The anticoccidial effect of narasin against Eimeria flavescens, E. intestinalis, E. magna, E. perforans and E. stiedai was tested in experimentally infected rabbits. The drug was highly effective in reducing oocyst output and in preventing clinical signs. Maximum activity as measured by oocyst discharge was achieved with feed levels from 12 to 24 p.p.m.; there was no influence on oocyst sporulation. Optimal weight gain and food consumption were obtained with levels between 8 and 12 p.p.m.; at higher levels there was an adverse effect on growth, food intake, relative liver and heart weights and on serum potassium and glucose.

The genus Myriocladus Okada is considered to be a synonym of the genus Dendrogaster Knipowitsch.

The female and male of a new species, D. tasmaniensis sp.nov. from the sea-star Allostichaster polyplax (Müller and Troschel), are described.

Appendages interpreted as antennae are present in the female, male and ascothoracid larva of the new species.

A brief account is given of the manner in which the larva of D. tasmaniensis sp.nov. enters and escapes from the host.

The development of granulomatous reactions against moulting nymphal pentastomids (Porocephalus crotali) in the tissues of rat and mouse intermediate hosts is described. Adipose tissue and lungs are favoured sites for encystment accounting for 70% of larvae. Six moults separate the primary larva from the final infective stage which first appears about 80 days post-infection (p.i.) and is fully infective by day 120. Larvae, and particularly their cast cuticles, are the foci of granulomatous reactions characterized by an intense eosinophilia. During ecdysis, large numbers of eosinophils permeate the entire lesion but, significantly, degranulation is limited to the underside of cast cuticles where the resultant debris is endocytosed by macrophage/epithelioid cells. A pronounced asymmetry in the granulomatous lesion, evident even in the earliest cysts, results from the accumulation of individual epithelioid granulomas associated with cuticle fragments close to the ventral side of the developing parasite; each is circumscribed by fibrosis. External to this region are extensive tracts of tissue composed of mature plasma cells. Particularly in rats, large numbers of partially degranulated mast cells ( = globule leucocytes) also surround cuticle granulomas, and mast cell granules can accumulate within macrophages and fibroblasts. Inflammation slowly subsides once the infective stage is attained. This 1 cm-long larva resides in a thin, fibrotic, C-shaped cyst and can remain viable for years: uniquely this instar retains its last moulted cuticle as a protective sheath. Nymphal instars II-VI feed predominantly upon eosinophils but we do not yet know whether this requirement is obligate.

Thymidylate synthase, dihydrofolate reductase and dUTPase specific activities were found to remain at a high and constant level in crude extracts from adult worms of Trichinella spiralis, as well as from muscle larvae of both Trichinella spiralis (isolated 1–24 months after infection) and Trichinella pseudospiralis (isolated 5·5–13 months after infection). The results obtained with Trichinella pseudospiralis muscle larvae isolated with the use of pepsin did not differ from those obtained when pepsin was not used. No thymidine kinase activity could be detected in muscle larvae of either species and thymidine phosphorylase could be found only in T. pseudospiralis larvae isolated without the use of pepsin. Muscle larvae of both species contained orotidylate phosphoribosyl transferase activity, pointing to a possibility of 5-fluorouracil activation. Uridine phosphorylase, another enzyme involved in 5-fluorouracil anabolism, was also present in T. pseudospiralis muscle larvae. Results of comparative studies on inhibition of purified T. spiralis and rat thymidylate synthases by substrate (4-thio-5-fluoro-dUMP, 2-thio-5-fluoro-dCMP and N4-hydroxy-dCMP) and cofactor (ZD 9331) analogues indicated only dUMP analogues to show feeble selectivity towards the parasite enzyme. A hypothesis is discussed, assuming high expression of thymidylate synthase in muscle larvae to be connected with their cells being arrested in the cell cycle.

A technique is described of the use of two infections in the rat which respond to the anthelmintics acting on a very wide range of gastro-intestinal nematodes. It can be used as a routine screening test for anthelmintics or for the study of variations in the degree of activity caused by alterations in formulation or presentation.

