A genomic library of Babesia caballi DNA was constructed in the plasmid vector pUC13. The specificity of the clones for B. caballi was established by the lack of hybridization to Babesia equi, Babesia bovis, Babesia bigemina and equine DNA. Two probes, pBC11 and pBC191, were isolated that could detect 0·25 ng and 0·125 ng of B. caballi DNA, corresponding to a parasitaemia of 0·12% and 0·06% respectively. pBC191 could detect B. caballi parasites in the blood of an experimentally infected horse as well as in naturally infected horses.

Experiments were conducted in guinea-pigs to elucidate the parameters affecting the development of protective immunity against Schistosoma japonicum induced by a cryopreserved, irradiated schistosomula vaccine such as the number of immunizations, route of injection and the use of adjuvants. Results obtained indicated that the cryopreserved, irradiated schistosomula vaccine was effective by either intradermal or intramuscular injection. One intradermal injection with BCG adjuvant resulted in an average worm reduction of 50·24%, only a little lower than that of a non-cryopreserved, irradiated vaccine, 53·55%, with no statistically significant difference between the two. By intramuscular injection the worm reduction was lower (max. 40%) whether given with or without adjuvants or in 1 or 2 injections.

This paper provides an overview of research on the nervous system of parasitic platyhelminths. We have emphasized studies concerned with the physiological, pharmacological and biochemical nature of the major small molecule neurotransmitters of these parasites. We have attempted to provide a critical review of the work by focusing on important unresolved issues. Finally, we have focused on some recent work in our laboratory, using patch-clamp recording techniques and quantitative fluorescence cytometry, as an example of newer methods that will hopefully resolve some of the unanswered questions concerning the nervous system of these parasites.

The midgut epithelium of Glossina morsitans centralis, G. austeni, G. pallidipes, G. palpalis palpalis, G. p. gambiensis, G. fuscipes fuscipes, G. tachinoides and G. brevipalpis from ILRAD-bred colonies was examined, by electron microscopy, for the presence and distribution of Rickettsia-like organisms (RLOs). RLOs were present in the midgut epithelial cells of all non-teneral tsetse. In G. m. centralis, G. pallidipes and, to a much lesser extent, G. brevipalpis, RLOs were numerous and were present in all the specimens examined. RLOs were present in fewer numbers in the epithelial cells of tenerals of these three tsetse species. In contrast, RLOs occurred in very much lower numbers within the midgut cells of nonteneral G. austeni, G. p. palpalis, G. p. gambiensis, G. f. fuscipes and G. tachinoides; were not seen in every specimen, and were rarely observed in the midgut cells of teneral tsetse. The RLOs were typical rod-shaped bacteria with an inner and outer membrane, which occurred free within the host cell cytoplasm and appeared to cause no obvious pathology. The micro-organisms divided by binary fission and at least two distinct morphological forms plus a range of intermediate forms were seen in the midgut cells. A comparison of the presence and numbers of RLOs within the midgut cells and the midgut infection rates of both Trypanosoma congolense and T. b. brucei, both between Glossina species and also within the same stock of tsetse, clearly indicates that the ability of trypanosomes to establish and develop to mature infections is unlikely to be correlated solely with the presence of RLOs within the tsetse midgut.

Maintaining the puparial stage of successive generations of a population of tsetse 3 °C lower than normal reduced the numbers of rickettsia-like organisms (RLO) carried by emerging flies. The susceptibility of these flies to midgut infection with Trypanosoma congolense was also significantly reduced compared with control flies held at normal temperature. These results support the view that the relationship between RLO and susceptibility is quantitative – teneral flies with heavier RLO infections being more susceptible to trypanosome infection.

Quantitative and qualitative analysis of lipids has been performed on a rhoptry fraction purified from Toxoplasma gondii tachyzoites. The lipid to protein ratio was estimated to be 0·26. The cholesterol to phospholipid ratio was unusually high at 1·48. Phosphatidylcholine was the major phospholipid; phosphatidylserine, phosphatidylinositol and sphingomyelin were absent whereas significant amounts of phosphatidic acid and lysophospholipids were found. This pelicular composition could be related to functional involvement of the organelle in host-cell invasion.

A new model for the study of granuloma formation in the liver is described. Rats received an injection of 20 000 Schistosoma mansoni eggs in the portal vein and granuloma formation was evaluated at 3, 5 and 7 weeks post-injection. Liver collagen was estimated at the same time and serum procollagen III peptide, a marker of collagenesis, weekly. With this model, wherein the number of S. mansoni eggs and the time of injury are standardized, the effect of high levels of acute phase proteins especially α2-macroglobulin on granuloma formation was studied. It appeared that in rats with high levels of α2-macroglobulin the mean size of granulomas was significantly greater at 3 and 5 weeks compared with controls. In both groups an increase in liver collagen was observed during this period, reaching a peak at 5 weeks in the acute phase group. This model facilitates the study of the effects of S. mansoni eggs on granuloma formation.

