The helminth fauna of the genus Talpa in the Palaearctic Realm is reviewed. Several helminth species reported in Talpa spp. by a number of authors are discussed, with reference to host specificity, parasite biology, and host ethology, ecology and phylogeny. Twelve species of cestodes were found, two of which exhibit stenoxenous specificity (Staphylocystis bacillaris and Multitesticulata filamentosa). Only three species of trematodes, Ityogonimus lorum, Ityogonimus ocreatus and Combesia macrobursata, are exclusive parasites of Talpa spp. The largest group are nematodes, with 37 species. Species of Tricholinstowia are parasites of holarctic talpids and several species of distinct genera, such as Capillaria, Soboliphyme and Trichuris, are found only in Talpa spp. Only acanthocephalans of the genus Moniliformis have been reported in moles of the genus Talpa. On the basis of these helminthological findings, the close phylogenetic relationship between moles (Talpidae) and shrews (Soricidae) supports the separation of the ordinal levels Soricomorpha and Erinaceomorpha.

The nucleotide sequences of partial 18S, complete internal transcribed spacer region 1 (ITS1), complete 5.8S, complete ITS2 and partial 28S of ribosomal DNA (rDNA) and cytochrome c oxidase subunit 1 of mitochondrial DNA (MCOI) from five species of gnathostomes (G. spinigerum, G. doloresi, G. nipponicum, G. hispidum and G. binucleatum with the former four species being distributed in Japan and Asia) that cause human gnathostomiasis were compared by direct polymerase chain reaction cycle-sequencing. The nucleotide sequences of each region of the18S (613 bp), 5.8S (158 bp) and 28S (598 bp) rDNA from the five species were almost identical. The ITS1 region was different in length for the five species. The nucleotide sequences of each region of ITS2 and partial MCO1 regions were different among the five species. Therefore, these two regions can be used as genetic markers for identification of worms.

Contamination of soil with feline and canine ascarid eggs in eight playgrounds in Kırıkkale, Turkey was investigated monthly from February 2003 to January 2004. Dog faeces were also collected and all samples were examined using the zinc sulphate centrifugal flotation method. Eggs of Toxocara were observed in 5 of 8 (62.5%) of playgrounds examined and in 15.6% of 480 soil samples. The number of eggs varied from 1 to 11. Eggs were observed in soil samples collected in February, March to June, August and November, with embryonated eggs appearing in June and August. Eggs of Toxascaris leonina and Taenia spp. and oocysts of Isospora spp. were also found in 1.5%, 1.0% and 0.2% of soil samples, respectively. Of 26 samples of dog faeces collected, 7.7% were contaminated with Toxocara spp. and 11.5% with Taenia spp. The presence of Toxocara eggs in the city playgrounds and dog populations suggests a potential human health hazard due to toxocariasis.

Cercariae and adult Schistosoma mansoni were used to prepare, respectively, cercarial secretions (CS) and worm vomit (WoV). These were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to test the IgG-reactivity of sera obtained in an S. mansoni-endemic area of Burkina Faso. Among the egg-excreting individuals (n=240), 94.6% reacted positively with WoV, but only 62.9% with CS, thus suggesting a high diagnostic sensitivity of WoV, but not of CS. Among those individuals without detectable eggs in two Kato-Katz thick smears from different stool specimens (n=215), the respective percentages of positive IgG reactivity were 78.1% and 63.3%. These positive reactions in the absence of detectable eggs are interpreted in terms of limited sensitivity of parasitological stool examinations. Optical density values in ELISA with CS, but not with WoV, correlated negatively with age, which may reflect decreasing exposure to cercariae in older individuals.

Electrorotation is a non-invasive technique that is capable of detecting changes in the morphology and physicochemical properties of microorganisms. The first detailed electrorotation study of the egg (ovum) of a parasitic nematode, namely Ascaris suum is described to show that electrorotation can rapidly differentiate between fertilized and non-fertilized eggs. Support for this conclusion is by optical microscopy of egg morphology, and also from modelling of the electrorotational response. Modelling was used to determine differences in the dielectric properties of the unfertilized and fertilized eggs, and also to investigate specific differences in the spectra of fertilized eggs only, potentially reflecting embryogenesis. The potential of electrorotation as an investigative tool is shown, as undamaged eggs can be subjected to further non-destructive and destructive techniques, which could provide further insight into parasite biology and epidemiology.

Waterfowl from Lake Wanaka and the Waitaki Lakes watershed of New Zealand's South Island were surveyed to find natural hosts with adult echinostomes and schistosomes to provide sufficient numbers of eggs for laboratory studies. The Canada goose (Branta canadensis) was found to host an echinostome determined to be a New Zealand strain of Echinostoma revolutum. The New Zealand scaup (Aythya novaeseelandia) concurrently hosts three species of echinostomes (E. revolutum, Echinoparyphium cinctum and E. recurvatum) plus two species of avian schistosomes (Trichobilharzia sp. and Dendritobilharzia pulverulenta). In the Canada goose and the New Zealand scaup, adult gravid echinostomes predominated over juveniles. In other waterfowl surveyed, very few echinostomes were found, with juveniles predominating.

