Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-08T13:19:59.572Z Has data issue: false hasContentIssue false
  • English
  • Français

Detecting Trichinella infections using inverse microscopy and an improved larval counting technique

Published online by Cambridge University Press:  18 June 2013

G. Makrutzki ,
K. Riehn ,
A. Hamedy ,
D. Petroff ,
D. Hasenclever ,
H. Meiler  and
E. Lücker
G. Makrutzki*
Affiliation:
Institute of Food Hygiene, Faculty of Veterinary Medicine, University of Leipzig, Leipzig, Germany
K. Riehn
Affiliation:
Department of Nutrition & Home Economics, Hamburg University of Applied Science, Hamburg, Germany
A. Hamedy
Affiliation:
Institute of Food Hygiene, Faculty of Veterinary Medicine, University of Leipzig, Leipzig, Germany
D. Petroff
Affiliation:
Coordination Centre for Clinical Trials, Medical Faculty, University of Leipzig, Leipzig, Germany
D. Hasenclever
Affiliation:
Institute for Medical Informatics, Statistics and Epidemiology, Medical Faculty, University of Leipzig, Leipzig, Germany
H. Meiler
Affiliation:
Städtisches Veterinäramt, Hof, Germany
E. Lücker
Affiliation:
Institute of Food Hygiene, Faculty of Veterinary Medicine, University of Leipzig, Leipzig, Germany
*
* E-mail: gregor.makrutzki@uni-leipzig.de
Get access Rights & Permissions [Opens in a new window]

Abstract

Several methods for the detection of Trichinella in meat are legally prescribed in regulation (EC) No 2075/2005, which prescribes the magnetic stirrer method for pooled sample digestion (MSM) as the reference method. However, the MSM's multistage protocol requires several preparatory steps that seem to be accountable for the loss of larvae. Here we present a modified MSM (mMSM) based on: (1) an inversion of the optical path using inverse microscopy; and (2) a modified larval counting basin (mLCB, ‘Trichoview’). This enables one to examine samples of up to 40 ml and reduces the examination area from 72 to 10.3 cm2. Preparatory steps that might cause the loss of Trichinella larvae are eliminated from the new protocol. Correspondingly, the overall analytical time is reduced. In a direct and blinded comparison using 60 digest samples containing spiked vital Trichinella larvae (1–90 L1), both methods performed well for both small and large numbers of L1. However, 1278 of 1285 L1 (99.4%) were detected using the mMSM, while MSM recovered only 1225 L1 (95.3%). The improvement stems largely from samples with small numbers of L1: in all samples spiked with fewer than 10 L1, the recovery rate of mMSM was 100% compared to only 93% with MSM. Our data suggest that the use of the mMSM can improve the recovery rate by about 4% and therefore reduce the chances of a false-negative result in a sample containing 5 larvae by a factor of about 4.

