cDNAs encoding the immunodominant allergens tropomyosin and paramyosin were amplified from RNA extracted from the sheep scab mite Psoroptes ovis. The tropomyosin cDNA contained an open reading frame (ORF) of 852 bp which encoded a predicted protein with 98% and 97% identity to the house dust mite allergens Der f 10 and Der p 10 respectively. The complete paramyosin ORF generated by RT-PCR was 2625 bp in length and encoded an 875aa predicted protein of 102·6 kDa with 97%, 95% and 89% identity to the paramyosins of Dermatophagoides pteronyssinus (Der p 11), Sarcoptes scabiei and Blomia tropicalis (Blo t 11) respectively. Full length tropomyosin and truncated and full-length paramyosin were expressed as recombinant proteins. IgG and IgE in sera from sheep with a 6-week duration primary infestation of P. ovis did not detect either full-length or truncated recombinant paramyosin. IgG in both infested and naïve sheep sera detected recombinant tropomyosin, suggesting cross-reactivity to tropomyosin and to other invertebrate species to which the sheep may have been exposed. Staining with antibodies directed against tropomyosin and paramyosin was observed throughout sections of P. ovis. Staining was especially prevalent in the anterior sections of the mites, possibly associated with locomotory muscles in this region.

We have determined the molar content of guanine + cytosine (GC content) of DNA of the filarial nematode (Brugia malayi, Brugia pahangi and Dirofilaria imitis) and of the free-living soil nematodes Caenorhabditis elegans and have analysed the DNA for the presence of methylcytosine. Two independent methods, thermal denaturation and direct analysis of base content by HPLC following enzymatic hydrolysis, reveal that the GC content of filarial nematodes is 26–28%. We have been unable to find methylcytosine in the DNA of B. malayi.

Five novel species, Auerbachia scomberoidi n. sp., Auerbachia chaetodoni n. sp., Auerbachia caranxi n. sp., Coccomyxa colurodontidis n. sp. and Coccomyxa gobiodoni n. sp. are described from the gall bladders of marine teleosts. These species descriptions provide the first record of Auerbachia from Australian waters. Each species is characterized morphologically, including additional measurements for Auerbachia spp. and small subunit ribosomal DNA (SSU rDNA) sequences were determined for molecular phylogenetic analyses. All 5 species were each recovered from a single (and different) species of host. Phylogenetic analyses revealed a close genetic relatedness between members of Auerbachia and Coccomyxa. Based on these phylogenetic data, on obvious paraphyly displayed by the Myxidiidae and on priority, we propose the re-establishment of the family Coccomyxidae to house all species of the genera, Coccomyxa, Auerbachia and Globospora.

As part of a drug trial against bancroftian filariasis in the East Sepik Province of Papua New Guinea we measured the pre-treatment microfilarial densities of 2219 individuals. Mean levels generally increased with age in both sexes, with a tendency to plateau at the highest ages. However, there was a reduction among women of approximately reproductive age. Allowing for the tendency for aggregation to decrease with age, this reduction was statistically significant. However, a comparison of pregnant women and controls showed no evidence that the reduction is specifically related to pregnancy. Moreover, a simple differential equation model of microfilarial acquisition and loss suggests that age-specific patterns of exposure are also unlikely to be solely responsible. Therefore, we suggest that the observed reduction in microfilarial intensity may result from hormonal changes associated with female reproduction, possibly in combination with other factors.

We tested a theoretical model based on the physics of capillary flow and confirmed that anaemia accelerates blood intake in the yellow-fever mosquito, Aedes aegypti (L.). We also investigated the influence of anaemic blood on egg production of mosquitoes and found that it has a negative influence on fecundity. Based strictly on egg production and the physics of fluid intake, we propose that although anaemia associated with blood-borne parasites may be detrimental to mosquitoes that can engorge to repletion in one session, it may be beneficial to those interrupted before repletion because the greater quantity of the bloodmeal may compensate for its lower quality. Epidemiological consequences are discussed but require further inquiry.

