A revised digestion method, developed for efficiency and quality assurance, was validated for the detection of Trichinella larvae in pork and horse meat to meet requirements for food safety testing and facilitate access to international markets. The method consisted of a tissue homogenization step and a spin bar digestion procedure conducted at 45 degrees C to free larvae from muscle tissue, followed by two sequential separatory funnel steps to concentrate the larvae for detection using a stereomicroscope. Critical control points were determined for the method and monitored during testing. Under conditions of a defined protocol, test capacity was suitable for industrial applications, since multiples of up to 100 g of tissue could be analyzed at one time. The overall sensitivity of the test system depended on the size and origin of the sample taken from individual infected carcasses. Data from swine indicated that the currently accepted sample size of 1 g from individual carcasses consistently detected larval loads of > or =3 larvae per gram. Larval loads of 1.0 to 1.9 larvae per gram required 3- to 5-g samples of muscle tissue for reliable detection. Five-gram samples were considered optimal, because they consistently detected more tissues than 3-g samples, although the difference was not statistically significant. Tissue localization studies in experimental pigs indicated that the tongue and diaphragm were the tissues of choice for the most sensitive larval recovery. A system of analyst training, laboratory certification based on ISO guide 25, and on-site proficiency panel testing was used to ensure that external laboratories would consistently produce reliable test results. The system developed for pork was successfully modified for the testing of horse meat.
Repeated outbreaks of trichinellosis caused by the consumption of Trichinella-infected walrus (Odobenus rosmarus) meat, which have sometimes led to serious morbidity, have stimulated Inuit communities in Nunavik (northern Quebec), Canada, to develop an innovative trichinellosis prevention program. The program involves preconsumption testing of meat samples from harvested walrus at a regional laboratory and the rapid dissemination of the results of such testing to communities. Local health authorities in Inukjuak conducted an epidemiological investigation after testing identified Trichinella-positive walrus meat in September 1997. This report describes the events that occurred before, during, and after the trichinellosis outbreak and also documents how the prevention program contributed to successful resolution of the outbreak.
The complement fixation test (CFT), competitive enzyme immunoassay (CELISA), indirect enzyme immunoassay (IELISA) and fluorescence polarization assay (FPA) were evaluated for the detection of antibodies to Brucella abortus and Brucella suis biotype 4 in caribou (Rangifer tarandus caribou), elk (Cervus elapus), red deer (Cervus elapus), and reindeer (Rangifer tarandus tarandus). When combining the data the FPA and the CELISA were determined to be the most suitable tests for serodiagnosis of Cervidae. The overall actual sensitivity of the CFT and the IELISA was 100%. The overall actual sensitivity for the CELISA and FPA was 99%. The overall relative specificity of the CFT (including treatment of anti-complementary data as positive or negative for analysis), the CELISA, the IELISA and the FPA were 65%, 93%, 99%, 99%, and 99%, respectively. The specificities of the buffered plate agglutination test (BPAT), the CFT, the CELISA, the FPA and the IELISA for 55 elk vaccinated with B. abortus strain 19 and tested 4 mo post vaccination were 14%, 31%, 51%, 84%, and 2%, respectively. The FPA is the diagnostic test of choice because it has sensitivity and specificity values comparable to the CELISA; it has the capability to distinguish vaccinal antibody and antibody resulting from exposure to crossreacting organisms such as Yersinia enterocolitica 0:9 from antibody to Brucella spp. in most cases; it is technically simple to do; it is adaptable to field use and it is relatively inexpensive.
A novel Brucella sp. was isolated from lymph nodes of four ringed seals (Phoca hispida) collected near Pangnirtung (Baffin Island, Canada) in January and February 1995 and in one harp seal (Phoca groenlandica) collected near the Magdalen Islands (Gulf of St. Lawrence, Canada) in March 1996. Bacteriological characteristics were the same for all five isolates. The colonies were typical of Brucella spp., but took 2 to 5 days longer than the traditional species to appear on primary isolation media. Biotyping results did not match any of the known biovars of Brucella, but were similar to isolates of the genus Brucella previously reported from marine mammals inhabiting other areas of the northern hemisphere. This is the first confirmed report of brucellosis in marine mammals from Canada, and the first report of this organism in ringed and harp seals.
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