Complete sequences of ribosomal and mitochondrial genes of the giant liver fluke Fascioloides magna are presented. In particular, small subunit (18S) and internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene (rDNA), as well as cytochrome c oxidase subunit I (cox1) and nicotinamide dehydrogenase subunit I (nad1) of the mitochondrial DNA (mtDNA), were analyzed. The 18S and ITS sequences were compared with previously published sequences of the liver fluke Fasciola hepatica. Fixed interspecific genetic differences were determined that allow molecular differentiation of F. magna and F. hepatica using either the PCR-RFLP method or PCR amplification of species-specific DNA regions. Additionally, intraspecific sequence polymorphism of the complete cox1 and nad1 mitochondrial genes in geographically distinct F. magna populations was determined. Based on the sequence divergences, short (< 500 bp) variable regions suitable for broader biogeographical studies of giant liver fluke were designed.
Summary
Toxoplasma gondii is among the most studied parasites worldwide but there is not much information about it published in Ireland. The objectives of this study were to determine the seroprevalence of T. gondii in sheep, pigs, deer and chickens and the molecular detection of T. gondii DNA in muscle tissue. Serum samples were collected from these species at the time of slaughter at Irish abattoirs during 2007 and tested for anti‐T. gondii antibodies using a commercial semi‐quantitative latex agglutination test. Antibodies (titre ≥1 : 64) were found in 36% (105/292) sheep, 4.7% (15/317) pigs and 6.6% (23/348) deer. In chickens, 18% (65/364) had antibody titres, ranging between 1 : 5 and 1 : 1024. Significant (P ≤ 0.05) age‐related differences in seroprevalence were found in adult sheep (58.1%) and pigs (23.1%). Significant gender differences in seroprevalence was also found in sheep with more females (43%) than males (22.4%) being positive. However, when adjusted for age through logistic regression gender was no longer significant. Seroprevalence was also evaluated on farm locations grouped to NUTS level 3, but the prevalence was too low to draw any statistical conclusions. Using a nested PCR, the presence of T. gondii DNA was detected in diaphragm samples from 3.6% (3/83) sheep, 13.0% (3/23) pig and 4.2% (3/71) deer. Meat digestion liquids from a Trichinella spp. survey in pigs were also used for the first time to detect T. gondii. Toxoplasma gondii DNA was detected in 50% (10/20) of pooled samples. This is the first in depth study of T. gondii seroprevalence in animals in Ireland and a novel method, using digestion liquid from pooled diaphragm samples, for PCR detection in pigs is described.
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