Metazoan parasites of the sand smelt Atherina boyeri Risso, 1810 in Lake Iznik were studied. A total of 271 specimens of A. boyeri, 208 female and 63 male, mean ( ± SD) total length 8.4 ± 2.2 cm (range 3.1–12.2 cm) and mean total weight 4.7 ± 3.5 g (range 0.1–11.7 g) were examined between June 2008 and May 2009 at monthly intervals. In 33 fish no parasites were found. In the other fish, four parasite species were recorded: Diplostomum sp. (prevalence 39.13%, mean intensity 3.50 ± 3.03); Tylodelphys clavata (prevalence 74.16%, mean intensity 10.41 ± 14.89); Bothriocephalus cf. acheilognathi (prevalence 40.59%, mean intensity 31.83 ± 57.74); and Eustrongylides excisus (prevalence 6.64%, mean intensity 1.16 ± 0.39). The mean abundance of Diplostomum sp., T. clavata and E. excisus was higher during the winter and early spring months. In contrast, the mean abundance of B. cf. acheilognathi was higher in the summer and autumn months. This distribution of the parasites was related to a change in the environment of A. boyeri from the pelagic zone to deeper water at the end of autumn and a change in food supply.

Specimens (n= 41) of the amphisbaenid Amphisbaena wuchereri taken from a population in Minas Gerais state, south-eastern Brazil, were examined for gastrointestinal parasites. A single nematode species was found, Paradollfusnema amphisbaenia. This was a new host record for this nematode species. This parasite was encountered in the large intestine (prevalence of 100%), in the stomach (prevalence of 2%) and in the small intestine (prevalence of 7.3%). The intensity of infection ranged from 1 to 457 individual parasites per host and was positively correlated with body size of both male and female amphisbaenians. The discrepancy index (D) indicated that P. amphisbaenia tended to an even distribution in this host population. The nematode, which did not affect fat body mass, induced inflammatory infiltrations in the small intestine, indicating that the parasites might injure the host's organs.

Among about 50 nominal Paragonimus species, Paragonimus proliferus is rather a rare species, found only in Yunnan province, China, until our recent discovery of this species in Lai Chau province, northern Vietnam close to Yunnan, China. Here we add Quang Binh province, central Vietnam as a new endemic area of P. proliferus. Large excysted metacercariae found in mountainous crabs, Potamiscus tannanti, were morphologically identified as P. proliferus, which was confirmed further by molecular analyses. Second internal transcribed spacer (ITS2) sequences of the P. proliferus population in Quang Binh province were completely (100%) identical with those of P. proliferus populations in Lai Chau province, northern Vietnam and Yunnan province, China. However, cytochrome oxidase subunit 1 (CO1) gene sequences of Quang Binh population were significantly different (5.6%) from that of previously reported northern Vietnam and Yunnan, China populations. A phylogenetic tree revealed that all CO1 sequences of P. proliferus Quang Binh population formed a distinct group, which was clustered with northern Vietnam and Yunnan, China populations with the bootstrap value of 75%. This is the first record of the genetically variant population of P. proliferus, distribution of which is geographically remote from the previously reported endemic areas in the border between northern Vietnam and Yunnan, China, suggesting that P. proliferus may be much more widely distributed in the Indochina peninsula (or South-East Asia) than expected.

The 14.5 kDa fatty acid binding protein (FABP) was isolated from the crude extract of adult Fasciola gigantica worms. Polyclonal anti-FABP IgG was generated in rabbits immunized with prepared FABP antigen. Sandwich enzyme-linked immunosorbent assay (ELISA) was applied to detect coproantigen in stools and circulating Fasciola antigen (CA) in sera of 126 water buffaloes by using purified and horseradish peroxidase (HRP)-conjugated anti-FABP IgG. Sandwich ELISA sensitivity was 96.97% and 94.95%; while specificity was 94.12% and 82.35% for coproantigen and CA detection, respectively. However, sensitivity and specificity of the Kato–Katz technique was 73.74% and 100%, respectively. The diagnostic efficacy of sandwich ELISA was 96.55% and 93.1% for coproantigen and CA detection, respectively. In contrast, the diagnostic efficacy of the Kato–Katz technique was 77.59%. In conclusion, these results demonstrate that the purified 14.5 kDa FABP provides a more suitable antigen for immunodiagnosis of early and current bubaline fascioliasis by using sandwich ELISA.

