Adults and plerocercoids of a new species of Heteronybelinia (Trypanorhyncha, Tentaculariidae) were recovered from the skate Sympterygia bonapartii (Rajiformes: Rajidae) and two species of teleosts, Raneya brasiliensis (Ophidiiformes: Ophidiidae) and Nemadactylus bergi (Perciformes: Cheilodactylidae), respectively. Heteronybelinia mattisi n. sp. differs from its congeners in its possession of a tentacular armature consisting of hooks with slight differences in size and shape on opposite surfaces of the tentacle (uncinate with rounded bases on the bothrial surface and uncinate with elongate bases on the antibothrial surface), without a characteristic basal armature, hooks increasing in size toward the tip of the tentacles, and pars bothrialis slightly overlapping with bulbs. The description of Heteronybelinia is emended regarding the distribution of testes to include species without postovarian testes (H. palliata and H. mattisi), and without testes anterior to cirrus sac on the poral side (H. robusta and H. mattisi). Host data are summarized for all 15 valid species of Heteronybelinia. Host specificity of adults and plerocercoids of Heteronybelinia is variable among species, the adults being more host specific than the plerocercoids. With the exception of H. mattisi, the definitive hosts of all species of Heteronybelinia for which adults are known include carcharhiniform sharks. Heteronybelinia mattisi seems to have oioxenous specificity for its definitive batoid host, S. bonapartii, and to be more specific for the intermediate hosts than most of its congeners.

Proteolytic activities in extracts from the plant-parasitic nematodes Heterodera glycines and Meloidogyneincognita were examined for their abilities to digest three FRET-modified peptide substrates representing members of the large FMRFamide-like peptide (FLP) family in nematodes. Included were sequences distributed across all nematode species (KSAYMRFa and KHEYLRFa) and a sequence confined to a narrow range of plant-parasitic nematodes (KHEFVRFa). Species variations were observed among substrate affinities, reaction rates and effect of protease inhibitors. Km values for KHEYLRFa (1.48 ± 0.34 μm) and KSAYMRFa (2.13 ± 0.24 μm) in H. glycines were each lower (P< 0.05) than those for the same substrates in M. incognita (5.26 ± 1.30 μm and 3.90 ± 0.61 μm, respectively). The Km of KHEFVRFa was lower (P< 0.05) in M. incognita (5.83 ± 0.36 μm) than in H. glycines (11.01 ± 1.26 μm). Reaction rates (Vmax/min/μg) for KHEYLRFa were the same for both species, but KSAYMRFa and KHEFVRFa digestion rates were each nearly twofold higher (P< 0.05) in M. incognita than in H. glycines. Digestion of KSAYMRFa was strongly inhibited in both species by 4-(2-aminoethyl)-benzenesulfonyl-fluoride-HCl (AEBSF) and EDTA, but M. incognita was more sensitive (P< 0.05) to inhibition. AEBSF and EDTA (both at 1 mm) inhibited M. incognita activity 62.3% and 36.6% more, respectively, than H. glycines activity. Serine protease inhibition differed significantly (P< 0.05) between the two species. Maximum inhibition of M. incognita (76%) occurred at 1.85 mm AEBSF while maximum inhibition of H. glycines was 40% at 1.19 mm AEBSF.

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.

Bivitellobilharzia nairi was first recorded from an Indian elephant (Elephas maximus) in Berlin. Infections with this parasite have become increasingly important in E. maximus maximus populations in Sri Lanka. The present work is the first morphological description of this schistosome from Sri Lanka. A number of adult worms were recovered from a dead Asian elephant near the elephant orphanage, Pinnawala, in Sri Lanka. The observed clinical features of the infected elephant included emaciation, subventral oedema and anaemia. Post-mortem results indicated that the liver was enlarged and adult schistosomes were found in the blood vessels of the liver parenchyma. The total number of worms recovered from a portion of the liver was 129,870, which is an average of 22 worms per 100 g of liver. The present study uses both light microscopic and scanning electron microscope (SEM) techniques for the morphological and topographical characterization of this parasite and to permit comparison with other species of schistosomes. Morphologically, these worms correspond very well to the description of B. nairi by Dutt & Srivastava (1955). Moreover, it is clear that B. nairi is a distinctive species easily differentiated from other schistosomes. The SEM study of the tegument of male worms shows that the surface of B. nairi is smoother than in other schistosomes.