The organization of Ascaris motoneurones and nervous system is summarized. There is an anterior nerve ring and associated ganglia, main dorsal and ventral nerve cords which run longitudinally, and a small set of posterior ganglia. Cell bodies of motoneurones are found in the ventral nerve cord and occur in 5 repeating ‘segments’; each contains 11 motoneurones. Seven morphological types of excitatory or inhibitory motoneurone are recognized.

Each Ascaris somatic muscle cell is composed of the contractile spindle; the bag region, containing the nucleus; the arm; and the syncytial region, the location of neuromuscular junctions. The resting membrane potential of muscle is approximately — 30 mV and shows regular depolarizing, Ca-dependent ‘spike potentials’ superimposed on smaller Na+- and Ca2+-dependent ‘slow waves’ and even slower ‘modulation waves’. The membrane shows high Cl- permeability. Adjacent cells are electrically coupled so that electrical activity in the cells is synchronized. Acetylcholine (ACh) and γ-aminobutyric acid (GABA) affect the electrical activity. Bath-applied ACh increases membrane cation conductance, depolarizes the cells, alters the frequency and amplitude of spike potentials and produces contraction. Bath-applied GABA increases Cl- conductance, decreases spike activity and causes hyperpolarization and muscle relaxation.

The extra-synaptic ACh receptors on the bag region of Ascaris muscle can be regarded as a separate subtype of nicotinic receptor. ACh and anthelmintic agonists (pyrantel, morantel, levamisole) produce a dose-dependent increase in cation conductance and membrane depolarization which is blocked by tubocurarine, mecamylamine but not by hexamethonium. The potency, of GABA agonists, with the exception of sulphonic acid derivatives, correlates with the vertebrate GABAa receptor. The potency of antagonists does not. Thus, bicuculline, securinine, pitrazepine, SR95531 and RU5135 are potent vertebrate GABAa antagonists but have little effect on GABA receptors. The potency order of the arylaminopyridazine GABA antagonists: SR95103, SR95132, SR42666, SR95133, SR95531, SR42627 and SR42640 at the Ascaris GABA receptors contrasts with that at vertebrate GABAa receptors. It has been suggested that the receptor is referred to as a GABAn receptor.

Patch-clamp studies show that ACh activates a non-selective cation channel which has a main conductance of 40–50pS and apparent mean open time of 1·3 ms; a smaller channel of 20–30 pS with a similar open-time is also activated. Pyrantel and levamisole also produce openings with similar conductances and open-times. GABA activates a Cl- channel with a main state conductance of 22 pS and an apparent mean open duration of 32 ms; conductance states of 10 and 15 pS are also seen. Piperazine similarly activates this channel but the mean open-time is shorter (14 ms). Ivermectin in high doses, is an antagonist which reduces the GABA channel conductance and Popen; it does, however, open ‘small’ Cl- channels when applied to the outside surface of membrane. These channels have a conductance of 9–15 pS and very long open times (> 100 mS). 5-HT does not have a direct effect on membrane potential or conductance but acts on cAMP levels and glycogen metabolism. Dopamine, octopamine and AF1 may act as neurotransmitters or neuromodulators.

This study examines the effects of host age and sex on the frequency distribution of Brugia malayi infections in the human host. Microfilarial (mf) counts for a large data base on the epidemiology of brugian filariasis in Shertallai, Kerala, South India are analysed. Frequency distributions of microfilarial counts partitioned by age are successfully described by zero-truncated negative binomial distributions, fitted by maximum likelihood. This analysis provides estimates of the proportion of mf-positive individuals who are identified as negative due to sampling errors, allowing the construction of corrected mf age–prevalence curves, which indicate that the observed prevalence may under-estimate the true figures by between 18 and 47%. There is no evidence from these results for a decrease in the degree of over-dispersion of parasite frequency distributions with host age, such as might be produced by the acquired immunity to infection. This departure from the pattern in bancroftian filariasis (where there is evidence of such decreases in over-dispersion; Das et al. 1990) is discussed in terms of the long history of filariasis control (and consequently low infection prevalence) in Shertallai.

Treatment of BALB/c or MF1 mice with cyclosporin A (CsA) around the time of infection with Schistosoma mansoni conferred almost complete protection. The migration kinetics of L-[75Se]selenomethionine-labelled infective cercariae were investigated by compressed tissue autoradiography. Similar levels of skin penetration were achieved by cercariae in control and drug-treated individuals. CsA arrested 87–94% of the worms in the skin and ultimately all of these died in this site. Few worms (7–14%) migrated from the skin to the lungs and none completed migration to the liver. Nevertheless, the autoradiograms revealed a limited degree of lateral cutaneous migration by the worms present in the skins of CsA-treated mice. Results of perfusion recovery experiments carried out during the course of infection reinforced the tracking data.

