In this paper, we describe the occurrence of a microsporidian parasite in female-biased populations of an intertidal amphipod, Corophium volutator (Pallas), at mudflat sites in the Bay of Fundy, Canada. Sequence data for the parasite's 16S rDNA indicate that it is a novel microsporidian species. This parasite was found principally in female host gonads, indicating that it might be a vertically transmitted, sex-distorting microparasite. At 4 sites each sampled in early and mid-summer, parasite prevalence varied from 0 to 21%. In the lab, infected mothers gave rise to more female-biased broods, than did uninfected mothers. Infection was not associated with size of females or with lowered survivorship of their young. Surprisingly, infected mothers actually had higher fertility controlling for body length than did uninfected mothers. Taken together, our results suggest that this novel microsporidian is likely a feminizing microparasite and is a contributing factor to local and widespread sex ratio distortion in C. volutator.

Nematodes represent one of the most abundant and species-rich groups of animals on the planet, with parasitic species causing chronic, debilitating infections in both livestock and humans worldwide. The prevalence and success of the nematodes is a direct consequence of the exceptionally protective properties of their cuticle. The synthesis of this cuticle is a complex multi-step process, which is repeated 4 times from hatchling to adult and has been investigated in detail in the free-living nematode, Caenorhabditis elegans. This process is known as moulting and involves numerous enzymes in the synthesis and degradation of the collagenous matrix. The nas-36 and nas-37 genes in C. elegans encode functionally conserved enzymes of the astacin metalloprotease family which, when mutated, result in a phenotype associated with the late-stage moulting defects, namely the inability to remove the preceding cuticle. Extensive genome searches in the gastrointestinal nematode of sheep, Haemonchus contortus, and in the filarial nematode of humans, Brugia malayi, identified NAS-36 but not NAS-37 homologues†. Significantly, the nas-36 gene from B. malayi could successfully complement the moult defects associated with C. elegans nas-36, nas-37 and nas-36/nas-37 double mutants, suggesting a conserved function for NAS-36 between these diverse nematode species. This conservation between species was further indicated when the recombinant enzymes demonstrated a similar range of inhibitable metalloprotease activities.

The two species of bed-bug, Cimex lectularius and C. rotundatus, are found in temperate and tropical regions respectively. They are very similar anatomically, and have the same habits.

Physiologically, the two species have many resemblances. Their water balance is similar; they are very little affected by humidity if there is plenty of food available. The thermal death-points are identical.

At temperatures below 30° C., C. lectularius is able to breed more rapidly and more successfully than C. rotundatus. Between 30 and 35° C. both species behave alike, and above 35° C. C. rotundatus may be the more efficient species. Under most conditions, C. lectularius is the more resistant to starvation and desiccation.

Results indicate that although C. lectularius is more fitted for life in temperate countries, if introduced into the tropics it should be able to carry on at least as well as C. rotundatus.

Studies of geographical distribution indicate that C. rotundatus has spread all over the tropics, but has never established itself outside them. C. lectularius has spread all over the temperate world, and into the ports in some tropical countries. Experimental work suggests that it may become widespread in those parts of the tropics into which it is introduced.

Studies of the cuticle of Aspiculuris tetraptera with both the light and the electron microscopes have shown that there are three basic layers, namely the cortex, which consists of an outer and an inner layer, the matrix layer and the fibre layer; the latter is made up of three layers. There is, in addition, a thin osmiophilic superficial membrane on the surface of the cuticle; this membrane is seen only with the electron microscope.

It is shown that the major structural protein of the cuticle, although showing no axial periodicity, is collagen; the collagen is associated with hyaluronic-acid- and chondroitin-sulphate-containing mucopolysaccharides.

Evidence is presented for the existence of two centres of protein synthesis (collagen fibrogenesis) in the cuticle of A. tetraptera, Syphacia obvelata and Ascaris lumbricoides. These centres are located in the inner cortex and the hypodermis. The cuticle is thus very active metabolically.

