The prophylactic efficacy of autoclaved Leishmania donovani (ALD) and autoclaved L. major (ALM) – a heterologous vaccine developed against cutaneous leishmaniasis (used as a reference vaccine), along with BCG – was evaluated against L. donovani in hamsters (Mesocricetus auratus). Animals were immunized with triple doses (21 days apart) of either ALD or ALM (1·0 mg) with or without BCG (0·1 mg) and challenged 21 days later with 1×106L. donovani amastigotes intracardially. Animals immunized with ALM+BCG and ALD+BCG yielded 94·3% and 86·1% parasite inhibition respectively in comparison to the BCG only and unvaccinated controls. Fifty and 33·3% of the vaccinated animals (ALM+BCG and ALD+BCG respectively) were completely devoid of parasites when tested on day 45 post-challenge (p.c.) and survived till the experiment was terminated. The mean survival of ALM+BCG and ALD+BCG groups (animals harbouring parasites) was longest (168 and 139 days respectively). No significant increase in anti-leishmanial antibody level (ELISA) was noticed in ALD+BCG and ALM+BCG groups whereas it increased progressively in the rest of the experimental groups. The lymphoproliferative responses to PHA and Con A, of the 2 vaccinated groups were comparable to that of normal controls on day 45 p.c. The study suggests that ALD along with BCG can offer substantial protection against visceral leishmaniasis in hamsters.

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 secretory cycle has been revealed in the intestinal epithelial cells of Fasciola hepatica. The basal regions of tall cells contain mitochondria-like bodies which stain deeply with basic dyes. These granules become numerous and are then arranged in vertical arrays of variable height which extend into the middle region of their cells. The distal halves of the epithelial cells contain numerous granules or droplets which have no affinity for basic stains and tend to swell, with consequent disarray of the secretory products. In the terminal parts of the cells vacuole-like formations appear, usually one to each cell, and eventually the lumen of the caecum becomes occluded by a froth-like mass of vacuoles. When food enters the caecum in small amount, the vacuoles are broken down and some secretion is liberated and mixed with the incoming homogenate. Meanwhile the vacuole-like structures encroach more deeply into the cell and eventually the residual secretion is discharged. New cell boundaries are being established near the nuclear zone and after complete discharge of secretion the distal parts of the cells consist of aggregates of long cytoplasmic filaments, which form a kind of fringe on the free borders of the now much shortened cells. These delicate cytoplasmic fringes, are abraded, probably by the physical action of streams of food, and typical short cells in caeca which contain food have much shorter fringes no more than one-half the height of the shallow cells. Ultimately all traces of such residual cytoplasm remaining after completion of the secretory cycle disappear. Short cells containing few granules are presumed to increase in height subsequently by regeneration of cytoplasm and, possibly, nuclear changes also, until they attain the form seen in cells commencing the secretory cycle. Such changes are especially difficult to observe because of variability in the heights of cells normally forming the epithelium. The especially tall cells which are seen, isolated or in small groups, at the height of the secretory cycle—the so-called pyramidal cells—are believed to be merely cells which have had their secretory activities delayed; ultimately, they are denuded in the same way as other cells during the secretory cycle. The type of secretory activity disclosed is therefore of the apocrine type, which is considered appropriate to the suctorial method of feeding employed by the fluke.

1. Gametocytes of Plasmodium gallinaceum emerged from the erythrocytes and the male gametocytes exflagellated normally in vitro, in normal or inactivated horse serum, or in rabbit plasma.

2. Gametocytes, washed and suspended in normal horse serum, completed their sexual development and produced oocysts in mosquitoes.

3. Hyperimmune chick plasma, as compared with normal chick plasma, did not significantly affect the time of onset or the frequency of exflagellation.

4. In a solution, isotonic to bird blood and adjusted to approximately pH 8, containing NaCl, KC1, CaCl2, Na2HPO4, MgSO4, NaHCO3 and Tris buffer, the in vitro development of the gametocytes was normal, and the numbers developing were comparable to those in horse serum, or in plasma or serum from the normal host. The development of the gametocytes was inhibited by the omission of Na or HCO3 ions, but the omission of Mg, K, Ca, SO4 or HPO4 ions had no effect.

5. Further experiments showed that the gametocytes emerged from the corpuscles and the males exflagellated normally in a solution containing only Na, C1 and HCO3 ions. Other combinations of ions were tested, but although gametocytes emerged from the corpuscles in some of these solutions, exflagellation either did not take place or did so infrequently, and the gametes produced were feeble in their movements. Within the range of inorganic salts tested, Na, HCO3 and C1 ions constituted the minimal requirement for the normal in vitro development of the male gametocytes.

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.

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.

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.

Laboratory hamsters (Mesocricetus auratus) were infected with Echinostoma revolutum (Trematoda). Hamsters developed marked and sometimes fatal echinostomiasis. Clinical signs included progressive unthriftiness, watery diarrhoea and weight loss. Packed cell volume, haemoglobin and red blood cell counts increased in those animals with severe diarrhoea. No increase in eosinophils was noted in peripheral blood smears. Examination of tissues of infected animals showed wide individual variations related to the intensity of infection. The histopathological responses of hamsters to the parasite showed erosion of intestinal villi with lymphocytic infiltration being the primary response. Periportal lymphocytic infiltration and focal hepatic necrosis were observed in livers from some infected animals. The findings of this study are discussed in terms of variations in pathogenicity of E. revolutum and as a model for the study of echinostomiasis.

