Artificial Digestion

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E Pozio - One of the best experts on this subject based on the ideXlab platform.

  • the occurrence of trichinella species in the cougar puma concolor couguar from the state of colorado and other regions of north and south america
    Journal of Helminthology, 2017
    Co-Authors: Mason V Reichard, Maria Interisano, Gianluca Marucci, Kenneth A Logan, Marc Criffield, Jennifer E Thomas, J M Paritte, D M Messerly, E Pozio
    Abstract:

    : Trichinella species are zoonotic nematodes that infect wild carnivores and omnivores throughout the world. We examined the prevalence and species of Trichinella infections in cougars (Puma concolor couguar) from Colorado, United States. Tongues from cougars were examined by pepsin-HCl Artificial Digestion to detect Trichinella spp. larvae. The species or genotype of individual worms was identified by multiplex polymerase chain reaction (PCR). Trichinella spp. larvae were detected in 17 of 39 cougars (43.6% (28.7-59.5%)). Five of the cougars (12.8%) were infected with T. murrelli, 3 (7.7%) were infected with T. pseudospiralis, and 1 (2.6%) had Trichinella genotype T6. Trichinella spp. larvae from eight cougars were not identified at the species level, due to degraded DNA. The high prevalence of Trichinella spp. in cougars from Colorado and reports of the parasite in other populations of Puma spp. suggest that this large predator is a key mammalian reservoir.

  • Comparison of three Artificial Digestion methods for detection of non-encapsulated Trichinella pseudospiralis larvae in pork.
    Veterinary parasitology, 2008
    Co-Authors: K Nöckler, E Pozio, S Reckinger, I Szabó, C Maddox-hyttel, J Van Der Giessen, I Vallée, P Boireau
    Abstract:

    In a ring trial involving five laboratories (A, B, C, D, and E), three different methods of Artificial Digestion were compared for the detection of non-encapsulated Trichinella pseudospiralis larvae in minced meat. Each sample panel consisted of ten 1g minced pork samples. All samples in each panel were derived from a bulk meat preparation with a nominal value of either 7 or 17 larvae per g (lpg). Samples were tested for the number of muscle larvae using the magnetic stirrer method (labs A, B, and E), stomacher method (lab B), and Trichomatic 35 (labs C and D). T. pseudospiralis larvae were found in all 120 samples tested. For samples with 7 lpg, larval recoveries were significantly higher using the stomacher method versus the magnetic stirrer method, but there were no significant differences for samples with 17 lpg. In comparing laboratory results irrespective of the method used, lab B detected a significantly higher number of larvae than lab E for samples with 7 lpg, and lab E detected significantly less larvae than labs A, B, and D in samples with 17 lpg. The lowest overall variation for quantitative results (i.e. larval recoveries which were outside the tolerance range) was achieved by using the magnetic stirrer method (22%), followed by the stomacher method (25%), and Trichomatic 35 (30%). Results revealed that T. pseudospiralis larvae in samples with a nominal value of 7 and 17 lpg can be detected by all three methods of Artificial Digestion.

  • trichinella spiralis and trichinella pseudospiralis mixed infection in a wild boar sus scrofa of germany
    Veterinary Parasitology, 2006
    Co-Authors: Karsten Nöckler, S Reckinger, E Pozio
    Abstract:

    A wild boar (Sus scrofa) from the island Usedom in Mecklenburg-Western Pomerania (north-east Germany) was detected as Trichinella-positive during routine meat inspection. Encapsulated and non-encapsulated larvae were detected in the muscle tissue by trichinoscopy. In the diaphragm, 922 larvae per g were detected by Artificial Digestion. Muscle larvae displayed two different sizes of about 700 and 1100 μm. By a multiplex PCR analysis, larvae with a large size were identified as Trichinella spiralis, whereas those of a smaller size were identified as Trichinella pseudospiralis. This is the first finding of a mixed infection of T. spiralis and T. pseudospiralis in a naturally infected animal and it supports the tendency of more frequent detection of the non-encapsulated species T. pseudospiralis in Europe.

