A study was carried out on the gastrointestinal helminth communities of blackspot seabream, Pagellus bogaraveo, from the Portuguese mainland and island waters. Fifteen helminth parasite species were detected, including the following new records in this host: the digeneans Accacladocoelium petasiporum, Brachyenteron helicoleni,Glomericirrus macrouri, Lecithocladium excisum, Lepocreadium album and Pachycreadium carnosum; the nematode Camallanus sp. and the acanthocephalan Rhadinorhynchus pristis. Infection levels were generally low. Samples from mainland waters had a significantly higher prevalence of digeneans than those from the islands, probably due to the greater availability of invertebrate intermediate hosts on the continental slope. The helminth community of P. bogaraveo from Azores was diverse but mostly composed of species that infected no more than a single host, whereas in Madeira the helminth community was very depauperate. Most helminth species exhibited niche preferences within the gastrointestinal tract of the host. Higher functional richness and evenness were obtained in helminth communities of P. bogaraveo from mainland waters, indicating a balanced use of available resources, which is suggestive of more stable gastrointestinal helminth communities in these waters.

Anisakis infection parameters were studied in horse mackerel (Trachurus trachurus) taken from two areas of northern Morocco (Tetouan and Tangier), which showed a mean prevalence of 54.9%. Identification of the ITS1–5.8S–ITS2 fragment by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) showed A. pegreffii to be the dominant species; no A. simplex s.s. were detected. The presence of A. pegreffii in horse mackerel was not influenced by the sex (P= 0.46) or catch area (Atlantic versus Mediterranean, P= 0.52) of the fish, but was significantly related to their length, weight, liver weight and gonad weight, and to the season of their capture (P< 0.05). A low prevalence (8.6%) and mean intensity (1.0) was detected in the muscle, probably related to the reduced ability of A. pegreffii to penetrate muscle. The risk of the presence of A. pegreffii in the muscle was fivefold higher in fish caught during the summer than during any other season. Susceptible members of the human population can minimize the risk of infection by avoiding the consumption of larger horse mackerel specimens during the summer.

Male dimorphism of the subfamily Ostertagiinae (Nematoda: Trichostrongylidae) is a well-known phenomenon, and two or more morphotypes of a single species have previously been described as different species. Two Spiculopteragia spp., S. houdemeri (syn. S. yamashitai) and S. andreevae (syn. Rinadia andreevae) recorded in Asian cervids and wild bovids, are considered to represent major and minor morphs of S. houdemeri, respectively, based solely on their co-occurrence in the same host individual along with monomorphic females. In this study, males of morph houdemeri ( = S. houdemeri) and morph andreevae ( = S. andreevae) as well as females with three different vulval ornamentations were collected from sika deer (Cervus nippon) and Japanese serows (Capricornis crispus) distributed on the mainland of Japan. Morphologically characterized worms were subjected to molecular genetic analyses based on the internal transcribed spacer region of the ribosomal RNA gene and a partial region of the cytochrome c oxidase subunit I gene of mitochondrial DNA. Of 181 collected sika deer, 177 (97.8%) and 73 (40.3%) deer harboured males of morphs houdemeri and andreevae, respectively. Worm numbers of the former morph were found to range between 1 and 444 per individual, whereas only 1–25 worms per individual were detected for the latter morph. Five out of six serows harboured 47–71 or 2–9 males of morphs houdemeri and andreevae per individual, respectively. Females with one or two vulval flaps were predominant, but there was a substantial presence of flapless females in both host species. All the morphs of male and female adults had an identical genetic background, thus directly confirming the morphological polymorphism of S. houdemeri.