Type
Research Papers
Information
Journal of Helminthology , Volume 88 , Issue 4 , December 2014 , pp. 453 - 458
Copyright
Copyright © Cambridge University Press 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Alban, L., Pozio, E., Boes, J., Boireau, P., Boué, F., Claes, M., Cook, A.J., Dorny, P., Enemark, H.L., van der Giessen, J., Hunt, K.R., Howell, M., Kirjusina, M., Nöckler, K., Rossi, P., Smith, G.C., Snow, L., Taylor, M.A., Theodoropoulos, G., Vallée, I., Viera-Pinto, M.M. & Zimmer, I.A. (2011) Towards a standardised surveillance for Trichinella in the European Union. Journal of Preventive Veterinary Medicine 99, 148–160.Google Scholar
Anonymous, (2004) Commission Regulation (EC) no 854/2004 of 29 April 2004 laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption. Official Journal of the European Union L139, 206–319.Google Scholar
Anonymous, (2005) Commission Regulation (EC) no 2075/2005 of December 2005 laying down specific rules on official controls for Trichinella in meat. Official Journal of the European Union L338, 60–82.Google Scholar
ECDC (European Centre for Disease Prevention and Control) (2011) Annual Epidemiological Report 2011. Reporting on 2009 surveillance data and 2010 epidemic intelligence data. Stockholm, ECDC.Google Scholar
EFSA (European Food Safety Authority Panel on Biological Hazards) (2005) Opinion of the Scientific Panel on biological hazards (BIOHAZ) on the ‘Request for an opinion on the feasibility of establishing Trichinella-free areas, and if feasible on the risk increase to public health of not examining pigs from those areas for Trichinella spp.’doi:10.2903/j.efsa.2005.277.Google Scholar
Forbes, L.B. & Gajadhar, A.A. (1999) A validated Trichinella digestion assay and an associated sampling and quality assurance system for use in testing pork and horse meat. Journal of Food Protection 62, 1308–1313.Google Scholar
Forbes, L.B., Parker, S. & Scandrett, W.B. (2005) Comparison of modified digestion assay with trichinoscopy for the detection of Trichinella in meat. Official Journal of the European Union 338, 14–16.Google Scholar
Gamble, H.R. (1999) Factors affecting the efficiency of pooled sample digestion for the recovery of Trichinella spiralis from muscle tissue. International Journal of Food Microbiology 48, 73–78.CrossRefGoogle ScholarPubMed
Gamble, H.R., Bessonov, A.S., Cuperlovic, K., Gajadhar, A.A., van Knapen, F., Nöckler, K., Schenone, H. & 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, 393–408.Google Scholar
Kapel, C.M.O. (2005) Changes in the EU legislation on Trichinella inspection – new challenges in the epidemiology. Veterinary Parasitology 139, 189–194.Google Scholar
Köhler, G. (1978) Untersuchungen mit der Stomachermethode im Vergleich zu anderen direkten Verfahren beim Nachweis der Trichinellose des Schweines. Fleischwirtschaft 59, 1258–1264.Google Scholar
Mayer-Scholl, V.A., Reckinger, S. & Nöckler, K. (2009) Ringversuch zum Nachweis von Trichinellen in Fleisch (2008). Fleischwirtschaft 89, 110–114.Google Scholar
Mayer-Scholl, V.A., Reckinger, S. & Nöckler, K. (2010) Ringversuch zum Nachweis von Trichinellen in Fleisch (2009). Fleischwirtschaft 90, 174–178.Google Scholar
Mayer-Scholl, V.A., Reckinger, S. & Nöckler, K. (2011) Ringversuch zum Nachweis von Trichinellen in Fleisch (2010). Fleischwirtschaft 91, 127–130.Google Scholar
Mayer-Scholl, V.A., Reckinger, S. & Nöckler, K. (2012) Ringversuch zum Nachweis von Trichinellen in Fleisch (2011)Available at websitehttp://www.bfr.bund.de/cm/343/ringversuch-zum-nachweis-von-trichinellen-in-fleisch-2011.pdf (accessed accessed 15 January 2013).Google Scholar
Murrell, K.D. & Pozio, E. (2011) Worldwide occurrence and impact of human trichinellosis, 1986–2009. Emerging Infectious Diseases 17, 2194–2202.Google Scholar
Prost, E. & Nowakowski, Z. (1990) Sicherer Nachweis von Trichinella spiralis mit der Digestionsmethode. Fleischwirtschaft 70, 593–595.Google Scholar
Riehn, K., Hasenclever, D., Petroff, D., Nöckler, K., Mayer-Scholl, A., Makrutzki, G. & Lücker, E. (2013) Trichinella detection: identification and statistical evaluation of sources of error in the magnetic stirrer method for pooled sample digestion. Veterinary Parasitology 94, 106–109.Google Scholar
Rossi, P. & Pozio, E. (2008) Guidelines for the detection of Trichinella larvae at the slaughterhouse in a quality assurance system. Annali dell Istituto Superiore di Sanita 44, 195–199.Google Scholar