Internal transcribed spacer (ITS) rDNA sequences of three Nematodirus species from naturally infected goats or sheep in two endemic provinces of China were analysed to establish an effective molecular approach to differentiate Nematodirus species in small ruminants. The respective intra-specific genetic variations in ITS1 and ITS2 rDNA regions were 0.3–1.8% and 0–0.4% in N. spathiger, 0–6.5% and 0–5.4% in N. helvetianus, and 0–4.4% and 0–6.1% in N. oiratianus from China. The respective intra-specific variations of ITS1 and ITS2 were 1.8–4.4% and 1.6–6.1% between N. oiratianus isolates from China and Iran, 5.7–7.1% and 6.3–8.3% between N. helvetianus samples from China and America. For N. spathiger, compared with samples from China, sequence differences in ITS1 rDNA were 0.3–2.4% in isolates from America, 0.3–2.9% in New Zealand and 2.1–2.4% in Australia. Genetic variations in ITS2 rDNA of N. spathiger were 0–0.4% between samples from China and America, and 0–0.8% between samples from China and New Zealand. Using mutation sites, polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and specific PCR techniques were developed to differentiate these three Nematodirus species. The specific PCR assay allowed the accurate identification of N. oiratianus from other common nematodes with a sensitivity of 0.69 pg and further examination of Nematodirus samples demonstrated the reliability of these two molecular methods.

Parasitic nematodes of the family Mermithidae were found to be infecting the introduced European earwig Forficula auricularia (Dermaptera: Forficulidae) in Dunedin, South Island, New Zealand. Adult females were later collected from various garden plants while depositing eggs. These mermithid specimens were identified morphologically as Mermis nigrescens Dujardin, 1842. A genetic distance of 0.7% between these specimens and a M. nigrescens isolate from Canada (18S rRNA gene), suggests that they have diverged genetically, but there are currently no available comparable sequences for the European M. nigrescens. Two additional nuclear fragments were also amplified, the 28S rRNA and the ribosomal DNA first internal transcribed spacer (ITS1), providing a basis for future studies. Bearing in mind the morphological similarity with other reported M. nigrescens and the lack of sequence data from other parts of the world, we retain the name M.nigrescens, and suggest that the species may be found to represent a complex of cryptic species when more worldwide data are available. Herein, we present a brief description of the post-parasitic worms and adult females, along with an inferred phylogeny using 18S rRNA gene sequences.

The rhadinorhynchid Sclerocollum saudii Al-Jahdali, 2010 was found in the intestine of its type host, Siganusrivulatus Forsskål & Niebuhr, 1775, a siganid fish permanently resident in a lagoon within the mangrove swamps found on the Egyptian coast of the Gulf of Aqaba (between 28°7′N and 28°18′N). Larval forms of this acanthocephalan (acanthors, acanthellae and cystacanths) were only found in Megaluropus agilis Hoek, 1889 (Crustacea: Gammaridae), a benthic amphipod abundant on algae and seagrasses in the lagoon. So, this life cycle of S. saudii was elucidated under semi-natural conditions: embryonated eggs of S. saudii were directly ingested by the amphipod and hatched in its intestine; the released acanthor penetrated the intestinal epithelium in 12–18 h to reach the connective tissue serosa, where it remained for about 3 days, then penetrated the intestinal wall and remained attached to its outer surface for 4 days. It then detached and dropped free in the amphipod haemocoel and transformed into an oval acanthella, growing for 16 days to reach the cystacanth stage. The cystacanth at 46 days post-infection was infective to fish (excysted in its intestine as an active juvenile). Male and female juveniles reached maturity 17 and 23 days post-infection. Recently copulated females first appeared 26 days post-infection and all females seemed to be copulated at 28 days post-infection; partially and fully gravid females first appeared 31 and 35 days post-infection. Mature males and fully gravid females started to die off naturally 31 and 43 days post-infection and were totally expelled from the fish intestine by 42 and 52 days post-infection. The cycle was completed in 89 days and is similar to other known palaeacanthocephalan life cycles, but has its own characteristics.

Aspidogastreans are commonly found infecting freshwater and marine molluscs, teleosts fishes and freshwater turtles. The subclass comprises four families – Rugogastridae Schell 1973, Stichocotylidae Faust & Tang 1936, Multicalycidae Gibson & Chinabut 1984 and Aspidogastridae Poche 1907 – and it is characterized by the presence of a ventral adhesive disc divided into rows of alveoli. In the current work, using light and scanning electron microscopy and molecular approaches, a new species of Aspidogastridae of the genus Rohdella Gibson & Chinabut, 1984, is described as a parasite of Colomesus psittacus in Brazil. The new taxon is distinguishable by the presence of oesophageal glands, teguments covered by ciliated papillae, and the position and shape of the hermaphroditic duct. The present work describes the third species of the genus Rohdella, thereby adding new morphological and molecular data regarding Aspidogastridae.

