The present study aimed to evaluate the ovicidal activity (type 3 effect) of VC1 and VC4 isolates of Pochonia chlamydosporia in a solid medium and the action of a crude extract of P. chlamydosporia against eggs of Ascaridia galli. To evaluate ovicidal activity in culture medium, 1000 A. galli eggs were plated on Petri dishes containing 2% water-agar with grown fungal isolates (VC1 or VC4) and without fungus (control group) and were examined at 1, 3 and 5 days post-inoculation (assay A). Then, to test the action of crude extracts of P. chlamydosporia (VC1 or VC4), 500 eggs of A. galli were plated on Petri dishes of 4.5 cm diameter with 5 ml of fungal filtrate from each tested isolate. The control group consisted of 500 eggs of A. galli with 10 ml of distilled water on each Petri dish (assay B). Fungal isolates were effective (P < 0.01) at destroying these eggs, showing a type 3 effect at the studied intervals. On the other hand, the crude extract of isolates (VC1 or VC4) reduced the number of A. galli eggs in the treated group compared with the control group by 64.1% and 56.5%, respectively. The results of the present study show that P. chlamydosporia is effective at destroying eggs of A. galli and could therefore be used in the biological control of nematodes.

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.

Wild isolates of the egg-parasitic fungi Paecilomyces lilacinus and Verticillium chlamydosporium, obtained from the organic environment of Durg, Chhattisgarh, India, were subjected to screening for in vitro growth using different media types, range of incubation temperature and pH, and their predatory activity to the eggs of Fasciola gigantica and Gigantocotyle explanatum. Maximum growth of P. lilacinus was obtained in corn-meal agar compared to any other media types. The preferred medium for growth of V. chlamydosporium was corn-meal agar, followed by potato-dextrose agar. After initial growth for 16 h of incubation, no growth was observed in water agar for both the fungi. Six different temperatures – 4°C, 10°C, 18°C, 26°C, 34°C and 40°C – were used to observe growth profiles of the fungi in corn-meal agar medium. While no and very little growth of P. lilacinus and V. chlamydosporium was observed at 4°C and 10°C, respectively, growth profiles of both the fungi were optimal at 26–40°C. A range of pH (pH 4–8) supported growth of both P. lilacinus and V. chlamydosporium. Full-grown plates of the fungi baited with viable eggs of F. gigantica and G. explanatum revealed that V. chlamydosporium was more vigorous in its egg-parasitic ability compared to P. lilacinus. Distortion of the eggs started on day 2–3 of egg baiting in culture plates of V. chlamydosporium, with complete distortion by day 7. On the contrary, P. lilacinus exhibited very limited egg-parasitic ability and some of the baited eggs even showed development of miracidia.

A survey for slug-associated nematodes in five locations of East and West Flanders in Belgium revealed the presence of one new and six known slug-parasitic nematodes, Agfa flexilis (Dujardin, 1845), Alloionema appendiculatum (Schneider, 1859), Angiostoma dentiferum (Mengert, 1953), Angiostoma limacis (Dujardin, 1845), Angiostoma norvegicum (Ross et al., 2017) and Phasmarhabditis hermaphrodita (Schneider, 1859). Angiostoma norvegicum and P. hermaphrodita are recorded for the first time in Belgium. The six known species are documented by light microscopy (LM) microphotographs and informative DNA sequences. Angiostoma gandavensis n. sp. (Angiostomatidae), discovered from arionid slugs, is described based on light microscopy, scanning electron microscopy (SEM) and molecular data. Based on analyses of D2D3 expansion segment of 28S and 18S rDNA sequences, this new species is found to be related to A. limacis, A. norvegicum, A. margaretae (Ross et al., 2011) and A. milacis (Ivanova and Wilson, 2009). The new species can be distinguished from these others based on morphological characters such as the distinctive mucronate structures at the tail tip of both sexes, presence of lateral ala, reflexed female ovaries and the number and arrangement pattern of male genital papillae.