Of the fifteen valid species of mammalian blood-flukes so far known, six have been originally described from India. Montgomery (1906) described Schistosoma indicum, S. spindalis and Ornithobilharzia bomfordi (Montgomery, 1906) Price, 1929. Datta (1932) found that nasal granuloma of cattle is due to a new schistosome, which was subsequently obtained and described as Schistosoma nasalis by Rao (1933). Rao & Ayyar (1933) obtained a new schistosome from pigs and described it as S. suis. Mudaliar & Ramanujachari (1945) described S. nairi n.sp., from an elephant, which was transferred to the genus Ornithobilharzia by Bhalerao (1947). Another new species of blood-fluke has now been found in Indian buffaloes and cattle. While studying the larval trematode fauna of the local aquatic snails, large numbers of a new schistosome cercaria were obtained from nine out of three hundred specimens of Limnaea luteola examined during November 1950. When the cercariae were administered to two guinea-pigs, a buffalo-calf and a bull-calf, they developed into an interesting blood-fluke referable to the genus Ornithobilharzia Odhner, 1912. Subsequently, three out of thirteen buffaloes examined post-mortem were found to be naturally infected with the same parasite. The morphology of the adult and of its cercaria is briefly described in this paper.

While human schistosomiasis is hardly a problem in India, schistosomiasis in domestic animals is a problem of considerable importance. Blood-flukes infect almost all the species of Indian domestic animals, and often seriously undermine their health and usefulness. Seven species of schistosomes are known to occur in Indian mammals, viz. Schistosoma indicum Montgomery (1906a), S. spindalis Montgomery (1906b), S. incognitum Chandler (1926), S. nasalis Rao (1933), Bivitellobilharzia nairi (Mudaliar & Ramanujachari, 1945) Dutt & Srivastava (1955), Orientobilharzia dattai (Dutt & Srivastava, 1952) Dutt & Srivastava (1955), and Orientobilharzia bomfordi (Montgomery, 1906b) Dutt & Srivastava (1955). The reported occurrence of Schistosoma bovis (Sonsino, 1876) Blanchard (1895) in India needs confirmation.

Primary infections of Heligmosomoides polygyrus were studied in adult male CD1 mice fed on isoenergetic synthetic diets containing either 2% or 20% protein (casein). A slight reduction in food intake was observed during infection, and this was unaffected by diet. Protein deficiency was also found to have no effect upon worm establishment. Evidence was found, though, to suggest that the worms have a more severe effect upon the host in protein-deficient mice. Hypoalbuminaemia was observed due to diet and also infection, and this was synergistic. There was an increase in non-albumin plasma protein during infection which was not effected by diet. The circulating urea concentration was reduced during protein deficiency, as was the level of essential amino acids, but non-essential amino acid levels were raised. An increase in the levels of urea and essential amino acids was observed at day 4 of infection in mice on the 20% diet, but not on the 2% diet. Splenic atrophy occurred during infection, but the splanomegaly that occurred early during infection was proportionally greater in protein-deficient mice. These results are discussed in terms of the pathophysiology of both protein deficiency and infection, and comparisons are made with human malnutrition syndromes.

The parasitic female of Strongyloides venezuelensis keeps invading the epithelial layer of the host intestinal mucosa. Upon invasion, it adheres to the surface of the intestinal epithelial cells with adhesion molecules secreted from the mouth. It has been demonstrated that S. venezuelensis are expelled from the intestine because mucosal mast cells inhibit the attachment of adult worms to the mucosal surface. In the present study, we generated specific antibodies against secreted adhesion molecules to investigate their function in vivo, because these molecules have been demonstrated only in vitro in spite of the importance in the infection processes. A mouse monoclonal antibody specific to S. venezuelensis adhesion molecules inhibited the attachment of adult worms to plastic dishes and the binding of adhesion molecules to rat intestinal epithelial cells. Immunohistochemical study revealed that adhesion molecules were produced by oesophageal glands and were continuously secreted in vivo to line the wall of the tunnels formed by adult worms in the intestinal mucosa. Our findings indicate that adhesion molecules play essential roles in the infection processes of S. venezuelensis in the host intestine.

