble, HR (2005) Muscle distribution of sylvatic and domestic Trichinella larvae in production animals and wildlife. Veterinary Parasitology 132(1–2), 101–105.10.1016/j.vetpar.2005.05.036CrossRefGoogle ScholarPubMed
Konecsni, K, Scheller, C, Scandrett, B, Buholzer, P and Gajadhar, A (2017) Evaluation of the PrioCHECK™ Trichinella AAD Kit for the digestion and recovery of larvae in pork, horse meat and wild meat. Veterinary Parasitology 243, 267–271.10.1016/j.vetpar.2017.07.004CrossRefGoogle ScholarPubMed
Leclair, D, Forbes, LB, Suppa, S and Gajadhar, AA (2003) Evaluation of a digestion assay and determination of sample size and tissue for the reliable detection of Trichinella larvae in walrus meat. Journal of Veterinary Diagnostic Investigation 15(2), 188–191.10.1177/104063870301500217CrossRefGoogle ScholarPubMed
Lin, Z, Cao, J, Zhang, H, Zhou, Y, Deng, M, Li, G and Zhou, J (2013) Comparison of three molecular detection methods for detection of Trichinella in infected pigs. Parasitology Research 112(5), 2087–2093.10.1007/s00436-012-3267-1CrossRefGoogle ScholarPubMed
Makrutzki, G, Riehn, K, Hamedy, A, Petroff, D, Hasenclever, D, Meiler, H and Lücker, E (2014) Detecting Trichinella infections using inverse microscopy and an improved larval counting technique. Journal of Helminthology 88(4), 453–458.10.1017/S0022149X13000424CrossRefGoogle Scholar
Mayer-Scholl, A, Pozio, E, Gayda, J, Thaben, N, Bahn, P and Nöckler, K (2017) Magnetic stirrer method for the detection of Trichinella larvae in muscle samples. Journal of Visualized Experiments 3(121), 55354.Google Scholar
Mostafa, E, Hassan, SS, Ahmed, M (2020) Comparison of different techniques and stains for direct diagnosis of Trichinella spiralis. Microbes and Infectious Diseases 1 (3), 199–208.Google Scholar
Nöckler, K, Reckinger, S, Szabo, I, Maddox-Hyttel, C, Pozio, E, van Der Giessen, J, Vallee, I and Boireau, P (2009) Comparison of three artificial digestion methods for detection of non-encapsulated Trichinella pseudospiralis larvae in pork. Veterinary Parasitology 159(3–4), 341–344.10.1016/j.vetpar.2008.10.075CrossRefGoogle ScholarPubMed
Noeckler, K, Pozio, E, van der Giessen, J, Hill, DE and Gamble, HR (2019) International Commission on Trichinellosis: Recommendations on post-harvest control of Trichinella in food animals. Food and Waterborne Parasitology 14, e00041.10.1016/j.fawpar.2019.e00041CrossRefGoogle ScholarPubMed
Othman, AA, Abou Rayia, DM, Ashour, DS, Saied, EM, Zineldeen, DH and El-Ebiary, AA (2016) Atorvastatin and metformin administration modulates experimental Trichinella spiralis infection. Parasitology International 65(2), 105–112.10.1016/j.parint.2015.11.001CrossRefGoogle ScholarPubMed
Pozio, E (2007) World distribution of Trichinella spp. infections in animals and humans. Veterinary Parasitology 149(1–2), 3–21.10.1016/j.vetpar.2007.07.002CrossRefGoogle Scholar
Pozio, E and Zarlenga, DS (2013) New pieces of the Trichinella puzzle. International Journal for Parasitology 43(12–13), 983–997.10.1016/j.ijpara.2013.05.010CrossRefGoogle ScholarPubMed
Riehn, K, Hasenclever, D, Petroff, D, Nöckler, K, Mayer-Scholl, A, Makrutzki, G and Lücker, E (2013) Trichinella detection: identification and statistical evaluation of sources of error in the magnetic stirrer method for pooled sample digestion. Veterinary Parasitology 194(2–4), 106–109.10.1016/j.vetpar.2013.01.031CrossRefGoogle ScholarPubMed
Rostami, A, Gamble, HR, Dupouy-Camet, J, Khazan, H and Bruschi, F (2017) Meat sources of infection for outbreaks of human trichinellosis. Food Microbiology 64, 65–71.10.1016/j.fm.2016.12.012CrossRefGoogle ScholarPubMed
Różycki, M, Korpysa-Dzirba, W, Bełcik, A, Bilska-Zając, E, Kochanowski, M, Karamon, J, Sroka, J and Cencek, T (2022) Validation of the magnetic stirrer method for the detection of Trichinella larvae in muscle samples based on proficiency tests results. Foods 11(4), 525.10.3390/foods11040525CrossRefGoogle ScholarPubMed
Vignau, ML, Guardis, MD, Risso, MA and Eiras, DF (1997) Comparison between two methods for diagnosis of trichinellosis: trichinoscopy and artificial digestion. Memórias do Instituto Oswaldo Cruz 92, 585–587.10.1590/S0074-02761997000500004CrossRefGoogle ScholarPubMed
Wang, N, Bai, X, Tang, B, Yang, Y, Wang, X, Zhu, H, Luo, X, Yan, H, Jia, H, Liu, M and Liu, X (2020) Primary characterization of the immune response in pigs infected with Trichinella spiralis. Veterinary Research 51(1), 17.10.1186/s13567-020-0741-0CrossRefGoogle ScholarPubMed