Haemonchus contortus is a haematophagous gastrointestinal nematode (GIN) that causes severe anaemia and even death in small ruminants, negatively impacting the economic viability of farms. Traditionally, this parasite has been controlled with chemical compounds; however, inadequate use of these types of products has favoured the emergence of anthelmintic resistance. Therefore, it is necessary to search for alternatives for GIN control. Previous studies have reported the anthelmintic activity of edible mushroom extracts against H. contortus. A recent study reported that a fraction constituted of different fatty acids and β-sitosterol isolated from the basidiomata of the edible mushroom Pleurotus djamor ECS-123 has ovicidal and larvicidal activity against H. contortus. Thus, this study aimed to assess the anthelmintic activity of the pure molecules: pentadecanoic acid, palmitic acid, β-sitosterol, stearic acid and linoleic acid. For this purpose, an egg-hatching inhibition test was carried out in which the compounds were evaluated individually and in combination at a final concentration of 20 mg mL−1. Furthermore, larval mortality was assessed using a combination of the five commercial compounds previously mentioned at different concentrations (1.25–20 mg mL−1). Palmitic acid and stearic acid, in some combinations, inhibited H. contortus egg hatching by 100%. On the other hand, in the larval mortality test, the combination of the five compounds showed dose-dependent behaviour, and 100% mortality was obtained 24 h post-incubation. Pure molecules and their combinations have anthelmintic-like activity against the eggs and larvae of H. contortus.

Fifty-nine of 60 (98%), 6-month-old male golden hamsters, Mesocricetus auratus, fed 15 (group A). 50 (group B), or 200 (group C) metacercarial cysts of Echinostoma caproni (Digenea: Echinostomatidae) were infected 7–34 days postexposure. The mean number of worms recovered in groups A, B and C were 9, 10, and 50, respectively. The percentage recovery was significantly different between group A (63%) and groups B (21%) and C (23%). The intestine was divided into three equal regions (I, II, III). Worms from group A were located in segments II and III of the small intestine whereas worms from groups B and C were distributed in all three segments. The body area, ovarian and testicular areas of worms from group A were greatest, followed in decreasing order by body and gonadal areas of worms from groups B and C. Echinostoma caproni eggs were found in the faeces of all the hamsters examined from groups A, B and C by days 9, 10 and 11, respectively. Physical damage occurred at the site of attachment of the echinostome. Pathological observations indicated the presence of enlarged lymphatic nodules with lymphocytes being the primary cellular infiltrate at the site of parasite attachment.

Using the local strains of Schistosoma bovis and Fasciola gigantica, it was shown that Sudanese zebu calves with mature primary infections of F. gigantica were highly resistant to challenge with S. bovis cercariae, and vice versa. Liver enzyme tests showed that, in both cases, the primary infections had caused some liver damage. Primary infection with irradiated S. Bovis cercariae, which did not cause significant liver damage, did not protect significantly against challenge with F. gigantica.

Spermatogenesis and the structure of the spermatozoon of the frog intestinal fluke, Ganeo tigrinum has been examined by electron microscopy. Each testis is covered by a tunica consisting of sustentacular cells and fibrous material. Internally the testis is lined by undifferentiated spermatogonial cells, with cytoplasm rich in free ribosomes. The arrangement of the diverse spermatogonic cell populations is similar to those studied in other digeneans. During spermatogenesis the elongation and spiralling of nuclear materials occurs synchronously in the late spermatid stage. The G. tigrinum spermatozoon appears to be biflagellate due to the presence of two filaments. Between the two filaments is an elongated nucleus; these elements fuse to form the monopartite shaft of the sperm. The spermatozoon of G. tigrinum is characterized by the presence of an unusually long nucleus that even surpasses the length of the mitochondrion. At almost half the length of the spermatozoon there is sharp rotation between first axial filament, mitochondrion and the nucleus. Both the axial filaments show 9+1 arrangement of axonemal units throughout the spermatozoon.

For two years fortnightly malacological samplings were carried out to collect Lymnaea truncatula (Mollusca; Basommatophora) at five points in the upper and middle Porma river basin, León, Spain. The highest numbers of snails were collected in September, May and November. Of the 5486 molluscs examined, 11·41% harboured F. hepatica (Trematoda; Digenea) with an average intensity of 20·14. In general, the values of both infection prevalence and intensity increased with the size of the snails. It was in October when the highest figures for each parameter mentioned above were detected (18·73% and 28·48, respectively). The chi-square test showed statistically significant differences in relation to the infection prevalence among the groups of molluscs established according to: their length; the months in which they were collected; the sampling localities; monthly average ambient temperature; precipitation during the collection. Similarly, statistically significant differences were detected in the intensity of the infection among the groups of molluscs previously established, except for that based on the values of precipitation. Generally, parasites were found in the same snail at different stages of development. It seems that most mollusc infections occur in February–March and at the end of summer–beginning of autumn periods. The highest rate of rediae with mature cercariae ready to be shed were detected between September and December. Metacercariae in the grass samples were also observed at the end of autumn. For this reason, this period could be considered as the most suitable for infection of the definitive hosts to take place.