  • Sylvatic trichinellosis in Texas.
    Parasite, 2001
    Co-Authors: Danny B. Pence, G. La Rosa, F. Mancini Barbieri, M. Amati, Adriano Casulli, E Pozio
    Abstract:

    Summary: There are no published reports of domestic or sylvatic trichinellosis in Texas. The aim of the present survey was to determine the presence of Trichinella species in selected representative species of potential wildlife reservoirs in southern Texas. In 1998-99, tongues of 211 wild mammals were collected in southern Texas: 154 coyotes (Canis latrans), three bobcats (Lynx rufus), 32 racoons (Procyon lotor), 1 3 opossum (Didelphis marsupialis), four ocelots (Leopardus pardalis) and five wild boars (Sus scrofa). Presence of Trichinella sp. larvae was investigated by Artificial Digestion and larvae of positive samples were identified at the species level by a multiple-polymerase chain reaction analysis. Nine (5.8 %) coyotes had trichinellosis ; in the muscles of seven of these coyotes, the larvae were identified as Trichinella murrelli. This is the first report of sylvatic trichinellosis in Texas.

Edoardo Pozio - One of the best experts on this subject based on the ideXlab platform.

  • A preliminary survey of Trichinella spp. in pigs raised under controlled housing conditions in Colombia: 2014-2016.
    Parasite, 2018
    Co-Authors: Jenny J. Chaparro-gutiérrez, Maria Angeles Gómez-morales, Edoardo Pozio, Anderson López, Jaime Mejía, Corina Zambrano, Diego Piedrahita, David Villar
    Abstract:

    A preliminary survey of Trichinella spp. infection was conducted in Colombian swine herds between 2014 and 2016. A total of 1,773 pigs reared on farms under controlled housing conditions and processed in 34 slaughterhouses were tested either by the Artificial Digestion of pooled muscle samples (n = 1,173) or by serology (n = 600). In addition, 550 rats trapped on 29 swine farm premises were also tested by Artificial Digestion. No positive pig samples were detected. Similarly, no Trichinella spp. muscle larvae were detected in rats. These results are in agreement with the lack of historical Trichinella infection reports in domestic and wild animals and humans in Colombia. However, a more extensive epidemiological investigation and a continuous surveillance program are needed to continue declaring swine herds in Colombia free of Trichinella infection.

  • UV-press method versus Artificial Digestion method to detect Anisakidae L3 in fish fillets: Comparative study and suitability for the industry
    Fisheries Research, 2018
    Co-Authors: Maria Angeles Gómez-morales, Cristina Martínez Castro, Marco Lalle, Rosa Fernández, Patrizio Pezzotti, Elvira Abollo, Edoardo Pozio
    Abstract:

    Abstract To screen the presence of Anisakidae third stage larvae (L3) in fish, fast methods such as the visual inspection and candling have been widely used by the industry over the last 50 years, and they are regulated by the European Parliament and the Council. These methods are ineffective to detect L3 embedded in fish muscles, consequently alternative methods, such as the Artificial Digestion (AD) and the UV-Press (UVP) are increasingly applied, but their performance needs to be evaluated. The aims of the present work were: 1) to compare the performance of AD and UVP. methods by a Ring Trial (RT) involving highly experienced laboratories; and 2) to evaluate the potential transferability of the best performing method to the industry by a collaborative study involving industrial partners (β-testing). For RT, each participating laboratory (n = 5) received 6 samples of 100 g of fish fillets spiked with 1 L3 (1 sample), 3 L3 (2 samples), 7 L3 (2 samples), and a negative control sample (without L3). In each positive sample, there were live Anisakis pegreffii L3 collected from a naturally infected fish. The result evaluation was based on the agreement between the number of reported and the number of spiked L3. No false positive sample was detected. The L3 number detected by the UVP method showed higher (90%) level of agreement with the number of spiked L3 than the number of L3 detected by the AD method (83.3%); however, no significant difference in terms of accuracy (p = 0.32) was detected when the two methods were compared. Moreover, considering only the presence/absence of L3 in the samples, the UVP reached 100% of accuracy and 100% of sensitivity; whereas, AD showed 98% of accuracy and 96% of sensitivity. The variability of the UVP method was lower than that of the AD method, indicating a better reproducibility. On the basis of the RT results, the UVP method was selected for the β-testing. Each industrial partner (n = 3) received 15 samples of 100 g of fish fillet spiked with 1 L3 (2 samples), 2 L3 (2 samples), 3 L3 (2 samples), 4 L3 (2 samples), 5 L3 (2 samples) and 6 L3 (2 samples) and three negative control samples (without L3). The number of L3 counted in 34 out of 45 samples (75.6%) by the UVP method was in agreement with the number of spiked L3. One company reached 93.3% of agreement; whereas the other two companies reached an agreement of 66.7%. Two false negative results were found; whereas, no false positive results were obtained. Moreover, at the industrial level, considering only the presence/absence of larvae in the samples, the UVP reached 97% of accuracy, 94.4% of sensitivity, and 100% of specificity. However, the UVP method, in spite of its accuracy, needs further investigations to provide new time −temperature combinations that could allow a reduction of the testing time and its integration in the fishing deck.