Toxocara spp. eggs require a period of time under appropriate environmental conditions to become infective to definitive and paratenic hosts. Temperature and humidity are important factors known to affect the levels of development in soil. We aimed to investigate whether the eggs of T. canis could embryonate in dog hair under controlled conditions of temperature and humidity and, if so, to what degree. No previous work had been carried out on embryonation in hair under controlled conditions. Soil samples exposed to the same conditions as the hair samples were considered a suitable comparison in order to investigate differing levels of development. Development at two temperatures (10°C and 20°C) and the addition of water to samples was investigated over a period of 8 weeks. Importantly, we demonstrated that unembryonated T. canis eggs are capable of development in hair under controlled conditions. The rate of development is lower than that observed in soil, but remains biologically significant in terms of the overall numbers of potentially infective embryonated eggs present. Temperature is responsible for the rate of embryonation while moisture is essential for encouraging development and maintaining egg viability in general. In light of these findings the transmission of Toxocara spp. as a result of direct contact with well-cared-for owned dogs seems unlikely, but should not be ignored.

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.

The bivalve Perumytilus purpuratus is a common species that is widely distributed throughout rocky intertidal zones in Chile. This bivalve is the first intermediate host for three trematode species: one bucephalid (an undetermined species) and two fellodistomids (Proctoeces lintoni and one undetermined species). A few studies based on morphological comparisons, experimental infection and molecular analyses have been performed to ascertain the taxon (at least at the family level) to which these trematodes belong; yet, there remains no clarification about the specific identity of these trematodes. Therefore, in this study, we compared the V4 region nucleotide sequences of the 18S rRNA of these three sporocyst species, classified as morphotypes, found in P. purpuratus and nine adult trematode species from intertidal fishes that are likely definitive hosts for these parasites. The sequences from two of the sporocyst morphotypes matched with adult trematodes from the intertidal fish: type 1 sporocyst was similar to Prosorhynchoides carvajali (Bucephalidae), with a mean genetic divergence of 0.78%, and type 2 sporocyst was similar to Proctoeces sp. (but not P. lintoni), with 0% genetic divergence. The third species (type 3 sporocyst) was classified to the family Fellodistomidae; however, the sequence from this species differed greatly from the three other fellodistomid species documented in the marine fish of Chile and from other fellodistomids in public databases. Moreover, this morphotype has a particular cercarial morphology that greatly differs from other fellodistomid species described thus far. Therefore, this intriguing trematode remains a mystery.

Investigations were undertaken using light and transmission electron microscopy to clearly delineate the morphology of the cercomer, i.e. the protective envelopes and tail appendage, in cysticercoids of Microsomacanthus paraparvula, which develop in the haemocoel of the caddisworm Grensia praeterita (Insecta: Trichoptera). Two protective envelopes, the exocyst and endocyst, were identified. The non-cellular exocyst is found to consist of granular material and of thin, dense membrane-like layers, which are located parallel to each other. The exocyst of the mature metacestode tightly adjoins the outer surface of the endocyst, containing prospective parts (the scolex and the neck), except for the areas at its poles. A long tail appendage is located outside the exocyst. Evidence was found to indicate the existence of active synthetic processes occurring in the tail appendage. Non-cellular exocysts are widely distributed within metacestodes of the families Hymenolepididae and Dilepididae, and, presumably, are formed by means of glandular secretions from the oncosphere, given the early appearance of non-cellular exocysts in ontogeny.

The aim of the present study was to investigate the relationship between nematode infection and the ontogeny and diet of the lizard Tropidurus torquatus from a rocky outcrop in the state of Minas Gerais, south-eastern Brazil. Eighty-nine of 110 lizards examined (81.9%) harboured nematodes. Two nematode species were identified, namely, Physaloptera lutzi in the stomach and Parapharyngodon bainae in the intestine, with prevalence values of 67.3 and 60.0%, respectively. The lizard diet was composed mainly of ants, other hymenopterans, beetles and flowers of the species Centrosema coriaceum (Fabaceae). Host body size was positively correlated with nematode abundance, with adults more heavily parasitized than juveniles. The consumption of C. coriaceum had a negative effect on the abundance of both nematode species, suggesting that this plant may possess anthelmintic properties. The probability of a higher worm burden in adult hosts is likely linked with a longer exposure time to infective stages. Beetles, ants and hymenopterans appear to be the main intermediate hosts for P. lutzi. In general, ontogeny and diet composition determine the structure of the helminth community in this species of lizard.

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.