Morphometric analysis of Schistosoma mansoni male worms obtained from AKR/J and Swiss mice was carried out. Rodents infected by the intraperitoneal route with 80 cercariae of the schistosome (LE strain) were killed by cervical dislocation at 45 and 60 days post-infection and both peritoneal lavage and perfusion of the portal system were performed for the recovery of adult worms. Characteristics including total body length, the distance between oral and ventral suckers, extension of testicular mass and the number of testes were considered in the morphological analysis. Changes that occurred in S. mansoni recovered from the peritoneal cavity or from the portal system of AKR/J and Swiss mice included total body length and reproductive characteristics. Significant morphometric alterations were also observed when worms recovered from the portal system of both strains of mice were compared with the schistosomes obtained from hamsters (Mesocricetus auratus), the vertebrate host in which the LE strain had been adapted and maintained by successive passages for more than four decades. The present results reinforce the idea that S. mansoni has high plastic potential and adaptive capacity.

To better evaluate the usefulness of female Haemonchus specimens for specific identification, we undertook a detailed analysis of the morphology of a collection of worms obtained from cattle and sheep in shared pastures. Based on the results, we also more precisely evaluated the host-specificity of Haemonchus contortus, H. placei and H. similis occurring sympatrically in a farm located in the western region of São Paulo State, Brazil. A synlophe analysis was employed to identify the three species of Haemonchus. In cattle, the predominant species was H. similis (90.9%), followed by H. placei (9.1%). With the exception of one H. placei specimen, only H. contortus was found in sheep. The longest body length was found for H. placei specimens, followed by H. contortus and then H. similis. It was possible to distinguish H. similis females from H. contortus and H. placei on the basis of vulval structure. The synlophe analysis proved to be very useful for identification of H. contortus, H. placei and H. similis in epidemiological studies involving different species of ruminants in the same pastures. The finding that H. placei and H. similis were adapted to cattle and that H. contortus was adapted to sheep also confirmed the high host-specificity of the three nematodes species.

A new genus of the Monogenea, Teraplectanum n. g., is proposed for two new species of diplectanids found on the gills of Terapon theraps Cuvier collected off Carey Island, Peninsular Malaysia. The genus is based on a unique arrangement of the male reproductive system. In the new species spermatozoa stored in the seminal vesicle and secretions stored in the prostatic reservoir are transferred into, and mixed to form semen within, a special sclerotized auxiliary piece (SAP), and not within the copulatory tube as occurs in the majority of monogeneans. Teraplectanum species also possess a unique sclerotized vaginal loop through which the vaginal tube passes en route from the vaginal pore to the seminal receptacle. The two new species are Teraplectanum crassitubus n. sp. (type species) and T. angustitubus n. sp. They differ from each other mainly in the morphology of their copulatory tube: in T. crassitubus, the proximal region of this tube is thicker compared to the slender proximal region in T. angustitubus, although in both cases the tube tapers and twists distally. Of the known diplectanid species, only Diplectanum undulicirrosum Zhang et al., 2000 (currently considered incertae sedis) possesses such sclerotized hard parts, which suggests the same type of arrangement of the male reproductive system. Consequently, D. undulicirrosum is re-assigned to this new genus as Teraplectanum undulicirrosum (Zhang et al., 2000) n. comb. The copulatory tube of T. undulicirrosum is similar to the slender, undulating copulatory tube of T. angustitubus but does not taper distally as in the latter species.

The Angiostrongylus lungworms are of public health and veterinary concern in many countries. At the family level, the Angiostrongylus lungworms have been included in the family Angiostrongylidae or the family Metastrongylidae. The present study was undertaken to determine the usefulness and suitability of the nuclear 18S (small subunit, SSU) rDNA sequences for differentiating various taxa of the genus Angiostrongylus, as well as to determine the systematics and phylogenetic relationship of Angiostrongylus species and other metastrongyloid taxa. This study revealed six 18S (SSU) haplotypes in A. cantonensis, indicating considerable genetic diversity. The uncorrected pairwise ‘p’ distances among A. cantonensis ranged from 0 to 0.86%. The 18S (SSU) rDNA sequences unequivocally distinguished the five Angiostrongylus species, confirmed the close relationship of A. cantonensis and A. malaysiensis and that of A. costaricensis and A. dujardini, and were consistent with the family status of Angiostrongylidae and Metastrongylidae. In all cases, the congeneric metastrongyloid species clustered together. There was no supporting evidence to include the genus Skrjabingylus as a member of Metastrongylidae. The genera Aelurostrongylus and Didelphostrongylus were not recovered with Angiostrongylus, indicating polyphyly of the Angiostrongylidae. Of the currently recognized families of Metastrongyloidea, only Crenosomatidae appeared to be monophyletic. In view of the unsettled questions regarding the phylogenetic relationships of various taxa of the metastrongyloid worms, further analyses using more markers and more taxa are warranted.