Trichinellosis is an important helminthic food-borne zoonosis, which is caused by nematodes of the genus Trichinella. Although, Trichinella spp. has been detected frequently in Iranian wildlife, this parasitic infection is not considered a major public health problem. This is largely because Islamic codes forbid consumption of pork meat in this country. However, knowledge about this zoonotic pathogen is important because human trichinellosis has been documented in countries where most of the population is Muslim. The aims of the present work were to investigate whether Trichinella spp. was still circulating in wildlife of the Khuzestan Province (south-west Iran) about 30 years after the first investigation, to identify the aetiological agent at the species level by molecular analyses, and to review the literature on Trichinella spp. in animals of Iran. During the winter 2009–2010, muscle samples from 32 road-killed animals (14 dogs and 18 jackals, Canis aureus) were collected. Muscle samples were digested and Trichinella sp. larvae were isolated from two jackals. The Trichinella sp. larvae have been identified as Trichinellabritovi by molecular analyses. These results confirm that T. britovi is the prevalent species circulating in wild animals of Iran.

A new entomopathogenic nematode in the genus Heterorhabditis is described from South Africa, from two singular isolates found 1000 km from each other, from beneath a fig tree and in a citrus orchard, respectively. Morphological and molecular studies indicate both isolates to be the same and a new undescribed Heterorhabditis species. Comparison of sequences of the internal transcribed spacer (ITS) rDNA and the D2D3 region of the 28S rDNA gene with available sequences of other described species within the genus, indicate the two isolates as a new species. Phylogenetic analysis of the sequence data concerned placed the new species, H. noenieputensis n. sp., closest to H. indica and H. gerrardi in the indica-group. The new species, H. noenieputensis n. sp., is distinguished from other species in the genus by a combination of several morphological traits of the males and the infective juveniles (IJs). The new species differs from all other species previously described, as regards the body length of the IJs, except for H. indica and H. taysearae, in which the IJ is smaller. The IJ also differs from that of H. indica in the length of the oesophagus, the body diameter, the length of the tail and the E%. In addition, males of H. noenieputensis n. sp. differ from their closest relative, H. indica, in the position of the excretory pore, SW% and D%; and from H. gerrardi in the length of the oesophagus and SW%. The seventh pair of genital papillae of H. noenieputensis n. sp. are normally developed, while for H. indica they are often branched or swollen at the base, while 8 and 9 are usually absent in both species.

The present study considers the influence of the trophic status of three Kashmir Himalayan lakes on the patterns of helminth infracommunities in populations of three species of fish during 2006 to 2008. Data were collected from three lakes of differing trophic status in the Kashmir Himalayas, namely Anchar, a hyper(eu)trophic lake; Dal, a eutrophic lake; and Manasbal, a meso(eu)trophic lake. Three species of fish examined included the native fish Schizothorax niger Heckel and two exotic species – Carassius carassius (Linnaeus) and Cyprinus carpio Linnaeus. The analysis of data showed a clear habitat effect on the abundance pattern of helminth species, thus revealing lake-specific differences in parasite infracommunities of both S. niger and C. carassius. Helminth infracommunity richness was the highest in host populations from the Anchar lake compared to other two lakes. Low values in the Manasbal lake emphasize the low diversity of their helminth infracommunities. On the other hand, there was no observed pattern of community structure in the case of C. carpio in the three lake sites. However due to bias in sampling there was no distinct effect of fish body size on parasite infracommunity structure, although the present results do show that fish parasite data can be meaningful in diagnosing changes in the trophic condition of eutrophic lakes.

Several factors may influence the structure of parasite communities in amphibian hosts. In this study, we describe the helminth parasites of three allopatric populations of the frog Leptodactylus podicipinus and test whether host size and sex were determinants of the structure and composition of the helminth communities. One hundred and twenty-three anurans were collected from three different study sites within the Pantanal wetlands and surveyed for helminth parasites. We found 14 helminth taxa: 7 species of nematodes, 4 species of trematodes, 1 species of cestodes, 1 species of acanthocephalan and one unidentified cyst. Host sex and size did not cause significant differences in helminth abundance or richness. The structure of helminth communities from the three study sites varied in terms of species composition, abundance and diversity. Six out of 14 helminth taxa were found in the three localities. Among those, the nematodes Cosmocerca podicipinus and Rhabdias sp., the trematode Catadiscus propinquus and the helminth cyst showed significant differences in mean abundances. We suggest that such differences found among the three component communities are driven by biotic and abiotic factors operating locally. Moreover, these differences stress the importance of local conditions, such as hydrologic characteristics and landscape composition, on helminth community structure.

