The epidemiology of toxocariasis in humans in Mexico has been poorly explored. There is a lack of information about toxocariasis in Tepehuanos, an indigenous ethnic group in Durango State in northern Mexico. Therefore, the presence of anti-Toxocara immunoglobulin (Ig) G antibodies was determined in 126 rural Tepehuanos using an enzyme-linked immunoassay. Socio-demographic, clinical and behavioural data of the participants were also obtained. Of the 126 Tepehuanos assessed (mean age 32.46 ± 17.36 years), 33 (26.2%) had anti-Toxocara IgG antibodies. Multivariate analysis showed that Toxocara seropositivity was associated with unemployment (students and housewives) (odds ratio (OR) = 3.1; 95% confidence interval (CI) 1.25–7.47). Other socio-demographic and behavioural characteristics, including age, educational level, contact with animals or soil, consumption of unwashed raw fruits and vegetables or untreated water, were not associated with Toxocara seropositivity. Clinical data were similar in seropositive and seronegative Tepehuanos. These results indicate that Toxocara exposure is common among Tepehuanos but Toxocara does not appear to impact on the health of the population. This is the first report of toxocaral infection in Tepehuanos, and of an association of toxocariasis in adults with unemployment. Further research is needed to elucidate the routes of transmission of Toxocara in Tepehuanos, including the role of hygiene practices, canine and feline contacts, paratenic hosts and soil contamination with infective Toxocara eggs.

Two new species of Syphacia (Nematoda: Oxyuridae) are described from endemic murids of Sulawesi, Indonesia: Syphacia(Syphacia)taeromyos sp. n. and S.(S.)paruromyos sp. n. parasitic in the caecum of Taeromys celebensis and Paruromys dominator, respectively. They are readily distinguished from all of the congeners recorded from Indonesia–Australian regions by having a round cephalic plate, vesicular lateral alae in the male, posteriorly positioned excretory pore in the male, and/or lacking cervical alae. Syphacia (S.) paruromyos differs from S. (S.) taeromyos by having a whip-like tail appendage in the male, longer relative distance between excretory pore and vulva, and larger eggs. The round cephalic plate in both sexes and developed vesicular lateral alae in the male are morphological traits common in endemic Syphacia species hitherto known from Sulawesi murids, suggesting that they have derived from a common ancestor and evolved with their hosts in the isolated insular environment.

Fascioliasis is an important food-borne parasitic disease caused by the two trematode species, Fasciola hepatica and Fasciola gigantica. The phenotypic features of fasciolid adults and eggs infecting buffaloes inhabiting the Central Punjab area, Pakistan, have been studied to characterize fasciolid populations involved. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardized measurements. Since it is the first study of this kind undertaken in Pakistan, the results are compared to pure fasciolid populations: (a) F. hepatica from the European Mediterranean area; and (b) F. gigantica from Burkina Faso; i.e. geographical areas where both species do not co-exist. Only parasites obtained from bovines were used. The multivariate analysis showed that the characteristics, including egg morphometrics, of fasciolids from Central Punjab, Pakistan, are between F. hepatica and F. gigantica standard populations. Similarly, the morphometric measurements of fasciolid eggs from Central Punjab are also between F. hepatica and F. gigantica standard populations. These results demonstrate the existence of fasciolid intermediate forms in endemic areas in Pakistan.

Entomopathogenic nematodes (EPN) were evaluated for their potential use as biological control agents against Phlyctinus callosus, the banded fruit weevil (BFW). The susceptibility of larvae and adults to EPN was evaluated using 400 infective juveniles (IJ) per insect after 4 days in 24-well bioassay trays. The nematode isolates used were all able to infect BFW, although the larvae were found to be more susceptible than were the adults. The percentage mortality for BFW larvae ranged from 41 to 73% and for BFW adults from 13 to 45%. The most effective isolate, SF41 of Heterorhabditis zealandica, was used to investigate the effect of vertical movement of nematodes in sand and sandy loam soil, at specified concentration and temperature. A higher (82.2 ± 0.084%) percentage mortality rate was obtained with the sandy loam soil, than with the use of sand (67.5 ± 0.12%). The LD50 and LD90 values after 4 days of incubation were 96 and 278 IJ/50 μl, respectively. Nematodes were inactive below 15°C, with the highest mortality of 74 ± 0.081% for BFW larvae recorded at 25°C. Heterorhabditis zealandica was able to complete its life cycle successfully in sixth-instar BFW larvae after a period of 22 days. The study showed BFW larvae not to be as susceptible to nematode infection as they need a high concentration (400 IJ/larva) and 4 days to give effective control.