  • Detection of Trichinella murrelli in coyotes (Canis latrans) from Oklahoma and North Texas.
    Veterinary Parasitology, 2011
    Co-Authors: Mason V Reichard, Kathryn E Tiernan, Kelsey L Paras, Maria Interisano, Michael H Reiskind, Roger J Panciera, Edoardo Pozio
    Abstract:

    We determined the prevalence and mean intensity of Trichinella sp. infection in coyotes from six counties in Oklahoma and one in northern Texas. Tongues from 77 coyotes were examined using histology and Artificial tissue Digestion. Histological examination showed a prevalence of 3.9% (3 of 77) whereas the prevalence was 6.5% (5 of 77) based on Artificial Digestion of 5.0 g of muscle from coyote tongues. One sample was positive for Trichinella sp. on histology but negative by Artificial Digestion. Combining data from both diagnostic techniques showed that six of 77 (7.8%) coyotes were infected with Trichinella spp. The mean intensity of Trichinella sp. larvae ranged from 0.2 to 66.2 with an average of 16.0 larvae per gram (LPG) of tongue. Genotyping results demonstrated that the coyotes were infected with Trichinella murrelli. This is the first report of T. murrelli infection in coyotes in Oklahoma. T. murrelli had previously been isolated from coyotes in Texas.

  • Trichinella spp. infection in horses of Romania: serological and parasitological survey.
    Veterinary Parasitology, 2009
    Co-Authors: Robert Blaga, Vasile Cozma, Călin Mircea Gherman, Edoardo Pozio, Carmen M Cretu, Alina Draghici, Karsten Noeckler, Christian M O Kapel, Ion Dida, Pascal Boireau
    Abstract:

    Herbivorous animals are usually, by virtue of their diet, outside the major transmission cycles of Trichinella spp. However, since 1975, the year of the first report of human trichinellosis caused by the consumption of infected horse meat, the domestic horse has appeared as a novel vector of Trichinella spp. infection to humans, with 15 outbreaks documented in France and Italy. Romania, one of the main countries exporting horses into the European Union (EU), experienced a dramatic increase of Trichinella spp. infection in both domestic pigs and humans in the 1990s. Some Trichinella spiralis-infected horses were exported to the EU during this period. The aim of this study was to evaluate the prevalence of Trichinella spp. infections in horses from Romania using both direct and indirect tests. Of 3000 serum samples tested in 2001, none were positive by ELISA using three different Trichinella antigens (crude; excretory/secretory, ES; stg-BSA antigens). Of 2992 serum samples tested in 2002, 17 (0.56%) showed optical density values higher than the cut-off in an ELISA using ES antigens and one was confirmed by western blot (WB). Four of the 17 ELISA positive horses, including the horse with a confirmed serology by WB, were subjected for intensive meat examination at slaughter, but no Trichinella spp. larvae were detected. Further, no Trichinella spp. larvae were detected by trichinelloscopy and Artificial Digestion of 25,838 horses slaughtered in Alexandria and Timisoara between 2001 and 2004. The false positive results obtained by serology confirm the previous work on the unreliability of serology for detection of Trichinella spp. infection in horses. Furthermore, the lack of detection of Trichinella spp. infected horses by Artificial Digestion, suggests a very low prevalence of infection in horses in Romania.