I wish to acknowledge with thanks the facilities afforded me at the Molteno Institute during the course of these studies by the Director, Dr P. Tate. My thanks are also due to Dr D. L. Lee for his encouragement and helpful discussions during the investigations.

We need a fuller understanding of the feeding habits of fleas on rodents, both to help in the study of the epidemiology of plague and also in carrying out laboratory experiments on the same subject. In an earlier paper (Buxton, 1938) I described a ‘synthetic mouse hole’ in which I kept a rodent with a population of fleas (Xenopsylla cheopis) under controlled conditions of climate and feeding for periods of many days; with this I obtained some measure of the progeny which resulted under a variety of conditions. There was evidence that the mouse ate a number of the fleas, and I never controlled this variable, which was erratic and sometimes very large. We attempted to avoid the destruction of the fleas by feeding them on a baby mouse, which was changed at intervals, so that there was always one in the experimental vessel. In a postscript to the above paper it was shown that the baby mouse does not consume the fleas, almost all of which are alive when the experiment has lasted a week; in that period the adult mouse may eat about half or more of the fleas. Further experiments are in progress on the factors which determine the destruction of fleas by the mouse, and on the proportion of fleas which are actually found on the mouse under different conditions of temperature and so forth. In the present note, I shall confine myself to studying the baby mouse, as a host of X. cheopis. All the experiments here described were carried out between 1937 and 1939, though it has not till now proved possible to tabulate and consider the results. The execution of the work was in the hands of Mr S. Smith, whose help is acknowledged.

Larval Digenea infecting Littorina rudis, L. littorea and L. obtusata around Portavogie harbour, Co. Down, were examined. High levels of infection were found among the littorinids collected from sites on the harbour shore, though levels of infection varied somewhat with position on the shore. There was a direct relationship between size of the snail host and likelihood of infection. There was a negative association between certain pairs of helminth species in the same host, particularly where the rediae of Cryptocotyle lingua or Himasthla sp. were involved. Infection rates in gastropods declined away from the harbour and it is concluded that the high infection rate centres around the harbour were probably due to the increased concentration of definitive hosts, mainly seagulls, attracted to this area by dumping of raw fish offal. The infection of Pholis gunnellus with the metacercariae of C. lingua was also examined. Older fish were more heavily infected than younger fish but no direct relationship was found between high snail infection rates and high P. gunnellus infection rates.

Although many interesting observations have been made upon gametogenesis in Haemosporidiidea many fundamental problems await solution. Our knowledge of the nuclear changes occurring during the development of the gametes and the fertilization of the female gamete is still incomplete. Little is known of the factors which control gametocyte production, though there is evidence that in some species, i.e. Leucocytozoon simondi and Haemoproteus columbae in the duck, it is influenced by the physiological stage of the host. The type of host and the method of transmission have also been shown to affect the numbers of gametocytes formed in some species. No correlation has been found in Plasmodium, the genus in which this subject has been studied extensively, between acquired immunity and gametocyte formation. Although a broad relationship has been demonstrated in Plasmodium between the density of gametocytes in the blood and the oocyst rate and density assessed in susceptible mosquitoes, hosts heavily infected with apparently ripe gametocytes may prove non-infective to susceptible mosquitoes. In birds heavily infected with P. gallinaceum, serum taken at the peak of parasitaemia has been shown to affect adversely the infectivity of gametocytes to mosquitoes. Pamaquin and primaquine, in small doses, inhibit the development of gametocytes completely, whereas proguanil may not prevent the formation of oocysts though it inhibits their subsequent development. High densities of gametocytes have been observed in patients treated with proguanil or sulphamethazine. An actual in crease in the number of gametocytes produced, as compared with the untreated control strain, was observed in strains of P. gallinaceum in process of developing resistance to sulphadiazine, proguanil or 2:4-diamino-6:7-diisopropylpteridine as a result of treatment with those drugs.