Sixty Epinephelus areolatus were examined for metazoan fish parasites in Indonesia, off Segara Anakan lagoon, Java and in Balinese waters. The study revealed 21 different parasite species, and 14 new host and locality records. The anisakid nematodes Anisakis typica and, for the first time in Indonesia, Anisakis sp. HC-2005 were identified by using molecular methods. Ecological parameters were calculated for both sites off the anthropogenically influenced Segara Anakan lagoon and the relatively undisturbed reference site at the southern Balinese coast. The fish from Segara Anakan demonstrated a significantly higher enzymatic activity (Hepatosomatic index) and a significantly reduced number of heteroxenous gut helminths (e.g. the digenean Didymodiclinus sp., the nematode Raphidascaris sp. and the acanthocephalan Serrasentis sagittifer). Other regional differences for E. areolatus included ecto-/endoparasite ratio, endoparasite diversity, the parasite species composition and prevalence of infection of the respective parasite species. We applied the stargraph method to visualize observed regional differences using grouper parasites as biological indicators for the sampled coastal ecosystems at both sampling sites.

Microphallid trematodes are common parasites in marine snails and crustacean hosts at Kuwait Bay. The larval stages of two microphallids, Maritrema eroliae and Probolocoryphe uca, are difficult to differentiate morphologically. In this study, two PCR-based techniques were established for quick and accurate discrimination between the larval stages of the two microphallid species, employing restriction fragment length polymorphism (PCR-RFLP) and species-specific primers. Both techniques utilized nucleotide differences in the second internal transcribed region (ITS2) of the ribosomal DNA (rDNA) in the two species. For the PCR-RFLP technique, restriction enzyme AvaII was selected and it generated different restriction profiles among the two microphallids. In addition, species-specific primers were prepared for each microphallid species that amplified distinctive fragments. Both techniques showed that the larval stages of the two microphallid species can be identified accurately. However, direct PCR amplification using species-specific primers was more advantageous than the PCR-RFLP technique since it allowed rapid and specific discrimination between the two species. This technique provides a useful tool that can be used in future studies for the study of the distribution of microphallid species and their definitive hosts at different localities of Kuwait Bay.

The helminth communities of wild murid rodents were investigated in Luang Prabang and Champasak province, Lao PDR. Thirteen species of rodents (404 individuals) were infected by 19 species of parasites (2 trematode, 3 cestode, 14 nematode species). Four of the recorded helminth species (Echinostoma malayanum, Raillietina sp., Hymenolepis diminuta and H. nana) are known to cause potential zoonotic helminthiases of medical importance in the South-East Asian region. Individual helminth infection was significantly higher in the wet season. Habitat significantly influenced individual helminth species richness and individual helminth abudance, with a decrease of individual helminth species richness and individual helminth abundance from forest habitat to agricultural and human settlement habitats. The reduction of helminth diversity and abundance is discussed in relation to the ongoing increase of human influence on habitats in Lao PDR.

A total of 850 pre-adult Galba truncatula (shell height, 4 mm), originating from four French snail populations differing in shell height at the adult stage (from 6.5 to 12 mm), were individually subjected at 20°C to single-miracidium infections with Fascioloides magna. At day 75 post-exposure, the surviving snails were dissected, and rediae and cercariae were counted. Snail groups differed in shell growth during the experiment: from 1.8 ± 0.4 mm in group A up to 4.0 ± 1.1 mm in group D. The prevalence of F. magna infection, the numbers of free rediae and cercariae significantly increased together with increasing growth of infected snails during the experiment. Group A produced 1–6 first-generation rediae per snail and the mean daughter redia production ranged from 7.5 second-generation rediae (when a single first generation per snail developed) to 2.3 (6 first-generation rediae per snail). In contrast, up to ten first-generation rediae were noted in group D, and each mother redia gave daughter rediae with averages ranging from 1.5 (ten first-generation rediae per snail) to 13 (a single first generation per snail). In conclusion, the development of F. magna in G. truncatula exhibited both inter- and intrapopulation variability, where the development of rediae and cercariae was positively correlated with snail growth.