Between 2005 and 2007, the presence of Onchocerca flexuosa (Wedl, 1856) was discovered and investigated in 110 red deer (Cervus elaphus) shot in the Riaño Regional Hunting Reserve, in the province of León (north-western Spain). Nodules containing O. flexuosa were located in the dorsal region and flanks of the deer. These were collected and measured, and some adult parasites were extracted from the nodules and identified by morphology and by obtaining mitochondrial 12S rDNA sequences, which were identical to those of previously published sequences for O. flexuosa. Some nodules were prepared for histology, embedded in paraffin, sectioned and stained with haematoxylin–eosin. Histologically, the worms were found in several compartments separated by an infiltrated fibrous tissue. These compartments were inhabited by several females and males, surrounded by a fibrous capsule. A total of 85.45% (95% confidence interval (CI): 78.86–92.04%) of red deer were parasitized, with a mean intensity of 9.53 ± 12.27 nodules/host, ranging between 1 and 74 nodules/deer. Significant differences in prevalence and intensity of infection were found between young and adult red deer, and also between seasons. However, no significant differences between males and females were observed. Five hundred and ninety-seven nodules were measured (15.81 ± 3.94 mm) and classified by sizes into small ( < 10 mm), medium (10–20 mm) and large (>20 mm). No relation was found between the size of the nodules and the time of infection. The high values found in the studied parameters show that northern Spain is an area of high-intensity infection for deer.

A redescription of the aspidogastrean trematode Lobatostoma kemostoma (MacCallum & MacCallum, 1913) is provided based on examination of type material, voucher specimens available at institutional collections and newly collected specimens from the type host, Trachinotus carolinus (Linnaeus, 1766), off the Angra dos Reis, Brazil. Partial sequence of its 28S rDNA is given. Lobatostoma kemostoma can be distinguished from the other eight species of the genus by: (1) the hindbody being longer than the ventral disc; and (2) the cephalic lobes having the same shape and size. Features observed for the first time include the number and detailed arrangement of alveoli on the ventral disc, the terminal genitalia and the ovarian complex. New illustrations and morphometric data are given. In the present survey the host specificity in the genus, the validity of some morphological features and the occurrence of an Indian species of Lobatostoma from off the Brazilian coast are discussed. A lectotype of L. kemostoma from the type series deposited at United States National Parasite Collection is also designated.

The composition and diversity of parasite communities and intestinal components, as well as infra-community structure, were assessed in eels Anguilla anguilla, from Mar Menor, a permanent Mediterranean hypersaline coastal lagoon. Data were used to determine whether this helminth community differs in composition and structure from that of eels in lagoons with lower salinity regimes and higher freshwater inputs. A total prevalence of 93% was detected. Specifically, parasites were identified as Deropristis inflata, Bucephalus anguillae, Contracaecum sp., Anguillicoloides crassus and two plerocercoid larvae belonging to the order Proteocephalidae, the marine species representing 91% of the isolated helminths. In the total community, digenetic trematodes were the dominant group of helminths, and D. inflata, an eel specialist, dominated both the component community and the infra-community. Richness and diversity were low but similar to those reported in other saline lagoons, and maximum species per eel did not exceed four. At the infra-community level, higher abundance than in other brackish or marine Mediterranean environments was detected. The findings provide further evidence of the similarity in composition and structure of helminth communities in eels from various Mediterranean coastal lagoons. Moreover, salinity-dependent specificities are well supported and reflect the life history of individual eels.

Lymphatic filariasis is a parasitic disease of tropical countries. This is a disfiguring and painful disease contracted in childhood, but the symptoms become apparent only in later years. Diagnosis of filarial infection is very crucial for the management of the disease. The main objective of this study was to develop a filarial antigen-based immunological assay for the diagnosis and surveillance of the disease. Monoclonal and polyclonal antibodies were raised to the recombinant protein Brugia malayi vespid allergen homologue (VAH). Capture enzyme-linked immunosorbent assay (ELISA) was standardized utilizing various combinations of antibodies and evaluated with serum samples of endemic normal (EN, n= 110), microfilaraemic (MF, n= 65), chronic pathology (CP, n= 45) and non-endemic normal (NEN, n= 10) individuals. Of the 230 samples tested, VAH capture assay detected circulating antigen in 97.91% of bancroftian and 100% of brugian microfilaraemic individuals, and 5% of endemic normal individuals, comparable to the earlier reported SXP-1 antigen detection assay. However, the combination of VAH and SXP-1 (VS) capture ELISA was found to be more robust, detecting 100% of microfilaraemic individuals and with higher binding values. Thus an antigen capture immunoassay has been developed, which can differentiate active infection from chronic infection by detecting circulating filarial antigens in clinical groups of endemic areas.