  • Trichinella T6 and Trichinella nativa in Wolverines (Gulo gulo) from Nunavut, Canada
    Parasitology Research, 2008
    Co-Authors: Mason V Reichard, Luigi Torretti, Timothy A. Snider, Jason M. Garvon, Gianluca Marucci, Edoardo Pozio
    Abstract:

    Infection of Trichinella spp. is common among animals in the Canadian Arctic. We determined the prevalence of Trichinella spp. infection in wolverines ( Gulo gulo ) from Nunavut, Canada. Diaphragms from 41 wolverines were examined by Artificial Digestion. Trichinella spp. larvae were detected in 36 (87.8%) examined animals. Trichinella T6 was detected in 33 (91.7%), Trichinella nativa in only one (2.8%), and a mixed Trichinella T6 and T. nativa infections were detected in two (5.6%) wolverines. This is the first report of Trichinella spp. infection in wolverines from Nunavut and the first report of sympatric Trichinella T6 and T. nativa in any host. The high prevalence of Trichinella spp. infection in combination with the natural history of wolverines suggests that the mustelid may be a key species in the natural cycle of these parasites in Arctic and Subarctic areas.

Karsten Nöckler - One of the best experts on this subject based on the ideXlab platform.

  • A study on the suitability of inactivated Trichinella spiralis larvae for proficiency samples.
    Veterinary Parasitology, 2013
    Co-Authors: Anne Mayer-scholl, S Reckinger, Karsten Nöckler
    Abstract:

    The consumption of raw or undercooked Trichinella infected meat, especially pork and horse meat, can have important implications for public health. Therefore each animal carcass from a Trichinella susceptible species intended for human consumption must be examined for Trichinella. Laboratories carrying out testing of official control samples must undergo a quality assurance program and should regularly participate in proficiency testing schemes. To date, Trichinella proficiency samples are prepared with live larvae, which, as a level 2 pathogen, require specific shipping and disinfection procedures. Therefore, the suitability of using inactivated Trichinella larvae as proficiency samples was tested. We found that Trichinella larvae treated with 2% formaldehyde for 24h had lost their infectivity and showed a comparable recovery rate to naive larvae after Artificial Digestion, albeit with a prolonged sedimentation time.

  • Surveillance systems for status monitoring of Trichinella-free declared pig farms: concepts and their confidence for freedom from disease
    Berliner und Munchener tierarztliche Wochenschrift, 2012
    Co-Authors: S. Gross, Matthias Greiner, Anne Mayer-scholl, Annemarie Käsbohrer, Lüppo Ellerbroek, Karsten Nöckler, Christine Müller-graf
    Abstract:

    Trichinella surveillance data in Germany show for indoor housed pigs hardly any cases. Nevertheless, obligatory testing is in place for each slaughtered pig. According to EU legislation systematic Trichinella testing can be replaced by a risk-based surveillance system if the risk of Trichinella infection in fattening pigs is negligible. The probability to detect a positive herd (herd sensitivity) was taken as an indicator for the effectiveness of the surveillance. Four different diagnostic methods: a) Digestion method, b) E/S-ELISA, c) Western Blot, and d) ELISA sequentially combined with Western Blot, were compared regarding herd sensitivity and specificity for different herd and sample sizes and different levels of prevalence. In a further step three potential surveillance systems were compared with regard to their suitability for herd classification: (i) classical Trichinella examination by Artificial Digestion method, (ii) ELISA screening followed by classical Trichinella examination and (iii) ELISA screening followed by Western Blot. Results show that: 1) testing by the Artificial Digestion method (i) provides only low sensitivity of detection for positive herds at present levels of prevalence despite perfect specificity. 2) The ELISA alone provides a high sensitivity of detection even at low sample sizes but at the cost of a very low herd specificity, converging towards zero at increasing sample sizes. In surveillance system (ii), a large number of farms would still need to be tested with the classical Digestion method, as they would be misclassified as positive by the ELISA. 3) The Western Blot as well as ELISA screening followed by Western Blot offer a high probability of correct herd classification. The latter diagnostic system appears to be the most suitable for a risk based surveillance (iii) and provides--despite reduced sample sizes--a higher probability for a correct herd classification than the traditional Trichinella examination.

