Factors affecting the efficiency of pooled sample digestion for the recovery of Trichinella spiralis from muscle tissue

1999 ◽  
Vol 48 (1) ◽  
pp. 73-78 ◽  
Author(s):  
H.R Gamble
Keyword(s):  
Muscle Tissue ◽  
Trichinella Spiralis ◽  
Factors Affecting ◽  
Sample Digestion ◽  
Pooled Sample

Detection of Trichinellosis in Pigs by Artificial Digestion and Enzyme Immunoassay

1996 ◽  
Vol 59 (3) ◽  
pp. 295-298 ◽  
Author(s):  
H. R. GAMBLE
Keyword(s):  
Enzyme Immunoassay ◽  
Trichinella Spiralis ◽  
Federal Regulations ◽  
Testing Methods ◽  
Digestion Method ◽  
Inspection Method ◽  
Sample Digestion ◽  
Artificial Digestion ◽  
Digestion Methods ◽  
Pooled Sample

The objective of this study was to demonstrate the reliability of current and proposed methods for the inspection of swine and other species for infection with the parasite Trichinella spiralis. Five groups of pigs were infected with doses of 2500, 500, 100, 50, and 20 T. spiralis larvae to establish moderate and low-level infections. Pigs were bled periodically during the study for samples to be tested by enzyme immunoassay (EIA). At the conclusion of the study, pigs were slaughtered and tissues collected for analyses of worm burdens and for comparison of digestion testing methods. Comparisons of pooled sample digestion methods were made using inspection methods prescribed by European Union Directives and the USDA, Code of Federal Regulations. Pooled sample digestion testing using 1-g samples was effective for detecting pigs with larval densities of >10 larvae per gram (LPG) of tissue but only partially effective for pigs with infections of <3 LPG. Pooled sample digestion testing using 5-g samples detected all pigs with infection levels >1 LPG. The EIA detected all T. spiralis-infected pigs, but did not detect infections in some pigs until 49 days after inoculation. These results demonstrate that the pooled sample digestion method using a 5-g sample size is the most effective inspection method for reducing the risk of human exposure to T. spiralis in pork.

scholarly journals Prevalence of Trichinella britovi in muscle tissue samples from foxes from Podkarpackie Voivodeship, Poland

2016 ◽  
Vol 72 (11) ◽  
pp. 681-685
Author(s):  
Jan Wiśniewski ◽  
Zbigniew Bełkot ◽  
Blanka Orłowska ◽  
Janusz Bogdan ◽  
Mirosław Welz ◽  
...  
Keyword(s):  
Muscle Tissue ◽  
Vulpes Vulpes ◽  
Tissue Sample ◽  
Reference Method ◽  
Red Foxes ◽  
Tissue Samples ◽  
Sample Digestion ◽  
Two Samples ◽  
Pooled Sample

The aim of the study was to detect Trichinella larvae in muscle tissue samples of red foxes (Vulpes vulpes) living in Podkarpackie Voivodeship and to determine their species composition. Muscle tissue samples had been collected from a total number of 139 red foxes culled during 2011-2012 in Podkarpackie Voivodeship and studied for the presence of Trichinella. Ten-gram muscle tissue samples were collected from each animal, combined into a fifty-gram pooled sample and then tested for the presence of Trichinella by the reference method for pooled sample digestion with a magnetic stirrer. Whenever a pooled sample gave a positive result, the suspected animals were resampled, and each ten-gram muscle tissue sample was tested separately. Trichinella larvae were identified to species by Multiplex PCR. The studies revealed the presence of one Trichinella larva in a pooled sample composed of five individual samples from foxes culled in the following locations: Leszczowate in Bieszczadzki District (one sample), Szczutków in Lubaczowski District (two samples), Podemszczyzna in Lubaczowski District (one sample) and Strzyżów in Strzyżowski District (one sample). No Trichinella larvae were detected during the test carried out on individual samples that composed the pooled sample. Numerous Trichinella larvae were found in samples from 4 foxes coming from Berezka and Glinka in Leski District, Dybawka in Przemyski District, and Golcowa in Brzozowski District. All Trichinella larvae isolated from the foxes belonged to the species T. britovi. The study confirmed the role of foxes as an important vector of Trichinella larvae circulating in the habitat of humans and animals. T. britovi is a species occurring most frequently in the fox population.

Trichinella detection: Identification and statistical evaluation of sources of error in the magnetic stirrer method for pooled sample digestion

2013 ◽  
Vol 194 (2-4) ◽  
pp. 106-109 ◽  
Author(s):  
Katharina Riehn ◽  
Dirk Hasenclever ◽  
David Petroff ◽  
Karsten Nöckler ◽  
Anne Mayer-Scholl ◽  
...  
Keyword(s):  
Statistical Evaluation ◽  
Sample Digestion ◽  
Sources Of Error ◽  
Magnetic Stirrer ◽  
Pooled Sample

scholarly journals Accumulation of α-2,6-sialyoglycoproteins in the muscle sarcoplasm due to Trichinella sp. invasion

2019 ◽  
Vol 14 (1) ◽  
pp. 470-481
Author(s):  
Rositsa Milcheva ◽  
Pavol Janega ◽  
Peter Celec ◽  
Svetlozara Petkova ◽  
Zuzana Hurniková ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Tissue ◽  
Nurse Cell ◽  
Trichinella Spiralis ◽  
Helix Pomatia ◽  
Skeletal Muscle Tissue ◽  
Vicia Villosa ◽  
Subsequent Formation ◽  
Arachis Hypogea ◽  
Adaptive Capabilities