DEC-fortified salt has been used successfully as a principal public health tool to eliminate lymphatic filariasis (LF) in China and, less extensively, in several other countries. Studies from 1967 to the present conducted in Brazil, Japan, Tanzania, India, China, and Taiwan involving administration of DEC salt for 18 days to 1 year, have shown this intervention to be effective for both bancroftian and brugian filariasis, as measured by reductions in both microfilarial density and positivity, and in some studies through reduction in mosquito positivity rates as well. Furthermore, studies suggest specific advantages from using DEC salt, including lack of side effects, particularly for bancroftian filariasis, and ability to reduce prevalence below 1% when used in conjunction with standard regimens of DEC tablets. However, use of DEC salt as a control tool suffers from a concern that health authorities might find it difficult to manage a programme involving a commodity such as salt. In the past decade, the very successful global efforts to eliminate iodine deficiency through universal salt iodization have demonstrated that partnership with the salt industry can be both successful and effective as a public health tool. Use of DEC salt can be most successfully implemented in areas in which (a) there is adequate governmental support for its use and for elimination of filariasis, (b) filariasis-endemic areas are clearly defined, (c) political leaders, health officials and the salt industry agree that DEC salt is an appropriate intervention, (d) the salt industry is well-organized and has known distribution patterns, (e) a successful national salt iodization effort exists, (f) a monitoring system exists that ensures adequacy of salt iodine content during production and that can also measure household coverage, and (g) measurement of impact on transmission of LF with the new antigen or filarial DNA detection methods can be established. There are advantages and disadvantages of using DEC-fortified salt compared with other interventions for LF elimination programmes, but rather than being considered as a ‘competing’ intervention, DEC salt should be seen as an additional option. Indeed, it is likely that many countries will derive maximal benefit from the synergistic effects of combining different intervention strategies in their national programmes to eliminate lymphatic filariasis.

The use of DNA markers to track the development of anthelmintic resistance in parasites of livestock would allow informed choices for the management of this important problem. We describe a genetic mapping approach for the discovery of DNA markers for anthelmintic resistance in Haemonchus contortus. We crossed a multi-drug resistant field isolate of H. contortus with a well-characterized laboratory strain susceptible to 4 drug classes. The F2 were separately selected with 5 anthelmintics from 4 drug classes, producing drug-resistant populations carrying gene variants derived from both the field isolate and the laboratory strain. Individual F2 worms were analysed using amplicon length polymorphisms (ALPs). We looked for field isolate alleles over- or under-represented in F2 populations compared to the unselected F2 and/or the laboratory strain. The data we obtained suggest that marker association can be used to link neutral markers with resistance, but also that more markers and perhaps more inbred laboratory strains would make the procedure more likely to succeed.

In order to determine how much of the variability in parasite assemblages is driven by differences in composition or in abundance we used multivariate dispersions (average distance from infracommunities to their size class centroid in the multivariate space) as a measurement of β-diversity in infracommunities of Conger orbignianus, applying a set of dissimilarity measures with different degrees of emphasis on composition versus relative abundance information. To evaluate comparatively the rate of such changes, we also analysed the effect of host size by regressing differences in β-diversity among size classes against differences in mean fish size. Multivariate dispersions varied along an ontogenetic gradient, its significance depending on the measurement used. Larger fish showed higher richness and abundance; however, smaller fish displayed lower variations in abundance but higher in composition. This could be caused by stochastic encounters at low densities due to the overdispersion of parasites in previous hosts. As fish grow, the composition of their parasite assemblages becomes homogenized by repeated exposure, with abundance thus arising as the main source of variability. Both variables act at different rates, with the exponential decay in the compositional variability as differences in fish size increase being about twice as steep as the decay in abundance variability, indicating that compositional homogeneity is reached faster than abundance heterogeneity as fish grow. Discerning between both variables is crucial in order to understand how community structure is formed by size-dependent variability of host populations.

Several major groups of parasitic helminths (Monogenea, Digenea and Nematoda) possess two surfaces that are potentially absorptive in nature. These are an external surface, a tegument in the platyhelminths and cuticle in the nematodes, and the intestine. This paper discusses the relative contributions of these absorptive surfaces in the nutrition of these parasitic helminths. There are many factors that determine the availability of, and a parasite's ability to absorb nutrients via either of these surfaces, and this review discusses individually some of the more important morphological, physiological and environmental factors affecting the potential nutritional roles of these surfaces. It is clear from such a summary of previous studies that the intestines and teguments (cuticles) of helminth parasites can each serve an important nutritional role. However, insufficient data make it impossible at this time to determine the relative nutritional roles of these surfaces in any single parasitic helminth.

This study established the mechanisms by which Microphallus primas cercariae penetrate the crab Carcinus maenas in which they form metacercarial cysts. Light and electron microscopy were used to investigate cercarial features and to follow the fate of cercariae released in close proximity to crabs. It was shown that cercariae were carried in respiratory currents into crabs' branchial chambers where each enveloped itself in a transparent penetration cyst on the gill lamellae. When cercariae were present the number of respiratory current reversals performed by crabs increased. Using an ‘artificial branchial chamber’ it was possible to observe how cercariae attached to crab gills during breaks in current flow that preceded each current reversal. Inside the penetration cysts the now tail-less larvae used their stylets to pierce holes through which they levered themselves into underlying haemolymph channels in the gills. Histochemical tests demonstrated that the penetration cysts were products of glands in the cercariae and that penetration of the crabs was achieved by mechanical means. The importance of crab respiratory current reversals to the success of cercarial penetration is discussed as it represents the exploitation by a parasite of a host behavioural response to an unrelated stimulus.