Trypanosoma rangeli is a protozoan parasite, which does not cause disease in humans, although it can produce different levels of pathogenicity to triatomines, their invertebrate hosts. We tested whether infection imposed a temperature-dependent cost on triatomine fitness using T. rangeli with different life histories. Parasites cultured only in liver infusion tryptose medium (cultured) and parasites exposed to cyclical passages through mice and triatomines (passaged) were used. We held infected insects at four temperatures between 21 and 30 °C and measured T. rangeli growth in vitro at the same temperatures in parallel. Overall, T. rangeli infection induced negative effects on insect fitness. In the case of cultured infection, parasite effects were temperature-dependent. Intermoult period, mortality rates and ecdysis success were affected in those insects exposed to lower temperatures (21 and 24 °C). For passaged-infected insects, the effects were independent of temperature, intermoult period being prolonged in all infected groups. Trypanosoma rangeli seem to be less tolerant to higher temperatures since cultured-infected insects showed a reduction in the infection rates and passaged-infected insects decreased the salivary gland infection rates in those insects submitted to 30 °C. In vitro growth of T. rangeli was consistent with these results.

The effect of the protostrongylid nematode, Elaphostrongylus rangiferi, on the fecundity and survival of an intermediate snail host, Arianta arbustorum, was studied. Over an experimental period of 12 weeks, infected snails had a significantly lower egg production than uninfected control snails. No significant difference in survival was found between infected and uninfected adult snails. Infected juvenile snails showed a consistent decrease in survival with increasing mean density of parasites. A linear relationship was found between snail instantaneous death rate over a 28–day interval and the mean number of parasites/snail. When calculated for 4–day intervals, the instantaneous death rate was highest between 8 and 12 days post–infection. It is concluded that the most pathogenic phase of this parasite is during the first moult.

Two autogenic (Triaenophorus crassus and T. nodulosus) and four allogenic (Diphyllobothrium latum, D. dendriticum, D. ditremum and Schistocephalus solidus) larval cestode species were found in 13 out of 31 fish species studied from the Bothnian Bay, NE Baltic. Gasterosteus aculeatus was the most heavily infected fish with 4 larval cestode species; for two of them (D. ditremum and S. solidus) the three-spined stickleback was found to be the required fish intermediate host. Among allogenic cestode species, those restricted to different definitive host species segregated their larval population in relation to the fish host, while, for example, D. ditremum and S. solidus, both maturing in fish-eating birds, had the highest percentage of co-occurrences. D. dendriticum, which had the widest range of definitive hosts, was found in the greatest number (8) of fish species and co-occurred with all other species found except T. crassus. The two autogenic species totally segregated their larval population from each other although they both require pike as definitive host. The ecological and evolutionary relationships behind the patterns found for larval cestodes are discussed.

The behaviour of Eimeria acervulina in relation to macrophage activity in chickens was investigated by in vitro and in vivo techniques, with special consideration of the excystation of sporozoites and the fate of intact oocysts and sporocysts inoculated parenterally.In vitro: macrophages obtained from peritoneal washings phagocytosed mechanically released sporocysts or previously excysted sporozoites after 30 minutes incubation.In vivo: phagocytic activity of macrophages against intact oocysts and sporocysts in diffusion chambers implanted subcutaneously and intraperitoneally was observed as early as 4 h after implantation. Changes in the oocyst wall (corrugation, thinning, indentation and increased. fragility) as well as activation and excystation of sporozoites in situ was observed 24 h after implantation. Interaction in vivo between the invasive sporozoite and the phagocytic cell was recorded. Disintegration of the parasite was noticed 72 h after implantation.

The larval stages of the celery fly, Acidia heraclei, have been described, and it is shown that this larva agrees with other biontophagous dipterous larvae in having the pharynx devoid of ventral ridges. The transparency of the larvae permits the internal anatomy to be seen clearly in the living larva, and by this means the structure of the perispiracular glands is clearly revealed.

The braconid Adelura apii occurs as a parasite of Acidia heraclei larvae, and its first. stage larva is described in detail. This larva is densely hairy, has a long, curved, hairy, tail-like appendage and, by the more rapid growth of the ventral surface, it develops a dorsal curvature which obscures the true orientation so that the true dorsal surface appears externally to be ventral. In these respects the first stage larva of Adelura apii resembles that of A. gahani described by de la Baume-Pluvinel. The later larval stages of A. apii, of which there are at least two, are naked, lack the tail-like appendage and do not differ from the normal type of parasitic hymenopterous larvae.