The internal transcribed spacer (ITS-1 and ITS-2) regions and the 5·8S ribosomal RNA gene of 2 Perkinsus spp. (G117 and H49) originating from the softshell clam, Mya arenaria, of the Chesapeake Bay were cloned and sequenced to obtain evidence for their genetic divergence. A high level of heterogeneity in both regions, probably resulting from deletions, insertions, and base substitutions, was evident from alignments of the sequences of the 2 isolates with published sequences of other Perkinsus spp. The isolate G117 and other Perkinsus spp. were highly divergent (13–26% and 19–20% sequence divergence in ITS-1 and ITS-2, respectively). These regions in the isolate H49 and Perkinsus marinus were similar (99·07% and 99% for ITS-1 and ITS-2, respectively). Evidence obtained from a phylogenetic analysis using the aligned sequences suggests that G117 and H49 belong to 2 distinct species of Perkinsus. The isolate G117 possibly belongs to an as yet undescribed species of Perkinsus, and H49 belongs to the species P. marinus. The conclusions drawn from the genetic analysis of H49 and G117 are supported by previously reported morphological characteristics (McLaughlin & Faisal, 1998b). Isolates H49 and G117 originated from the same molluscan species demonstrating that at least 2 different species of Perkinsus can co-exist in 1 host.

It is generally recommended to perform multiple stool examinations in order to improve the diagnostic accuracy when assessing the impact of mass drug administration programmes to control human intestinal worm infections and determining efficacy of the drugs administered. However, the collection and diagnostic work-up of multiple stool samples increases costs and workload. It has been hypothesized that these increased efforts provide more accurate results when infection and drug efficacy are summarized by prevalence (proportion of subjects infected) and cure rate (CR, proportion of infected subjects that become egg-negative after drug administration), respectively, but not when these indicators are expressed in terms of infection intensity and egg reduction rate (ERR). We performed a meta-analysis of six drug efficacy trials and one epidemiological survey. We compared prevalence and intensity of infection, CR and ERR based on collection of one or two stool samples that were processed with single or duplicate Kato-Katz thick smears. We found that the accuracy of prevalence estimates and CR was lowest with the minimal sampling effort, but that this was not the case for estimating infection intensity and ERR. Hence, a single Kato-Katz thick smear is sufficient for reporting infection intensity and ERR following drug treatment.

Biomphalaria glabrata embryonic (Bge) cells have been shown to be a valuable in vitro cellular model for the study of snail host–parasite interactions. They both promote the growth and differentiation of various trematode species including Schistosoma mansoni, and Echinostoma caproni and share some morphological and functional features with circulating haemocytes. As an approach to investigate snail genes potentially regulated following exposure to trematode excretory–secretory (ES) products, we compared gene expression profiles of Bge cells exposed to saline solution, or saline solution containing ES products from S. mansoni or E. caproni, two trematode species parasitizing B. glabrata. Following differential display RT-PCR analysis we characterized 23 differentially displayed cDNAs and we focussed on the 5 cDNAs showing sequence similarity to known genes for expression validation. Using RT-PCR, we confirmed that ES products from S. mansoni and E. caproni differentially affect the expression levels of 4 out of the 5 transcripts. These partial transcripts corresponded to novel B. glabrata sequences, and showed significant sequence similarity to genes coding for (i) cytochrome C, (ii) methyl-binding proteins, (iii) glutamine synthetases, and (iv) protease inhibitors from the Kunitz family. The possible significance of these gene expression changes in host–parasite molecular interactions is discussed.

The structure of the cysticercoid of Raillietina cesticillus has been studied with the aid of light and electron microscopes. While the ultrastructure of the superficial parts of the scolex closely resembles that of other cestodes, the fine structure of the cyst wall differs from both these. The arrangement and functions of the tissues in the cyst wall are discussed and interpreted with reference to the tissues of the scolex of the cysticercoid of R. cesticillus and to the tissues of other cestodes. R. cesticillus cysticercoids, subjected to various histochemical tests, were shown to contain proteins and carbohydrate materials.