Excretory/secretory antigens (ES) of larval Taenia solium were obtained by maintaining the bladder worms in Medium 199 for 3 days. Analsysis by SDS-PAGE showed that ES antigens consisted of at least 19 polypeptides, with Mrranging from 14–116 kDa. Analytical isoelectric focusing revealed eight bands with acidic pI. An immunocytolocalization study using the peroxidase method demonstrated the presence of ES epitopes on the tegument of the wall of the spiral canals of bladder worms. The specificity of ES antigens was evaluated by EITB, ELISA and FAST-ELISA using antisera against the common parasites of Chinese pigs and man. ES antigens cross-reacted with the antiserum against larval T. hydatigena of pigs. However, these antigens were generally more specific in diagnosing human cysticercosis. Three host-like molecules with molecular masses 43, 58 and 66 kDa were present in the ES products.

Infections by gastrointestinal nematodes negatively affect small ruminant health and at the same time cause substantial economic losses worldwide. Because resistance to conventional anthelmintic compounds is growing, target studies evaluating the effectiveness of alternative ingredients of botanical origin on gastrointestinal nematodes are needed. In this study, we evaluated the in vitro anthelmintic activity of Ruta chalepensis L. extracts on the third-stage larvae of sheep gastrointestinal nematodes. A methanol extract showed the highest anthelmintic activity, with an EC50 = 0.10 ± 0.06 mg/ml after 96 h, while the essential oil had an EC50 = 1.45 ± 1.22 mg/ml after 48 h. Moreover, three secondary metabolites of the essential oil, i.e. 2-decanone, 2-nonanone and 2-undecanone, showed EC50 values of 0.07 ± 0.06, 0.25 ± 0.29 and 0.88 ± 0.73 mg/ml at 24 h, respectively. The present study indicated that the R. chalepensis methanol extract, the essential oil and its metabolites 2-decanone, 2-nonanone and 2-undecanone showed promising anthelmintic activity on gastrointestinal nematodes.

A half a century ago, R. T. Leiper described a new species belonging to the genus Fasciola from specimens which he collected from hippopotami at the Murchison Falls, on the Victoria Nile in Uganda, in the summer of 1907. This species, Fasciola nyanzae, was found in the bile ducts of the liver, and in most instances the specimens were somewhat macerated owing to the length of time that had elapsed before the animal could be recovered from the water. Nevertheless, the characteristic shape of these liver flukes and the restriction of the branched testes to the anterior third of the body made it possible to recognise them as being different from all other species of the genus known at that time.

Co-expression of Trichinella spiralis macrophage migration inhibitory factor (TsMIF) with T. spiralis cystatin-like domain protein (TsMCD-1) in a DNA vaccine induces a Th1 immune response and partial protection against T. spiralis infection. The present study evaluated whether co-expression of mouse ubiquitin (Ub) with TsMIF and TsMCD-1 might improve the immune response against T. spiralis infection. Groups of BALB/c mice were immunized twice at 2-week intervals with 100 μg of plasmid DNA encoding either a TsMIF–TsMCD-1 fusion protein (pVAX1-Tsmif-Tsmcd-1) or an Ub-co-expressing triple fusion protein Ub–TsMIF–TsMCD-1 (pVAX1-Ub-Tsmif-Tsmcd-1). Control animals were immunized with pVAX1-Ub or blank vector plasmid. Specific antibody levels (IgG, IgG1, IgG2a, IgG2b, IgM, IgA, IgE) against the recombinant protein TsMIF–TsMCD-1, serum cytokines (interferon (IFN)-γ, interleukin (IL)-4, IL-5, transforming growth factor (TGF)-β1 and IL-17), CD4+/CD8+ T cells and cytotoxic T lymphocyte (CTL) responses were monitored. Challenge infection was performed 2 weeks after the second immunization and worm burden was assayed at 35 days post-challenge. Antibody responses induced by pVAX1-Ub-Tsmif-Tsmcd-1 were significantly lower than for TsMIF-TsMCD-1, but the vaccine induced increased levels of Th1 cytokine (IFN-γ) and increased T-cell cytotoxicity. The reduction of worm burden (37.95%) following immunization with pVAX1-Ub-Tsmif-Tsmcd-1 was significantly greater than that induced by the pVAX1-Tsmif-Tsmcd-1 vaccine (23.17%; P< 0.05).