This study describes polymorphism in Theileria annulata, an intracellular protozoan parasite of bovine leucocytes and red blood cells. Fifty-three different stocks of T. annulata, isolated from 17 sites (districts) in Tunisia, have been characterized by anti-parasite monoclonal antibody (MAb) reactivity, glucose phosphate isomerase (GPI) isoenzyme electrophoresis, and Southern blotting with two genomic DNA probes. There appears to be considerable diversity amongst T. annulata stocks from Tunisia, no two isolates being identical, even those from animals on the same farm. Two distinct antigenic populations were detected by MAb 7E7. They were defined by negative and positive cells in the indirect fluorescent antibody test. The percentage of positive cells in different isolates ranged between 0 and 100%. The population variation seen by GPI analysis and DNA probes was greater; 7 different GPI phenotypes were identified amongst the stocks studied, while DNA probes T. annulata Tunis (TaT) 17 and 21 detected up to 5 different variants. The majority of isolates were shown to contain more than one parasite population, the number of variants per isolate ranging from 1 to 4. No correlation between particular parasite phenotypes or genotypes and their geographical site of isolation was observed. Selection of parasite populations in vivo and in vitro is also discussed.

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.

For the diagnosis of imported malaria, optical or immunochromatographic methods are known to be less sensitive and less specific than PCR-based methods, which are conversely more complicated and time-consuming. An original strategy, based upon the sequential use of a multiplex competitive real-time PCR detecting Plasmodium falciparum or Plasmodium spp. infection, followed by, if necessary, a single real-time PCR for species identification, was therefore performed and then tested versus conventional PCR in routine conditions. Conventional PCR has been used since October 1999 in the Department of Parasitology, University Hospitals in Toulouse, as a 2nd line diagnostic method. Out of 183 patients tested, 48 were found to be harbouring a falciparum infection by conventional microscopy, 60 by conventional PCR and 60 by multiplex competitive real-time PCR. Nine further patients had a non-falciparum infection, and concordant species identifications were obtained by both conventional PCR and single real-time PCR. The major value of PCR-based methods, when compared to microscopical techniques, was to ascertain the negativity of a suspect sample. Moreover, real-time PCR allows simplification of the operating procedure, with a diagnosis being made within 2 h.

Cadmium and lead concentrations were compared in tissues of cutlassfish, Trichiurus lepturus L., its intestinal nematode Hysterothylacium sp. type MB larvae, and in water from the same location in the Sea of Oman. Metal accumulation in hosts, parasites and sea water was measured by ICP-OES. Hysterothylacium larvae from the intestinal lumen and visceral cavity showed much higher metal concentrations than in host tissues or sea water. Statistical analyses revealed no significant differences in metal accumulation between infected and uninfected hosts. Cadmium concentration in the host muscle was lower than in intestine, liver and gonad tissues. The mean concentrations of lead and cadmium in nematodes were 289·03 and 81·5 times higher than in host intestine, 188·4 and 225 times higher than in host muscle, 108·6 and 65·3 times higher than in host gonads, 70·5 and 19·5 times higher than in host liver and 3351 and 148 times higher than in sea water. The results show the value of this and possibly related nematodes as bioindicators of heavy metals and their potential use in environmental studies.

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.

Toxoplasma gondii infections are common in humans and animals worldwide. Wild and domestic avian species are important in the epidemiology of T. gondii infections because felids prey on them and excrete millions of oocysts in the environment, disseminating the infection. Herbivorous birds are also excellent sentinels of environmental contamination with T. gondii oocysts because they feed on the ground. Toxoplasma gondii infections in birds of prey reflect infections in intermediate hosts. Humans can become infected by consuming undercooked avian tissues. Here, the authors reviewed prevalence, persistence of infection, clinical disease, epidemiology and genetic diversity of T. gondii strains isolated from turkeys, geese, ducks, ratites and avian species (excluding chickens) worldwide 2009–2020. Genetic diversity of 102 T. gondii DNA samples isolated worldwide is discussed. The role of migratory birds in dissemination of T. gondii infection is discussed.

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.

1. The hatching of N. battus under laboratory conditions was investigated. A high hatch of ripe eggs was obtained on exposure to constant temperatures in the approximate range 8–15° C., and very little hatching took place at temperatures outside this range.

2. These, and other published results can be explained in terms of a hypothesis transferred from the study of the breaking of insect diapause. This hypothesis supposes that two processes underly the completion of hatching—a cold process with low temperature optimum and a warm process with a higher temperature optimum and which can only proceed as the cold process is completed.