The author wishes to thank the Principal and Governors of West Ham College of Technology, a constituent college of the North-east London Polytechnic, for their general and financial support of this work. Thanks are also due to the Directors and Governors of the several institutions whose electron microscopes the author used. The work described here forms part of a thesis approved by the University of London for the degree of Ph.D.

A 24-week randomized double blind intervention trial was conducted on adult female tea pluckers from an estate in Bangladesh to investigate the impact of iron supplementation and anthelmintic treatment on changes in ferritin and haemoglobin levels as well as on prevalence and intensity of helminth infections. A total of 553 women were randomly assigned to 1 of 4 intervention groups: group 1 received iron supplementation on a weekly basis, group 2 received anthelmintic treatment at the beginning and half way through the trial, group 3 received both iron supplementation as group 1 and anthelmintic treatment as group 2, and group 4 was a control group and received placebos for both iron supplementation and anthelmintic treatment. Prevalence and intensity of helminth infections (egg counts/g stool) of Ascaris lumbricoides, Trichuris trichiura and hookworms significantly fell in the 2 groups receiving anthelmintic treatment and there were some reductions in the 2 groups not receiving anthelminthic treatment. Haemoglobin and haematocrit concentrations increased significantly in the iron supplemented groups with smaller increases in the anthelmintic only group. All women showed a decrease in serum ferritin levels post-trial with greater losses in the 2 dewormed groups. Significant negative associations were found between hookworm egg counts and ferritin levels and Trichuris trichiura egg counts and haemoglobin concentration.

We studied the pattern of infection and the inter-annual variation and individual factors affecting the infection of 2 species of nasopharyngeal bot flies, Cephenemyia auribarbis and Pharyngomyia picta (Diptera: Oestridae), in a population of Iberian red deer (Cervus elaphus hispanicus) from south central Spain (10 annual periods between 1990 and 2003). Mean prevalence±S.E.95%CI of infection was 35·19±4·24% (n=486). The frequency distribution of the parasites was markedly aggregated (K: 0·213, mean abundance±S.D.: 5·49±12·12). Parasite load of Oestridae peaked at calf and subadult age groups and declined thereafter, which suggests that acquired immunity may be acting. In common with other host–parasite relationships, male hosts were found to have higher prevalence and abundance levels than females. The prevalence of P. picta was positively affected by the presence of C. auribarbis whereas the intensity of infection of P. picta was negatively affected by the presence of C. auribarbis. Intensity of P. picta in concomitant infections with C. auribarbis was lower than in pure P. picta infections, whilst the intensity of C. auribarbis infections did not change. This provides good evidence of interspecific competence, which could be dealt with by parasites by means of asynchronous life-cycles and different maturation periods. Weather also affects the dynamics and transmission rates of these parasites. Previous annual rainfalls positively affected the level of infection with oestrids. Yearly autumn rainfalls affected positively P. picta, possibly due to an effect on the pupal stage survival. Infection of Oestridae affected body condition in calves and subadults, suggesting that oestrids could have sublethal effects on Iberian red deer. Future research is needed to investigate the effect of parasites on the dynamics of the Iberian red deer.

Theileria annulata (Ta)-infected leucocytes are able to disseminate in scid mice. The dose of virulent parasites of the Ta-Ode line required to achieve quantifiable dissemination was found to be 2×10 cells given i.p. Dissemination was higher on day 11 post-inoculation than on day 18. The attenuated Ta-Ode cells were found to disseminate very poorly compared to their virulent progenitors, which correlates with a marked reduction in matrix metalloproteinase (MMP) expression. A daily i.p. injection of mice with BB94, a synthetic inhibitor of MMPs, almost completely ablated dissemination compared to controls. This provides strong evidence that metastasis of Theileria annulata macroschizont-infected host cells is mediated by host MMPs induced by the parasite. This has important implications for explaining a number of pathological features of tropical theileriosis in cattle.

