In the present study, the nematicidal activity of a Moringa oleifera ethyl acetate leaf extract against the eggs and larvae of Haemonchus contortus and Nacobbus aberrans, nematodes of agricultural importance, was evaluated. The experimental design for the evaluation of the effects against both nematodes consisted of eight treatments (n = 4). Distilled water, Tween (4%) and a commercial anthelmintic agent (ivermectin, 5 mg/mL) were used as controls, and for treatments 4–8, the concentrations of the extract were 20, 10, 5, 2.5 and 1.25 mg/mL, respectively. Readings were taken at 12 h and 24 h for N. aberrans and 48 h and 72 h for H. contortus post-treatment under an optical microscope (10× and 40×). The data obtained were analysed by analysis of variance through a completely randomized factorial design using the SAS V9 program. The results show that, for H. contortus egg hatching, 85.88% inhibition was obtained at a concentration of 20 mg/mL at 48 h, while for third-stage larva (L3) mortality, the highest percentage was 68.19% at 1.25 mg/mL at 72 h. In the case of N. aberrans, the greatest inhibition of egg hatching was 90.69% at 5 mg/mL at 12 h post-treatment, and for larval mortality, it was 100% at 10 mg/mL at 24 h post-treatment. The main major compounds identified by qualitative analysis and by gas chromatography coupled to mass spectrometry were 9,12,15-octadecatrienoic acid, (Z,Z,Z)-, n-hexadecanoic acid and 2,4-di-tert-butylphenol, and the minor compounds included phytol, γ-sitosterol and α-tocopheryl acetate. It was demonstrated that the ethyl acetate leaf extract of M. oleifera Lam. shows great potential for combating agricultural nematodes.

Host-seeking behaviour and how a parasite identifies the correct host to infect remains a poorly understood area of parasitology. What is currently known is that host sensation and seeking behaviour is formed from a complex mixture of chemo-, thermo- and mechanosensory behaviours, of which chemosensation is the best studied. Previous studies of olfaction in parasitic nematodes suggested that this behaviour appears to be more closely related to target host and infection mode than phylogeny. However, there has not yet been a study comparing the chemotactic and temperature-dependent behaviours of very closely related parasitic and non-parasitic nematodes. To this end, we examined the temperature-dependent and chemotactic responses of the Strongyloidoidea superfamily of nematodes. We found differences in temperature response between the different species and within infective larvae. Chemotactic responses were highly divergent, with different attraction profiles between all species studied. When examining direct stimulation with fur, we found that it was insufficient to cause an attractive response. Overall, our results support the notion that olfactory sensation is more closely related to lifestyle and host range than phylogeny, and that multiple cues are required to initiate host-seeking behaviour.

Carbon dioxide (CO2) is an important sensory cue for many animals, including both parasitic and free-living nematodes. Many nematodes show context-dependent, experience-dependent and/or life-stage-dependent behavioural responses to CO2, suggesting that CO2 plays crucial roles throughout the nematode life cycle in multiple ethological contexts. Nematodes also show a wide range of physiological responses to CO2. Here, we review the diverse responses of parasitic and free-living nematodes to CO2. We also discuss the molecular, cellular and neural circuit mechanisms that mediate CO2 detection in nematodes, and that drive context-dependent and experience-dependent responses of nematodes to CO2.

Strongyloides venezuelensis is a parasitic nematode that infects rodents. Although Strongyloides species described to date are known to exhibit parthenogenetic reproduction in the parasitic stage of their life cycle and sexual reproduction in the free-living stage, we did not observe any free-living males in S. venezuelensis in our strain, suggesting that the nematode is likely to depend on parthenogenetic reproduction. We confirmed by cytological analysis that S. venezuelensis produces eggs by parthenogenesis during the parasitic stage of its life cycle. Phylogenetic analysis using nearly the full length of 18S and D3 region of 28S ribosomal RNA gene suggested that S. venezuelensis is distantly related to another rodent parasite, namely Strongyloides ratti, but more closely related to a ruminant parasite, Strongyloides papillosus. Karyotype analysis revealed S. venezuelensis reproduces with mitotic parthenogenesis, and has the same number of chromosomes as S. papillosus (2n = 4), but differs from S. ratti (2n = 6) in this regard. These results, taken together, suggest that S. venezuelensis evolved its parasitism for rodents independently from S. ratti and, therefore, is likely to have a different reproductive strategy.