Different species of Cassia plant are widely available in India and are commonly used either for their laxative, antimicrobial or antibacterial activity. In the present study the effectiveness in vitro of the crude alcoholic extracts of three species, namely Cassia alata, C. occidentalis and C. angustifolia, in the early paralysis and mortality of the fowl tapeworm Raillietina tetragona at concentrations ranging from 5 to 80 mg/ml was investigated. Time of paralysis and death were monitored frequently. Immediately after paralysis the tapeworms were processed for electron microscopic studies. While the untreated or control parasites survived for 81.93 ± 5.85 h, the parasites treated with C. alata took less time (1.68 ± 0.27 h) to be paralysed, followed by those treated with C. angustifolia (2.95 ± 0.29 h). Although C. occidentalis took more time (4.13 ± 0.31 h) to paralyse, in combination with either C. alata or C. angustifolia the time taken to paralyse became shorter. All the plant-treated parasites showed irrevocable changes in the scolex and proglottids as compared with the control, and these observations are comparable with those obtained with praziquantel. These results indicate that the three plants tested can be claimed to have anthelmintic activity in addition to their known properties, both when used individually and in combination. Further investigations will be required to evaluate their mechanism of action.

The present paper describes the taxonomy of two new monogeneans, namely, Thaparocleidus longiphallus sp. n. and T. siloniansis sp. n., based on morphological, morphometric and molecular biological analysis, collected from the fish Wallago attu (Bloch & Schn.) and Silonia silondia (Ham.), respectively, at Meerut, UP, India. Genetic characterization of the two new species is based on sequence analyses of the rDNA 28S gene using neighbour-joining and maximum-parsimony techniques. These methods are congruent in depicting T. longiphallus sp. n. and T. siloniansis sp. n. as closely related species, but distinct from each other and forming a subclade with other species of the genus Thaparocleidus Jain, 1952. Secondary-structure models of the large subunit rDNA of the two species were also predicted using a combined comparative and thermodynamic approach. Molecular morphometric and phylogenetic relationships of the isolates of the Thaparocleidus species are discussed in detail.

Ethanol extracts of 25 plant species were screened for anthelmintic effects against Haemonchus contortus. Ethanol extracts of each plant were used at various concentrations (10, 20 and 30%) to treat 10-day faecal cultures, incubated at 27°C with control cultures which were treated with ethanol for 48 h. Five plants with high efficacies (Ananas comosus, Aloe ferox, Allium sativum, Lespedeza cuneata and Warburgia salutaris) were selected from the first screening for further investigation using ethanol, dichloromethane and water extracts at four concentrations (2.5, 5, 10 and 20%). Ethanol was the most effective solvent. Larval counts decreased with increasing extract concentrations, of which 10 and 20% had similar effects. Lespedeza cuneata caused more than 70% mortality at all concentrations. However, there remains a need to assess in vivo efficacy of these plants.

Infectivity of Himasthla elongata cercariae to mussels, their second intermediate hosts, and resistance by these hosts to infection were assessed on the basis of the cercariae's ability to encyst in mussel haemolymph in vitro. A series of experimental in vivo infections of mussels with batches of cercariae, each batch released from a different single infected mollusc and referred to as a clone (due to their shared genotype), demonstrated that the results of the in vitro tests corresponded to the actual indices of infectivity/susceptibility of the parasites and their hosts. Most cercarial clones had high infectivity, with a few clones having very high or, at the other extreme, very low infectivity. A similar pattern was revealed in mussel resistance to cercarial infection. Most of the molluscs tested were moderately susceptible to cercarial infection, but at each extreme a small fraction (less than 10%) displayed very high or very low susceptibility. It was shown that there were no totally compatible or totally incompatible ‘cercaria clone/mussel’ combinations. Results obtained are compared with the data on intra-population variability using the characters parasite infectivity/host compatibility for trematode/mollusc–first intermediate host associations. Results are made relevant to actual infection levels in mussel settlements at the White Sea.

The development of molecular methods is becoming a promising field in the analysis of parasite fauna in wildlife species. This is especially useful in the case of parasite species where developmental larval stages are difficult to assess using standard methods. In this study we screened for the presence of parasitic nematodes infecting lacertid lizards from the Iberian Peninsula and Balearic islands using nematode-specific 18S ribosomal RNA gene primers on host tissue samples. Sequencing of positive samples revealed the presence of different genera of nematodes. The detection of Strongyloides, a monoxenous genus reported for the first time in Podarcis lilfordi is probably the result of the amplification of larval stages present in the host circulatory system. Two spirurid nematodes, Synhimantus and a new unidentified clade, were also found, suggesting that reptiles might be paratenic hosts of several spirurid species. This study demonstrates the benefits of using specific molecular markers on tissue samples to identify infecting larval stages of nematodes, otherwise difficult to assess using traditional screening methods.