Host fertility is often curtailed as a result of parasitic infection. The hypothesis that this may confer an adaptive advantage upon the symbionts if nutrients are directed from reproduction and made available for host/parasite maintenance is explored. The suggestion is made that an understanding of the mechanisms underlying the pathophysiology of fecundity reduction may shed light upon the evolutionary implications of this strategy for both parasite and host. To illustrate this the down-regulation of egg production is explored with reference to a particular model system, the association between metacestodes of the rat tapeworm, Hymenolepis diminuta and the mealworm beetle, Tenebrio molitor. Several aspects of host reproductive behaviour and physiology are affected by infection in this association, including vitellogenesis. Metacestodes directly inhibit the fat body synthesis of vitellogenin in a stage-specific, density-dependent manner. This inhibition is likely to be orchestrated by a modulator molecule, produced by the parasite. In the ovarian follicles, juvenile hormone III binding to a specific follicular membrane-binding protein is inhibited in infected beetles, resulting in the down-regulation of a cascade of events which enables vitellogenin to pass into the developing oocyte. Data to support the proposed existence of a parasite-induced antigonadotrophin, of host origin, are discussed. Evidence that similar mechanisms operate in Plasmodium-iniected anopheline mosquitoes and Onchocerca-infected blackflies is presented in support of the possibility that a parasite-induced reduction in host reproductive fitness is an adaptive strategy and an assessment of who is manipulating whom is made.

To investigate whether the stress response of European eels infected with Anguillicola crassus is influenced by environmental pollutants, experimentally infected eels were exposed to Cd and/or to 3,3′, 4,4′, 5-pentachlorobiphenyl (PCB 126). Serum cortisol and glucose concentrations of these eels were monitored over a period of 103 days and were compared with data from infected, unexposed eels as well as with data from uninfected eels. Additionally, the levels of cortisol were correlated with concentrations of Anguillicola-specific antibodies. All eels showed an initial increase of the cortisol levels until day 63. This general elevation of plasma cortisol is most likely due to handling stress, as all eels were repeatedly netted and afterwards inoculated with a feeding tube. At the end of the exposure period eels which were infected and those which were infected and simultaneously exposed to Cd and PCB showed significantly higher levels than the controls. The general course of serum glucose levels in eels resembled that of cortisol. Accordingly, Spearman correlation analysis revealed that an increase in serum cortisol concentrations is correlated with rising levels of glucose. With respect to immune-endocrine interactions a significant negative correlation between cortisol and anti-A. crassus antibodies was found. Our data show that A. crassus is the most potent stressor for European eels among the treatments tested within this study. This is important in terms of ecotoxicological studies as the main effects are caused by parasites rather than chemicals. Accordingly, effects of parasites on the physiological homeostasis of organisms must be considered in ecotoxicology. From the parasitological point of view our results suggest that probably as part of an unbalanced host-parasite interaction A. crassus evokes a strong cortisol response in A. anguilla, thereby suppressing the immune response which in turn enables the parasite to establish. The parasite-induced stress response in the newly adopted European eel might be one of the factors which contributes to the extremely effective colonizing strategy of A. crassus.

Trypanosomiasis is a major veterinary problem over much of sub-Saharan Africa and is frequently associated with undernutrition. There is growing evidence that nutrition can have a profound effect on the pathophysiological features of animal trypanosomiasis. These features include anaemia, pyrexia, body weight changes, reduced feed intake and diminished productivity including reduced draught work output, milk yield and reproductive capacity. Anaemia is a principal characteristic of trypanosomiasis and the rate at which it develops is influenced by both protein and energy intakes. Pyrexia is associated with increased energy demands for maintenance which is ultimately manifested by reductions in voluntary activity levels and productivity. Weight changes in trypanosomiasis are markedly influenced by the levels of protein intake. High intakes allow infected animals to grow at the same rate as uninfected controls providing energy intake is adequate whilst low energy levels can exacerbate the adverse effects of trypanosomiasis on body weight. Reductions in feed intake are less apparent in animals which are provided with high protein diets and where intake is limited by the disease animals will often exhibit preferential selection of higher quality browse. Further studies are required to evaluate the minimum levels of protein and energy supplementation required to ameliorate the adverse effect of trypanosomiasis, the nature and quality of protein supplement to achieve these benefits and the influence these have on digestive physiology.

