The proteinases of Trichomonas vaginalis have been analysed by electrophoresis in polyacrylamide gels containing denatured haemoglobin. Seven bands of activity were detected indicating multiple proteinases. All of the enzymes were stimulated by 1 mM dithiothreitol and had inhibitor sensitivities characteristic of cysteine proteinases. The enzymes differed significantly, however, with respect to pH optima and relative sensitivities to inhibitors.

The development of microgamonts was studied in the small intestine of a 2-day-old cat after oral inoculation with 107 bradyzoites of Sarcocystis hirsuta. The cat was given bradyzoites in 2 doses at 2 h apart and killed 4 h later. Gamonts were seen in parasitophorous vacuoles of goblet cells in the epithelium of the small intestine. All stages of microgametogony, from the single nucleated meront to fully developed gametes, were seen. Microgamonts measured 6·9 × 6·0 μm and macrogamonts measured 6·0 × 5·0 μm.

Seven microfilaraemic and five amicrofilaraemic cats which had been repeatedly infected with Brugia pahangi were challenged along with normal cats 28, 14 and 1 day before autopsy. The lymphatics of the amicrofilaraemic cats contained no female adult worms originating from the repeat infections and only two adult males (both from the same cat). Only 5·2 % of the worms in the control cats were recovered from the amicrofilaraemic cats. Most of the challenge worms were killed in the first 24 h. The microfilaraemic cats all contained fertile adult male and female worms derived from the repeated infection but in such low numbers as to indicate considerable resistance to infection. Compared to their controls 26·4% of the challenge worms were recovered. Analysis of the life-cycle stages recovered showed that in both groups there was attrition of all stages and that although a number of worms reached L5 these were all killed later in the amicrofilaraemic cats.

In order to determine whether a given H-2 haplotype has similar effects on responses to schistosomiasis mansoni on different genetic backgrounds, mice of 2 pairs of congenic strains (H-2b and H-2k on BALB/c and C57BL/10 backgrounds) were infected. Worm burdens, mortality, splenomegaly, tissue and faecal egg counts, and antibody titres to worm and egg antigens were measured. The genetic background had a major effect on the genesis of splenomegaly, on the deposition of eggs in the spleen, the maximum faecal egg count, the antibody titre to egg and worm antigens and the rate of generation of antibody response. The H-2 haplotype was shown to consistently influence the maximum faecal egg count and the antibody titres. Worm burden was not influenced by genetic differences between strains and mortality differences were not significant. The data presented here indicate that the effect of the major histocompatibility complex on responses to infection is greatly influenced by the genetic background on which it is expressed.

The number of schistosomula in lungs was determined by compressed organ autoradiography at intervals up to 14 days after exposure of mice to 75Se-labelled cercariae by tail immersion. Probit analysis of compressed lung autoradiogram focus counts, expressed as percentages of initial infection level, yielded estimates of the average time of arrival, peak accumulation in the lungs and average time of departure of schistosomula: 4·5±0·87, 6·3 ± 0·45 and 11±0·58 days, respectively. At peak accumulation 92±3·5 % of the initial number of schistosomula were found in the lungs. It thus appears that little or no significant attrition of schistosomula occurred in the skin and, instead, that most of the 50–70% of penetrant cercariae that fail to reach adulthood are lost somewhere between the pulmonary and hepatic phases of development. Loss of 75Se label from schistosomula during the first 14 days was exponential, with an average half-life of 4·5±0·81 days. However, the high sensitivity of autoradiography tended to compensate for this rather rapid rate of label loss. It was pointed out that autoradiographic detection of schistosomula as discrete loci of radioactivity can also be expected to overcome the problem posed by the accumulation in such tissues as liver and kidney of 75Se label that has become separated from larvae.

The technique of tail amputation is utilized as a method for interrupting the migration process of Schistosoma mansoni schistosomula from the skin of Fischer rats infected by exposure of the tail to cercariae. The yields of schistosomula recovered from the lungs at different times post-infection are compared, using rats with or without tail amputation. Residence times of schistosomula in skin and lungs, as well as their transit time and efficiency of migration between these sites, are estimated. At least one-third of the infecting cercariae migrate from skin to lung in rats. Amputation of the tail on days 4 or 5 post-infection isolates a definable number of schistosomula in the lung and their migration to the portal circulation can be followed. The kinetics of this migration in rats and mice is compared and a significant difference is revealed.