The indicator value (Ind Val) method which combines measures of fidelity and specificity has been used in a study on wild boar parasites in Corsica during 2001–2003. Because of its resilience to changes in abundance, IndVal is a particularly effective tool for ecological bioindication. The Ind Val method showed how season can influence the occurrence of parasite species in the wild boar, and also identified parasites as bioindicators relative to host age. The randomization test identified five parasite species having a significant indicator value for the season (the ticks, Hyalomma aegyptium and Rhipicephalus sanguineus, the louse, Haematopinus suis and the nematodes Globocephalus urosubulatus and Ascaris suum and two indicator species of an age class (the nematodes G. urosubulatus and Metastrongylus sp.). Data on species composition and infection levels would help improve the monitoring and management of parasitism in Suidae populations.

Anisakid nematodes belonging to the Anisakis simplex complex are highly prevalent in several fish species off the coast of Portugal and are an important zoonotic problem in the Iberian Peninsula. Two reproductively isolated sibling species of the Anisakis simplex complex were identified from Pleuronectiformes inhabiting the Portuguese coast using restriction fragment length polymorphism (RFLP). Recombinant genotypes corresponding to presumptive Anisakis simplex sensu stricto and Anisakis pegreffii hybrids were also detected by this technique, as well as the species Anisakis typica. Although 25 species of Pleuronectiformes were investigated, Anisakis spp. larvae were only found in seven: Arnoglossus imperialis, Arnoglossus laterna, Lepidorhombus boscii, Citharus linguatula, Platichthys flesus, Dicologlossa cuneata and Solea senegalensis. The occurrence of hybrids in relatively sedentary fishes such as the Pleuronectiformes suggests that the Portuguese coast may constitute an area of hybridization and, therefore, is of particular interest for the study of the process of hybridization and speciation for these anisakids.

The plant-parasitic cyst nematode Heterodera glycines requires a host plant to complete its life cycle, which involves hatching of infective juveniles that parasitize through root entry. A laboratory population of H. glycines grown on soybean, Glycine max, undergoes a sharp increase in maturity between 5 and 6 weeks in culture, as measured by the proportion of eggs containing well developed pre-hatch juveniles (late development eggs) versus eggs without visible juveniles (early development eggs). The median percent of eggs classified as late development, representing all samples taken from 4 to 7 weeks in culture, was 61%. For all samples taken up to 5 weeks, 80% scored below the median. In samples taken after 5 weeks, 15% scored below the median. This shift in population maturity was accompanied by a significant increase (P < 0.01) in the number of hatched juveniles present in each sample. There was also a significant increase (P < 0.02) in amount of FaRP-like peptide detected by specific ELISA. Total FaRP levels increased from 0.18 ± 0.07 fMol FLRFamide equivalents per ng protein in early development eggs to 0.40 ± 0.17 in late development eggs. The level remained high in hatched juveniles. HPLC/ELISA detected as many as nine potential FaRPs in H. glycines, two of which were specifically increased (P < 0.005) in hatched juveniles. The association of FaRPs with maturing eggs and the possible involvement of these neuropeptides with juvenile hatching and motility are discussed.

Karyotypes of three dilepidid species: Molluscotaenia crassiscolex, Anomotaenia bacilligera and Dilepis undula, which have not been recorded previously, were studied using conventional Giemsa staining and comparative karyometric analysis. Twelve small biarmed chromosomes were observed in mitotic cells of M. crassiscolex, 16 biarmed chromosomes of gradually decreasing size were found in cells of A. bacilligera, while 18 elements were characteristic for D. undula. These data, together with information available in literature, prove the heterogeneity and possible polyphyletic nature of the family Dilepididae.

To estimate prevalence of amphistomiasis and its association with bovine Fasciola gigantica infections in Zambia, 709 faeces of cattle presented for slaughter from three major cattle rearing areas of Central, Southern and Western provinces were analysed. The prevalence rate of amphistomes was 51.6%. Egg counts per gram (EPG) of faeces ranged from 0 to 385 with a mean (± SEM) of 11.96±1.07. The origin of the cattle had a significant influence (P<0.001) on the prevalence rate. Fasciola gigantica infections accounted for 46.7% of the cattle examined. The mean EPG count (± SEM) was 6.3±0.66 with a range of 0 to 223. A total of 34.6% were mixed infections while single Fasciola and amphistome infections represented 12.1% and 17.1%, respectively. Significantly more cattle (63.8%) were infected with either single or both trematode infections (P<0.0001) than not. Mixed trematode infections were highest in the Southern province (80.0%) while the lowest were recorded in Central province (16.3%). A positive correlation (r2=0.0428) was present in mixed infections. In order to minimize losses, epidemiology and cross-resistance of amphistomiasis and fascioliasis should be studied in different ecological regions of Zambia to formulate efficient control programmes.