The helminth fauna of Agouti paca (Linnaeus, 1766) has seldom been studied. In this paper, we report an unusual mixed infection of Echinococcus vogeli Rausch & Bernstein, 1972 and Calodium hepaticum (syn. Capillaria hepatica Bancroft, 1863) in free-ranging paca from a forested region in Acre (Brazil). Gross morphological examination revealed that paca liver contained multiple spherical to subspherical white or translucent lesions, which were isolated or frequently contiguous and partially covered by Glisson's capsule. Microscopic examination revealed unilocular cystic structures that contained abundant brood capsules in which numerous protoscolices budded from the inner surface. The protoscolices possessed rostellar hooks (33–41 μm in length), a morphological characteristic of the blade and calcareous corpuscles that is consistent with the metacestode E. vogeli. The diagnosis of C. hepaticum infection was based on the morphology and morphometry of the egg-shaped ellipsoids with bipolar plugs (44.8 ± 1.9 μm (length) × 24.4 ± 2.0 μm (width)) and liver histopathology. This finding expands the known range of C. hepaticum hosts in South America and, to the best of our knowledge, it is the first case of a mixed infection of E. vogeli and C. hepaticum. Furthermore, our data provide evidence that wild animal meat may be a source of C. hepaticum infection.

Dirofilariosis is considered to be the arthropod vector-borne disease with the fastest spread in Europe. Slovakia belongs to new endemic regions for canine and human infections. This paper reports the fourth human case, where diagnosis was confirmed using computed tomography (CT), histological and molecular examinations – for the first time in this endemic region. The epidemiological history of this case indicated the autochthonous origin, and infection manifested by intense swelling in the periocular region, where a subcutaneous nodule was localized. Microscopic analysis of cross-sections of the surgically removed nodule confirmed the presence of a single male worm of Dirofilaria repens, indicated by the external cuticular ridges, the robust muscle cells and a single male sex organ seen as one tube beside the intestine. Considering that the worm morphology was partially damaged, molecular study was performed using DNA isolated from formalin-fixed and paraffin-embedded tissue sections of the worm. In polymerase chain reactions (PCR) a set of primers specific for D. repens DNA for the CO1-encoding gene amplified the expected 246 bp product using a high concentration of DNA template. Our diagnostic approach, involving molecular techniques, showed that identification of D. repens from excised tissues is possible even when the morphology of the worm and DNA are damaged during tissue processing. It is expected that the spread of this disease will continue due to climatic changes in central Europe. This warrants higher awareness among clinicians, who will initially be approached by patients, and better co-operation with parasitologists in newly endemic countries.

Trichinella zimbabwensis has been found naturally infecting crocodiles (Crocodylus niloticus) in Zimbabwe, Mozambique, Ethiopia and South Africa, as well as monitor lizards (Varanus niloticus) in Zimbabwe. The reports on natural infections were mostly accidental rather than structured surveys and involved very few animals. Previous surveillance studies in South Africa reported a 38.5% prevalence of T. zimbabwensis among wild crocodiles tested from the Mpumalanga province and Kruger National Park (KNP). No studies have been conducted to date on the geographical distribution and occurrence of T. zimbabwensis in wild crocodiles and varans in countries in southern Africa. Recent outbreaks of pansteatitis in crocodile populations of the KNP, South Africa, provided an opportunity to conduct a more structured survey aimed at elucidating the occurrence and distribution of T. zimbabwensis in culled wild crocodile populations within the KNP. Results from this study showed that T. zimbabwensis occurred in 10 out of 12 culled crocodiles form the KNP. The results also showed that the natural distribution of T. zimbabwensis in crocodiles includes all the major river systems in the KNP. The predilection sites of larvae in muscles followed a different pattern in naturally infected crocodiles compared to observations in experimentally infected mammalian hosts.

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.

Experimental infections of two South American lymnaeid populations with Fascioloides magna were carried out to determine whether these snails may sustain larval development of this digenean and, if so, to quantify their potential for cercarial production. The reference group was a French population of Galba truncatula infected and raised according to the same protocol. According to the internal transcribed sequence (ITS)-1 segment of their genomic rDNA, these South American populations were identified as Lymnaea neotropica (origin, Argentina) and Lymnaea viatrix var. ventricosa (origin, Uruguay). In the snail groups followed for cercarial shedding, longer prepatent periods and lower numbers of shed cercariae were noted in South American lymnaeids. In other snails dissected at day 65 post-exposure, the redial and cercarial burdens of F. magna found in the bodies of L. neotropica and L. v. ventricosa were significantly lower than those noted in G. truncatula. Compared to the total cercarial production noted in the dissected snails, the percentage of cercariae that exited from snails was 51.3% for G. truncatula, 32.2% for L. neotropica and 46.8% for L. v. ventricosa. The two South American species of snails can thus be considered as potential intermediate hosts of F. magna.