Stray cats live in high-density colonies in urban areas and pose a health hazard to household cats and humans. In Portugal, information on the parasitic fauna of stray cats is limited and relies mostly on results from faecal analysis. The present survey aimed to determine the prevalence, diversity and intensity of parasites in stray cats from the urban area of Lisbon by means of parasitological necropsy. Internal organs were collected from 162 cats captured in different areas of the city and systematically subjected to parasitological dissection. Helminths were identified by macro- and microscopic examination and protozoa by faecal floatation and sedimentation techniques. The overall prevalence of parasites was 90.7% (95% confidence interval (CI): 85.3–94.6%). A total of 12 parasite species was recorded: Cystoisospora felis (14.2%), Cystoisospora rivolta (46.3%), Sarcocystis sp. (1.2%), Ancylostoma tubaeforme (19.1%), Toxocara cati (38.3%), Ollulanus tricuspis (30.9%), Aelurostrongylus abstrusus (12.4%), Eucoleus aerophilus (0.6%), Taenia taeniaeformis (3.1%), Dipylidium caninum (53.1%), Joyeuxiella pasqualei (15.4%) and Diplopylidium nölleri (3.7%). Overall mean species richness was 2.36 ±  1.52. Helminth mean intensity was highest for O. tricuspis (285.8), followed by D. caninum (42.4), J. pasqualei (14.4), A. tubaeforme (8.1) and T. cati (5.9). The prevalence and variety of parasites found in our sampling are substantially higher than the numbers previously reported in Portugal. Some of the parasites, including T. cati and A. tubaeforme, are zoonotic, which emphasizes the need for parasite control strategies based on demographic containment of stray cat populations in urban areas to promote public health protection.

Predation by the mite Lasioseius penicilliger was studied on three nematode species, i.e. infective larval stages (L3) of Haemonchus contortus and adults of Panagrellus redivivus and Rhabditis sp. Experiments were carried out in 5.5-cm diameter Petri dishes containing 2% water-agar over a period of 5 days. Batches of up to 1500 third-stage larvae (L3) of H. contortus and 1000 adult nematodes of P. redivivus and Rhabditis sp. were exposed to five mites in separate Petri dishes. Upon contact, each mite used its pedipalp and legs to identify and hold its prey and then used its chelicerae to feed upon the prey. Predation by L. penicilliger was chance dependent but mites became aggregated around any injured/damaged prey, thereby suggesting some form of chemoperception. The rate of predation on the three species of nematodes was high but L3 of H. contortus and adult Rhabditis sp. were preferred.

A rapid dot immunogold filtration assay (DIGFA) was adopted for specific immunodiagnosis of human cerebral angiostrongyliasis, using purified 31-kDa glycoprotein specific to Angiostrongylus cantonensis as diagnostic antigen and protein A colloidal gold conjugate as antigen–antibody detector. A total of 59 serum samples were assayed – 11 samples from clinically diagnosed patients with detectable A. cantonensis-specific antibody in immunoblotting; 23 samples from patients with other related parasitic diseases, i.e. gnathostomiasis (n= 8), cysticercosis (n= 5), toxocariasis (n= 2), filariasis (n= 4), paragonimiasis (n= 2) and malaria (n= 2); and 25 samples from normal healthy subjects. The sensitivity and specificity of DIGFA to detect anti-A. cantonensis specific antibodies in serologically confirmed angiostrongyliasis cases, were both 100%. No positive DIGFA was observed in cases with other parasitic diseases, and the healthy control subjects. The 3-min DIGFA is as sensitive and specific as the 3-h immunoblot test in angiostrongyliasis confirmed cases that revealed a 31-kDa reactive band. The gold-based DIGFA is more rapid and easier to perform than the traditional enzyme-linked immunosorbent assay (ELISA). The test utilizing purified A. cantonensis antigen is reliable and reproducible for specific immunodiagnosis of human infection with A. cantonensis – thus can be applied as an additional routine test for clinical diagnostic support. Large-scale sero-epidemiological studies in endemic communities in north-east Thailand are under way to evaluate its usefulness under field conditions.