AbstractThe sialylation of the glycoproteins in skeletal muscle tissue is not well investigated, even though the essential role of the sialic acids for the proper muscular function has been proven by many researchers. The invasion of the parasitic nematode Trichinella spiralis in the muscles with subsequent formation of Nurse cell-parasite complex initiates increased accumulation of sialylated glycoproteins within the affected area of the muscle fiber. The aim of this study is to describe some details of the α-2,6-sialylation in invaded muscle cells. Asynchronous invasion with infectious T. spiralis larvae was experimentally induced in mice. The areas of the occupied sarcoplasm were reactive towards α-2,6-sialic acid specific Sambucus nigra agglutinin during the whole process of transformation to a Nurse cell.The cytoplasm of the developing Nurse cell reacted with Helix pomatia agglutinin, Arachis hypogea agglutinin and Vicia villosa lectin-B4 after neuraminidase pretreatment.Up-regulation of the enzyme ST6GalNAc1 and down-regulation of the enzyme ST6GalNAc3 were detected throughout the course of this study. The results from our study assumed accumulation of sialyl-Tn-Ag, 6`-sialyl lactosamine, SiA-α-2,6-Gal-β-1,3-GalNAc-α-Ser/Thr and Gal-β-1,3-GalNAc(SiA-α-2,6-)-α-1-Ser/Thr oligosaccharide structures into the occupied sarcoplasm. Further investigations in this domain will develop the understanding about the amazing adaptive capabilities of skeletal muscle tissue.

Sensitivity of Artificial Digestion and Enzyme Immunoassay Methods of Inspection for Trichinae in Pigs

1998 ◽  
Vol 61 (3) ◽  
pp. 339-343 ◽  
Author(s):  
H. R. GAMBLE
Keyword(s):  
Sample Size ◽  
Enzyme Immunoassay ◽  
Trichinella Spiralis ◽  
Serological Testing ◽  
Digestion Method ◽  
Artificial Digestion ◽  
Eu Directives ◽  
Digestion Methods ◽  
Pooled Sample ◽  
The Eu

Forty-seven pigs were infected with varying doses of Trichinella spiralis and tested for evidence of infection by serology, using an enzyme immunoassay (EIA), and by artificial digestion methods. Using a 1-g sample, as prescribed in accordance with European Union (EU) directives, the sensitivity of the pooled-sample artificial digestion method was between three and five larvae per gram (LPG) of tissue. Using a 5-g sample size, in accordance with methods described in the U.S. Code of Federal Regulations, and as required for the inspection of horses exported to the EU, the sensitivity of the test was increased to approximately 1 LPG. Serological testing by EIA detected pigs with as few as 0.02 LPG, but detection times varied from 4 to 8 weeks after infection. Mean postinoculation times for detection by serology were 32 to 42 days. On the basis of these results, it is clear that digestion testing using a 5-g sample size is the only method of those tested here that is completely reliable for detection of trichinae infection at a level that will protect public health. Both digestion testing using a 1-g sample and EIA have drawbacks. However, EIA remains a highly effective tool for epidemiological purposes and for monitoring trichinae infection on the farm.

Development of A Real-Time PCR Assay for the Detection of Trichinella Spiralis in Muscle Tissue of Swine and Derivatives

2015 ◽  
Vol 36 (2) ◽  
pp. 282-287 ◽  
Author(s):  
S. Quintana ◽  
M. Recavarren ◽  
E. Scialfa ◽  
I. Viera ◽  
M. Rivero ◽  
...  
Keyword(s):  
Real Time ◽  
Muscle Tissue ◽  
Real Time Pcr ◽  
Trichinella Spiralis ◽  
Pcr Assay

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

2013 ◽  
Vol 88 (4) ◽  
pp. 453-458 ◽  
Author(s):  
G. Makrutzki ◽  
K. Riehn ◽  
A. Hamedy ◽  
D. Petroff ◽  
D. Hasenclever ◽  
...  
Keyword(s):  
Recovery Rate ◽  
False Negative ◽  
Reference Method ◽  
Optical Path ◽  
False Negative Result ◽  
Sample Digestion ◽  
Counting Technique ◽  
Large Numbers ◽  
Analytical Time ◽  
Pooled Sample

AbstractSeveral 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.

BLOOD SERUM AND TISSUE FREE AMINO ACID CONCENTRATIONS OF GILTS FED DL- OR L-METHIONINE

1972 ◽  
Vol 52 (1) ◽  
pp. 155-161
Author(s):  
J. E. KNIPFEL ◽  
B. D. OWEN ◽  
D. A. CHRISTENSEN
Keyword(s):  
Amino Acid ◽  
Blood Serum ◽  
Free Amino Acid ◽  
Muscle Tissue ◽  
Free Amino ◽  
Short Term ◽  
Factors Affecting ◽  
Amino Acid Requirements ◽  
Amino Acid Concentrations ◽  
Term Storage

A semipurified ration containing 0.55% DL-methionine was fed to two gilts from each of two litters, and 0.55% L-methionine was fed to an additional two gilts from each litter. Duodenal, liver, and serum methionine levels were higher when DL-methionine was fed, indicating differences in the abilities of the test gilts to utilize the D- and L-isomers. Serum and muscle methionine levels were higher for litter 2, suggesting possible methionine requirement differences between the two litters. Histidine, isoleucine, and leucine appeared to accumulate in muscle, serum, and liver, respectively, suggesting some form of short-term storage. The accumulation of histidine in muscle tissue was thought to partially explain the fluctuations often associated with serum histidine levels that have not been explainable in terms of diet. A need for further research into factors affecting amino acid requirements was indicated.

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