Fish-borne zoonotic trematodes (FZTs) can cause major human health problems. The aim of this study was to quantify the transmission of parapleurolophocercous cercariae to common carp (Cyprinus carpio) and to study the effect of the density of cercariae and the density of fish on transmission with respect to the volume of water and surface area of the bottom. Fish were kept individually either as controls (n= 91) or were exposed to 250 cercariae in tubes with a volume of 25, 50, 100, 250 or 500 ml water (n= 190) with a surface area of 4, 12, 21, 30 or 49 cm2 (n= 195). The dose to which the fish were exposed was kept constant. Infection occurred in 94–100% of fish, with a mean of 15–18 metacercariae per fish and the proportion of FZTs established at 0.06–0.07 metacercariae per cercariae per fish. Neither the prevalence of infection with FZTs nor the number of metacercariae per fish nor the proportion of FZTs established were significantly associated with differences in the density of cercariae or the density of fish per ml water or per cm2 surface area. Thus, it was concluded that the transmission of cercariae to fish is independent of density.

Trichuris trichiura and Trichuris suis parasitize (at the adult stage) the caeca of humans and pigs, respectively, causing trichuriasis. Despite these parasites being of human and animal health significance, causing considerable socio-economic losses globally, little is known of the molecular characteristics of T. trichiura and T. suis from China. In the present study, the entire first and second internal transcribed spacer (ITS-1 and ITS-2) regions of nuclear ribosomal DNA (rDNA) of T. trichiura and T. suis from China were amplified by polymerase chain reaction (PCR), the representative amplicons were cloned and sequenced, and sequence variation in the ITS rDNA was examined. The ITS rDNA sequences for the T. trichiura and T. suis samples were 1222–1267 bp and 1339–1353 bp in length, respectively. Sequence analysis revealed that the ITS-1, 5.8S and ITS-2 rDNAs of both whipworms were 600–627 bp and 655–661 bp, 154 bp, and 468–486 bp and 530–538 bp in size, respectively. Sequence variation in ITS rDNA within and among T. trichiura and T. suis was examined. Excluding nucleotide variations in the simple sequence repeats, the intra-species sequence variation in the ITS-1 was 0.2–1.7% within T. trichiura, and 0–1.5% within T. suis. For ITS-2 rDNA, the intra-species sequence variation was 0–1.3% within T. trichiura and 0.2–1.7% within T. suis. The inter-species sequence differences between the two whipworms were 60.7–65.3% for ITS-1 and 59.3–61.5% for ITS-2. These results demonstrated that the ITS rDNA sequences provide additional genetic markers for the characterization and differentiation of the two whipworms. These data should be useful for studying the epidemiology and population genetics of T. trichiura and T. suis, as well as for the diagnosis of trichuriasis in humans and pigs.

The prevalence of the digenean Plagiorchis sp. was investigated in a natural wood mouse population (Apodemus sylvaticus) in a periaquatic environment. Classical identification was complemented with the use of molecular differentiation to determine prevalence and verify species identity. Use of the complete ITS1-5.8S rDNA-ITS2 and partial 28S rDNA gene sequences have confirmed that the species reported at this location was Plagiorchis elegans and not Plagiorchis muris as reported previously. This underlines the difficulties in identification of these morphologically similar parasites. Plagiorchis elegans is typically a gastrointestinal parasite of avian species but has also been reported from small mammal populations. Although the occurrence of this digenean in A. sylvaticus in the UK is rare, in the area immediately surrounding Malham Tarn, Yorkshire, it had a high prevalence (23%) and a mean worm burden of 26.6 ± 61.5. The distribution of P. elegans followed a typically overdispersed pattern and both mouse age-group and sex were determined to be two main factors associated with prevalence. Male mice harboured the majority of worms, carrying 688 of 717 recovered during the study, and had a higher prevalence of 32.4% in comparison to only 8.7% in the small intestine of female mice. A higher prevalence of 43% was also observed in adult mice compared to 14% for young adults. No infection was observed in juvenile mice. These significant differences are likely to be due to differences in the foraging behaviour between the sexes and age cohorts of wood mice.

In Cuba, only two lymnaeid snails, Galba cubensis and Pseudosuccinea columella, with different ecology and distribution patterns, are intermediate hosts for Fasciola hepatica. The compatibility of these two species as hosts was analysed through their rates of infection, the production of rediae and survivorship when exposed to F. hepatica miracidia. Ten populations of G. cubensis, eight of P. columella collected from various habitats and six isolates of F. hepatica sampled in slaughterhouses from different localities were tested. Our results clearly demonstrate that G. cubensis is a more compatible host for F. hepatica in Cuba when compared with P. columella. However, the role that P. columella may have in fascioliasis transmission under certain conditions should not be disregarded. Variation in infectivity among isolates of F. hepatica were also observed and may explain why some regions in Cuba are more commonly subjected to fascioliasis outbreaks.