Our knowledge that the infective larvæ of certain parasitic nematodes are capable of actively boring into intact skin and of thus gaining entry into the host dates from the epoch-making discovery of Looss, who, in 1897, accidentally infected himself with Ancylostoma duodenale, the mature larvae of which entered through the skin of his hand (Looss, 1911, pp. 456−459).

(1) A survey was made of the trematode and nematode parasites of Apodemus sylvaticus, Clethrionomys glareolus and Sorex araneus from the islands of Raasay, Ulva and Scalpay.

(2) The following species have been recorded:—

A. sylvaticus:

Trematodes—

Lyperosomum villa

Brachylaemus recurvum

Nematodes—

Capillaria muris-sylvatici

Heligmosomum glareoli*

Syphacia stroma

Two species of Proctoeces Odhner, 1911 have been described in marine organisms from Chile: P. humboldti George-Nascimento & Quiroga (1983), parasitizing the gonads of keyhole limpets (Fissurella spp.), and P. chilensis Oliva (1984), an intestinal parasite of Sicyases sanguineus (Teleostei); both species were subsequently considered as P. lintoni Siddiqi & Cable (1960). To assist in the resolution of the taxonomic identification of Proctoeces species in marine organisms from Chile, phylogenetic studies using DNA sequences from the V4 region of the SSU rRNA gene were performed. Several specimens of P. lintoni were isolated from keyhole limpets (Fissurella spp.) and clingfish (S. sanguineus) from Bahia San Jorge (23°40′S) and Bahia Concepción (36°50′S). Phylogenetic analyses were conducted using three different approaches: a neighbour-joining (NJ), a maximum likelihood (ML) and Bayesian inference (BI). The phylogenetic analysis confirms that specimens of Proctoeces obtained from keyhole limpets and those specimens from the clingfish are in fact the same species. We prefer to consider our specimens as Proctoeces cf. lintoni, as the morphology of Proctoeces appears to be of doubtful value and genetic information about P. lintoni Siddiqi & Cable (1960) is not available. In addition, our results strongly suggest that there are at least three species in this genus.

A substantial resistance to a heterologous challenge with Fasciola hepatica was demonstrated in calves harbouring primary patent Schistosoma bovis infections of an age of 10 weeks at challenge, the mean worm burden being reduced by 29·9% (p < 0025) in calves primarily infected with S. bovis as compared with that of the challenge control group. The heterologous resistance was also evidenced by less pronounced liver tissue damage and lower serum gammaglutamyl-transpeptidase values in the S. boms-infected animals compared with the challenge control group.

Mice infected with Nematospiroides dubius were incapable of responding normally to i.p. or i.v. challenge with SRBC. The HA and PFC response to SRBC in infected animals was characterized by a severe depression of antibody to SRBC on day 4 and a reduced HA peak titre during the following week. The greatest depression of the response to SRBC was associated with an interval of 14 days between infection and the administration of antigen, suggesting that a particular stage of the parasite contributed significantly to immunodepression during this critical period. It was proposed that a combination of parasite induced damage to the intestine, release of parasite secretory/excretory products and loss of appetite by the host produced trauma during which the host was incapable of responding normally.

However, mice given low-level and long-standing infections also showed reduced responses to SRBC, although these animals were not severely depressed. It is possible that this generalized weakening of host immunocompetence is the inevitable consequence of a parasite mechanism which operates more specifically to suppress the expression of homologous immunity at the intestinal level.

During the spring, summer and autumn of 1967, 1968 and 1969, 109 rabbits were collected from two localities in Northumberland. Post mortem worm counts indicated that six species of helminths were present, Trichostrongylus retortaeformis, Passalurus ambiguus, Strongyloides papillosus, Nematodirus battus, N. filicollis and Cittotaenia denticulata.

T. retortaeformis was recovered from both localities, an overall infection rate of 88% being recorded.

P. ambiguus and C. denticulata were found at only one of the localities which would indicate that these species have a locally discontinuous distribution and an explanation for this has been discussed.

Both Nematodirus battus and N. filicollis which are normally sheep parasites were recorded for the first time from wild rabbits. The numbers of rabbits infected and size of infection were small and in consequence the rabbit can not be considered an important reservoir for these species.

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