A yeast-like fungus occurs as a parasite in the blood of Acidia heraclei larvae. It is always found associated with existing or abortive infection of the larvae with Adelura apii. Dense mycelial masses sometimes occur in the gut of A. apii pupae and are probably derived from the yeast cells parasitic in the host larvae. It is suggested that this is a unique case of a fungus parasitic in a host larva (Acidia heraclei) undergoing part of its development in a parasitic braconid (Adelura apii), adult females of which transmit the fungus to the host larva during oviposition.

In the present paper, the phylogeographies of two monogenean species, Pseudokuhnia minor and Kuhnia scombri, on the same species of host, Scomber japonicus, were studied. Fragments of the mitochondrial cytochrome c oxidase subunit 1 gene were sequenced for 264 individuals of P. minor and 224 individuals of K. scombri collected from 10 localities along the coast of China. Genetic diversity of K. scombri was higher than that of P. minor, which may imply that P. minor has a lower evolution rate and/or is a younger species. The neighbour-joining (NJ) trees of both parasites were comprised of two clades without association to sample sites, which is the signature of remixing populations following past division. Analyses of molecular variance and pairwise fixation index revealed different genetic structures for the populations of these two closely related species along the coast of China: P. minor without significant genetic structure, while K. scombri has some genetic differentiation. Both neutrality tests and mismatch distribution suggested that the populations of these two species of parasites experienced population expansion in the late Pleistocene era due to the glacial–interglacial cycles induced by climatic oscillations.

A strain of the cattle piroplasm Babesia divergens isolated from a fatal human infection was propagated in the Mongolian gerbil through 150 semi-continuous intraperitoneal passages. The infection was normally fatal; death, accompanied by profuse haemoglobinuria and debilitation, occurred as early as 44 h after intraperitoneal inoculation of heavily parasitized blood with precipitous drops in red blood cell and platelet counts. The average maximum parasitaemia achieved increased on continuous passage reaching 80% by the 150th stage. Twenty-four hours after infection erythrophagocytosis and splenic congestion were apparent by light and electron-microscopical examination and by 48 h hepatic necrosis, renal tubular damage with haemoglobin cast accumulation and ischaemic necrosis of ileal mucosa had developed. Gerbils were highly susceptible to small numbers of parasites when the inoculum was either fresh parasitized blood in high dilution or erythrocytes concentrated from animals showing minimal parasitaemia. Animals inoculated with parasites preserved in dimethyl sulphoxide at low temperatures usually developed fatal infections. However, occasionally animals suffered at most a low grade parasitaemia subsequent to recovery with parasite elimination. These animals were immune to further challenge, and no chronic infections developed. A. field strain of B. divergens isolated locally from a case of bovine redwater behaved similarly to the human strain on continuous passage in gerbils.

Two-dimensional polyacrylamide gel electrophoresis was used to examine the differences in total protein composition between two motile stages and two sedentary stages of the potato cyst-nematode, Globodera rostochiensis. Using a sensitive silver stain, 542 reproducible protein spots were distinguished. A list of these spots is presented, showing their apparent molecular weights, estimated isoelectric points, and occurrences in the different developmental stages. When the protein patterns were compared, 401 spots were found to change their presence or size in one or more of the four developmental stages. It is therefore estimated that during the post-embryonic development of G. rostochiensis, 74% of the polypeptides undergo modulation of their synthesis, or are affected by protein degradation or modification. In the motile stages several abundant proteins were present, which disappeared or decreased in concentration in the sedentary stages. Some of these proteins are presumably muscle proteins, and their modulation may illustrate the degeneration of body-wall musculature in the sedentary stages. It is concluded that the potato cyst-nematode has a very dynamic protein metabolism.

