The male intromittent apparatus of Ergenstrema labrosi and Ligophorus angustus, monogenean parasites of the thick-lipped grey mullet Chelon labrosus, consists of a slender flexible penis tube with an expanded proximal region and whose distal end passes freely through a canal in an accessory sclerite; the tube is surrounded by a sleeve of longitudinal muscle and the apparatus works in a manner generally similar to an engineer's Bowden cable. There is a substantial difference in the form of the accessory sclerite in the two species. In both parasites there is a slender sclerotized vagina which, in L. angustus, is short and straight or curved but in E. labrosi is about 7 times longer and forms a spiral. The special role of the sclerotization of the distal part of the vagina may be to resist perforation of the wall by the exceedingly fine penis tube and the consequent discharge of sperms into the parenchyma.

Isospora rastegaievae undergoes its asexual development in the intestinal tract of hedgehogs by endodyogeny. Using light- and electron-microscopy it was demonstrated that daughter organisms were formed by typical endodyogeny as described for different coccidia.

Feulgen-pararosaniline(SO2) staining was performed on stages in the life-cycle of Isospora (Toxoplasma) gondii, Eimeria tenella, Sarcocystis cruzi and Plasmodium berghei. The fluorescence emission of the stained DNA in nuclei of these stages was examined and compared with absorption microscopy measurements at 560 nm (green light) of the same specimens. Accurate identification of single cells, and especially discrimination between young schizonts and young gamonts was difficult after Feulgen staining. To overcome this problem preparations were first stained with Giemsa and the cells of interest precisely located with the aid of an England finder. The same preparations were then hydrolysed and stained with Feulgen—pararosaniline(SO2), after which the previously identified cells were investigated. The DNA distribution after Feulgen staining corresponded with the shape of nuclei after Giemsa staining. DNA was present in all investigated stages of the four parasites, including macrogamonts of I. (T.) gondii and E. tenella and peripheral blood gamonts of P. berghei. Macrogamonts could be recognized, even at a stage at which they can hardly be differentiated from young schizonts in Giemsa-stained preparations, by their typical distribution pattern of Feulgen fluorescence. Feulgen fluorescence was more granular and confined to the peripheral region of the nucleus, leaving a distinct nucleolus unstained. The horseshoe-shaped nuclei typical of macrogamonts could also be observed in some second generation merozoites of E. tenella, indicating that these merozoites are already sexually differentiated. The relationship between the present cytochemical observations and the ultrastructure of the four investigated parasites is discussed.

Plasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200000 and 180000 were detected only after extraction with deoxycholate or SDS.

The endogenous development of the Houghton (H) strain of Eimeria mitis (= mivati) was compared with the life-cycle of a precocious (HP) line derived from the H strain. In both parasites 4 generations of schizonts which developed in epithelial cells were observed: the 1st and 2nd were found in the crypts and the 3rd and 4th in the villi. Gametocytes and zygotes occupied epithelial cells at the tips of the villi. The onset of gametogony normally coincided with the maturation of 4th-generation schizonts. The infection was confined initially to an area of the gut extending from the jejunum to the ileo-caecal junction but 3rd-generation merozoites and subsequent stages were also found in the caeca and rectum. The life-cycle of the precocious line was shorter than that of the parent strain. Gametocytes appeared to develop from 3rd-generation as well as from 4th-generation merozoites. Also, sporozoites of the precocious line transformed to trophozoites before those of the parent strain. First-generation schizonts of the HP line tended to be smaller and to contain fewer merozoites than those of the H strain. The differences between the life-cycles of the two parasites account for the lower reproductive potential of the precocious line.