The feeding behaviour of Glossina morsitans morsitans infected with Trypanosoma vivax, T. congolense or T. brucei was studied and compared with that of uninfected control tsetse. The following parameters were measured. (1) Mean number of feeds taken in 20 consecutive days; (2) mean rate of blood meal intake in μg/sec of the total days fed; (3) mean weight of blood meal intake/day of the total days fed; (4) mean total blood meal engorgement in 20 days; (5) mean number of probes/day of the days fed; (6) mean frequency of the multiple probe feeds and (7) mean number of pre-feeding probes of the multiple probe feeds. Evidence was found to suggest that the rosettes of T. vivax and T. congolense in the proboscis, and colonies of T. brucei parasites in the salivary glands, did not significantly alter the feeding behaviour of the vector.

The numbers, distribution in the small intestine, sexual development and growth (dry weight) of 5-week-old Moniliformis dubius (Acanthocephala) were investigated experimentally in adult, female CFHB rats fed on theoretically isoenergetic diets containing known amounts of fructose in combination with either maize-oil fatty acids or maize oil and two concentrations of casein. There was no obvious development of M. dubius when there was no fructose in the host's diet. In contrast, estimated consumption by the host of as little as about 2 g of fructose during the 5-week infection period was accompanied by marked sexual dimorphism and weight gain in most of the M. dubius present. The dry weights of M. dubius of both sexes were positively correlated with fructose concentrations ranging from 0 to 2·5 % (w/w) in the diets containing fatty acids. Significant, but not substantial, increases in M. dubius dry weight were observed as the dietary fructose concentration was raised to 12 % (w/w). Similar trends were observed when the fructose was offered to the infected rats with maize oil, but in general, fructose added to the fatty-acid based diets supported most M. dubius growth. Differences in the distribution pattern of the worms in rats fed on the fatty-acid or maize-oil based diets were observed and their possible significance is discussed.

The immunogenicity of the secretory exo-antigen (EXA) originating in the stichosome of Trichuris muris has been investigated. High levels of immunity to challenge were produced using EXA (100 μg) by subcutaneous administration. Vaccination with a range of doses of EXA in Freunds' Incomplete Adjuvant (FIA) showed a dose response; 1 μg of EXA reduced the worm burden by 50% and doses of 10 μg or more reduced the worm burden by 80–90% at Day 9 post-infection. Worms recovered from vaccinated mice were significantly smaller than those from controls. A precipitin line identifiable with EXA was shown to immunize mice against T. muris challenge by vaccination. The protective antigen contained in EXA has been partially isolated using gel filtration, ion exchange filtration and isoelectric focusing and has been shown to be a protein of 30000 molecular weight having a pi of 6·8–7·3.

Prior to scolex-retraction, the tegumentary syncytial cytoplasm of the presumptive rostellar region of Hymenolepis diminuta (from Tenebrio molitor kept at 26 °C) is indistinguishable from that of the rest of the cysticercoid. At 1 day after scolex-retraction differentiation of the rostellum has commenced, the tegumentary cytoplasm containing a small number of membrane-bound, ovoid, electron-dense granules which are absent from all other tegumentary regions of the metacestode. By 3 days after scolex-withdrawal there is a substantial increase in the number of ovoid granules within the rostellar tegumentary syncytium and the Golgi systems of the rostellar cytons are highly secretory. At this stage, although the cyst wall tissues are not fully developed, the metacestode is infective. This early and rapid development of the rostellar region presumably enables the ‘adult’ condition to be readily attained. In older metacestodes there is a progressive accumulation of ovoid granules within the rostellar tegumentary cytoplasm, accompanied by a decrease within the rostellar cytons. At 23 days following scolex-retraction the rostellar syncytium has the appearance of that of the adult tapeworm. The rostellar cytons also produce tegumentary discs and vesicles and are therefore regarded as homologous to the other tegumentary cytons of the metacestode.

The fine structure of the nervous system in the scolex and neck region of Hymenolepis nana has been investigated by transmission electron microscopy. A description of the gross neuroanatomy in these regions of the worm is presented. The ganglia, commissures and nerve cords consist of an incomplete cortex of nerve cell bodies, and a core of nerve fibres. A delimiting sheath or capsule is absent. The nerve cell bodies contain a single nucleus with a single nucleolus, mitochondria, many ribosomes, Golgi complexes and vesicles formed within the Golgi cisternae. Numerous sub-surface cisternae are present beneath the outer plasma membrane of the nerve cell body, and the inner surfaces of these cisternae are studded with ribosomes. Some of the cisternae run tangentially into the cytoplasm of the perikaryon, particularly in the vicinity of the Golgi complexes; both sides of these cisternae are studded with ribosomes. From each neuronal perikaryon arise one or more neurites that contain neurotubules, mitochondria, ribosomes and electron-lucent or dense-cored vesicles. Five types of vesicle have been distinguished on the basis of their size and content. The neurites are unmyelinated and form synapses in the neuropile; the synapses possess features typical of those where mechanical strength is of importance. Three types of sensory receptors have been observed in H. nana, two ciliated and one non-ciliated; the latter typically form double or triple nerve endings which terminate within the tegument.