Simple mathematical models are developed to examine the influence of variability in host susceptibility to infection, on the dynamics of host–parasite population interactions. When hosts differ in their innate susceptibility (at birth), to infection by a specific parasite, the average susceptibility of the host population as a whole may show systematic changes through time. Such patterns may arise as a result of demographic factors associated with the interaction between host and parasite populations, in the absence of inheritance mechanisms (a genetic component) or acquired resistance (an immunological component). The general significance of this observation is discussed in terms of the coevolution of host–parasite associations.

The problem of anthelmintic resistance prevents efficient control of parasites of livestock and may soon compromise human parasite control. Research into the mechanisms of resistance and the quest for diagnostic tools to aid control has required research that focuses on field resistance. On the other hand, resistant worms, including those kept in the laboratory, provide useful tools for studying drug action, especially at neuromuscular targets in worms. While the needs and directions of these research aims overlap, this review concentrates on research on drug targets. In this context, resistance is a useful tool for site of action confirmation. For example, correlations between molecular expression studies and resistance assays conducted on whole worms can strengthen claims for sites of anthelmintic action. Model systems such as Caenorhabditis elegans have been very useful in understanding targets but give a limited picture as it is now clear that resistance mechanisms in this worm are different from those in parasites. Accordingly, research on parasites themselves must also be performed. Resistant isolates of the sheep nematode parasite Haemonchus contortus are the most widely used for this purpose as in vivo, in vitro, physiological and molecular studies can be performed with this species. Neuromuscular target sites for the anthelmintics levamisole and ivermectin are the best studied and have benefited most from the use of resistant worm isolates. Resistance to praziquantel and the newer chemical groups should provide new tools to explore targets in the future.

The filarial nematodes Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus represent major public health problems in the Tropics. Effective diagnosis of infection with these parasites is required both for administration of drugs to infected individuals and for monitoring of control programs. However parasitological diagnosis is associated with a number of problems including frequently inadequate sensitivity, long pre-patency of infection and inconvenience for patients. For these reasons there has been considerable effort expended in developing other forms of diagnosis, in particular immunoassays for measuring antibody and circulating parasite antigen as well as molecular-biology-based assays for detecting parasite DNA. This article reviews the progress and achievements obtained to date. The latter include the development of ELISAs employing recombinant antigen for detection of antibody to O. volvulus which have both high sensitivity and specificity, the commercial availability of immunoassays to measure circulating antigen in W. bancrofti infection and the generation of specific DNA-based detection systems for all three parasites.

In three experiments, the chief hosts, sheep, were withheld from a grazing until 0–3 weeks after the normal spring peak of female tick activity. This did not give rise to any appreciable prolongation of female tick activity into the normal summer offseason. More than half the female ticks actually available during the entire spring season were unfed when activity came to an end. This was due to the withholding of hosts. Hence that aspect of the ‘two-brood’ theory which claims that activity ends in spring, because all the available ticks are fed, is untenable as the complete explanation of bimodal annual activity.

Increase in sheep-stocking density, over the normal for a grazing, lowers the average infestation per sheep for that season at least.

The experimental data suggests the following:

(i) Delay in stocking a grazing with sheep results in a slightly higher-than-normal population of active female ticks which is fairly quickly absorbed after reintroduction of sheep.

(ii) A varying proportion of the tick population becomes active each week throughout the tick season, increasing up to the peak and decreasing thereafter.

(iii) The average duration of individual activity in a natural population of female ticks is probably not more than 1 month, perhaps less.

Practical applications of the findings to hill sheep farming are indicated.

Methyl benzoquate at 20 ppm or clopidol at 250 ppm in the diet show true coccidiostatic activity in infections with Eimeria tenella in the chick. Lerbek, a synergistic mixture yielding 8·35 ppm methyl benzoquate and 100 ppm clopidol, produced a partially coccidiocidal effect; relapses following restricted periods of medication were less severe, and suppression of parasite development following delayed medication was quicker and more effective than with either component of the mixture used alone.

