Previous research has indicated that supplementing an apparently adequate diet with additional protein improves both host resistance and resilience in lambs infected with Haemonchus contortus. The present study tested the influence of supplementation with non-protein nitrogen (urea). Helminth-naive Hampshire Down lambs were given an apparently adequate basal diet or a diet supplemented with urea. The lambs were then infected with Haemonchus contortus for 10 weeks. Supplementation with urea had no discernible effect on resistance to infection; faecal egg counts, worm burdens, worm lengths and mean number of eggs per adult female worm did not differ between the 2 groups. However, lambs on the supplemented diet showed better resilience; they had greater packed red cell volumes, higher plasma albumin concentrations and increased liveweight gain compared to lambs on the basal diet. The loss of appetite following infection was less in lambs fed the urea-supplemented diet. The observed effect of urea supplementation was seemingly due to greater food consumption as well as the better diet.

The effects of dietary urea supplementation and of a 10-week trickle infection regime, simulating chronic exposure to Haemonchus contortus, on the zymogenic population of the abomasa of Hampshire Down lambs was examined. At necropsy a variety of parameters including plasma pepsinogen concentrations, the wet weights of abomasal fundic mucosal pieces and the amounts of pepsinogen contained in them, were assessed. Tissue pepsinogen concentration was measured as the total, acid-stable proteolytic activity present in mucosal homogenates, as well as immunohistochemically. The immunohistochemical findings were quantified using computer-aided image analysis. Elevation of plasma pepsinogen concentrations in infected animals was of borderline significance (P=0·06). The fundic mucosae of infected animals were heavier (P<0·02) than those of control animals, but there was no overall change in the pepsinogen content of tissues. Immunohistochemistry revealed that infected animals had increased numbers of zymogenic cells, due to mucous cell hyperplasia and the adaptation of cells to produce both mucins and pepsinogen. The pepsinogen content of chief cells, the major source of pepsinogen in uninfected animals, was reduced in infected lambs. Image analysis confirmed that at a mid-point of the mucosa of infected animals there was increased pepsinogen-specific immunoreactivity that corresponded with areas of mucosal hyperplasia. Mucous cell hyperplasia might therefore allow the maintenance of pepsinogen secretion in infected animals even if chief cell output is reduced.

A one-year study of gastro-intestinal parasitism in a free-ranging population of maras at Whipsnade Wild Animal Park, UK, revealed a strong relationship between membership of social units and both intensity and prevalence of infection. The mara, a hystricomorph rodent from southern Argentina, has a social organization including both monogamy and communal denning of the young, an apparently unique combination among mammals. From October 1992 to September 1993, strongyloid parasite loads were estimated from faecal egg counts. A minimum adequate model was fitted to the data using the Genstat statistical package. This showed that family membership had a highly significant effect on the intensity of egg shedding in faeces, and a significant effect on the prevalence of infection. After controlling for both extrinsic environmental and intrinsic demographic factors, homogeneity of infection was greater within than between families and adult pairs.

Ascaris suum contains a large number of FMRFamide-related peptides (FaRPs) of which KNEFIRFamide (AF1), KHEYLRFamide (AF2) and KSAYMRFamide (AF8, also called PF3) have been extensively studied and are known to exert actions on somatic muscle strips of the worm. In the present study, the effects of AF1, AF2 and AF8 on the activity of the vagina vera of female A. suum have been examined in vitro. The vagina vera is a muscular tube connecting the uterus and vagina uteri to the gonopore and is probably involved in regulating egg output. The tissue exhibited spontaneous, rhythmic contractions in vitro, which were modulated by each of the FaRPs tested. The effects of each of the peptides were qualitatively and quantitatively different, and in each case were reversible. AF1 (1 μM) caused a biphasic response in the form of a transient lengthening of the preparation, followed by a shortening; contractions were initially inhibited but resumed 5 min post-addition of the peptide. Lower concentrations ([les ]0·1 μM) induced a less marked effect, with rhythmic contractions returning 5 min post-addition. AF2 and AF8 reduced contraction frequency at concentrations [ges ]0·1 μM. Both peptides also caused the tissue to shorten, although the effects of AF8 on baseline tension were inconsistent. The apparent potencies of AF1 and AF8 on contraction frequency of the vagina vera were 10-fold greater than AF2 and, unlike their actions on A. suum somatic body wall muscles, the actions of AF1 and AF2 were qualitatively different. Indeed, the effects of each of these FaRPs on the vagina vera were markedly different from those observed on the somatic muscle.