In July 2009 and July 2010 (two dry periods separated by an atypically large flood in the Pantanal wetland of Brazil), 34 and 33 specimens of the sardine fish Triportheus nematurus were collected, respectively, for the study of the metazoan parasite community of this species. Parasite ecological and community descriptors were calculated for both host samples, and possible similarities were tested statistically. Five species of metazoan parasites were identified, four of which were common to both host samples. A total of 61 metazoan parasites were collected from all fish hosts (17 specimens in July 2009 (mean: 0.5 ± 0.66 parasites/fish) and 44 specimens in July 2010 (mean: 1.33 ± 1.41 parasites/fish)). The nematode Procamallanus hilarii and the monogenean Anacanthorus sp. were the most prevalent and abundant species in 2009 and 2010, respectively. The mean total abundance and species richness were significantly higher in 2010. Parasite communities in both samples of T. nematurus were characterized by species with low prevalence, abundance, mean total abundance and species richness, thus indicating low parasite diversity. Significant differences in the prevalence and abundance of P. hilarii and Anacanthorus sp. between the two samples allowed the discrimination of infracommunities, which were united in two distinct groups. This appears to be the first evidence that the peculiar hydrological dynamics of the southern Pantanal wetland (Brazil) exert an important influence over the structure of the parasite community.

In this cross-sectional study, the associations between helminth infections and ABO blood group, anaemia and undernutrition were investigated in 480 febrile outpatients who visited Dore Bafeno Health Centre, southern Ethiopia, in December 2010. Stool specimens were processed using the Kato–Katz method and examined for intestinal helminth infections. Haemoglobin level was measured using a HemoCue machine and blood group was determined using an antisera haemagglutination test. Nutritional status of the study participants was assessed using height and weight measurements. Among the study participants, 50.2% were infected with intestinal helminths. Ascaris lumbricoides (32.7%), Trichuris trichiura (12.7%), Schistosoma mansoni (11.9%) and hookworm (11.0%) were the most frequently diagnosed helminths. The odds of infection and mean eggs per gram of different intestinal helminth species were comparable between the various blood groups. Among individuals who were infected with intestinal helminth(s), the mean haemoglobin level was significantly lower in individuals harbouring three or more helminth species and blood type AB compared to cases with double or single helminth infection and blood type O, respectively. The odds of being underweight was significantly higher in A. lumbricoides and T. trichiura infected individuals of age ≤ 5 and ≥ 20 years, respectively, when compared to individuals of the matching age group without intestinal helminths. In conclusion, infection with multiple intestinal helminths was associated with lower haemoglobin level, which was more severe in individuals with blood type AB. Future studies should focus on mechanisms by which blood group AB exacerbates the helminth-related reduction in mean haemoglobin level.

Helminth parasites use antioxidant defence strategies for survival during oxidative stress due to free radicals in the host. Accordingly, tissue-dwelling filarial parasites counteract host responses by releasing a number of antioxidants. Targeting these redox regulation proteins together, would facilitate effective parasite clearance. Here, we report the combined effect of protective immune responses trigged by recombinant Wuchereria bancrofti thioredoxin (WbTRX) and thioredoxin peroxidase (WbTPX) in an experimental filarial model. The expression of WbTRX and WbTPX in different stages of the parasite and their cross-reactivity were analysed by enzyme-linked immunosorbent assay (ELISA). The immunogenicity of recombinant proteins and their protective efficacy were studied in animal models when immunized in single or cocktail mode. The antigens showed cross-reactive epitopes and induced high humoral and cellular immune responses in mice. Further, parasite challenge against Brugia malayi L3 larvae in Mastomyscoucha conferred significant protection of 57% and 62% against WbTRX and WbTPX respectively. The efficacy of L3 clearance was significantly higher (71%) (P <  0.001) when the antigens were immunized together, showing a synergistic effect in multiple-mode vaccination. Hence, the study suggests WbTRX and WbTPX to be attractive vaccine candidates when immunized together and provides a tandem block for parasite elimination in the control of lymphatic filariasis.