Few studies have examined vertebrate models of invasive species to explore parasite release as a proposed mechanism through which host species might become invasive. In this study, we examined evidence for parasite release in invasive American bullfrogs (Rana catesbeiana/Lithobates catesbeianus) from five sites in Victoria, British Columbia, Canada. We examined helminth species richness, as well as the prevalence, intensity and abundance of lung and kidney fluke infections. These flukes are expected to impose costs on host survival, growth and reproductive output. We compared measures of these parasite taxa with bullfrogs from Ontario and New Brunswick where they are endemic. Helminth species richness in bullfrogs from the Victoria sites was lower than in Ontario bullfrogs, but comparable to reported indices for other endemic populations. The prevalence of lung flukes (Haematoloechus spp.) in bullfrogs from Victoria was twice as high as was observed in the Ontario bullfrogs, and higher than has been reported from other endemic locations. In four of the five study sites in Victoria, numbers of Echinostoma spp. kidney cysts were lower than observed in endemic populations; however, the fifth site had uncharacteristically high numbers of cysts. In this study, there did not appear to be clear evidence to support parasite release using either parasite species numbers, or infection by specific parasite taxa. Instead, the invasive bullfrogs demonstrated high parasite species richness and high levels of infection for parasites known to be harmful to their hosts.

This study aimed to evaluate the helminth parasites of Geophagus proximus from the São José dos Dourados River, a tributary of Paraná River, Ilha Solteira Reservoir, São Paulo State, Brazil. From May 2006 to May 2007, 116 G. proximus specimens were examined and seven different taxa of helminth were collected and identified: proteocephalidean plerocercoids (Cestoda); metacercariae of Austrodiplostomumcompactum, Clinostomum heluans and Clinostomum sp. (Trematoda); and Raphidascaris (Sprentascaris) hypostomi, and larvae of Raphidascaris sp. and Contracaecum sp. (Nematoda). All parasites presented the typical aggregated pattern of distribution, as well as the presence of a high number of larval stages, an absence of influence of the host sex and seasonality upon community parameters, as well as a correlation between species richness and host body weight. Moreover, with the exception of A. compactum metacercariae, all helminths found in this study are reported for the first time in G. proximus.

Trehalose 6-phosphate (T6P) synthase (TPS; EC 2.4.1.15) was isolated from muscles of Ascaris suum by ammonium sulphate fractionation, ion-exchange DEAE SEPHACELTM anion exchanger column chromatography and Sepharose 6B gel filtration. On sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE), 265-fold purified TPS exhibited a molecular weight of 66 kDa. The optimum pH and temperature of the purified enzyme were 3.8–4.2 and 35°C, respectively. The isoelectric point (pI) of TPS was pH 5.4. The studied TPS was not absolutely substrate specific. Besides glucose 6-phosphate, the enzyme was able to use fructose 6-phosphate as an acceptor of glucose. TPS was activated by 10 mM MgCl2, 10 mM CaCl2 and 10 mM NaCl. In addition, it was inhibited by ethylenediaminetetra-acetic acid (EDTA), KCl, FeCl3 and ZnCl2. Two genes encoding TPS were isolated and sequenced from muscles of the parasite. Complete coding sequences for tps1 (JF412033.2) and tps2 (JF412034.2) were 3917 bp and 3976 bp, respectively. Translation products (AEX60788.1 and AEX60787.1) showed expression to the glucosyltransferase-GTB-type superfamily.

During a survey of edible Crustacea for recovery of infective stages (metacercariae) of potential helminthozoonoses of trematode origin in north-east India, the crab species Barytelphusa lugubris mansoniana, collected from suspected foci of lungfluke infection in Meghalaya and Assam, was found to harbour metacercarial cysts that were different from the earlier reported infection, in which the lungfluke Paragonimus was confirmed to be implicated. Using morphological criteria, this metacercaria was identified as Microphallus indicus Mukherjee & Ghosh, 1967 of the trematode family Microphallidae. The present study extends the previous work by providing molecular characterization of this parasite using ribosomal internal transcribed spacer regions (rDNA ITS1 and ITS2) and the partial large ribosomal subunit DNA, lsr. These target regions were amplified by polymerase chain reaction (PCR) using trematode universal primers and sequenced. In BLAST analysis the query sequences were found close to members of Microphallidae and closest to the genus Microphallus.

