Following recent suggestions that a peaked host age–parasite abundance curve, concomitant with a decline in the degree of dispersion of parasites in the older age classes of hosts, can provide evidence of parasite-induced host mortality, the changes in mean abundance and over-dispersion of metacercarial stages of Diplostomum spathaceum, D. gasterostei, Tylodelphys clavata and T. podicipina in relation to fish age were studied in a field locality. The mean parasite burden of D. spathaceum, D. gasterostei and T. clavata increased with host age and the maximum mean burden was found in the oldest hosts. The variance to mean ratio also increased in D. gasterostei, but decreased in the oldest hosts in D. spathaceum and T. clavata. It is concluded that this decrease could be due to parasite-induced host mortality but could equally be due to death of parasites within the host or to changes in infection rate or could be a reflection of the small sample size of the oldest fish. The mean burden of T. podicipina declined gradually with host age, but the variance to mean ratio remained constant and it is concluded that this could be explained by death of the parasites within the host. None of these data or data from other localities provided clear and unambiguous evidence of host mortality induced by heavy infections of any of the four species, although they are consistent with such mortality and do not refute such a possibility. It is concluded that it may be just as difficult to detect and unequivocally demonstrate parasite-induced host mortality in metacercarial digenean–fish host systems as in any other parasite–host systems.

Temperature-dependent survival and reproduction of Gyrodactylus bullatarudis Turnbull, 1956 were examined at 17, 21, 25·5, 27·5 and 30 °C on isolated guppies under conditions where parasite density dependence and host responses were minimized. Parasites had the longest life-span (5·5 days) at 21 °C, the highest average fecundity (1·73 offspring) at 25·5 °C and the highest instantaneous per capita birth rate (0·543/parasite/day) at 27·5 °C. The intrinsic rate of increase of the parasite population was maximum (0·230/parasite/day) at 27·5 °C. Data obtained from this study indicate that G. bullatarudis are not able to survive at 30 °C.

The filarial worm Litomosoides carinii showed a rapid uptake of glucose during in vitro incubation. This uptake proceeded linearly with time, and was significantly higher under aerobic compared to anoxic conditions. Under an atmosphere of nitrogen the worms converted glucose almost quantitatively to lactate, whereas in the presence of oxygen appreciable quantities of acetate, acetoin and CO2, in addition to lactate, were formed. Although aerobically only 73% of the carbohydrate carbon could be accounted for by the latter products as well as by a net glycogen synthesis, attempts to identify other compounds presumed to be derived from glucose metabolism have been unsuccessful. The complete sequence of the glycolytic enzymes was detected in particulate-free cytosolic extracts of the filarial worm. With the exception of 6-phosphofructokinase, all glycolytic enzyme activities were considerably higher than those reported for rat liver. In addition, L. carinii possesses the entire set of enzymes catalysing the eight successive reaction steps of the tricarboxylic acid cycle. On a mitochondrial protein basis, the specific activities of these enzymes were similar to those present in rat liver. Various enzymatic activities of the mitochondrial respiratory chain were detected in the parasite. These include low levels of NADH and cytochrome c oxidases, but a high activity value for NADH dehydrogenase. Cell-free extracts and the mitochondrial fraction of the worms were found to exhibit an enzyme capable of catalysing the decarboxylation of pyruvate. Since this activity was stimulated 5- to 20-fold by the cofactors known to be required by the pyruvate dehydrogenase complex of other animal cells, pyruvate decarboxylation and thus acetate formation in the parasite may be mediated by an enzyme similar to, or identical with, the pyruvate dehydrogenase system. Isotopic carbon balance studies and experiments in which substrates specifically labelled with 14C were employed showed that substrate carbon can to some extent enter into respiratory CO2. From these and the enzymatic analyses it is suggested that complete oxidation of carbon substrate may be of relevance as an energy-conserving pathway in the filarial worm.

Haemolymph free amino acids (HFAAs) in adult Tenebrio molitor varied in concentration with beetle age. Between 9 and 30 days post-emergence the concentration range was 34–94 mM with a peak occurring in 15-day-old insects. Much of this age-related variation was due to changes in the concentration of a single amino acid, proline. An analysis of concentrations of individual HFAAs in 15-day-old beetles, following infection with metacestodes of Hymenolepis diminuta, revealed that changes in female insects are more marked than those occurring in males. In the former, changes occurred in the concentration of 9 amino acids (isoleucine, leucine, arginine, serine, threonine, tyrosine, phenylalanine, proline and alanine/citrulline) whereas in the latter only 4 amino acids were affected (threonine, glycine, histidine and arginine). It is suggested that H. diminuta may interact with the endocrine system of its beetle host.

The effects of metacestodes of Hymenolepis diminuta on haemolymph proteins of Tenebrio molitor are restricted to female hosts. In beetles aged 15 days post-emergence, and harbouring 12-day-old metacestodes, haemolymph concentration is 46·7% higher than in non-infected animals and similar effects are found in longer standing infections. Electrophoresis of haemolymph revealed the presence of 13 bands. Densitometric analysis showed that only band 2/3 was significantly elevated in infected hosts although band 7/8 also showed an increase. These bands were also present in egg homogenates and are thought to be vitellogenins. It is therefore proposed that the excess protein found in infected beetles is a female-specific protein resulting from an interaction between the parasite and the host endocrine system.