Adults of Clinostomum spp. are digenetic trematodes found in fish-eating birds, reptiles and occasionally mammals, including humans. Freshwater snails serve as first intermediate hosts and many fish species and amphibians as second intermediate hosts. To date, amphibian hosts of Clinostomum metacercariae include members of urodele and anuran families in North America, but no data are available on infections of European amphibians, including newts. In this study, we characterize infections of Clinostomum complanatum metacercariae in four smooth (Lissotriton vulgaris) and 18 Italian crested newts (Triturus carnifex) from an artificial pond located in a protected area in Tuscany, Italy. Parasites were surgically removed from the infected newts and identified both morphologically and using sequences of a mitochondrial gene, cytochrome c oxidase I, and the ribosomal markers, internal transcribed spacers. This is the first record of C. complanatum in European newts and, more generally, in amphibians in Europe.

Levels of calcium in the haemolymph and reserves in the shell of Biomphalaria glabrata experimentally infected by Angiostrongylus cantonensis were determined for the first time. At the same time, histochemical analyses of the digestive gland of infected and uninfected snails were performed to better understand the possible changes in metabolism of calcium in these organisms. After 1, 2 and 3 weeks of infection, the snails were dissected for collection of haemolymph and separation of tissues. The highest calcium concentrations in the haemolymph were found 2 weeks after infection, with a 39.61% increase in relation to the respective control group. However, there was a significant reduction in the concentration of this ion in the haemolymph of infected snails after 1 week of infection in relation to the uninfected specimens. In parallel, intense hypocalcification was shown in the shell of infected snails 1 and 2 weeks after infection, differing significantly in relation to the respective control groups. Morphological changes in the digestive gland of infected snails were also observed, confirming the role of this ion as an important element in the parasite encapsulation process.

Human alveolar echinococcosis (AE) is caused by the accidental ingestion of Echinococcus multilocularis eggs. Early detection is essential as surgical resection is the only treatment for a complete cure. However, details are unclear about changes in the antibody response during the initial stages of infection, yet such information is useful for early serodiagnosis. Therefore, a long-term investigation was performed into the time course of the antibody response before ‘positive’ detection. Patient sera were used for enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) analysis using crude antigens extracted from E. multilocularis protoscoleces. Rats were experimentally infected with AE and similarly analysed by ELISA and WB. Among the markers for diagnoses, the 18 kDa band (main marker) appeared last in the preoperative stages and disappeared first after operation in a WB test. Although the 18 kDa antigen can be useful as a marker for AE diagnosis, it cannot contribute to the detection of some patients before the 18 kDa band appearance. To avoid misdiagnosis, different diagnostic antigens such as the 26–28 and 7–8 kDa bands should also be considered. These bands tend to appear earlier than the 18 kDa band and thus offer the potential for early detection of AE. We first observed changes in the antibody response in a relatively early stage after infection in human AE cases. Notably, changes in the antibody response of two intermediate species were similar. These findings provide valuable information for the early detection of human AE cases in the future.

Because the digenetic trematode fauna of Nepal is poorly known, we began to search for schistosomes in and around Chitwan National Park (CNP) of southern Nepal. Both domestic and wild Indian elephants (Elephus maximus) are present, and we found one of two dung samples from wild elephants and 1 of 22 (4.5%) dung samples from domestic elephants to be positive for schistosome eggs. The morphology of the eggs and both cox1 and 28S sequences derived from the eggs/miracidia were consistent with Bivitellobilharzia nairi, reported here for the first time from Nepal. Also, 7 of 14 faecal samples from the Asian or greater one-horned rhinoceros (Rhinoceros unicornis) contained viable eggs indistinguishable from those of B. nairi. This identification was confirmed by comparison with both cox1 and 28S sequences from B. nairi eggs/miracidia derived from Nepalese and Sri Lankan elephants. This represents the first sequence-verified identification of a schistosome from any species of rhinoceros, and the first verified occurrence of a representative of Bivitellobilharzia (a genus of ‘elephant schistosomes’) in mammals other than elephants. Our work suggests that elephants and rhinos share B. nairi in CNP, even though these two members of the ‘charismatic megafauna’ belong to unrelated mammalian families. Their shared life style of extensive contact with freshwater habitats likely plays a role, although the snail intermediate host and mode of definitive host infection for B. nairi have yet to be documented. This report also supports Bivitellobilharzia as a monophyletic group and its status as a distinct genus within Schistosomatidae.