The aim of the present investigation was to determine whether temporal variation in environmental factors such as rainfall or temperature influence long-term fluctuations in the prevalence and mean abundance of the nematode Mexiconema cichlasomae in the cichlid fish Cichlasoma uropthalmus and its crustacean intermediate host, Argulus yucatanus. The study was undertaken in a tropical coastal lagoon in the Yucatan Peninsula (south-eastern Mexico) over an 8-year period. Variations in temperature, rainfall and monthly infection levels for both hosts were analysed using time series and cross-correlations to detect possible recurrent patterns. Infections of M. cichlasomae in A. yucatanus showed annual peaks, while in C. urophthalmus peaks were bi-annual. The latter appear to be related to the accumulation of several generations of this nematode in C. urophthalmus. Rainfall and temperature appear to be key environmental factors in influencing temporal variation in the infection of M. cichlasomae over periods longer than a year together with the accumulation of larval stages throughout time.

The aim of this study was to elucidate transcriptional changes in the parasitic nematode Teladorsagia circumcincta upon encountering either naïve or immune ovine hosts. Pools of 100 000 exsheathed 3rd- stage T. circumcincta larvae were exposed in vitro to either an immune or naïve ovine abomasal environment, RNA was extracted from the larvae and sequenced using the Roche 454 platform. Each sample produced approximately 82 000 reads that assembled to give approximately 5500 Isotigs (contigs). The two sequence datasets were clustered together to give a total of 6969 clusters of which 18 were differentially expressed (P<0·001) between the two groups. Clusters with a predominance of reads in larvae exposed to the immune abomasal environment encoded homologues of peptidyl-glycine alpha-amidating monooxygenase, heat shock-protein 16-2 and IDA-1, a tyrosine phosphatase-like receptor protein. Clusters with a predominance of reads in the naïve environment encoded homologues of cytochrome b, EGg Laying defective family member 21 and NADH dehydrogenase subunit 5. Gene ontology analyses indicated that larvae exposed to the immune environment showed an increase in expression of genes involved in ‘carbon utilization’, ‘response to stimulus’ and ‘developmental process’. These data suggest that T. circumcincta modulates gene expression in response to the immune status of the host.

Infection of sheep with the abomasal nematode Teladorsagia circumcincta can cause a relative protein deficiency and reduce growth rate in growing lambs. A key event appears to be the destruction of junctions between epithelial cells. If the infection is heavy or prolonged, this leads to increased mucus production, hyperplasia, decreased acid production, gastrinemia, inappetance and pepsinogenemia. The severity of the infection depends upon the extent of concurrent infection, the nutritional status of the host and genetically controlled variation in the ability to mount protective immune responses.

Citrus orchards in the Nile Delta and middle Egypt were surveyed for predacious soil mites and parasitic nematodes. There were 16 species of predacious mites and the citrus parasitic nematode, Tylenchulus semipenetrans Cobb. was recorded everywhere. In the medium-heavy soil of the Nile Delta orchards, the general predators or decomposer mites like uropodids were most common. In the light-medium soil of El-Fayoum in middle Egypt, the specialized predator mites like gamasids were most common. In localities where gamasids were common, negligible infestations of citrus parasitic nematodes were reported. Regarding Egypt as a whole, the uropodid mites Uroobovella krantzi (Zaher & Afifi) and Nenteria hypotrichus (El-Borolossy & El-Banhawy) represented about 50% of the sampled mites. The remaining mites were represented by 14 mesostigmatid species, the most common being Gamasiphis pulchellus (Berlese). At least 6% of orchards suffered injurious nematode infestations, 13% critical infestations and 22% moderate infestations, and the remaining 59% were free or with negligible infestations.