The development of the egg-shell of the trichuroid nematode, Trichuris muris, was observed by light and electron microscopy. Initial stages of shell formation occurred immediately following sperm entry into the oocyte in the spermatheca of the female worm. The external vitelline layer separated from the oolemma, reticulate oocyte granules were discharged into the resultant perivitelline space and stored glycogen contributed to the formation of the chitinous shell. A lipid layer developed between the inner surface of the chitinous layer and the oolemma. The fully formed egg-shell thus comprised an external vitelline layer, a middle chitinous layer and an inner lipid layer. The outer vitelline layer resembled a unit membrane with strands of particulate material attached to its outer surface. The middle chitinous layer was composed of chitin microfibrils encased in a protein coat; the sheets of microfibrils were arranged as parallel lamellae forming a helicoidal architecture. The inner lipid layer contained a variable number of loosely arranged strands of electron-opaque and less-opaque material.

The larval output of Angiostrongylus cantonensis at different initial densities in rats was examined throughout the reproductive life-span of the worms using a modified dilution technique. With an initial density of 2 worms the mean duration of larval output was 414·2 days. The output was at a level above 50000 larvae/female/day from 15 to 45 weeks after infection. The mean total output was 18·9×106 larvae/female. The larval output/female decreased with an increase in the worm burden and also with the passage of time after infection. The longevity of host rats also decreased with increasing worm density and many of the heavily infected rats died early during infection. These results suggest that worm density has an effect on the fecundity of this parasite, and that the effects may be due to fibrous changes in host lung tissue but not changes in the physiology of the worms. It is also suggested that density-dependent effects on fecundity play a role in the population dynamics of both the parasite and host as regulatory mechanisms in the field.

Immunity to Hymenolepis nana was adoptively transferred to recipient mice of 4 strains using mesenteric lymph node cells (MLNC), spleen cells (SC), and/or serum taken from infected donor mice. Donor cells capable of transferring immunity appeared in the mesenteric lymph nodes and spleen at an early stage (5 days) after infection. Immunity was most effectively transferred with MLNC and immune serum did not enhance this effect. It is suggested that effective immunity to H. nana involves both antibody and T cell-mediated inflammatory events.

Adult Onchocerca volvulus worms obtained by enzyme digestion from nodules of infected Mexicans were radio-isotope labelled by the chloramine-T or Bolton–Hunter methods. No antigenic determinants were detected in extracts of worms labelled by the chloramine-T method but 3 antigens were detected in extracts of the Bolton–Hunter labelled worms. Two were present in such small amounts that it was impractical to investigate them further, but a major component of mol. wt 20 kDa was purified by gel filtration and used in a serological survey of inhabitants of villages in Southern Mexico. Using the 20 kDa antigen, which is superficially located on both sexes of O. volvulus, sera from both non-endemic and endemic regions were analysed by radio-immunoprecipitation of this antigen. In Southern Mexico, the average sensitivity of the test was 92%, and the specificity 98%. Whilst the 20 kDa antigen did not detect antibodies in the sera of Trinidadians infected with Wuncheria bancrofti or Mansonella ozzardi, this antigen detected high levels of antibodies in Indians exposed to W. bancrofti.

Three groups of stocks of Trypanosoma brucei ssp, defined by the criteria of host, human serum resistance and place of isolation as T. b. gambiense, T. b. brucei (Nigeria) and T. b. brucei/rhodesiense (non-gambiense, Uganda) were screened for electrophoretic variation at 20 enzyme loci. One enzyme (Peptidase C) was found to differentiate all T. b. gambiense stocks from the other T. brucei stocks and, taken together with specific variants of 5 other enzymes, could be used to unambiguously define T. b. gambiense stocks. Using a population genetics approach, the frequencies of the different variants in the three groups of stocks were estimated and from them the average similarity and difference between groups were measured using the statistics of genetic identity (I) and genetic distance (D). These results show firstly, that T. b. gambiense is more different from the other two groups than they are from each other and, secondly, that the values of I and D obtained are consistent with T. b. gambiense constituting a sub- or sibling species of T. brucei. Three domestic animal isolates from Zaire and Cameroun were also screened for enzyme variation and two of these identified as T. b. gambiense, thereby establishing the existence of an animal reservoir host. In parallel, these stocks were tested for human serum resistance resulting in the same identification. Studies of the antigen repertoires and antigen gene structure were carried out by other workers on all the T. b. gambiense stocks reported here and the same conclusions reached as to the identification and ability to discriminate this subspecies from other groups of T. brucei stocks. The results presented here are discussed in relation to other published data on enzyme variation in T. brucei.