The reproduction of Eimeria vermiformis in different strains of phenotypically normal mice and in mice with various immunological characteristics or defects was compared. In some strains of phenotypically normal mice there were marked differences in oocyst production, both in terms of numbers and in the duration of patency, allowing the strains to be classified as resistant or susceptible to infection with E. vermiformis. These differences were apparent only in primary infections; both types of strain were equally resistant to reinfection. Amongst the strains of mice with immunological deficiencies, reproduction of the parasite was greatest in the athymic (nu/nu) mutants and these mice were completely susceptible to reinfection. Strains of mice with lowered or defective antibody production, or with defective neutrophils and low NK cell activity (bg/bg) were more susceptible than the relevant controls to primary infection but all developed substantial immunity to reinfection. Asplenic (Dh/+) mutants were remarkably resistant to infection.

We have performed restriction endonuclease digestion of nuclear DNAs, combined with gel electrophoresis and molecular hybridization to characterize different Trypanosoma brucei brucei stocks and to identify trypanosome species. Cloned DNA complementary to the messenger RNAs (mRNAs) for different variable surface glycoproteins (VSGs) of T. b. brucei stock LUMP 227 was used to examine differences in genomic DNAs from T. b. brucei stocks and from various other trypanosome species. These cDNA reagents differentiated the stocks of T. b. brucei and also distinguished the trypanosome species. Our data also show that a T. b. brucei nuclear DNA fragment used as a probe in molecular hybridization analysis provides an appropriate marker in determining trypanosome species. We have extended our analysis of trypanosome nuclear DNA by developing a simple and rapid spot test, for the identification of trypanosome species, which can be carried out using crude materials such as isolated parasites, infected whole blood and tsetse saliva. The procedure is species-specific, sensitive to the level of 104 organisms and is suitable for use under field conditions. Thus, with appropriate sequences for hybridization, the procedure has obvious applications for the diagnosis of African trypanosomiasis.

Following inoculation of Trypanosoma brucei into large mammals by the tsetse fly a local skin reaction, the ‘chancre’, develops due to trypanosome proliferation. We have cannulated the afferent and efferent lymphatics of the draining lymph node in goats and examined the onset of a cellular reaction, the emigration of the parasite from the chancre and the development of both antigenic variation and the specific immune response. The chancre first became detectable by day 3 post-infection, peaked by day 6 and then subsided. Lymphocyte output increased 6- to 8-fold by day 10 and the number of lymphoblasts increased 50-fold in this period. Both then declined. Trypanosomes were detected in lymph 1–2 days before the chancre, peaked by days 5–6, declined during development of the chancre and then peaked again. The bloodstream population appeared by days 4–5 and displayed different kinetics from that in lymph. Recirculation of parasites through the lymphatics ensued. Lymph-borne trypanosome populations were highly pleomorphic. Parasites in lymph expressed firstly a mixture of the Variable Antigen Types (VATs) which are found characteristically in the tsetse fly, this being followed by a mixture of other VATs. The two groups overlapped in appearance. In the bloodstream the same sequence of events occurred although 2 or 3 days later. The specific antibody response, as measured by radioimmunoassay and agglutination, arose within a few days of the first detection of each VAT. Activities appeared first in the lymph and then in plasma.

Fibronectin determinants were revealed in a soluble extract of Schistosoma mansoni adult worms by standard immunodiffusion and immunoelectrophoresis techniques. The same target molecules were demonstrated on the parasite's surface using a binding assay with [125I] anti-fibronectin. The use of fluorescein- and peroxidase-conjugated antibodies confirmed the above observations and provided a fairly precise means for locating the cross-reacting antigen on the worm surface. An in vitro cytotoxic assay using inactivated (56°C, 2 h) anti-fibronectin rabbit immune serum and guinea-pig serum as a source of complement was developed. In these conditions, anti-fibronectin exerted cytotoxic activity against lung schistosomula and adult worms but not skin schistosomula. The ultrastructural damage induced by anti-fibronectin in the presence of complement was studied using transmission electron microscopy. The results suggest a close association between fibronectin determinants and S. mansoni membrane.