A total of 124 specimens of the tufted duck, Aythya fuligula, and 63 greater scaup, A. marila, were examined for digenean parasites. Both duck species, which overwinter in a coastal lake connected with the southern Baltic (north-west Poland) were found to support Amblosoma exile, Cyathocotyle prussica,Paracoenogonimus ovatus, Australapatemon minor, Cotylurus cornutus, Echinoparyphium recurvatum, Echinostoma revolutum and Notocotylus attenuatus. In addition, the tufted duck hosted Hypoderaeum conoideum, Bilharziella polonica, Neoeucotyle zakharovi, Renicola mediovitellata, Psilochasmus oxyurus, Psilostomum brevicolle and Cryptocotyle concava; Echinostoma nordiana occurred in the greater scaup only. The two duck species differed significantly in the intensity and abundance of their digenean infection. Aythya marila harboured higher intensity levels and a wider assemblage of digeneans than A. fuligula, and this was likely to be due to differences in the pre-wintering exposure of the duck species to infective stages of these freshwater digeneans. Digenean communities in both duck species, strongly dominated by E. recurvatum, were relatively similar in their structure. No significant sex-dependent differences in digenean infections were revealed, except for the infection with N. attenuatus in A. fuligula. Similarly, there were no significant age-dependent differences (adult versus immature birds) in digenean infections, except for that with N. attenuatus in A. fuligula. The structural similarity between digenean communities in the two duck species is most likely an effect of overlapping diets based on freshwater molluscs, components of the digenean transmission pathway to definitive hosts.

Hydatidosis is an important public health problem in several parts of Iran. The aim of this molecular study is to investigate Echinococcus granulosus genotypes as the causative agents of hydatidosis in the south-west of Iran (Khuzestan province). In this study, isolates of 334 hydatid cysts were collected from the liver and lungs of 141 sheep, 104 cattle, 84 goats and 5 human cases. DNA was extracted and examined by nested polymerase chain reaction (PCR) of ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1) and restriction fragment length polymorphism (RFLP)-PCR. In addition, fragments of genes coding for ITS1 were sequenced. The results of RFLP-PCR analysis revealed the presence of the G1 genotype in all human, cattle, goat and sheep isolates. Furthermore, no camel strain (G6) was detected among all samples in the regions studied. The molecular findings indicate that the predominant genotype involved in E. granulosus transmission in south-west Iran is the common sheep strain (G1 genotype), which occurs in human, cattle, sheep and goat populations. In conclusion, these results may have important implications for hydatid disease control in the areas studied.

The aim of the present study was to detect changes in the structure of the helminth parasite infracommunities in Salminus brasiliensis (Cuvier 1816) from the floodplain of the upper Paraná River after construction of the Porto Primavera Hydroelectric Plant. A total of 126 fish in the period before the dam's construction and 56 specimens 10 years after this event were analysed. Three species of parasites were collected before the construction of the dam: Prosthenhysteraobesa Diesing, 1850 (Digenea), Cladocystis intestinalis Vaz, 1932 (Digenea) and Monticellia coryphicephala Monticelli, 1892 (Cestoda), and one nematode species in the larval stage, whose identification was not possible. After dam construction, the following helminth parasites were found: C. intestinalis, M. coryphicephala, Octospiniferoides incognita, Contracaecum spp. larvae and Contracaecum sp. type 2 larvae Moravec, Kohn & Fernandes 1993. The diversity of helminth parasites measured by the Brillouin diversity index (HB) differed significantly between the pre- and post-dam periods (mean HB = 0.069 and HB = 0.2, respectively; P= 0.0479; Mann–Whitney U test). The parasite community of S. brasiliensis before the construction of the dam showed concentration of dominance (C) of P. obesa (C = 0.38), while there was no concentration of dominance of any species of parasite (C = 0.22) after the dam's construction. Before the Porto Primavera dam the relative condition factor of fish was 1.0; after the dam's construction it was 0.93 (P <  0.0001; Mann–Whitney U test). This study records the disappearance of the species P. obesa and suggests that there has been local extinction of this parasite. The results show that the anthropic influence on natural systems is interfering with the welfare and health of S. brasiliensis, reflected by its fauna of helminth parasites.

Opisthorchiasis, caused by the liver fluke Opisthorchis viverrini, a food-borne trematode, is an important public health problem; however, only a single drug, praziquantel is available. We investigated tribendimidine–praziquantel combinations against O. viverriniin vitro and in vivo. The IC50 values of 0.16 μg/ml and 0.05 μg/ml were determined for praziquantel and tribendimidine, respectively, against adult O. viverriniin vitro. When O. viverrini was exposed to both drugs simultaneously (using a drug ratio based on the IC50 (1:3.2)) a synergistic effect was calculated (combination index (CI) at the IC50= 0.7). A similar result was observed when drug addition in vitro was spaced by the respective half-lives of the drugs (a CI of 0.78 at the IC50 for tribendimidine followed by praziquantel and a CI of 0.47 at the IC50 for praziquantel followed by tribendimidine). In vivo median-effect dose (ED50) values of 191 mg/kg and 147 mg/kg were calculated for praziquantel and tribendimidine, respectively. Low to moderate worm burden reductions (38–62%) were observed in O. viverrini infected hamsters when both drugs were administered simultaneously or on subsequent days, pointing to antagonistic effects in vivo. Further studies are necessary to understand the striking differences between the in vitro and in vivo observations using combinations of praziquantel and tribendimidine on O. viverrini.