The value of tongue and meat inspection as diagnostic tools for porcine cysticercosis was assessed in 65 Zambian village pigs by comparing the results with carcass dissections. In addition, the intensity of infections, distribution and viability of cysts in infected pigs were measured. Five pigs (7.7%) were positive on tongue examination, while routine meat inspection showed 12 (18.5%) positives. However, carcass dissections detected cysticerci in 31 (47.7%) pigs. The range in number of cysticerci was 1 to 14,662 per carcass. Cysticerci were distributed throughout the carcass with the highest concentration in the heart, tongue and hind legs. In one animal 13 viable cysts were detected only in the brain. Fourteen pigs had more than 100 viable cysts, six between 2 and 100, and four had single cyst infections. Seven animals harboured only calcified cysts. These findings demonstrate the serious shortcomings of routine detection methods for porcine cysticercosis. While the specificity of tongue palpation and meat inspection was 100%, these tests failed to detect the infection in 83.9% and 61.3% of infected pigs, respectively.

The immunological, haematological and enzymatic responses to the inoculation in pigs of 100,000 embryonated eggs of Toxocara canis were studied. Fifteen females were inoculated and three remained as controls. Haematological values were analysed from day 7 p.i. until day 126 p.i. In the inoculated group, white blood cells were raised on day 14 p.i. and eosinophil values on days 7, 14, 21, 35 and 49 p.i. showing significant differences compared with controls (P<0.05). Absolute eosinophil counts (per ml) presented two rises, the first on days 7, 14 and 21 p.i. and the second on days 35 and 49 p.i. Blood biochemistry was maintained within normal values. Serological examination by ELISA to determine antibody levels against Toxocara canis L2/L3 excretory–secretory (ES) antigens showed values higher than the positive cut-off (1:32) from day 7 p.i. and until the end of the study on day 126 p.i., presenting two peaks: one on day 28 p.i. and the second covering days 49 to 56 p.i. Western blots of sera of inoculated animals presented, from day 7 p.i., two polypeptide bands of 55 and 70 kDa MW and, from day 56 p.i., an additional band of 120 kDa MW, all of which persisted until the end of the study. Immunological responses were sustained over time. No direct correlation was observed between the rise in eosinophils and antibody titres. To validate the conclusions, more studies are required on the polypeptide bands.

Plasma etching and ashing for demonstrating the three-dimensional ultrastructure of the internal organs of helminths is described. Adult worms of the cestode Caryophyllaeides fennica were dehydrated through an ethanol series, critical point dried (Polaron E3000) and sputter coated with 60% gold-palladium (Polaron E5100) and glued to a standard scanning electron microscope (SEM) stub positioned as required for ashing. After initial SEM viewing of worm surfaces for orientation, stubs were placed individually in the reactor chamber of a PT7150 plasma etching and ashing machine. Worms were exposed to a radio frequency (RF) potential in a low pressure (0.2 mbar) oxygen atmosphere at room temperature. The oxidation process was controlled by varying the times of exposure to the RF potential between 2 to 30 min, depending on the depth of surface tissue to be removed to expose target organs or tissues. After each exposure the oxidized layer was blown from the surface with compressed air, the specimen sputter-coated, and viewed by SEM. The procedure was repeated as necessary, to progressively expose successive layers. Fine details of organs, cells within, and cell contents were revealed. Ashing has the advantage of providing three dimensional images of the arrangement of organs that are impossible to visualize by any other procedure, for example facilitating testes counts in cestodes. Both freshly-fixed and long-term stored helminths can be ashed. Ashing times to obtain the desired results were determined by trial so that some duplicate material was needed.

Adult male worms of Trichobilharzia sp. recovered from a pied wagtail (Motacilla alba) in the Czech Republic were found to belong to a new species of the genus. The finding of Trichobilharzia sp. in a passeriform bird in Europe represents an important discovery, as only anseriform birds have thus far been reported as final hosts of the European Trichobilharzia species.

Hard structures of helminths have often been used for taxonomic identification but are usually not clearly defined when treated with conventional methods such as ammonium picrate-glycerin for monogeneans and glycerin for nematodes. The present study reports a rapid and simple technique to better resolve the hard parts of selected monogeneans and nematodes using 5–10% alkaline sodium dodecyl sulphate (SDS). In comparison with established methods, SDS-treated worms become more transparent. In monogeneans treated with SDS, clear details of the hooks, hook filaments, anchors, bars and the sclerotized copulatory organs could be observed. In SDS-treated nematodes, spicules and ornamentations of the buccal capsules could be clearly seen.