The anthelmintic properties of tanniferous plants and of their secondary metabolites represent one possible alternative to chemotherapy that is currently being explored as a means of achieving sustainable control of gastrointestinal nematodes in ruminants. Previous in vivo and in vitro results suggest that tanniferous plants can have direct anti-parasitic effect against different stages of nematodes. However, the mode of action of the bioactive plant compounds remains obscure. The objectives of the current study were (1) to examine the hypothesis that extracts of tanniferous plants might interfere with the exsheathment of third-stage infective larvae (L3); (2) to assess the role of tannins in the process by examining the consequence of adding an inhibitor of tannins (polyethylene glycol: PEG) to extracts. The effects of 4 tanniferous plant extracts on exsheathment have been examined on L3 of Haemonchus contortus and Trichostrongylus colubriformis. Artificial exsheathment was induced in vitro by adding hypochloride solution to larval suspension. The evolution of exsheathment with time was measured by repeated observations at 10-min interval for 60 min. The selected plants were: genista (Sarothamnus scoparius), heather (Erica erigena), pine tree (Pinus sylvestris), and chestnut tree (Castanea sativa), with tannin contents ranging from 1·5 to 24·7% of DM. Extracts of a non-tanniferous plant (rye grass, tannin content: 0·3% of DM) were included in the assay as negative controls. The extracts were tested at the concentration of 600 μg/ml and the effects were compared to the rate of exsheathment of control larvae in PBS. No statistical differences in the pattern of exsheathment was observed after addition of rye grass or genista extracts for both nematode species and with heather extracts for T. colubriformis. In contrast, pine tree extracts on larvae of both species and heather extracts with H. contortus induced a significant delay in exsheathment. Last, contact with chest nut extracts led to a total inhibition of the process for both nematodes. These results suggest that extracts of tanniferous plants might affect a key process in the very early stages of larval invasion of the host. In most cases, the addition of PEG led to a total or partial restoration towards control values. This suggests that tannins are largely involved in the inhibitory process. However, other secondary metabolites may also interfere with the process that would help to explain some of the differences in response observed between the two nematode species.

In theory, the age at which maturation occurs in parasitic nematodes is inversely related to pre-maturational mortality rate, and cross-species data on mammalian nematodes are consistent with this prediction. Immunity is a major source of parasite mortality and parasites stand to gain sizeable fitness benefits through short-term adjustments of maturation time in response to variation in immune-mediated mortality. The effects of thymus-dependent immune responses on maturation in the nematode parasites Strongyloides ratti and Nippostrongylus brasiliensis were investigated using congenitally thymus-deficient (nude) rats. As compared with worms in normal rats, reproductive maturity of parasites (presence of eggs in utero) in nude rats occurred later in S. ratti but earlier in N. brasiliensis. Immune-mediated differences in maturation time were not associated with differences in worm length. Thymus-dependent immunity had no effect on pre-maturational mortality. Results are discussed in relation to theoretical expectations and possible explanations for the observed patterns in parasite maturation.

A polymorphic set of 14 kDa excretory–secretory (E–S) antigen-encoding cDNAs, with similarity to a previously characterized 15 kDa E–S antigen of Haemonchus contortus, was cloned from Cooperia punctata. Five cDNAs encoding predicted proteins of 70–80% identity were sequenced. Nucleotide sequence data reported in this manuscript are available in the GenBankTM databases under the accession numbers AF352718, AF352719, AF352720, AF352721, AF352722, AF352723, AF352724, AF352724, AF352726 and AF352727. Genomic analyses of individuals proved the existence of three 14 kDa E–S antigen-encoding genes, excluding that the differences reflected polymorphisms between individuals in a population. Southern blots indicated the presence of additional members of this gene family. Thus, despite the fact that heterologously expressed C. punctata 14 kDa E–S products are shown to be recognized by immune sera, potential pitfalls in the development of a recombinant vaccine are presented by this genetic diversity. Vaccine design could be further rationalized by knowledge of the function, and possible redundancy in function, of the E–S products which is presently lacking. The limitations encountered in assigning a function to the 14/15 kDa family of E–S proteins that is thus far unique to the trichostrongyloid nematodes are discussed.

