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Many trematode groups have a long history of systematic revision, which can make parasite identification a difficult task. The trematode parasites of muskrats are no exception. Here, we highlight the systematic issues associated with trematodes of muskrats (Ondatra zibethicus). Then, we demonstrate the utility of using both morphological and molecular tools to identify these parasites. Morphological examinations of specimens from muskrats (n = 63) first suggested that at least 4 genera were present including Echinostoma, Wardius, Quinqueserialis, and Notocotylus. For the latter 3 groups, the 28S region verified this assessment. For echinostomes, ND1 sequences revealed at least 5 genetic lineages. A particular lineage, Echinostoma trivolvis lineage b, predominated in both prevalence and intensity of infection. Molecular sequences provided a more accurate estimate of echinostome diversity in the muskrats and further support the idea that E. trivolvis is a species complex. Future studies will focus on whether there are differences in host specificity among the E. trivolvis lineages. In addition, this study has provided initial sequences that will help verify the life cycles of Wardius, Quinqueserialis, and especially, Notocotylus. By linking molecular, morphological, and life history information, we can better understand parasite diversity.

Findings provided strong evidence that anthelmintic resistance is a serious problem on small ruminant farms throughout the southeastern United States. Owing to the frequent movement of animals among regions, the prevalence of resistance in other regions of the United States is likely to also be high. Consequently, testing of parasite eggs for anthelmintic resistance should be a routine part of parasite management on small ruminant farms.

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The most challenging obstacles to testing products for their anthelmintic activity are: (1) establishing a suitable nematode in vitro assay that can evaluate potential product use against a parasitic nematode of interest and (2) preparation of extracts that can be redissolved in solvents that are miscible in the test medium and are at concentrations well tolerated by the nematode system used for screening. The use of parasitic nematodes as a screening system is hindered by the difficulty of keeping them alive for long periods outside their host and by the need to keep infected animals as sources of eggs or adults when needed. This method uses the free-living soil nematode Caenorhabditis elegans as a system to screen products for their potential anthelmintic effect against small ruminant gastrointestinal nematodes, including Haemonchus contortus. This modified method uses only liquid axenic medium, instead of agar plates inoculated with Escherichia coli, and two selective sieves to obtain adult nematodes. During screening, the use of either balanced salt solution (M-9) or distilled water resulted in averages of 99.7 (± 0.73)% and 96.36 (± 2.37)% motile adults, respectively. Adult worms tolerated DMSO, ethanol, methanol, and Tween 80 at 1% and 2%, while Labrasol (a bioenhancer with low toxicity to mammals) and Tween 20 were toxic to C. elegans at 1% and were avoided as solvents. The high availability, ease of culture, and rapid proliferation of C. elegans make it a useful screening system to test plant extracts and other phytochemical compounds to investigate their potential anthelmintic activity against parasitic nematodes.

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