An investigation was carried out on the distribution and biodiversity of steinernematid and heterorhabdtid entomopathogenic nematodes (EPN) in nine regions of Italy in the period 1990–2010. More than 2000 samples were collected from 580 localities and 133 of them yielded EPN specimens. A mapping of EPN distribution in Italy showed 133 indigenous EPN strains belonging to 12 species: 43 isolates of Heterorhabditis bacteriophora, 1 of H. downesi, 1 of H. megidis, 51 of Steinernema feltiae, 12 of S. affine, 4 of S. kraussei, 8 of S. apuliae, 5 of S. ichnusae, 3 of S. carpocapsae, 1 of S. vulcanicum, 3 of Steinernema ‘isolate S.sp.MY7’ of ‘S. intermedium group’ and 1 of S.arenarium. Steinernematids are more widespread than heterorhabditids and S. feltiae and H. bacteriophora are the most commonly encountered species. Sampling sites were grouped into 11 habitats: uncultivated land, orchard, field, sea coast, pinewood, broadleaf wood, grasslands, river and lake borders, caves, salt pan and moist zones; the soil texture of each site was defined and the preferences of habitat and soil texture of each species was assessed. Except for the two dominant species, S. feltiae and H. bacteriophora, EPN occurrence tends to be correlated with a specific vegetation habitat. Steinernema kraussei, H. downesi and H. megidis were collected only in Sicily and three of the species recently described – S. apuliae, S. ichnusae and S. vulcanicum – are known only from Italy and seem to be endemic.

Helminths use several strategies to evade and/or modify the host immune response, including suppression or inactivation of the host antigen-specific response. Several helminth immunomodulatory molecules have been identified. Our studies have focused on immunosuppression induced by the roundworm Ascaris suum and an A. suum-derived protein named protein 1 from A. suum (PAS-1). Here we assessed whether PAS-1 is an excretory/secretory (E/S) protein and whether it can suppress lipopolysaccharide-induced inflammation. Larvae from infective eggs were cultured in unsupplemented Dulbecco's modified Eagle medium (DMEM) for 2 weeks. PAS-1 was then measured in the culture supernatants and in adult A. suum body fluid at different time points by enzyme-linked immunosorbent assay (ELISA) with the monoclonal antibody MAIP-1. Secreted PAS-1 was detected in both larval culture supernatant and adult body fluid. It suppressed lipopolysaccharide (LPS)-induced leucocyte migration and pro-inflammatory cytokine production, and stimulated interleukin (IL)-10 secretion, indicating that larval and adult secreted PAS-1 suppresses inflammation in this model. Moreover, the anti-inflammatory activity of PAS-1 was abolished by treatment with MAIP-1, a PAS-1-specific monoclonal antibody, confirming the crucial role of PAS-1 in suppressing LPS-induced inflammation. These findings demonstrate that PAS-1 is an E/S protein with anti-inflammatory properties likely to be attributable to IL-10 production.

Onchocerca volvulus is a tissue-dwelling, vector-borne nematode parasite of humans and is the causative agent of onchocerciasis or river blindness. Natural infections of BALB/c mice with Litomosoides sigmodontis and of cattle with Onchocerca ochengi were used as models to study the immune responses to O. volvulus-derived recombinant proteins (OvALT-2, OvNLT-1, Ov103 and Ov7). The humoral immune response of O. volvulus-infected humans against OvALT-2, OvNLT-1 and Ov7 revealed pronounced immunoglobulin G (IgG) titres which were, however, significantly lower than against the lysate of O. volvulus adult female worms. Sera derived from patients displaying the hyperreactive form of onchocerciasis showed a uniform trend of higher IgG reactivity both to the single proteins and the O. volvulus lysate. Sera derived from L. sigmodontis-infected mice and from calves exposed to O. ochengi transmission in a hyperendemic area also contained IgM and IgG1 specific for O. volvulus-derived recombinant proteins. These results strongly suggest that L. sigmodontis-specific and O. ochengi-specific immunoglobulins elicited during natural infection of mice and cattle cross-reacted with O. volvulus-derived recombinant antigens. Monitoring O. ochengi-infected calves over a 26-month period, provided a comprehensive kinetic of the humoral response to infection that was strictly correlated with parasite load and occurrence of microfilariae.