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.

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.

Recombinant ribosomal DNA sequences were amplified by PCR and used as probes to perform a fingerprint analysis of total DNA from different Entamoeba histolytica isolates. RFLPs obtained with one of the probes, R-1, support previous proposals that pathogenic and non-pathogenic E. histolytica are closely related, yet genotypically distinct. Another probe, R-2, while not distinguishing between the two forms of E. hystolytica, was able to differentiate between them and E. moshkovskii, which has morphologically identical cysts and trophozoites. A third probe, BR-1, identified strain-specific RFLPs.

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.

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.

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.

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.

The integrity of the hepatic portal vasculature was examined, relative to the resistance to Schistosoma mansoni observed in 68°0 of 129/Ola mice. The passage of microspheres to the lungs, following their injection via the superior mesenteric vein, indicated the presence of shunts in the majority of both naive and infected mice. There was a negative association between shunting of microspheres to the lungs and paucity of liver worms at 28/35 days post-infection. Schistosomula accumulated in the livers of resistant mice at a slower rate than in susceptible animals, and after day 21 relocated to the lungs. Many lung schistosomula injected via the superior mesenteric passed immediately to the lungs; the shunts thus greatly reduce the probability of trapping in the liver. Some parasites migrated back from the lungs, successfully lodged in the liver and began to feed on blood. Latex infusion demonstrated the location of large intrahepatic connections between the portal and hepatic veins. We suggest that as these liver worms grow, migrating upstream into progressively larger vessels, they reach the connections, pass out of the hepatic portal system, and relocate to the lungs. The presence of the natural shunts thus accounts for the resistant status of the mice.

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.

Scanning and transmission electron microscopy of Trypanosoma congolense epimastigotes attached to a plastic substratum shows them to elaborate a complex flagellum filament system and plaque with a highly organized structure. Non-ionic detergent extraction of these cells shows that the resulting cytoskeletons remain attached to the plaque. The subpellicular corset of microtubules can be removed by salt or Ca+2 treatment leaving the axoneme, paraflagellar rod, associated filaments and the plaque. Neither of these treatments therefore removed the plaque-associated material from the substratum. Analysis of these fractions by SDS–polyacrylamide gel electrophoresis reveals an abundant 70 kDa protein that is highly enriched in the salt extracted ‘minimal plaque’ structures and appears likely to be a major constituent of this structure. These studies reveal that the complex filament and microtubule systems of the cytoskeleton involved the attachment of trypanosomes to substrata and have established a method of biochemical fractionation of the structures and components involved.

One of the most common causes of calf diarrhoea is the parasite Cryptosporidium parvum. Two longitudinal studies were carried out on a dairy farm Scotland to determine the prevalence of Cryptosporidium species and subtypes in a group of calves and to determine whether dams were a possible source of calfhood infection. Fecal samples were collected from 25 calves from birth to 12 months in the first year. In the second year, fecal samples were collected from pregnant cows (n = 29) and their calves (n = 30) from birth to 6 months. The samples were tested for Cryptosporidium and speciated. Cryptosporidium parvum-positive samples were subtyped by GP60 fragment analysis. All calves in both studies shed Cryptosporidium during the study period. Cryptosporidium parvum was the predominant species detected in calves ⩽6 weeks of age and at 6 months of age, C. bovis and C. ryanae were detected in calves older than 4 weeks of age but ⩽6 months of age. The prevalence of Cryptosporidium was higher in younger animals than in older animals. GP60 subtyping revealed two subtypes in calves on this farm (IIaA15G2R1 and IIaA19G2R1) that differed in frequency by age. Adult cattle also shed C. parvum, of four gp60 genotypes.

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.

A quick and simple method of purifying the histones from Plasmodium falciparum culture supernatant or from infected erythrocytes is described. The proteins were present only in preparations rich in P. falciparum nuclear material and were soluble at acid pH and in strongly anionic detergents. Four proteins, of 14–18 kDa were identified as the P. falciparum core histones. N-terminal sequence analysis of the 16 and 18 kDa proteins and of a tryptic fragment of the protein mixture revealed strong homologies with deduced cDNA or protein sequences of histones H2A, H2B, and H3 of P. falciparum and other species. Antibodies raised against the proteins cross-reacted weakly with histones of other species and, on immunofluorescence, localized the proteins to schizont nuclei. Anti-P. falciparum histone antibodies were detected in sera of semi-immune human adults but these antibodies did not react with human histones on a Western blot. The large quantities of P. falciparum histone released and the chronic nature of malarial infection, together with the unusually high avidity of histones for ligands found in renal and vascular basement membrane, raise the question of a role for histones in the pathogenesis of malarial infection. Wesuggest that histones or histone±antibody complexes may contribute to disease pathology.

