International Commission on Trichinellosis: Recommendations for quality assurance in digestion testing programs for .

Effective performance of digestion testing methods for , and their use for the detection of infected animals and the prevention of human trichinellosis require system-wide incorporation of appropriate quality assurance (QA) practices. The recommendations of the International Commission on Trichinellosis (ICT) aim to facilitate reliable test results when laboratories operate within a quality management system (QMS) which includes: 1) a quality manual (or similar documentation of the QMS); 2) a validated test method with identified critical control points; 3) a training program; 4) procedures utilizing proficiency testing and other methods to confirm technical capability of analysts; 5) equipment calibration and maintenance; 6) standard operating procedures, related documentation and reporting; 7) procedures to enable continuous monitoring and improvements; and 8) regular internal and third party audits. The quality manual or similar documentation describes the QMS within a testing laboratory, and lists the QA policies and good laboratory practices.

TRANSMISSION DYNAMICS OF IN ARCTIC FOXES (): A LONG-TERM MARK-RECAPTURE SEROLOGIC STUDY AT KARRAK LAKE, NUNAVUT, CANADA.

Transmission dynamics of , a parasite of importance for wildlife and human health, are enigmatic in the Arctic tundra, where free-ranging wild and domestic felid definitive hosts are absent and rarely observed, respectively. Through a multiyear mark-recapture study (2011-17), serosurveillance was conducted to investigate transmission of in Arctic foxes () in the Karrak Lake region, Nunavut, Canada. Sera from adult foxes and fox pups were tested for antibodies to by using serologic methods, including the indirect fluorescent antibody test, direct agglutination test, and modified agglutination test.

Tongue has higher larval burden of Trichinella spp. than diaphragm in wolverines (Gulo gulo).

Trichinella is an important zoonotic parasite found in a range of wildlife species harvested for food and fur in Canada. We compared larval intensity from tongue and diaphragm, the best predilection sites in other animal species, from naturally infected, wild wolverines (Gulo gulo) (n = 95). Muscle larvae of Trichinella spp.

Development and validation of a duplex real-time PCR assay for the diagnosis of equine piroplasmosis.

Background: Equine piroplasmosis (EP) is an economically significant infection of horses and other equine species caused by the tick-borne protozoa Theileria equi and Babesia caballi. The long-term carrier state in infected animals makes importation of such subclinical cases a major risk factor for the introduction of EP into non-enzootic areas. Regulatory testing for EP relies on screening of equines by serological methods.

Pathology, clinical signs, and tissue distribution of in experimentally infected reindeer ().

is a zoonotic parasite found in vertebrates worldwide for which felids serve as definitive hosts. Despite low densities of felids in northern Canada, Inuit people in some regions show unexpectedly high levels of exposure, possibly through handling and consumption of Arctic wildlife. Free-ranging caribou () are widely harvested for food across the Canadian North, show evidence of seroexposure to , and are currently declining in numbers throughout the Arctic.

Multi-scale occupancy approach to estimate Toxoplasma gondii prevalence and detection probability in tissues: an application and guide for field sampling.

Increasingly, birds are recognised as important hosts for the ubiquitous parasite Toxoplasma gondii, although little experimental evidence exists to determine which tissues should be tested to maximise the detection probability of T. gondii. Also, Arctic-nesting geese are suspected to be important sources of T.

ESTIMATING TOXOPLASMA GONDII EXPOSURE IN ARCTIC FOXES (VULPES LAGOPUS) WHILE NAVIGATING THE IMPERFECT WORLD OF WILDLIFE SEROLOGY.

Although the protozoan parasite Toxoplasma gondii is ubiquitous in birds and mammals worldwide, the full suite of hosts and transmission routes is not completely understood, especially in the Arctic. Toxoplasma gondii occurrence in humans and wildlife can be high in Arctic regions, despite apparently limited opportunities for transmission of oocysts shed by felid definitive hosts. Arctic foxes (Vulpes lagopus) are under increasing anthropogenic and ecologic pressure, leading to population declines in parts of their range.