  • trichinella spiralis and trichinella pseudospiralis mixed infection in a wild boar sus scrofa of germany
    Veterinary Parasitology, 2006
    Co-Authors: Karsten Nöckler, S Reckinger, E Pozio
    Abstract:

    A wild boar (Sus scrofa) from the island Usedom in Mecklenburg-Western Pomerania (north-east Germany) was detected as Trichinella-positive during routine meat inspection. Encapsulated and non-encapsulated larvae were detected in the muscle tissue by trichinoscopy. In the diaphragm, 922 larvae per g were detected by Artificial Digestion. Muscle larvae displayed two different sizes of about 700 and 1100 μm. By a multiplex PCR analysis, larvae with a large size were identified as Trichinella spiralis, whereas those of a smaller size were identified as Trichinella pseudospiralis. This is the first finding of a mixed infection of T. spiralis and T. pseudospiralis in a naturally infected animal and it supports the tendency of more frequent detection of the non-encapsulated species T. pseudospiralis in Europe.

S Reckinger - One of the best experts on this subject based on the ideXlab platform.

  • A study on the suitability of inactivated Trichinella spiralis larvae for proficiency samples.
    Veterinary Parasitology, 2013
    Co-Authors: Anne Mayer-scholl, S Reckinger, Karsten Nöckler
    Abstract:

    The consumption of raw or undercooked Trichinella infected meat, especially pork and horse meat, can have important implications for public health. Therefore each animal carcass from a Trichinella susceptible species intended for human consumption must be examined for Trichinella. Laboratories carrying out testing of official control samples must undergo a quality assurance program and should regularly participate in proficiency testing schemes. To date, Trichinella proficiency samples are prepared with live larvae, which, as a level 2 pathogen, require specific shipping and disinfection procedures. Therefore, the suitability of using inactivated Trichinella larvae as proficiency samples was tested. We found that Trichinella larvae treated with 2% formaldehyde for 24h had lost their infectivity and showed a comparable recovery rate to naive larvae after Artificial Digestion, albeit with a prolonged sedimentation time.

  • Comparison of three Artificial Digestion methods for detection of non-encapsulated Trichinella pseudospiralis larvae in pork.
    Veterinary parasitology, 2008
    Co-Authors: K Nöckler, E Pozio, S Reckinger, I Szabó, C Maddox-hyttel, J Van Der Giessen, I Vallée, P Boireau
    Abstract:

    In a ring trial involving five laboratories (A, B, C, D, and E), three different methods of Artificial Digestion were compared for the detection of non-encapsulated Trichinella pseudospiralis larvae in minced meat. Each sample panel consisted of ten 1g minced pork samples. All samples in each panel were derived from a bulk meat preparation with a nominal value of either 7 or 17 larvae per g (lpg). Samples were tested for the number of muscle larvae using the magnetic stirrer method (labs A, B, and E), stomacher method (lab B), and Trichomatic 35 (labs C and D). T. pseudospiralis larvae were found in all 120 samples tested. For samples with 7 lpg, larval recoveries were significantly higher using the stomacher method versus the magnetic stirrer method, but there were no significant differences for samples with 17 lpg. In comparing laboratory results irrespective of the method used, lab B detected a significantly higher number of larvae than lab E for samples with 7 lpg, and lab E detected significantly less larvae than labs A, B, and D in samples with 17 lpg. The lowest overall variation for quantitative results (i.e. larval recoveries which were outside the tolerance range) was achieved by using the magnetic stirrer method (22%), followed by the stomacher method (25%), and Trichomatic 35 (30%). Results revealed that T. pseudospiralis larvae in samples with a nominal value of 7 and 17 lpg can be detected by all three methods of Artificial Digestion.