Experimental studies on the survival of Fasciola hepatica miracidia show no evidence that miracidial mortality varies with the pH of the medium, at least in the range 6·0–8·0. On the other hand, miracidial mortality is shown to vary with both the temperature of the medium and the age of the larvae. The mean expected life-span of the miracidium decreases from about 35 h at 6°C to about 6° h at 25° C. The Gompertz survival function provides a good description of the miracidial survivorship curves over the range of temperatures used, and we describe, a maximum likelihood method of estimating the mean values of the parameters of this function, together with their approximate 95% confidence limits.

Chimaerocestos prudhoei gen. et sp. nov. from Rhinochimaera atlantica in the North Atlantic off north-eastern Scotland represents a new family, the Chimaerocestidae, of tetraphyllideans and is the first record of a strobilate tapeworm from a holocephalan. The main taxonomic features are described and figured along with aspects of the ecology and mode of attachment to the gut mucosa. The lobed annular ovary and distribution of the vitellarium in two fields are characteristic features. About 15 eggs are deposited together in capsules. C. prudhoei is discussed in relation to other unusual fish tapeworms and the evolutionary biology of tapeworms, and to our scant knowledge of the holocephalans.

The development of the tegument and cercomer of Paricterotaenia paradoxa polycephalic larvae was examined using electron microscopy. Larvae are formed by budding from the inner surface of the tegument of the degenerating hexacanth embryo. A new secondary tegument formed around the larvae is probably produced from the original hexacanth sub-tegumental cells. Microvilli covering the surface of young larvae are converted directly into microtriches, as the larvae develop, by addition of electron-dense material to the proximal part of the microvillus. Remnants of the original microvillus are visible at the distal surface of each new microthrix, but they eventually degenerate. The cercomer homologue is represented by scattered follicular cells, bearing microvilli, lying just within the containing cyst wall. The continuity of tegumentary tissue from one developmental stage to the next is discussed.

Variations in the general protein and 40 enzymes of the 8 European pathotypes of the potato cyst nematode Globodera rostochiensis and G. pallida have been examined by isoelectric focusing (IEF) and cluster analysis. Differences in the protein patterns were observed and the presence of a major band at pH 8·0 allowed the two species to be discriminated. Extra isozymed of peptidase (EC 3.4. 11.*) and esterase (EC 3. 1. 1. 1) were also found in G. rostochiensis. Intra-species variation was seen within G. rostochiensis for superoxide dismutase (EC 1. 15. 1. 1) and within G. pallida for phosphoglucomutase (EC 2. 7. 5. 1) and glucose phosphate isomerase (EC 5. 3. 1. 9). The results suggest that the two species are very similar and the prospect for recognizing their pathotypes by IEF is discussed.

Fourth-stage larvae of Ascaris suum, recovered from piglets at 11–22 days post-infection (p.i.), were examined by light microscopy for changes in the somatic musculature. During this time the estimated total number of muscle cells in the body increases from approximately 600 to 21 000 cells. This non-eutelic development appears to occur by division of platymyarian muscle cells into coelomyarian cells, thereby increasing the number of muscle cells/quadrant from 5 to 85. The incompleteness of these divisions results in sarcoplasmic connections among muscle cells, and between muscle cells and nerve chords. At 11 days p.i., immediately after the 3rd moult, the variation in muscle cell length in a quadrant, with the longest cells being at the lateral chords, has been established. The average muscle cell length at this point is 0·101 mm, the longest is 0·164 mm. These values have increased to 0·365 mm and 1·122 mm, respectively, by 22 days p.i. At 18 days p.i. those cells in the posterior section of the larva are still the shortest in the body.