The treatment of Neospora caninum infection in the bovine host is still at an experimental stage. In contrast to the in vivo situation, a wide range of compounds have been intensively investigated in cell-culture-based assays. Tools to demonstrate efficacy of treatment have remained conventional including morphological and cell biological criteria. In this work, we present a molecular assay that allows the distinction between live and dead parasites. Live parasites can be detected by measuring the mRNA level of specific genes, making use of the specific mRNA available in live cells. The NcGra2 gene of N. caninum, which is known to be expressed in both tachyzoites and bradyzoites, was used to establish a quantitative real-time RT-PCR, for monitoring parasite viability. Validation of the system in vitro was achieved using Neospora-infected cells that had been treated for 2–20 days with 30 μg/ml toltrazuril. NcGRA2-RT-real time PCR demonstrated that a 10-day toltrazuril-treatment exerted parasitostatic activity, as assessed by the presence of NcGRA2-transcripts, whereas after a 14-day treatment period no NcGRA2-transcripts were detected, showing that the parasites were no longer viable. Concurrently, extended culture for a period of 4 weeks in the absence of the drug following the 14-day toltrazuril treatment did not lead to further parasite proliferation, confirming the parasiticidal effect of the treatment. This assay has the potential to be widely used in the development of novel drugs against N. caninum, with a view to distinguishing between parasiticidal and parasitostatic efficacy of given compounds.

The relationship between host dietary protein and intestinal helminth population biology during primary infection was studied using a mouse–T. muris experimental model. Uninfected CBA/Ca mice fed either a 4% (by mass) or 16% protein diet grew at a similar rate, but both suffered significant growth retardation when infected with T. muris. T. muris initial establishment and fecundity were unaffected by dietary protein content, but were significantly influenced by infection dose, declining at the higher level of infection. Dietary protein deficiency increased parasite survival at high infection levels. Heavily infected hosts fed the 16% protein diet were found to expel almost the whole of their infections around day 21 p.i. By contrast, this response appeared defective in hosts fed the lower 4% protein diet, resulting in the establishment and survival of large, chronic and patent infections. Mean faecal egg output per mouse increased with infection dose in animals fed a low protein diet, but was low and unrelated to infection exposure in hosts fed the 16% protein diet. It is concluded that low protein levels in diet, via alterations in host resistance, may have an important influence on the transmission of T. muris.

Within-host interactions among coinfecting parasites are common and have important consequences for host health and disease dynamics. However, these within-host interactions have traditionally been studied in laboratory mouse models, which often exclude important variation and use unnatural host–parasite combinations. Conversely, the few wild studies of within-host interactions often lack knowledge of parasite exposure and infection history. Here we exposed laboratory-reared wood mice (Apodemus sylvaticus) that were derived from wild-caught animals to two naturally-occurring parasites (nematode: Heligmosomoides polygyrus, coccidia: Eimeria hungaryensis) to investigate the impact of coinfection on parasite infection dynamics, and to determine if the host immune response mediates this interaction. Coinfection led to delayed worm expulsion and prolonged egg shedding in H. polygyrus infections and lower peak E. hungaryensis oocyst burdens. By comparing antibody levels between wild and colony-housed mice, we also found that wild mice had elevated H. polygyrus-IgG1 titres even if currently uninfected with H. polygyrus. Using this unique wild-laboratory system, we demonstrate, for the first time, clear evidence for a reciprocal interaction between these intestinal parasites, and that there is a great discrepancy between antibody levels measured in the wild vs those measured under controlled laboratory conditions in relation to parasite infection and coinfection.

Groups of mice were exposed to multiple bites by tsetse flies (Glossina morsitans morsitans) infected with a clone of Trypanosoma congolense spread over a period of 8 days. The mice were subsequently treated with Berenil 10 days after the first fly bite as were uninfected control mice. The group of mice which received 12–15 infectious fly bites on two occasions, 21 days apart, were subsequently resistant to infection when re-challenged by flies infected with the same clone of T. congolense. These mice were also immune to challenge by flies infected with a different bloodstream variable antigen type derived from this same stock. The immunity was stock-specific and directed against the metacyclic forms of the parasite, but was short-lived.

Objective. This paper assessed the potential of traditional Chinese medicine (TCM) preparations for using as environmentally acceptable and alternative commercial acaricides. Methods. 22 kinds of TCM, which contained abundant essential oils and showed insecticidal effects, were collected. Samples extracted with petroleum ether, ethyl acetate and methanol were tested against house dust mites Dermatophagoides farinae and their toxicity assessed. Results. The results showed that 3 TCM of Cinnamonum cassia, Eugenia caryophyllata and Pogostemon cablin have higher activity, and the parallel tests showed that the petroleum ether extract had higher activities (0·0046 mg/cm2, 0·005 mg/cm2 and 0·006 mg/cm2 respectively, 24 h, LD50) than the extracts of ethyl acetate and methanol. The acaricidal activity of the ethyl acetate extracts from C. cassia, P. cablin and Asarum sieboldii (0·00144 mg/cm2, 0·00347 mg/cm2 and 0·05521 mg/cm2 respectively, 24 h, LD50) were almost comparable to that of benzyl benzoate and dibutyl phthalate. However, the methanolic extracts of were less effective. Conclusions. This study shows the use of extracts with petroleum ether of C. cassia, P. cablin and E. caryophyllata as eco-friendly biodegradable agents for the control of the house dust mite.