The main goal of the current work was to develop and validate an in vitro fluke egg hatch test, as a method for the detection of albendazole (ABZ) resistance in the liver fluke, Fasciola hepatica. Fluke eggs (200/ml, n= 5) from six different isolates were used in the current experimental work. They were obtained from different geographical locations and named Cullompton (UK), CEDIVE (Chascomus, Argentina), INTA-Bariloche (Bariloche, Argentina), Rubino (Uruguay), Cajamarca (Perú) and Río Chico (Catamarca, Argentina). The fluke eggs were incubated (25°C) for a 12-h period in the presence of either ABZ or its sulphoxide metabolite (ABZ.SO) (5, 0.5 or 0.05 nmol/ml). Untreated eggs were incubated as a control. Incubated eggs (with or without drug present) were kept in darkness at 25°C for 15 days. Afterwards, the trematode eggs were exposed to daylight over a 2-h period. Hatched and unhatched eggs were evaluated using an optical microscope, and the ovicidal activity was assessed for each fluke isolate. A very low ovicidal activity ( ≤ 13.4%) was observed in the ABZ-resistant CEDIVE isolate for both ABZ and ABZ.SO. Conversely, in the INTA-Bariloche and Río Chico isolates, which are suspected to be susceptible to ABZ, ovicidal activities ≥ 70.3% were observed after incubation with ABZ at the lowest concentration tested (0.05 nmol/ml). This finding correlates with that previously described for the ABZ-susceptible Cullompton. Finally, the Cajamarca and Rubino isolates behaved as ABZ resistant, since no ovicidal activity was observed after eggs were incubated with ABZ at 0.5 nmol/ml. Considering the specific results obtained for each isolate under assessment, the egg hatch test described here may be a suitable method for detection of ABZ resistance in F. hepatica.

In spite of using aquatic snails as bioindicators for water pollution, little attention has been paid to the effect of parasitism upon the concentration of heavy metals (Al, Cd, Cu, Fe, Mn, Pb and Zn) in these organisms. The present study therefore aimed to compare the concentrations of heavy metals in trematode-infected Biomphalaria alexandrina collected from Kafer Alsheikh and Menofia provinces, Egypt, with uninfected snails from the same sites, in order to assess the effect of parasitism on the use of these snails as bioindicators. The concentrations of heavy metals in the soft parts and shells of snails were measured by flame atomic absorption spectrometry. The results showed that the heavy metal profile in snails infected with Echinostoma liei was very different from that in snails infected with Schistosoma mansoni. The total concentration of heavy metals in E. liei-infected snails collected from Kafer Alsheikh or Menofia province was greater than in uninfected snails. In contrast, the total concentration of heavy metals in S. mansoni-infected snails was reduced compared with uninfected snails. In conclusion, the status of snails with respect to parasitic infection must be taken into consideration when these snails are used as bioindicators.

A new species, Paradiplozoon ichthyoxanthon (Monogenea: Diplozoidae) is described from the gills of Labeobarbus aeneus (Cyprinidae) in the Vaal Dam and Vaal River Barrage in the Vaal River, South Africa. This species is the first in this genus from southern Africa; Paradiplozoon aegtyptense and Paradiplozoon ghanense have been described from northern Africa. The new species differs from all other Paradiplozoon species on the basis of the combination of morphological characteristics, namely disk-like haptor, eggs without filaments, clamp and central hook morphology and size of specimens. The species description is accompanied by a description of the reproductive organs. The internal transcribed spacer of the ribosomal gene (ITS2) was studied and the South African individuals formed a distinct clade, supporting the establishment of a new species.