The effect of the nematode Toxocara canis on social behaviour and anxiety levels of adult male outbred (LACA) mice was examined following infection with a single dose of 2000 ova. The actual number of larvae recovered from the brain of each individual mouse was determined after behavioural testing. The effect of the parasite on mouse behaviour was analysed by both the initial dose administered (i.e. infected versus control) and the degree of infection in the brain. There was substantial variation in the number of larvae recovered from the brains of the individual mice and the magnitude of behavioural change was associated with the level of infection. Examination of social behaviour for both analyses revealed that the infection reduced levels of aggressive behaviour and increased levels of flight and defensive behaviours. High infection in the brain induced the greatest degree of behavioural change which decreased in mice with lower infections. In contrast the analysis of anxiety levels in mice by initial dose administered revealed no difference between infected and control mice. Mice with low infection in the brain, however, displayed a greater level of risk behaviour by spending more time in the vicinity of a predator odour and in the light area of a light/dark paradigm than control or high infection mice. The results suggest that the behaviour of mice infected with T. canis is influenced by the number of larvae accumulated in the brain. This may have important consequences for the conclusions drawn on the effect of this parasite on murine behaviour.

Species richness and diversity of macroparasite assemblages were compared among American eels (Anguilla rostrata) from Nova Scotia, European eels (A. anguilla) from the United Kingdom (Kennedy, Bush & Aho, 1986; Esch et al. 1988; Kennedy, 1990, 1993), and Australian eels (A. reinhardtii) from Queensland (Kennedy, 1995). Community richness and diversity of the macroparasite fauna of American and European eels did not differ significantly for total parasite component communities, intestinal parasite component communities, and intestinal parasite infracommunities. The similarities in richness and diversity between the parasite communities of American and European eels are not surprising given the common, recent origin of these sister species. However, differences in species composition were noted between Nova Scotia and the UK. Both species of eels were infected by a nearly identical suite of specialists, but differences occurred in the species number and composition of generalist parasites. In addition, generalist species were rarely dominant in Nova Scotia, but commonly so in the UK. These differences can be attributed to the differences in the freshwater fish fauna and their parasites that occur between Nova Scotia and the UK. American and European eels are derived from a common ancestor and, whereas they have carried with them a common suite of specialist parasites during their brief period of independence, they acquired different suites of generalists apparently from their respective continental faunas after they diverged. In contrast, parasite communities of American and European eels were significantly less diverse and speciose than those of Australian eels regardless of scale (total component community, intestinal component community, intestinal infracommunity). These results support the notion that parasite communities have had more time to evolve and/or that tropical conditions have promoted parasite speciation in Australian eels.

Full-length cDNAs encoding cytosolic (SODc) and putative extracelluar (SODe) Cu/Zn superoxide dismutases (SODs) from the ovine gastrointestinal parasitic nematode Haemonchus contortus have been isolated and characterized. The predicted sequences of the H. contortus SODs showed strong homology to other helminth SODs, the highest level of sequence similarity was with those of the free-living nematode Caenorhabditis elegans. The predicted amino acid sequence of the putative extracellular form contained an N-terminal extension with the characteristics of a signal sequence including a potential signal peptidase cleavage site. Transcripts of both classes of Cu/Zn SOD were detected in all life-cycle stages examined. The cytosolic SOD mRNA was approximately 6-fold more abundant than that of the extracellular enzyme in adult parasites. Immunoblotting with antisera raised to in vitro-expressed parasite SODs revealed the presence of 2 proteins in extracts of adult H. contortus, with molecular masses of approximately 19·8 and 18 kDa. An additional protein of approximately 16·8 kDa was detected in adult ES material. Immunofluorescent staining showed Cu/Zn SOD was localized in the body wall musculature and the pharynx in adult worms and in the uterine tract of adult females. The immunogenic properties of recombinant H. contortus Cu/Zn SODs was assessed in a challenge infection experiment in lambs.

Although there has been a great deal of research effort within the last two decades on identifying the active components of the saliva of blood-sucking ticks, mosquitoes, biting flies, fleas and bugs, essentially neglected have been the human lice. Despite initial reports in the early part of this century suggestive of vasodilatory, anticoagulant and immunosuppressive properties of the saliva, for the next 50 years there were no biochemical studies on the active principles. Very recently, anatomical and biochemical studies have begun to characterize the bioactive molecules in lice saliva. The louse stocks a salivary vasodilator in excess over what is needed for a single bite, and injects similar amounts at each successive bite. The vasodilator in lice saliva appears to have different pharmacological properties than peroxidative, oxidative and maxidilan types of vasodilators reported from other blood-sucking insects. Possible anticoagulant activities have also been characterized. This belated, but welcome, interest comes at a time of resurgence of lice-born disease in certain parts of Africa, and of resistance to chemical control in Europe and North America.