Mice infected with Nematospiroides dubius generate weaker immune responses to concurrently administered sheep red blood cells (SRBC), than non-infected controls. The experiments described in this paper demonstrate that both adult and larval stages of N. dubius cause non-specific immunodepression of the response to SRBC. Mice which had been infected with larvae exposed to 25 krad. of irradiation, which prevents development to the adult luminal stages, produced as weak haemagglutination responses to SRBC as mice infected with normal worms even when SRBC were administered 6 weeks after infection. The removal of adult N. dubius by treatment with pyrantel 9, 11 and 15 days after infection with normal larvae did not restore the host's ability to respond to SRBC given on day 14. It was only when the mice had been without worms for 17 days that their capacity to respond normally to SRBC was restored. Mice infected with 60 or 400 transplanted adult worms produced depressed haemagglutination and plaque-forming responses to concurrently injected SRBC when compared with normal or sham-operated controls. The significance of these results is discussed in relation to the possible role of non-specific immunodepression in facilitating the survival of N. dubius in the host.

The effects of mitochondrial inhibitors on the in vitro development of Nippostrongylus brasiliensis have been studied in free-living and parasitic life-cycle stages. Mitochondrial inhibitors were chosen as being representative of established electron transport inhibitors and oxidative phosphorylation inhibitors and uncouplers of the classical mammalian respiratory chain. All mitochondrial inhibitors tested were highly effective in killing or retarding development of free-living stages of N. brasiliensis. Free-living stages were particularly susceptible to such inhibitors upon hatching of embryonated eggs to 1st-stage larvae. Concentrations of inhibitors effective against free-living stages were consistent with their level of inhibition against isolated mitochondria from embryonated eggs and 3rd-stage infective larvae. Results suggest an absolute requirement in the development of free-living stages for the mammalian-like respiratory chain and associated oxidative phosphorylation. Electron transport inhibitors were effective in retarding at least the initial development of 4th-stage larvae to adults, but only antimycin A and azide produced a lasting effect leading to worm death. Oxidative phosphorylation inhibitors and uncouplers were ineffective against developing parasitic stages of N. brasiliensis. Experiments on whole-worm respiration indicated that most electron transport inhibitors were able to penetrate the adult worm, but oxidative phosphorylation inhibitors were without effect on whole-worm respiration. Results suggest that the mammalian-like electron transport chain is a necessary requirement to adult N. brasiliensis, but oxidative phosphorylation in the adult worm may not be required for development and survival in vitro although it could be necessary to support the parasite in vivo.

Parasitic helminths belong to 3 separate phyla and there is always the danger of over-generalization. The various routes of anaerobic carbohydrate breakdown in parasitic helminth differ in their efficiencies and in their power output. The choice of end-product represents a compromise between these two conflicting forces. In addition, anaerobic pathways must satisfy the redox requirements of the tissues and provide a source of intermediates for synthetic reactions. Other considerations include the metabolic cost of excretion and the effect of end-products on protein structure and function. The different end-products may fulfil additional functions such as pH control, nitrogenous excretion, osmotic regulation, intracellular signalling and the suppression of host responses.

A complicating factor in parasitic helminths is the existence of strains with different biochemical characteristics, including marked variation in end-product formation. The various tissues of the same parasite can also produce different end-products and the pattern of end-product formation is influenced by a variety of extrinsic and intrinsic factors such as age, sex, length of incubation, pO2 and availability of substrates. The catabolic pathways of helminths thus show considerable functional adaptation.

There is, as yet, no satisfactory explanation as to why helminths do not make the maximum use of any oxygen available to them; and the contribution of oxidative processes to the overall energy balance of parasites probably varies from species to species.

The catabolic pathways of adult helminths are derived from the anaerobic pathways present in their free-living relatives. Two main trends are evident, homolactic fermentation and carbon dioxide fixation, the latter involving a partial reverse tricarboxylic acid cycle. In general, homolactic fermentation is found in blood and tissue parasites, carbon dioxide fixation in gut parasites. These two types of metabolism are, of course, in no way absolute, most homolactic fermentors fix carbon dioxide to a certain extent and many parasites which fix carbon dioxide also produce lactate. Parasitic helminths possess a wide range of different catabolic pathways, superimposed upon which is a high degree of functional plasticity.