Trypanosomatids represent the causative agents of major diseases in humans, livestock and plants, with inevitable suffering and economic hardship as a result. They are also evolutionarily highly divergent organisms, and the many unique aspects of trypanosome biology provide opportunities in terms of identification of drug targets, the challenge of exploiting these putative targets and, at the same time, significant scope for exploration of novel and divergent cell biology. We can estimate from genome sequences that the degree of divergence of trypanosomes from animals and fungi is extreme, with perhaps one third to one half of predicted trypanosome proteins having no known function based on homology or recognizable protein domains/architecture. Two highly important aspects of trypanosome biology are the flagellar pocket and the nuclear envelope, where in silico analysis clearly suggests great potential divergence in the proteome. The flagellar pocket is the sole site of endo- and exocytosis in trypanosomes and plays important roles in immune evasion via variant surface glycoprotein (VSG) trafficking and providing a location for sequestration of various invariant receptors. The trypanosome nuclear envelope has been largely unexplored but, by analogy with higher eukaryotes, roles in the regulation of chromatin and most significantly, in controlling VSG gene expression are expected. Here we discuss recent successful proteomics-based approaches towards characterization of the nuclear envelope and the endocytic apparatus, the identification of conserved and novel trypanosomatid-specific features, and the implications of these findings.

A genomic library of Entamoeba histolytica (pathogenic strain HM-1: IMSS) was screened to detect repetitive DNA clones other than those from the highly abundant ribosomal DNA (rDNA). One such clone (HMc) had a 2·3 kb insert which hybridized with the main genome and not the rDNA circle. Southern hybridization of E. histolytica genomic DNA, digested with EcoR I and probed with HMc, showed multiple bands. The banding pattern was identical in all axenic pathogenic strains tested. Differences, however, existed when the banding pattern of a pathogenic strain was compared with that of a non-pathogenic strain. HMc was present in about 25–30 copies per genome in strain HM-1: IMSS. Nucleotide sequence analysis of HMc revealed a partial open reading frame which hybridized with a 1·35 kb poly A+ transcript in Northern blots. The deduced amino acid sequence did not, however, show significant homology with known proteins. The HMc sequence was found only in E. histolytica as it hybridized with 5 different axenic strains of E. histolytica but did not recognize other closely related species of Entamoeba. It has thus the potential to be used as a species-specific DNA probe.

The surface coat of the infective larvae of the parasitic nematode Trichinella spiralis was characterized with respect to its biophysical properties, morphology and composition. Labelling of larvae with the fluorescent surface probe PKH26 was lost after activation (by incubation in mammalian medium containing trypsin and bile), or following pronase treatment. Electron microscopical examination revealed that pronase treatment resulted in the loss of an amorphous surface layer only, further demonstrating the specificity of PKH26 for the larval surface coat. Surface coat shedding was inhibited by sodium azide and carbonyl cyanide, or by incubation of larvae at 4°C, suggesting the shedding process required metabolic energy. Pre-labelled, unactivated larvae demonstrated continuous slow surface coat shedding and could be re-labelled with PKH26, indicating that the shed coat is replaced in these parasites. However, pre-labelled larvae which were activated failed to re-label with the probe, suggesting that activation provides an irreversible trigger for surface changes. PKH26, therefore, is a useful marker for larval activation. Examination of the shed coat material by scanning electron microscopy revealed 2 types of morphologies; one comprising thin multilaminate sheets and the other of amorphous material with ridges producing a fingerprint-like motif. Western- and lectin-blotting of the shed coat material demonstrated 2 prominent entities; a 90 kDa glycoprotein, which bound Datura stramonium agglutinin and was resistant to N- and O-glycanase treatment and a 47–60 kDa set of protein(s). Analysis of the surface lipids by electrospray mass spectometry revealed the presence of lysophosphatidic acid (lysoPA, C14[ratio ]2) and an unidentifiable component of 339·4 Da. These two lipids constituted 36·9% and 36% by mass of surface coat lipids respectively. The presence of lysoPA was confirmed by thin layer chromatography, which also detected phosphatidic acid (PA). The polar lipids detected in solvent rinses of intact parasites by electrospray mass spectrometry were PI (C48[ratio ]4), PE (C40[ratio ]4 and C38[ratio ]4), PS (C40[ratio ]4), lysoPC (C20[ratio ]2 and C18[ratio ]2) and lysoPA (C14[ratio ]2). These observations are discussed with respect to the role of the surface coat and its shedding in the T. spiralis host–parasite relationship.