The development, survival and infectivity of larvae of Nippostrongylus brasiliensis at 10 and 21 °C have been studied. In cultures developed for 1 weekat21 °C but stored at either 10 or 21 °C the mean recovery of larvae in 1·5 h Baermann sediment (46% and 42% of total eggs, respectively) and their infectivity (66% and 70% day 8 p.i. recovery, respectively) at 4 weeks of cultivation were similar. However, at 20 weeks of cultivation only a few larvae survived storage at 21 °C (1·4%) and the survivors showed very low infectivity (4·1%), whereas larvae stored at 10 °C retained most of their survival and infectivity rates (41% and 44%, respectively). In cultures developed and stored at 10 °C a lower mean proportion (17%) of the eggs was recovered as larvae at 4 weeks of cultivation. The mean infectivity rate of these larvae increased from 4 to 12 weeks of cultivation (18 % and 42 %, respectively) but was reduced at 20 weeks of cultivation (27 %). An increased proportion of adult female worms was recovered from rats infected with larvae from cultures showing poor development or survival and the fecundity of the adult females decreased in all groups as the storage time of the cultures was increased. Eggs from low fecundity adult worms showed normal development of larvae. The relation between survival, temperature in the habitat and the geographical distribution of the larvae of N. brasiliensis is discussed.

Glycosidase activities in the adults and juveniles of the lung fluke Paragonimus ohirai and P. westermani adults were demonstrated histo-chemically. For comparative studies, histochemical examination was also made on the adults of the liver fluke Fasciola hepatica. The enzymes examined were N-acetyl-β-glucosaminidase (EC 3.2.1.30), β-glucuronidase (EC 3.2.1.31), β-galactosidase (EC 3.2.1.23), α-glucosidase (EC 3. 2. 1. 20) and β-glucosidase (EC 3.2.1. 21). The distribution of β-glucosaminidase was similar in juveniles and adults. Strong reaction sites for the enzyme were the caecal brush border, tegument, sub-tegumental cells and testes. In contrast, no staining reaction occurred in the caeca of F. hepatica, although the tegument and parenchymal cells were weakly stained, β-glucuronidase activity was associated only with the luminal surface of the caeca in the juveniles. However, luminal contents also appeared stained and this might suggest that the activity in the caeca is not endogenous. β-galactosidase was localized in the caeca, sub-tegumental cells and tegument in both juveniles and adults. No reaction occurred for the other two enzymes, α- and β-glucosidase.

The development of acquired resistance in opisthorchiasis was studied in hamsters experimentally infected with Opisthorchis viverrini. The induction of protective immunity was attempted by first exposing adult female golden Syrian hamsters to 1,2 or 3 doses of infective metacercariae obtained from naturally infected cyprinoid fishes and then reinfecting them with 80 metacercariae. In other experiments, animals that were infected with 50 metacercariae were treated with praziquantel prior to being rechallenged in order to eliminate the flukes that had developed from the first infection. The effect of long-term chronic infections was also studied. Faecal egg counts were determined at weekly intervals from 4–5 weeks onwards. The animals were killed 2–3 months after the last infection for worm recovery, and terminal faecal egg output/g faeces/worm was calculated. The data showed that prior infection of animals with O. viverrini did not induce significant protective immunity against reinfection by the same parasite. Lack of protection was also noted in animals reinfected several times with small doses of metacercariae. However, under certain circumstances, prior infection could result in a significant reduction in the faecal egg output due to subsequent infection.

Rats and mice were infected with either normal metacercariae or metacercariae γ-irradiated at 3 krad. or 4 krad. Flukes were recovered at various times after infection and their teguments examined using a transmission electron microscope. In normal flukes, the secretory granules TO, Tl and T2 were all seen during tegumental development. The teguments of flukes from mice developed faster than the corresponding teguments in rats. TO granules were present from day 0 to day 10 post-infection (p.i.) in mouse flukes and from day 0 to day 14 p.i. in rat flukes. Tl granules first appeared in mouse flukes by day 4 p.i. but not until day 8 p.i. in rat flukes. T2 granules were seen in mouse flukes 2 days p.i. but not before 6 days p.i. in rat flukes. γ-Irradiation at 4 krad prevented normal tegumental development in flukes from both rats and mice. TO granules were present at all times in flukes from either host. Tl granules were produced in mouse flukes but their appearance was delayed until day 6 p.i. No significant production of T2 granules occurred in flukes from either host. Parasite survival was also affected by γ-irradiation and none of the flukes reached maturity. Flukes from rats died between 10 and 21 days p.i and flukes from mice died between 14 and 28 days p.i. γ-Irradiation of metacercariae at 3 krad. had an extremely variable effect on subsequent tegumental development in both rats and mice. Some flukes developed normally, some showed development associated with γ-irradiation at 4 krad, whilst some showed intermediate development.