  • trichinella spiralis and trichinella pseudospiralis mixed infection in a wild boar sus scrofa of germany
    Veterinary Parasitology, 2006
    Co-Authors: Karsten Nöckler, S Reckinger, E Pozio
    Abstract:

    A wild boar (Sus scrofa) from the island Usedom in Mecklenburg-Western Pomerania (north-east Germany) was detected as Trichinella-positive during routine meat inspection. Encapsulated and non-encapsulated larvae were detected in the muscle tissue by trichinoscopy. In the diaphragm, 922 larvae per g were detected by Artificial Digestion. Muscle larvae displayed two different sizes of about 700 and 1100 μm. By a multiplex PCR analysis, larvae with a large size were identified as Trichinella spiralis, whereas those of a smaller size were identified as Trichinella pseudospiralis. This is the first finding of a mixed infection of T. spiralis and T. pseudospiralis in a naturally infected animal and it supports the tendency of more frequent detection of the non-encapsulated species T. pseudospiralis in Europe.

Pascal Boireau - One of the best experts on this subject based on the ideXlab platform.

  • Seroprevalance of Trichinella Spp. in Wild Boars (Sus Scrofa) from Bihor County, Western Romania.
    Helminthologia, 2020
    Co-Authors: Z. Boros, Pascal Boireau, Călin Mircea Gherman, Isabelle Vallée, L. C. Panait, A. Chevillot, Vasile Cozma
    Abstract:

    The wild boar (Sus scrofa) has a wide geographical distribution and can be an important source of Trichinella spp. infection in humans in Romania. The objective of this study was to identify the presence of Trichinella spp. in the wild boar population in Bihor County, Romania. Eighty four plasma and diaphragm samples, collected from wild boars, were included in this study. Artificial Digestion, ELISA and Western blot were performed on these specimens. All diaphragm samples were negative for Trichinella larvae in Artificial Digestion, while in ELISA, 54 (64.2 %) plasma samples were positive and 6 (7.1 %) plasma samples were doubtful. Western blot was performed on 26 plasma samples from which only 6 (23.0 %) gave a positive result. Serological evidences indicate the presence of Trichinella spp. in wild boars from western Romania. Therefore, human consumers might be at risk to ingest Trichinella larvae, even in low numbers.

  • Trichinella spp. infection in horses of Romania: serological and parasitological survey.
    Veterinary Parasitology, 2009
    Co-Authors: Robert Blaga, Vasile Cozma, Călin Mircea Gherman, Edoardo Pozio, Carmen M Cretu, Alina Draghici, Karsten Noeckler, Christian M O Kapel, Ion Dida, Pascal Boireau
    Abstract:

    Herbivorous animals are usually, by virtue of their diet, outside the major transmission cycles of Trichinella spp. However, since 1975, the year of the first report of human trichinellosis caused by the consumption of infected horse meat, the domestic horse has appeared as a novel vector of Trichinella spp. infection to humans, with 15 outbreaks documented in France and Italy. Romania, one of the main countries exporting horses into the European Union (EU), experienced a dramatic increase of Trichinella spp. infection in both domestic pigs and humans in the 1990s. Some Trichinella spiralis-infected horses were exported to the EU during this period. The aim of this study was to evaluate the prevalence of Trichinella spp. infections in horses from Romania using both direct and indirect tests. Of 3000 serum samples tested in 2001, none were positive by ELISA using three different Trichinella antigens (crude; excretory/secretory, ES; stg-BSA antigens). Of 2992 serum samples tested in 2002, 17 (0.56%) showed optical density values higher than the cut-off in an ELISA using ES antigens and one was confirmed by western blot (WB). Four of the 17 ELISA positive horses, including the horse with a confirmed serology by WB, were subjected for intensive meat examination at slaughter, but no Trichinella spp. larvae were detected. Further, no Trichinella spp. larvae were detected by trichinelloscopy and Artificial Digestion of 25,838 horses slaughtered in Alexandria and Timisoara between 2001 and 2004. The false positive results obtained by serology confirm the previous work on the unreliability of serology for detection of Trichinella spp. infection in horses. Furthermore, the lack of detection of Trichinella spp. infected horses by Artificial Digestion, suggests a very low prevalence of infection in horses in Romania.