In PVG rats a single immunizing infection with Schistosoma mansoni cercariae exposed to 0, 5, 10 or 20 krad. gamma radiation failed to induce more than minimal resistance (17–29%) to challenge 4 weeks later, whereas 4 immunizations with 20 krad.-irradiated cercariae, over several months, induced substantial resistance (75%). In contrast, significant protection was induced in Fischer rats by a single immunization with unirradiated cercariae or cercariae irradiated with up to 80 krad. Comparable resistance was induced by unirradiated and by 2, 5 and 20 krad.-irradiated cercariae (67–74%) and lower levels by 10, 40 and 80 krad.-irradiated infections (57, 48 and 33%, respectively). Although the resistance induced by a single dose of 1000 20 krad.-irradiated cercariae could be boosted by a second (88%), further immunizations failed to enhance this resistance. Also, increasing the number of immunizing cercariae in single or multiple vaccinations from 1000 to 3000 failed to increase the resistance. While the resistance induced by 20 krad.-irradiated cercariae persisted undiminished for at least 25 weeks (67%), the resistance induced by normal cercariae declined to insignificant levels by 25 weeks (21%). Comparison of the migration and survival of unirradiated and of 20 and 40 krad.-irradiated cercariae revealed dramatic differences in their fate: parasites exposed to 40 krad. remained in the skin, while the majority of 20 krad.-irradiated parasites died in the lungs after a sojourn of at least 14 days. Thus, although skin schistosomula alone could induce significant protection, optimal resistance was induced only by parasites which migrated to the lungs or beyond. The immunizing potential of these older parasites was investigated by exposing rats to lung-and liver-stage larvae injected into the tail and mesenteric veins, respectively. Irradiated 4-day lung schistosomula induced comparable resistance (79%) to that induced by a complete unattenuated cercarial infection (78%), whereas less resistance was induced by irradiated and unirradiated 11-day-old liver worms (30 and 27%) and by 25-day-old pre-adult worms (48%).

Acquired resistance to Schistosoma japonicum was evident after the lung phase of migration of challenge schistosomula. ‘Trickle’ infections which gave similar primary worm burdens to single infections stimulated similar levels of resistance. There was a strong correlation between the size of the primary worm burden of individual mice and their ability to resist infection 8 weeks after exposure to S. japonicum. However, acquired resistance did not develop until 6 weeks, that is after the primary infection had become patent, and was maximal some 12 weeks after infection; thereafter resistance declined. The numbers of parasite eggs deposited in the tissues increased during acute infection (duration of 12 weeks or less) and tended to stabilize or decrease during chronic infection (longer than 12 weeks). There were good correlations between the resistance of mice to challenge and the numbers of eggs deposited in their livers, guts or lungs during acute infection, but these correlations were not evident during chronic infection. The inflammatory responses to tissue-bound eggs regress during chronic infection and this may influence the relationship between acquired resistance and egg burden. Comparable primary infections of S. japonicum or S. mansoni protected equally well against both heterologous and homologous challenges. However, unisexual infections of S. mansoni, which produced no overt pathology, failed to confer protection against a challenge of S. japonicum.

The skin phase of schistosomular migration was neither essential for the induction or expression of acquired resistance since mice given primary infections by the intraperitoneal route were as resistant to intra-peritoneal or percutaneous challenge as were mice given their primary infections percutaneously. Serum taken from resistant donors with acute or chronic infections conferred no ability to resist infection upon recipient mice. In two experiments mice that were deprived of their T-cells were as resistant to infection as intact mice and both types of host had comparable tissue egg loads and liver pathology. In a third experiment the immune-deprived mice had lower tissue egg counts than the intact mice and were less resistant to challenge. It is suggested that specific immune effector mechanisms may not be involved in acquired resistance to reinfection but rather that resistance is affected by the T-cell independent immunopathology induced by tissue-bound Schistosoma japonicum eggs.

When mice were infected with female cercariae of Schistosoma mansoni and male cercariae of Schistosomatium douthitti, many mixed pairs formed. The paired females were approximately the same size as those in unisexual infections, far smaller than females paired with S. mansoni males. Although the Sch. douthitti males possessed well-developed testes, sperm were not found in their female partners, which developed scanty vitelline glands and produced laterally spined eggs typical of S. mansoni. Such eggs yielded swimming miracidia infective to the snail host of S. mansoni, Biomphalaria glabrata, but not to the lymnaeid snail host of Sch. douthitti. Sporocysts arising from these miracidia were haploid and produced cercariae infective to mice. Parthenogenetically derived female cercariae in mice co-infected with either parthenogenetically derived male or normal diploid male S. mansoni developed to large adults of normal appearance, whose eggs yielded diploid miracidia and subsequent generations of normal diploid schistosomes. Parthenogenetically derived females co-infected with Sch. douthitti males also paired and produced some eggs containing viable miracidia, which gave rise once again to haploid sporocysts. These observations confirm previous suggestions that the stimulus for maturation in female S. mansoni is distinct from that for growth, and is independent of insemination and fertilization. It is concluded that both diploid and haploid S. mansoni females are capable either of parthenogenesis or of bisexual reproduction when appropriately stimulated.