Resistance against anthelmintics is widespread, particularly in parasitic nematode populations of small ruminants. Several new techniques or supplements have been developed or are under investigation. Biological control (BC) is one of these new methods. The net-trapping predacious fungus Duddingtonia flagrans produces thick walled resting spores, chlamydospores, which are able to survive passage through the gastrointestinal tract of cattle, horses, sheep and pigs. Under Danish climatic conditions it has been shown that the number of parasite larvae on pasture and the worm burden of the grazing animals is significantly reduced when animals are fed spores during the initial 2–3 months of the grazing season. Work with D. flagrans in France, Australia, USA, and Mexico has confirmed the strong BC potential of this fungus. Today much work is going into development of suitable delivery systems for grazing livestock worldwide. Ultimately, BC should be implemented in integrated parasite control strategies, both in conventional and organic livestock production.

The initial infection process of nematode-trapping fungi is based on an interaction between the trapping structure of the fungus and the surface of the nematode cuticle. A bioassay was designed to investigate the predatory response of several isolates of nematode-trapping fungi against 3 mutants of Caenorhabditis elegans (AT6, AT10 and CL261), which have been reported to differ in the reaction of their cuticle to antibodies and lectins. The bioassay was also applied to infective larvae of animal (Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus axei) and plant (Meloidogyne spp.) parasitic nematodes. Differences in trapping ability were most marked in the first 24 h, and were density dependent. Although the isolate of Arthrobotrys responded very rapidly in the first 24 h, Duddingtonia flagrans was generally the most effective isolate and Monacrosporium responded relatively poorly throughout all experiments. All the fungi tested trapped the srf mutants of C. elegans more efficiently than the wild type, and there were differences between the different srf mutants of C. elegans. Differences in trapping ability were also observed between different isolates of D. flagrans; similarly, differences in trapping behaviour were observed not only amongst the different species of plant-parasitic nematodes, but also between the sheathed and exsheathed larvae of the animal-parasitic nematodes.

Faecal nematode egg counts (FECs) were examined in lactating ewes from divergent flocks of Romney sheep which had been selectively bred from 1979 to 1996 for or against FEC in 4 to 7 month old lambs. Faecal samples were obtained from the ewes while under normal grazing management, 1 to 2 months after lambing in spring for each of 6 years between 1987 and 1996 (no. = 785 records; 298 animals). Analyses were carried out on loge (FEC + 100)–transformed data using animal-model maximum likelihood procedures, accounting for repeated records on ewes, within and between lactations. An examination of non-genetic effects indicated that there was no significant effect of age class of ewe on FEC but ewes which gave birth to single lambs had significantly lower post-parturient FECs than those bearing twins (back-transformed means of 184 v. 276 egg per g, respectively; P < 0·001). In relation to genetic effects, post-parturient FECs were significantly lower in ewes from the flock bred for low lamb-FEC than in their counterparts from the flock bred for high lamb-FEC, with ewes from the most recent birth years (1991 to 1994) showing a nine-fold difference (back-transformed means of 33 and 305 eggs per g faeces respectively; P < 0·001). This was equivalent to 69% of the divergence observed between loge (FEC + 100) in their lambs in the same years. Heritability and repeatability estimates for loge (FEC + 100) in ewes were 0·37 (s.e. 0·06) and 0·46 (s.e. 0·03) respectively. Genetic correlation estimates between a ewe's post-parturient loge (FEC + 200) and her loge (FEC + 100) as a lamb, based on analysis of (co)variance or realized responses, were 0·70 or 0·58 respectively. The phenotypic correlation between a ewe's loge (FEC + 100) and that of her lamb(s) in the same lactation was 0·29 (s.e. 0·06) (P < 0·001). From the results it is clear that substantial genetic changes in post-parturient FECs of breeding ewes can be induced through a correlated response to selective breeding for or against reduced FEC in lambs. This may have important implications for the epidemiology of nematode parasite infections in spring-born lambs, a possibility which is currently being investigated.