Clinostomum complanatum is a digenetic trematode that causes yellow grub disease in some fish species and also shows zoonotic potential by sporadically infecting humans. In this study, progenetic metacercariae of C. complanatum were obtained from the fish Trichogaster fasciatus, and were aseptically placed in conjunctival incisions made in the superior and inferior fornices of the eye of rabbits, which served as the experimental hosts. Worms were harvested without necropsy of the host on days 4 and 8 post infection, to observe in vivo transformation of the progenetic metacercariae into ovigerous adult worms. The worms appeared to cause minimal damage to the host although they were tenaciously attached. In vivo maturation was evident by the development of the vitellaria, enlargement of gonads, the presence of a large number of shelled eggs in a distended uterus and ramifications of the intestinal caeca. Obtaining mature ovigerous worms without sacrificing the host clearly gives the rabbit eye model an advantage over those described previously. Due to the relative advantage of the short time required for maturation and the prolific egg production by C. complanatum, it is suggested that this host–parasite system could be used as an excellent model for classroom teaching of trematode biology and to investigate the cues involved in in vivo transformation and host–parasite interactions.

Flukes were found in the bile ducts of domestic ducks (Anas platyrhynchos), necropsied in the Binh Dinh province of Central Vietnam. Following staining, morphological characteristics of the bird flukes were compatible with Opisthorchis viverrini, although some characteristics differed from those described in specimens collected from mammal hosts. Computation of the phylogenetic trees on the partial sequences of the second internal ribosomal spacer (ITS2) of the ribosomal DNA and cytochrome c oxidase subunit I (COI) markers of the mitochondrial DNA showed close similarity of the ‘bird’ Opisthorchis sp. with O. viverrini. We speculate that these bird flukes are O. viverrini that show intraspecies morphological and molecular variability compared to isolates from mammals. This demonstrates the complex epidemiological situation of opisthorchiasis in Vietnam and urges investigations on the potential of birds as a reservoir host of this zoonotic fluke.

Coproantigen test kits for Echinococcus spp. worms in dogs, designed for commercial use, were obtained from three different Chinese producers, and were compared with a laboratory kit using reagents from New Zealand. None of the three producers would provide details of their test validation. From a known set of dog faeces obtained at necropsy from infected and uninfected dogs, and from faeces collected from dogs necropsied in the field, results differed between the kits. For field material, the Tiankang kit showed the best specificity but lacked sensitivity. The Combined kit showed best sensitivity but lacked specificity. Results for the Haitai kit were intermediate. With samples from experimentally infected dogs, both the Haitai and Combined kits lacked sensitivity. Kits will need to be validated by the user before they can be relied on to predict progress in Echinococcus spp. control in China or in other countries.

Dichloromethane and aqueous fractions from leaves and stems of Piper arboreum Aubl., P. aduncum L., P. amalago L., P. crassinervium H.B. & K., P. diospyrifolium Kunth, P. hispidum Sw. and P. xylosteoides (Kunth) Steud. were tested against adult worms of Schistosoma mansoni. The in vitro activity was evaluated in terms of mortality, number of separated worms and number of worms with reduced motor activity. Most dichloromethane fractions from all Piper species showed moderate schistosomicidal activity, but aqueous fractions were not active. The dichloromethane fraction of P. amalago leaves (at 100 μg/ml) showed the highest activity, resulting in worm mortality, the separation of worm pairs and reduced motor activity. Chromatographic fractionation of the dichloromethane fraction of P. amalago leaves led to the isolation of its major compound, which was also tested against adults of S. mansoni. The isolated piperamide N-[7-(3′,4′-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl] pyrrolidine, at 100 μm, resulted in the mortality of all adult worms after 24 h of incubation. The findings suggest that species of Piper are potential sources of schistosomicidal compounds.

In the present study, the internal transcribed spacers (ITS) of ribosomal DNA (rDNA) of Oesophagostomum asperum and O. columbianum were amplified and sequenced. The ITS-1, 5.8S and ITS-2 rDNA sequences of O. asperum were 374 bp, 153 bp and 259 bp in length, respectively, and the corresponding sequences of O. columbianum were 259, 153 and 218 bp in length, respectively. Sequence differences in the ITS-1 and ITS-2 rDNA between the two Oesophagostomum species were 9.5–10.2% and 12.7–13.9%, respectively. Sequence differences in the ITS-1 and ITS-2 rDNA among members of the genus Oesophagostomum were 2.5–11.6% and 6.8–22.3%, respectively. Based on genetic markers in the ITS rDNA, an effective polymerase chain reaction (PCR) approach was developed to differentiate O. columbianum from O. asperum with a sensitivity of 0.2 ng/μl DNA. Since accurate characterization of parasites at different taxonomic levels is essential for population genetic studies and control of parasitosis, the present findings have important implications for studying epidemiology, taxonomy and population biology, as well as for the control of oesophagostomiasis.