Toxoplasma gondii exposure in arctic-nesting geese: A multi-state occupancy framework and comparison of serological assays.

The zoonotic parasite, Toxoplasma gondii, has a worldwide distribution and a cosmopolitan suite of hosts. In arctic tundra regions, the definitive felid hosts are rare to absent and, while the complete transmission routes in such regions have yet to be fully elucidated, trophic and vertical routes are likely to be important. Wild birds are common intermediate hosts of T.

Endoparasites in the feces of arctic foxes in a terrestrial ecosystem in Canada.

The parasites of arctic foxes in the central Canadian Arctic have not been well described. Canada's central Arctic is undergoing dramatic environmental change, which is predicted to cause shifts in parasite and wildlife species distributions, and trophic interactions, requiring that baselines be established to monitor future alterations. This study used conventional, immunological, and molecular fecal analysis techniques to survey the current gastrointestinal endoparasite fauna currently present in arctic foxes in central Nunavut, Canada.

A new multi-host species indirect ELISA using protein A/G conjugate for detection of anti-Toxoplasma gondii IgG antibodies with comparison to ELISA-IgG, agglutination assay and Western blot.

Toxoplasma gondii is a zoonotic protozoan parasite which can cause significant disease and losses in livestock and wild animals. It is increasingly recognized as an important foodborne pathogen in a broad range of food animals and products. Effective control strategies require rapid, reliable and cost-effective detection methods for large scale surveys and diagnostic applications in a broad range of warm-blooded animals.

Application of a qPCR assay with melting curve analysis for detection and differentiation of protozoan oocysts in human fecal samples from Dominican Republic.

A quantitative polymerase chain reaction assay with melt curve analysis (qPCR-MCA) was applied for the detection of protozoan oocysts in 501 human fecal samples collected in Dominican Republic. Samples were subjected to qPCR using universal coccidia primers targeting 18S rDNA to detect oocysts followed by MCA to identify oocyst species based on amplicon melting temperature. Putative positive samples were also tested by conventional PCR and microscopy.

Performance of commercial ELISA and agglutination test kits for the detection of anti-Toxoplasma gondii antibodies in serum and muscle fluid of swine infected with 100, 300, 500 or 1000 oocysts.

Serum and tissue fluid samples from experimentally infected swine were tested for antibodies to Toxoplasma gondii using both an indirect ELISA and a modified agglutination test (MAT) available commercially in kit form. Ten 8-9 week-old swine were fed meatballs containing 100, 300, 500 or 1000 T. gondii oocysts and three control animals were fed meatballs with no oocysts.

Serological evidence of Besnoitia spp. infection in Canadian wild ruminants and strong cross-reaction between Besnoitia besnoiti and Besnoitia tarandi.

Bovine besnoitiosis, caused by Besnoitia besnoiti, is considered to be emergent in Europe and responsible for severe economic losses due to the chronic and debilitating course of the disease but has not been reported in North America. Besnoitia tarandi is a related species and it has been reported in reindeer and caribou from different locations of the Arctic Pole, including North America. Diagnosis of clinical besnoitiosis is largely based on the recognition of dermal grossly visible tissue cysts of Besnoitia.

Validation of an immunohistochemical assay for bovine cysticercosis, with comparison to a standard histological method.

The larval stage (syn Cysticercus bovis) of the human tapeworm Taenia saginata causes cysticercosis in cattle, which has both aesthetic and food safety implications to consumers of beef. A monoclonal antibody-based immunohistochemical (IHC) assay developed to improve postmortem diagnosis of this parasite and a standard histological method were assessed to determine their fitness for intended use. Sections from 169 known-positive specimens of T.

Detection and differentiation of coccidian oocysts by real-time PCR and melting curve analysis.