We present a discrete time model for age-related changes in the mean and variance of the number of helminth parasites per host. We assess the degree of aggregation as the negative binomial parameter, k, and use the model to examine the influence of various factors on changes in aggregation with host age: discrete versus continuous infection; the degree of predisposition to infection; infection rate; parasite survival rate; and the variance in exposure to infective stages. The model can produce both increases and decreases in k with host age. However, with parameter values typical of many human helminth infections, a monotonic increase in k (decrease in aggregation) with age is predicted. With an age-dependent infection rate, convex relationships between k and age are possible. These predictions are consistent with data from field studies, but differ from those of previous models which have suggested that k is independent of host age in the absence of density dependence in parasite population dynamics. Differences between the models, and some difficulties in the interpretation of field data, are discussed.

Trypanosomes were studied from the blood of 6 species of small British fish caught in the River Lee. Morphologically the trypanosomes from the blood of Nemacheilus barbatulus L., Phoxinus phoxinus L., Cottus gobio L., Gobio gobio L., Gasterosteus aculeatus and Pungitus pungitus L. were indistinguishable. Cross-transmission experiments using syringe passage of culture forms and also the leech vector Hemiclepsis marginata showed that the trypanosomes were not host specific. The isoenzyme patterns of culture forms from N. barbatulus and P. phoxinus were identical for 11 enzymes studied. The trypanosomes from the 6 species of fish previously classified as separate species are, on the basis of these results, regarded as a single species, Trypanosoma cobitis (Mitrophanow, 1883).

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.

Artificially transformed schistosomula of Schistosoma mansoni develop a consistent but small protection against the lethal effects of antibody plus complement when cultured for 24 h in a chemically defined medium. In contrast, they become rapidly resistant to antibody plus complement, when cultured in the presence of a complex medium consisting of equal parts of heat-inactivated rabbit serum and Earle's/lactalbumin or in defined medium supplemented with small amounts of heat-inactivated rabbit serum. Sephadex G-200 gel filtration revealed that the protective factor in rabbit serum is a macromolecule with a molecular weight between 7 and 19S. Parasites cultured at 10 °C or in the presence of 200 μg of puromycin show less serum-induced protection against the lethal effects of antibody plus complement than do controls.

New biological species and high levels of inter- and intraspecific genetic divergence were discovered in an allozyme study of some North European members of the acanthocephalan genus Echinorhynchus (sensu lato), parasites of fish and malacostracan crustaceans. (i) A strong differentiation between the marine E. gadi and the fresh- and brackish-water E. salmonis (genetic identity I ≃ 0) supports a generic distinction between these taxa; however, the subdivision would not entirely concur with the concepts of Echinorhynchus (sensu stricto) and Metechinorhynchus suggested earlier. (ii) Samples of E. gadi from the Baltic, Norwegian and North Seas included three distinct, partially sympatric biological species (spp. I–III; I ≃ 0·5). (iii) E. bothniensis, previously only known from the northern Baltic Sea, represents a complex of freshwater taxa with an intermediate host relationship to the ‘glacial relict’ Mysis spp. and with a distributional and host analogy to the North American E. leidyi. A population in a northern lake in the Barents Sea basin is closely related to E. bothniensis of the Baltic area, but is probably specifically distinct; the divergence between these populations (I ≃ 0·6) is similar to that between their Mysis host species. (iv) Considerable intraspecific differentiation (Fst = 0·25), probably reflecting post-glacial population bottlenecks, was found between Baltic and nearby lacustrine E. bothniensis, and between Atlantic and Baltic E. gadi sp. I.

The understanding of the mechanisms whereby intracellular parasites counteract the microbicidal processes of macrophages has progressed considerably in recent years. Various factors contribute to intracellular parasite destruction; from a biochemical standpoint, particularly important is the oxidative burst triggered by phagocytosis and by macrophage ‘activation’, that leads to the generation of toxic metabolites of oxygen. At the ultrastructural level, fusion of the parasitophorous vacuole with surrounding lysosomes appears to be a pre-requisite for the final digestion and elimination of the infecting microorganisms. The counter-measures evolved by microorganisms to escape intracellular destruction are best illustrated by studies in vitro on the interaction of parasites of the Leishmania, Toxoplasma and Trypanosoma spp. with mononuclear phagocytes. Some microbes are able to inhibit the fusion of phagosomes with lysosomes, thus avoiding the potentially harmful action of lysosomal hydrolases. Other microorganisms are able to resist the effects of such enzymes, perhaps by secreting inhibitory substances. Others still avoid lysosomes by leaving the phagocytic vacuole, to reach the cytoplasmic matrix where their development is unhindered. Particularly critical is the capacity of certain parasites to subvert the lethal effects of the oxidative burst. This can be achieved either by failing to evoke this metabolic response, or by producing scavengers that can detoxify harmful oxygen metabolites. Intracellular death or survival will thus depend on a delicate balance between the potency of macrophage cidal mechanisms, and the efficacy of the protective measures evolved by the infecting agents.

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.