The life-cycles of two closely related cephalobaenid pentastomids, Raillietiella gehyrae and Raillietiella frenatus, which utilize geckos as definitive hosts and cockroaches as intermediate hosts, have been investigated in detail. Early development in the fat-body of cockroaches involves 2 moults to an infective, 3rd-stage larva which appears from 42–44 days post-infection. Complete development in geckos involves a further 5 moults in the case of males and 6 for females. Males mature precociously and copulation is a once-in-a-lifetime event which occurs around day 80 post-infection when both sexes are the same size but the uterus of the female is undeveloped. Sperm, stored in the spermathecae, is used to fertilize oocytes which slowly accumulate in the developing saccate uterus. Patency commences when the uterus carries approximately 4000–5500 eggs but only 25–36 % of these contain fully developed primary larvae. Since only mature eggs are deposited, we postulate that the vagina (?) of the female must be equipped with a selective filter that allows through large eggs but retains smaller, immature eggs. Thus the only limit on fecundity is the total number of sperms in the spermathecae and this is precisely the same factor that constrains egg production in the advanced order Porocephalida.

O-group bib, Trisopterus luscus caught on the cooling water intake screens of Fawley Power Station in March and April 1982 were infected (prevalence = 34·8%; intensity = 1·65/infected fish) with adult females of the pennellid parasitic copepod Lernaeocera lusci. The bib sample of 293 fish was divided, for analysis, into 5 length classes. In each of these the level of parasitization with L. lusci was broadly similar and in all 5 classes the parasite population was over-dispersed. Of the living copepods on T. luscus, 93% possessed egg strings while the remaining 7% consisted of earlier developmental stages. The majority of the parasites were attached to gill arches and here they exhibited distinct microhabitat and orientation preferences. A 3–2–4–1 descending order of gill arch utilization was apparent and copepods were non-randomly distributed along the dorso-ventral axis of gill arches with medial and ventral sectors being more heavily used than dorsal ones. An analysis was carried out on the body and egg string wet weights of parasites removed from fish harbouring different parasite densities. As copepod density increased neither the mean parasite body weight nor egg string weight altered significantly. Length/weight relationships of uninfected and infected fish were remarkably similar and thus provided no evidence that L. lusci has a marked effect on the condition of O-group T. luscus. The study demonstrated a significant positive association between infection with L. lusci and heavy infections with metacercarial cysts of Cryptocotyle lingua.

In the ideal situation, the development of new amoebicides, or more accurately anti-amoebic compounds which are compounds with activity against Entamoeba histolytica, should initially proceed with the study of parasite-specific metabolic pathways and their inhibition, followed by whole parasite in vitro studies, experimental in vivo models and finally clinical trial. However, there are considerable gaps in our knowledge which will be discussed below, and consequently many investigators consider that empirically selected compounds should be tested experimentally in addition to specifically designed compounds. Before clinical trials can begin, extensive examination of the candidate amoebicide in experimental animals is required in order to investigate possible toxicological hazards.

In addition to inhibiting the amoebic parasite, the drug has to reach the parasite in several different sites in the body, thus there is also a problem of pharmacokinetics and distribution. Prior to the discovery of the nitroimidazole class of amoebicides, the multi-site attack was solved by the use of several drugs, sometimes in sequence during the treatment of an individual case (Powell, 1972). The discovery of the nitroimidazole class of compound changed the situation dramatically and these have shown a clinical and parasitological effect against extra-intestinal and intestinal wall infections. The effect on intralumenal infection (that is mildly symptomatic or asymptomatic infections) of the large intestine is, however, less certain (Finegold, 1977; Spillman, Ayala & Sanchez, 1976).

Although the treatment of amoebiasis appears to be satisfactory at the present time, it is difficult to predict problems which might arise in the future, and therefore it is valuable to continue the pre-clinical development, especially the investigation of parasite metabolism, in order to define parasite-specific points of chemotherapeutic attack.

The chemotherapy of amoebiasis was reviewed comprehensively by Woolf (1963, 1965). The present account of the development of amoebicides therefore starts from the Woolfe reviews.