Agricultural ruminants usually harbour mixed infections of gastrointestinal nematodes. A specific diagnosis is important because distinct species can differ significantly in their fecundity and pathogenicity. Haemonchus spp. and Cooperia spp. are the most important gastrointestinal nematodes infecting ruminants in subtropical/tropical environments. In Brazil, C. punctata is more adapted to cattle than sheep. Additionally, C. spatulata appears to be more adapted to cattle, whereas C. curticei is more adapted to sheep. However, infection of sheep with C. punctata is common when cattle and sheep share the same pasture. Although morphological analyses have been widely used to identify nematodes, molecular methods can overcome technical limitations and help improve species-specific diagnoses. Genetic markers in the first and second internal transcribed spacers (ITS-1 and ITS-2, respectively) of nuclear ribosomal DNA (rDNA) have been used successfully to detect helminths. In the present study, the ITS-1 region was analysed and used to design a species-specific oligonucleotide primer pair to identify C. curticei. The polymerase chain reaction (PCR) product was sequenced and showed 97% similarity to C. oncophora partial ITS-1 clones and 99% similarity to the C. curticei sequence JF680982. The specificity of this primer pair was corroborated by the analysis of 17 species of helminths, including C. curticei, C. punctata and C. spatulata. Species-specific diagnosis, which has implications for rapid and reliable identification, can support studies on the biology, ecology and epidemiology of trichostrongylid nematodes in a particular geographical location.

Numerous eggs of capillariid nematodes have been found in coprolites from a wide range of hosts and in raptor pellets in archaeological samples from Patagonia. The structure and sculpture of the eggshell of these nematodes and their biometry are commonly used for identification. The aim of this study was to determine whether eggs of the genus Calodium with similar morphology, found in different archaeological samples from Patagonia, belong to the same species. For this purpose, capillariid eggs (N= 843) with thick walls and radial striations were studied by permutational multivariate analysis of variance (PERMANOVA). Eggs exhibiting similar shape and structure also showed similar biometry, regardless of the zoological origin of coprolites (P= 0.84), host diet (P= 0.19), character of the archaeological sites (P= 0.67) and chronology (P= 0.66). Thus, they were attributed to the same species. We suggest that an unidentified zoonotic species of the genus Calodium occurred in the digestive tract of a wide range of hosts in Patagonia during the Holocene and that both human and animal populations were exposed to this parasite during the Holocene in the study area.

Entomopathogenic nematodes (EPN) are lethal parasites of insects, used as biocontrol agents. The objectives of this work were to survey the presence of EPN in continental Portugal and to characterize the different species. Of the 791 soil samples collected throughout continental Portugal, 53 were positive for EPN. Steinernema feltiae and Heterorhabditis bacteriophora were the two most abundant species. Analysis of EPN geographical distribution revealed an association between nematode species and vegetation type. Heterorhabditis bacteriophora was mostly found in the Alentejo region while S. feltiae was present in land occupied by agriculture with natural vegetation, broadleaved forest, mixed forest and transitional woodland-shrub, agro-forestry areas, complex cultivated patterns and non-irrigated arable land. Although no clear association was found between species and soil type, S. feltiae was typically recovered from cambisols and H. bacteriophora was more abundant in lithosols. Sequencing of the internal transcribed spacer (ITS) region indicated that S. feltiae was the most abundant species, followed by H. bacteriophora. Steinernema intermedium and S. kraussei were each isolated from one site and Steinernema sp. from two sites. Phylogenetic analyses of ITS, D2D3 expansion region of the 28S rRNA gene, as well as mitochondrial cytochrome c oxidase subunit I (COXI) and cytochrome b (cytb) genes, was performed to evaluate the genetic diversity of S. feltiae and H. bacteriophora. No significant genetic diversity was found among H. bacteriophora isolates. However, COXI seems to be the best marker to study genetic diversity of S. feltiae. This survey contributes to the understanding of EPN distribution in Europe.