  • first identification of trichinella sp in golden jackal canis aureus in romania
    Journal of Wildlife Diseases, 2008
    Co-Authors: Robert Blaga, D Seucom, Vasile Cozma, Călin Mircea Gherman, Pascal Boireau
    Abstract:

    Larvae of Trichinella sp. were identified in a golden jackal (Canis aureus) from Romania by both trichinelloscopy and Artificial Digestion. The larvae were identified as Trichinella britovi using a multiplex polymerase chain reaction biotyping method. This is the first report of Trichinella sp. in a jackal in Romania.

  • Use of proficiency samples to assess diagnostic laboratories in France performing a Trichinella Digestion assay.
    Journal of Food Protection, 2007
    Co-Authors: Isabelle Vallée, Alvin A. Gajadhar, Brad Scandrett, Pauline Macé, Lorry B. Forbes, Benoit Durand, Pascal Boireau
    Abstract:

    Routine diagnosis of animal trichinellosis for food safety and trade relies on a method of Artificial Digestion to free Trichinella muscle larvae from meat for subsequent identification by microscopy. As part of a quality control system, the French National Reference Laboratory (NRL) initiated ring trials to determine the sensitivity of the test performed in the 72 routine diagnostic laboratories in France. A method was devised to obtain calibrated meat samples containing known numbers of capsules with Trichinella spiralis muscle larvae. This method was based on an incomplete Artificial Digestion of Trichinella-infected mice carcasses to allow the collection of intact Trichinella capsules. Capsules were placed into a meatball of 100 ± 2 g of pork and horsemeat to produce proficiency samples. Three categories of samples were prepared: small (3 to 5 capsules), medium (7 to 10), and large (12 to 15). The sensitivity was expressed as the percentage of muscle larvae recovered from each proficiency sample. Repr...

  • Use of proficiency samples to assess diagnostic laboratories in France performing a Trichinella Digestion assay
    Journal of Food Protection, 2007
    Co-Authors: Isabelle Vallée, Brad Scandrett, Pauline Macé, Benoit Durand, Lorry Forbes, Alvin Gajadhar, Pascal Boireau
    Abstract:

    Routine diagnosis of animal trichinellosis for food safety and trade relies on a method of Artificial Digestion to free Trichinella muscle larvae from meat for subsequent identification by microscopy. As part of a quality control system, the French National Reference Laboratory (NRL) initiated ring trials to determine the sensitivity of the test performed in the 72 routine diagnostic laboratories in France. A method was devised to obtain calibrated meat samples containing known numbers of capsules with Trichinella spiralis muscle larvae. This method was based on an incomplete Artificial Digestion of Trichinella-infected mice carcasses to allow the collection of intact Trichinella capsules. Capsules were placed into a meatball of 100 +/- 2 g of pork and horsemeat to produce proficiency samples. Three categories of samples were prepared: small (3 to 5 capsules), medium (7 to 10), and large (12 to 15). The sensitivity was expressed as the percentage of muscle larvae recovered from each proficiency sample. Reproducibility was tested with ring trials organized between two NRLs (France and Canada), and a reference sensitivity of 84.9% was established. National ring trials were then organized in France, with the 72 routine diagnostic laboratories each receiving four proficiency samples per session. After five sessions, an improvement in the digest test sensitivity was observed. Results at the fifth session indicated sensitivities of 78.60% +/- 23.70%, 81.19% +/- 19.59%, and 80.52% +/- 14.71% muscle larvae for small, medium, and large samples, respectively. This study supports the use of proficiency samples to accurately evaluate the performance of routine diagnostic laboratories that conduct Digestion tests for animal trichinellosis diagnosis.