3. It is not easy to apply these findings directly to field events, but the attempt to do so leads to the suggestion that the apparent synchrony of hatching and the presence of susceptible lamb hosts in the north-east of the British Isles may be partly fortuitous, and that in slightly different climatic circumstances many eggs might hatch in the autumn or during the winter.

I wish to thank Mr A. V. Cunningham of Threepwood, Galashiels, and Mr J. M. M. Cunningham of the Edinburgh School of Agriculture for their generous help in making sampling of their flocks possible, and Dr J. T. Stamp for advice on the presentation of this paper.

A panel of immunologically and biochemically defined monoclonal antibody probes has been used in conjunction with immunocytochemical techniques to localize target antigens in sections of different life-cycle stages of Trichinella spiralis. Monoclonals that immunoprecipitate surface components from adult worms, show reactivity with the surface but not with internal tissues of sectioned parasites. Reagents that immunoprecipitate radio-isotope labelled stage-specific surface components of muscle-stage larvae, however, react with the stichosome and gut lining of sectioned larvae, as well as with the surface. Monoclonal antibody probes that do not stain the surfaces of live, intact muscle larvae in immunofluorescence assays, but which immunoprecipitate solubilized surface glycoproteins, also show reactivity with cuticular and stichosomal antigens of sectioned larvae. The more powerful resolution provided by electron microscopy has localized the surface antigens to the epicuticle and the intestinal antigens to the brush-border microvilli. Of particular interest was the finding that antigens of muscle-stage larvae, known to confer protection upon recipient mice, also exist in the stichosome of adult parasites. This observation may shed some light on the fact that mice immunized with antigens from muscle-stage larvae show, in addition to reduced muscle larva burden, accelerated expulsion of adult worms. The implications of these data for stage specificity of immune responses to trichinosis are discussed.

The distribution of exposed and hidden lectin-binding glycoconjugates in adult Schistosoma mansoni worms was studied. Fluorochrome-conjugated lectins were allowed to react both with intact worms and with frozen sections of worms. The results show a selective reactivity of lectins with tegumental surface structures, the intestine, eggs and basement membranes. Strong binding of wheat germ agglutinin (WGA) was seen to the exposed surface of intact worms. Peanut agglutinin (PNA), Ricinus communis agglutinin (RCA) and concanavalin A (Con A) gave a weaker reaction. There was a preferential affinity of PNA and RCA to the spikes covering the tubercles of male worms whereas Con A gave a more uniform staining pattern. Internal structures exposed in frozen sections of worms gave distinct staining reactions. Binding sites for Con A were widely distributed throughout the worm. PNA produced a more selective binding and was the only lectin staining basement membranes as distinct structures. There was a strong selective staining of the gut with soybean agglutinin (SBA) and RCA. Eggs seen in large quantities inside female parasites reacted strongly with SBA and PNA and less intensely with RCA and WGA. The observed selectivity of lectin binding may reflect physiological function. The results may be important as a basis for using lectin affinity-chromatography for purification and characterization of worm constituents

The cell-mediated resistance of human leucocytes to Trypanosoma dionisii, a bat parasite related to T. cruzi, was investigated. Human peripheral blood neutrophils and monocytes were cytotoxic to T. dionisii as assessed by electron microscopy and by induction of 99mTc release from trypanosomes pre-labelled with [99mTc]pertechnetate. The enhancement of cytotoxicity by specific antiserum varied considerably from one individual to another. Neither blood lymphocytes nor blood eosinophils induced 99mTc release from T. dionisii. The trypanosomes were readily phagocytosed by neutrophils and monocytes even in the absence of added antiserum but the rate was enhanced when antiserum was present. Eosinophils also phagocytosed T. dionisii but only in the presence of antiserum. Investigation by electron microscopy revealed that T. dionisii is rapidly destroyed in the phagocytic vacuole of neutrophils and monocytes and by eosinophils. Phagocytosis, ultrastructural damage and induction of 99mTc release occurred more rapidly in neutrophils than in monocytes.

The present study determined the prevalence, hematological findings and genetic diversity of Bartonella spp. in domestic cats from Valdivia, Southern Chile. A complete blood count and nuoG gene real-time quantitative PCR (qPCR) for Bartonella spp. were performed in 370 blood samples from cats in Valdivia, Southern Chile. nuoG qPCR-positive samples were submitted to conventional PCR for the gltA gene and sequencing for species differentiation and phylogenetic analysis. Alignment of gltA gene was used to calculate the nucleotide diversity, polymorphic level, number of variable sites and average number of nucleotide differences. Bartonella DNA prevalence in cats was 18·1% (67/370). Twenty-nine samples were sequenced with 62·0% (18/29) identified as Bartonella henselae, 34·4% (10/29) as Bartonella clarridgeiae, and 3·4% (1/29) as Bartonella koehlerae. Bartonella-positive cats had low DNA bacterial loads and their hematological parameters varied minimally. Each Bartonella species from Chile clustered together and with other Bartonella spp. described in cats worldwide. Bartonella henselae and B. clarridgeiae showed a low number of variable sites, haplotypes and nucleotide diversity. Bartonella clarridgeiae and B. koehlerae are reported for the first time in cats from Chile and South America, respectively.