The total lipid content of the dry weight of whole cysts and 2nd-stage juveniles (J2) of Globodera rostochiensis was 17·1% in dry cysts, 20·9% in cysts soaked in distilled water (DW), 20·3% in cysts that had been in potato root diffusate (PRD) for 7 days, 7·3% in cysts that had been in PRD for 28 days and 29·2% for hatched J2. The fatty acid composition of the total lipid did not differ between dry cysts, cysts in DW and cysts in PRD for 7 days. However, major differences in the fatty acid composition of all lipid classes were found between rehydrated cysts in PRD and freshly hatched J2. After hatching, the degree of saturation and the percentage of monounsaturated fatty acids decreased and the percentage of polyunsaturated fatty acids increased considerably, especially in the free fatty acid fraction, where C20[ratio ]1 showed an 8-fold decrease and C20[ratio ]4 a 33-fold increase. There was a difference in the fatty acid composition of cysts in PRD for 7 days and cysts in PRD for 28 days (after most of the J2 had hatched); with increased time in PRD the percentage of polyunsaturated fatty acids increased and the percentage of monounsaturated fatty acids decreased in all lipid classes. Differences in the fatty acid profiles between cysts in PRD for 28 days and hatched J2 were found mainly in the free fatty acid and the non-acidic phospholipid fractions. The free fatty acid fraction of the cysts was dominated by monounsaturated fatty acids (52%), whereas the same fraction of hatched J2 contained large amounts of polyunsaturated fatty acids (60%). These polyunsaturated fatty acids, especially C20[ratio ]4, might play an important part in nematode–plant interactions during infection.

Metacestodes of Hymenolepis diminuta affect several aspects of female reproductive physiology in Tenebrio molitor and such effects are mediated via the endocrine system. The effects on male reproduction are less well known and were studied with respect to the Bean-Shaped Accessory Glands (BAGs). The size and wet and dry weight of BAGs from infected and uninfected beetles were compared and rose to a plateau from 0–6 days post-emergence in uninfected beetles but in infected individuals continued to increase in both size and weight. These effects were density independent. Glands from both infected and uninfected beetles were assayed for trehalase activity measured by its ability to convert the sugar trehalose to glucose. The activity of this enzyme, per mg wet weight, was not affected by the parasite. However, total activity per gland increased in infected males. Total protein content and electrophoretic profiles of BAGs from infected and uninfected individuals showed no change in profile but showed an increase in all protein subunits per gland over a broad molecular weight range.

In order to estimate the potential benefits of interventions against multiple geohelminth species in endemic areas, an improved understanding of the population biology of multiple infections is required. This paper presents a detailed analysis of the associations among Ascaris lumbricoides, Trichuris trichiura and hookworm infections in 1539 schoolchildren on Pemba Island, Tanzania, where 58% of the sampled children carried infections of all 3 parasites at the time of the study. Infection intensities of different species were positively correlated, and individuals with single-species infections had generally lower species-specific egg counts than individuals with multiple-species infections. There was no age- or sex-related clustering of infections. A weak clustering of intense infections among individuals with multiple-species infections was observed, which became more pronounced as the threshold defining an intense infection increased for each species. The results suggest that individuals with multiple species infections are likely to be at highest risk of geohelminth-related morbidity, not only because of the number of infections they harbour, but also because they generally carry heavier infections of each species.

Cryptic species, belonging to the 37 collar-spine Echinostoma group, were distinguished using nuclear rDNA ITS (884 bases) and mtDNA CO1 (257 bases) and ND1 (530 bases) sequences. Sequences were obtained from five 37 collar-spine species, Echinostoma trivolvis, E. paraensei, E. caproni, E. revolutum and E. sp.I, a parthenogenetic isolate from Africa. Three geographic isolates of E. caproni were compared. Average sequence divergence among the 37 collar-spine species range from 2·2% in the rDNA ITS through 8% for the CO1 and 14% for the ND1. In addition, genes were sequenced from 2 non 37 collar-spine species, E. hortense and an undescribed Australian species, E. sp. (Aus). For each gene, distances of terminals from a predicted ancestral sequence were calculated. These indicated that ND1 is diverging significantly faster than the other 2 regions. In the CO1 gene most substitutions are synonymous and saturation has been reached for the majority of pairwise comparisons. The ND1 gene exhibits greater pairwise divergence but less evidence of saturation due to weaker conservation of first and second codon positions. The ITS has no amino acid coding constraints and displays no evidence of saturation. Although all 3 regions successfully distinguished the nominal species, ND1 appears to be the most informative region for investigating relationships within the 37 collar-spine group.

