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Strepsiptera are an enigmatic order of insects with extreme sexual dimorphism which makes it difficult to "match-up" free-living adult males with parasitic conspecific females of the Stylopidia, and free-living females of the Mengenillidae using morphological characters. Species identification is further complicated for the Stylopidia because adult females are endoparasitic and neotenic. Therefore, we used DNA sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to confirm the species identity of adult strepsipterans that were morphologically identified as Stylops advarians. These specimens, collected from Saskatoon (Saskatchewan, Canada), included one adult male, and eight females, the latter of which had been collected from solitary bees (Andrena milwaukeensis). Also included in the analyses were three pools of first-instar larvae that had emerged from three of the females. The results of the molecular analyses revealed that all specimens had an identical cox1 sequence, and belonged to a clade, with total statistical support (bootstrap value of 100%), that contained specimens of S. advarians from New York and Maine (USA). Hence, the results were consistent with the morphological identification of S. advarians. This study demonstrates the usefulness of a molecular approach for the identification of endoparasitic adult female and larval strepsipterans, life cycle stages that lack significant morphological characters for species identification.

The phylogenetic relationships of the endemic (or largely endemic) Australasian trichostrongylin nematode families Herpetostrongylidae, Mackerrastrongylidae and Nicollinidae as well as endemic trichostrongylin nematodes currently placed in the families Trichostrongylidae and Molineidae were examined using the complete large subunit (28S) ribosomal RNA gene. The Herpetostrongylinae proved to be monophyletic. However, representatives of the Nicollinidae nested with the Herpetostrongylinae. The Mackerrastrongylidae was also a monophyletic group and included Peramelistrongylus, currently classified within the Trichostrongylidae. The Globocephaloidinae, currently considered to be a subfamily of the Herpetostrongylidae, was excluded from the family in the current analysis. Ollulanus and Libyostrongylus, included for the first time in a molecular phylogenetic analysis, were placed within the Trichostrongylidae. This study provided strong support for the Herpetostrongylidae (including within it the Nicollinidae, but excluding the Globocephaloidinae) and the Mackerrastrongylidae as monophyletic assemblages. Additional studies are required to resolve the relationships of the remaining endemic Australasian trichostrongylin genera.

Sequences of the first and second internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal (r) DNA were characterised for Labiosimplex australis, a parasitic nematode of macropodid marsupials from continental Australia and from island populations which have been isolated from mainland Australia for relatively short periods of time (<40,000 years). The results showed that the geographically isolated populations of L. australis on Kangaroo Island and Tasmania were genetically different from each other and from populations on the mainland. There were two unequivocal nucleotide differences sites within the ITS-1 rDNA sequence between the two island populations; however, the ITS-1 sequences of individuals from mainland populations contained one or both of these nucleotides. In contrast, L. australis from each island population had unique nucleotides in ITS-2 sequence that were not detected in any individual from the mainland. Although these results are consistent with the hypothesis that L. australis represents a single species, the genetic divergence in the ITS-2 sequences amongst individuals from different isolated populations suggests that the island populations of L. australis may be in the initial stages of allopatric speciation.

Genetic variation among 231 individuals of Opisthorchis viverrini from the Ban Phai District, Khon Kaen Province (Thailand) was examined at three polymorphic enzymes: enolase (ENOL), phosphoglucomutase (PGM), and triose phosphate isomerase (TPI). Four alleles were detected for TPI and PGM, whereas only two alleles were detected for ENOL. The inferred genotype frequencies for both TPI and ENOL were not significantly different from Hardy-Weinberg equilibrium. In contrast, the inferred genotype frequencies for PGM showed a significant departure from Hardy-Weinberg equilibrium, with a lack of heterozygous individuals. This heterozygote deficiency suggests non-random mating and/or potentially high self fertilization.

Multilocus enzyme electrophoresis (MEE) was used to genetically compare Moniezia spp. collected from sheep and cattle in southern Australia. Fifteen enzyme loci were established for the genetic characterisation of individual specimens of Moniezia, which represents a significant increase in the number of loci that was previously available for these cestodes. Another four enzyme loci were also identified as providing potential genetic markers. There were no differences in staining activity or banding patterns of different body regions of individual cestodes. Fixed genetic differences between Moniezia expansa and Moniezia benedeni were detected at 10 (77%) loci. Individuals that lacked the diagnostic morphological characters (i.e. the arrangement pattern of the interproglottidal glands) could be assigned to either morphospecies based on their MEE profiles. Genetic differences were also detected among individuals of the two morphospecies. One specimen of M. expansa had fixed genetic differences at 33% of enzyme loci when compared to all other specimens of M. expansa. Fixed genetic differences were also detected between the two groups of M. benedeni at 12 of 13 (92%) enzyme loci. This magnitude of genetic difference was greater than that detected between the two morphospecies. This provides evidence for the existence of cryptic species in the genus Moniezia.

Genetic variation in Opisthorchis viverrini adults originating from different locations in northeast Thailand and Laos, People's Democratic Republic (PDR), was examined using random amplified polymorphic DNA (RAPD) analyses. In an initial analysis, the genomic DNA of one fluke from each of ten localities was amplified using 15 random primers (10-mers); however, genetic variation among O. viverrini specimens was detected reliably for only four primers. A more detailed RAPD analysis using these four primers was conducted on ten individuals from nine localities. Considerable genetic variation was detected among O. viverrini from different geographical areas and among some individuals from the same collecting locality. Comparison of the RAPD profiles revealed that O. viverrini adults from Laos PDR were genetically distinct from those from northeast Thailand. The taxonomic significance of this finding needs to be explored in more detail. The RAPD markers established in the present study provide opportunities to examine the biology and epidemiology of O. viverrini and fish-borne trematodes within the region. Additionally, application of these markers in such studies could have important implications in relation to the prevalence of cholangiocarcinoma in different regions of Asia.

In this study, cDNAs encoding myosin from the parasitic nematode Haemonchus contortus were isolated and characterized. Several exhibited a considerable degree of sequence variation at the nucleotide and limited divergence at the amino acid levels within the various functional domains. The results suggest that the cDNAs isolated represented a single myosin heavy chain, which, by comparison with a number of other myosins, is inferred to represent a homologue of a muscle myosin (CeMHCA) of the free-living nematode Caenorhabditis elegans. The findings could have implications for investigating cytoskeletal dynamics and/or signalling pathways.

The nuclear ribosomal DNA (rDNA) region spanning the first (ITS-1) and second (ITS-2) internal transcribed spacers was sequenced for 15 taxa of ascaridoid nematodes. The length of the ITS-1 and ITS-2 sequences in the 15 taxa ranged from 392-500 bp and 240 348 bp, respectively. While nucleotide variation of 0-2.9% in the ITS-1 and/or ITS-2 sequences was detected within taxa where multiple samples were sequenced, significantly higher level of nucleotide difference (9.4-66.6%) was detected between the taxa, except for Ascaris suum and A. lumbricoides whose taxonomic status remains uncertain. These interspecific differences were linked with the considerable size differences (0-108 bp) in the rDNA spacers. Phenograms based on the genetic differences among the 15 taxa showed some concordance with previous classification schemes derived from morphological data.