Rapid and reliable detection and identification of coccidian oocysts are essential for animal health and foodborne disease outbreak investigations. Traditional microscopy and morphological techniques can identify large and unique oocysts, but they are often subjective and require parasitological expertise. The objective of this study was to develop a real-time quantitative PCR (qPCR) assay using melting curve analysis (MCA) to detect, differentiate, and identify DNA from coccidian species of animal health, zoonotic, and food safety concern.

A novel Ehrlichia genotype detected in naturally infected cattle in North America.

During a research investigation to determine if cattle from British Columbia (BC), Canada were infected with Anaplasma marginale or other related rickettsial blood parasites, a novel Ehrlichia genotype was revealed. Blood from seven BC source cattle was bioassayed by intravenous inoculation into naïve splenectomised calves. Additional splenectomised calves were used as uninoculated negative control or A.

Genetic diversity of equine piroplasms in Greece with a note on speciation within Theileria genotypes (T. equi and T. equi-like).

Equine piroplasms in Greece were studied using the reverse line blot hybridization (RLB) assay. Three genotypes consisting of two Theileria (T. equi and T.

A seroepidemiological study of exposure to Toxoplasma, Leishmania, Echinococcus and Trichinella in equids in Greece and analysis of risk factors.

The role of horses in the transmission of parasitic zoonoses either as a source of infection to vectors or through contamination of definitive hosts is gaining importance worldwide. For this reason sera from 773 equids including 753 horses, 13 mules and seven ponies in four regions of Greece were investigated by ELISA for the presence of IgG antibodies against Toxoplasma, Leishmania, Echinococcus and Trichinella. Anti-Toxoplasma antibodies were detected in all regions with an overall prevalence of 1.

Seroprevalence of equine piroplasms and host-related factors associated with infection in Greece.

Serum samples were collected from a total of 544 equids that included 524 horses, 13 mules, and 7 ponies from various regions of mainland Greece and were examined by competitive-inhibition ELISA (cELISA) to evaluate the level of exposure of Greek equids to Theileria (Babesia) equi and/or Babesia caballi, the causative agents of piroplasmosis. Association between seropositivity and host-related factors of species, gender, age, origin, activity and location were investigated. The overall seroprevalence was 11.

Detection of prohibited animal products in livestock feeds by single-strand conformation polymorphism analysis.

Single-strand conformation polymorphism (SSCP) analysis of amplicons produced from a mitochondrial DNA region between the tRNA(Lys) and ATPase8 genes was applied for the detection of animal product within livestock feeds. Identification of prohibited animal (cattle, elk, sheep, deer, and goat) and nonprohibited animal (pig and horse) products from North America was possible based on the differential display of the single-stranded DNA fragments for the different animal species on SSCP gels. This method allowed specific detection and identification of mixed genomic DNA from different animal species.

A 10-year wildlife survey of 15 species of Canadian carnivores identifies new hosts or geographic locations for Trichinella genotypes T2, T4, T5, and T6.

A survey of wild carnivores in Canada was conducted over a 10-year period to determine the prevalence and genotypes of Trichinella. Muscle samples collected from 1409 animals representing 15 hosts species were enzymatically digested to recover Trichinella larvae. Larvae were recovered from a total of 287 (20.

Detection and identification of Tetratrichomonas in a preputial wash from a bull by PCR and SSCP.

Motile Tritrichomonas foetus-like trichomonads were found during microscopic examination of a wet mount sample of a preputial wash collected from a bull. Staining of the organisms with a modified Wright-Giemsa stain revealed that several had four anterior flagella of unequal length instead of the three anterior flagella of equal length characteristic of T. foetus.

Trichinella diagnostics and control: mandatory and best practices for ensuring food safety.

Because of its role in human disease, there are increasing global requirements for reliable diagnostic and control methods for Trichinella in food animals to ensure meat safety and to facilitate trade. Consequently, there is a need for standardization of methods, programs, and best practices used in the control of Trichinella and trichinellosis. This review article describes the biology and epidemiology of Trichinella, and describes recommended test methods as well as modified and optimized procedures that are used in meat inspection programs.