A curious aspect of the evolution of the hypnozoite theory of malarial relapse is its transmogrification from theory into ‘fact’, this being of historical, linguistic, scientific and sociological interest. As far as it goes, the hypnozoite explanation for relapse is almost certainly correct. I contend, however, that many of the genotypically homologous, non-reinfection, relapse-like Plasmodium vivax recurrences that researchers ascribe to hypnozoite activation are probably hypnozoite-independent. Indeed, some malariologists are starting to recognize that homologous P. vivax recurrences have most likely been overattributed to activation of hypnozoites. Hitherto identified, non-hypnozoite, possible plasmodial sources of recurrence that must be considered, besides circulating erythrocytic stages, include parasites in splenic dendritic cells, other cells in the spleen (in addition to infected erythrocytes there), bone marrow (importantly) and the skin. I argue that we need to take into account the possibility of a dual or multiple extra-vascular origin of P. vivax non-reinfection recurrences, not arbitrarily discount it. The existence of a P. vivax reservoir(s) is a topical subject and one of practical importance for malaria eradication. Pertinent drug-associated matters are also discussed, as is the dormancy-related significance of clues provided by blood-stage-induced malarial infection.

MicroRNAs (miRNAs) are small, endogenous non-coding RNA molecules that regulate gene expression post-transcriptionally by targeting the 3′ untranslated region (3′ UTR) of messenger RNAs. Since the discovery of the first miRNA in Caenorhabditis elegans, important regulatory roles for miRNAs in many key biological processes including development, cell proliferation, cell differentiation and apoptosis of many organisms have been described. Hundreds of miRNAs have been identified in various multicellular organisms and many are evolutionarily conserved. Schistosomes are multi-cellular eukaryotes with a complex life-cycle that require genes to be expressed and regulated precisely. Recently, miRNAs have been identified in two major schistosome species, Schistosoma japonicum and S. mansoni. These miRNAs are likely to play critical roles in schistosome development and gene regulation. Here, we review recent studies on schistosome miRNAs and discuss the potential roles of miRNAs in schistosome development and gene regulation. We also summarize the current status for targeting miRNAs and the potential of this approach for therapy against schistosomiasis.

An amendment of the family Sinuolineidae (Myxosporea) is proposed in order to include a newly described genus Latyspora n. gen. The type species Latyspora scomberomori n. gen. n. sp. is a coelozoic parasite in the kidney tubules of Scomberomorus guttatus. In addition to the morphological and molecular characterization of L. scomberomori n. gen. n. sp., we also present novel SSU rDNA data on Sphaerospora testicularis, a serious parasite of Dicentrarchus labrax. Performed phylogenetic analyses revealed that both species cluster within the marine urinary clade encompassing the representatives with a shared insertion within their V4 SSU rRNA region and grouping according to the shape of their spores’ sutural line and their similar tissue tropism in the host. Sphaerospora testicularis is the closest relative to Parvicapsula minibicornis within the Parvicapsula subclade and L. scomberomori n. gen. n. sp. is the basal species of the Zschokkella subclade. The phylogenetic position of S. testicularis, outwith the basal Sphaerospora sensu stricto clade, and its morphology suggest it being a non-typical Sphaerospora. The sequence data provided on S. testicularis can help in future revisions of the strongly polyphyletic genus Sphaerospora. We recommend re-sequencing of several sphaerosporids as an essential step before such taxonomic changes are accomplished.