The ookinete is one of the most important stages of Plasmodium development in the mosquito. It is morphologically and biochemically distinct from the earlier sexual stages - gametocytes and zygote, and from the later stages - oocyst and sporozoites. Development to ookinete allows the parasite to escape from the tightly packed blood bolus, to cross the sturdy peritrophic matrix (PM), to be protected from the digestive environment of the midgut lumen, and to invade the gut epithelium. The success of each of these activities may depend on the degree of the biochemical and physical barriers in the mosquito (such as density of blood bolus, thickness of peritrophic matrix, proteolytic activities in the gut lumen etc.) and the ability of the ookinete to overcome these barriers. Ookinete motility, secretion of chitinase, resistance to the digestive enzymes, and recognition/invasion of the midgut epithelium all may play crucial roles in the transformation to oocyst. The overall sporogonic development of Plasmodium, therefore, depends on the results of the two-way manipulations between the parasite and the vector mosquito. Study of ookinete development and of the cellular and biochemical complexities of the mosquito gut may therefore lead to the design of novel strategies to block the transmission of malaria. This article reviews the intricate interactions between the parasite and the mosquito midgut in the context of development and transmission of Plasmodium parasites.

In most real-world contexts the sampling effort needed to attain an accurate estimate of total species richness is excessive. Therefore, methods to estimate total species richness from incomplete collections need to be developed and tested. Using real and computer-simulated parasite data sets, the performances of 9 species richness estimation methods were compared. For all data sets, each estimation method was used to calculate the projected species richness at increasing levels of sampling effort. The performance of each method was evaluated by calculating the bias and precision of its estimates against the known total species richness. Performance was evaluated with increasing sampling effort and across different model communities. For the real data sets, the Chao2 and first-order jackknife estimators performed best. For the simulated data sets, the first-order jackknife estimator performed best at low sampling effort but, with increasing sampling effort, the bootstrap estimator outperformed all other estimators. Estimator performance increased with increasing species richness, aggregation level of individuals among samples and overall population size. Overall, the Chao2 and the first-order jackknife estimation methods performed best and should be used to control for the confounding effects of sampling effort in studies of parasite species richness. Potential uses of and practical problems with species richness estimation methods are discussed.

The effects of a parasitic infection with the nematode Nippostrongylus brasiliensis on the degradation rates of cytoplasmic tRNA, rRNA and mRNA in rats have been investigated by measuring the renal excretion rates of the modified RNA catabolites N6-threoninocarbonyladenosine, pseudouridine and 7-methylguanine. Between days 9 and 13 post-infection when the expulsion of N. brasiliensis is usually the most pronounced, the degradation rates of the different RNA classes were significantly higher than in the control rats (P<0·05) by, on average, +24% (tRNA), +34% (rRNA) and +26% (mRNA). We suspect that the elevated degradation rates of RNA are related to an increased production of reactive oxygen species by the host during the expulsion of N. brasiliensis.

The most important function of any parasite is to secure transmission to new hosts. The gametocyte, the stage which has become developmentally committed to the sexual cycle, provides a critical link in the transmission of Plasmodium falciparum from the human host to the anopheline mosquito vector. It is therefore imperative that our determination to understand the biology of the gametocyte is greater than the technical obstacles which have resulted in the gametocyte being left very much out of the limelight by the intensive investigation of the asexual bloodstream parasite. Here we explore the areas of gametocyte biology which by nature of their relevance to control and pathology as well as basic biology, are the subjects of investigation in our laboratory. We also point out areas in need of particular attention.

Cytoplasmic incompatibility consists of sterility in cross matings, the crossing type being maternally inherited. It can be explained by the action of Wolbachia symbionts which are transmitted through the egg cytoplasm and leave an imprint on the sperm which prevents it fertilizing unless it is ‘rescued’ by the action of the same type of Wolbachia in the egg. Thus matings between infected males and uninfected females are sterile, but the reciprocal matings are fertile. Hence uninfected females are at risk of failing to transmit their uninfected cytoplasm if they cross mate, but infected females are at no such risk. Therefore natural selection favours the infected state and in two wild insect populations the infection has been observed spreading. If a gene for inability to transmit malaria could be introduced into Wolbachia and if this could be introduced into Anopheles (where these symbionts appear not to occur naturally), release of a limited number of such insects should trigger a process of displacement of malaria vectors, by the non-vector type. A simple model is used to demonstrate the limitations to this process which would be introduced by immigration.