The present study examined sequence variation in three mitochondrial DNA (mtDNA) genes, namely cytochrome c oxidase subunit 3 (cox3) and NADH dehydrogenase subunits 1 and 4 (nad1 and nad4), among Ascaridia galli isolates from different geographical localities in China. A portion of cox3 (pcox3), nad1 (pnad1) and nad4 (pnad4) genes were amplified by polymerase chain reaction (PCR) separately from adult A. galli individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of pcox3, pnad1 and pnad4 were 408 bp, 471 bp and 333 bp, respectively. The intraspecific sequence variations within A. galli were 0–1.7% for pcox3, 0–2.8% for pnad1 and 0–3.4% for pnad4. The A+T contents of the sequences were 67.16–67.65% (pcox3), 67.09–67.94% (pnad1) and 69.91–71.77% (pnad4). The interspecific sequence differences among members of the Ascaridida were significantly higher, being 13.2–30.9%, 12.8–29.0% and 15.1–34.1% for pcox3, pnad1 and pnad4, respectively. Phylogenetic analyses using combined sequences of pcox3, pnad1 and pnad4, with three different computational algorithms (Bayesian analysis, maximum likelihood and maximum parsimony), all revealed distinct groups with high statistical support. These findings demonstrated the existence of intraspecific variation in mitochondrial DNA (mtDNA) sequences among A. galli isolates from different geographical regions in China, and have implications for studying molecular epidemiology and population genetics of A. galli.

The parasite fauna of juvenile Dissostichus eleginoides, while they inhabit the Falkland's shelf, was examined, giving new detailed information on spatial, ontogenic and seasonal variations. A total of 24,943 parasites from 15 different taxa were found in the stomach of 502 individual fish. Parasite species composition and abundance allowed separation of toothfish by area between the north-west and south-east of the Falklands. The digenean, Elytrophalloides oatesi, and the nematodes, Hysterothylacium spp. and Anisakis spp., were the most common, all with a prevalence >20%. For some seasons ontogenic changes in abundance were significant in these three parasite taxa, and this is discussed in terms of ontogenic and seasonal changes in diet. Elytrophalloides oatesi and Hysterothylacium spp. showed spatial and seasonal differences in abundance with greater numbers in the warmer waters of the north-west and during the summer months. Differences in abundance of E. oatesi between the Falklands and other regions indicate its potential for use as a biological tag to study toothfish movements and population structure; however, more seasonal data would be required before this technique could be used.

Parasites commonly share their hosts with specimens of the same or different parasite species, resulting in multiple parasites obtaining resources from the same host. This could potentially lead to conflicts between co-infecting parasites, especially at high infection intensities. In Pool Los Juncos (Patagonia, Argentina), the amphipod Hyalella patagonica is an intermediate host to three parasites that mature in birds (the acanthocephalan Pseudocorynosoma sp. and larval stages of two Cyclophyllidea cestodes), in addition to a microsporidian (Thelohania sp.), whose life cycle is unknown, but very likely to be monoxenous. The aim of this study was to describe interactions between these parasite species in their amphipod host population. Amphipods were collected monthly between June 2002 and January 2004 to assess parasite infection. Infection prevalence and mean intensity were greatest in larger male amphipods for all parasite species. We also found a positive association between Thelohania sp. and both Pseudocorynosoma sp. and Cyclophyllidea sp. 1 infections, though Pseudocorynosoma sp. and Cyclophyllidea sp. 1 were negatively associated with each other. We conclude that contrasting associations between parasite species may be associated with competition for both food intake and space in the haemocoel.

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 nematode communities in spruce forests were compared with the short-term effects of forest damage, caused by windstorm, wildfire and management practices of forest soils. Soil samples were collected in June and October from 2006 to 2008 in four different sites: (1) forest unaffected by the wind (REF); (2) storm-felled forest with salvaged timber (EXT); (3) modified forest affected by timber salvage (wood removal) and forest fire (FIR); and (4) storm-felled forest where timber had been left unsalvaged (NEX). Nematode analysis showed that the dominant species in all four investigated sites were Acrobeloides nanus and Eudorylaimus silvaticus. An increase of A. nanus (35% of the total nematode abundance) in the first year in the FIR site led to the highest total abundance of nematodes compared with other sites, where nematode abundance reached the same level in the third year. In the FIR site bacterial feeders appeared to be the most representative trophic group, although in the second and third year, after disturbance, the abundance of this trophic group gradually decreased. In the NEX site, the number of nematode species, population densities and Maturity Index were similar to that recorded for the FIR site. In EXT and NEX sites, the other dominant species was the plant parasitic nematode Paratylenchus microdorus. Analyses of nematodes extracted from different forest soil samples showed that the highest number of species and diversity index for species (H'spp) were in the REF site. Differences between the nematode fauna in REF and other localities were clearly depicted by cluster analysis. The greatest Structure Index and Enrichment Index values were also in REF. In the EXT site, the number of nematode species, their abundance, H'spp and Maturity Index were not significantly different from those recorded in the reference site.