The recent development of analytical models of trypanosomiasis has increased the general applicability of models to the strategic control of the disease. An analysis of detailed data on tsetse abundance and infection rates and trypanosome prevalence in village-based trypanotolerant cattle over 4 years in The Gambia showed that seasonal patterns of abundance in Glossina morsitans-infested areas were consistent, and that the rates of trypanosome infection remained relatively unchanging. However, there were two distinct seasonal trypanosome prevalence patterns in cattle, with peaks occurring either in May/June/July or November/December. The peaks of trypanosome prevalence therefore occurred either 4 months before or after the times of peak challenge from G. morsitans, not 1 or 2 months after as predicted by analytical models. In G. palpalis-infested areas there was little seasonal variation in abundance or trypanosome infections, but peak trypanosome prevalence still occurred mostly in June/July. Despite the incongruity between the months of peak prevalence and challenge, the overall annual prevalence rates and tsetse challenge indices showed a significant linear relationship. It is concluded that existing analytical models need to be refined to take into account trypanotolerance and the various influences on the expression of this trait.

Many medically important diseases of man are caused by blood-sucking arthropods which serve as vectors for a wide range of viral, bacterial, protozoal and nematode infections (Table 1). Furthermore, serious economic losses are caused by the numerous arthropod parasites which infect domesticated animals (for examples, see Table 2). Among these the ixodid hard ticks are particularly important and it has been estimated that the global cost of hard tick infections is $7000 million per annum (F.A.O., 1984). Not surprisingly, there have been strenuous efforts to control infections caused by arthropods.

Several recombinant Trypanosoma cruzi proteins previously isolated were used as antigens to analyse antibody specificities present in sera from human infections. Some parasite proteins such as SAPA (Shed Acute Phase Antigen) are antigenic early after infection. Others, like antigens 1 and 30, are antigenic mainly during the chronic phase of the infection. To understand why different proteins are antigenic at different periods of infection, specificities of antibodies present in the sera of infected mice were compared with the antigens expressed by parasites collected directly from blood. Parasites collected during the acute parasitaemia peak expressed not only antigen SAPA, but also antigens 1 and 30. However, only antibodies against SAPA were frequently observed during the early period and also in the chronic phase of murine infection. Long-lasting antibodies against SAPA were detected regardless of the mouse and parasite strains used. Furthermore, all 8 recombinant clones detected in a T. cruzi expression library with pooled sera from acutely infected mice were homologous to the SAPA gene. These results show that even though parasites from the acute parasitaemia peak in mice may express simultaneously several proteins known to be antigenic, only antibodies against SAPA were consistently detected.

Pseudodiplorchis americanus accumulates its entire reproductive output in utero during hibernation of its host, Scaphiopus couchii. Infective larvae are released only during the toads' brief entry into water for breeding. In one host population studied prior to transmission, in S.E. Arizona, adult worms contained 0–326 infective stages; 19% of 178 worms produced 51% of the total larvae. Reproductive output was positively correlated with body length which was considered to reflect worm age; first-, second- and third-year worms produced means of 5, 43 and 96 larvae, respectively. Within second- and third-year cohorts, there was a density-dependent reduction in offspring numbers. Under controlled laboratory conditions, the mean rate of offspring accumulation during the first 6 months of development was significantly reduced only in burdens in excess of those found naturally (36–68 worms). In older parasites, a density-dependent reduction in the mean accumulation rate occurred in burdens of 20–32 worms, where 88% of parasites produced offspring at less than half the maximum rate. The asymmetry in individual reproductive output increased with competition and there were significant reductions in the median rates of offspring accumulation at smaller burdens (17–25 and 5–14 worms, in experimental and natural infections, respectively) than evident from decreases in the mean. Thus, individual variation is a more sensitive measure of competition than is the mean.

The results of studies on the schistosomulicidal activity of activated peritoneal and alveolar macrophages (pMø and aMø) from rats immunized with highly irradiated (50 krad.) Schistosoma japonicum cercariae are reported. The authors have examined the activation of these macrophages in terms of spreading, adhesion and ingestion of sheep erythrocytes and pinocytosis of horse-radish peroxidase. Using three criteria, peritoneal macrophages and alveolar macrophages from immunized rats and from rats intraperitoneally injected with BCG were significantly more active than those from normal rats or rats stimulated with 10% proteose-peptone or 1% sodium thioglycolate. A significantly higher percentage of adhesion and ingestion was obtained with the sheep erythrocytes that were co-opsonized by heat-inactivated rat anti-sheep erythrocyte serum and fresh normal rat serum. Schistosomulicidal effects were observed with macrophages from irradiated cercariae-immunized rats in two activation systems: in vitro activation in the presence of macrophage-activating factor (MAF), and in vivo activation by the intraperitoneal challenge with sonicated cercarial antigens.

At the cellular and molecular levels, the small and simpler nervous systems of invertebrates do not differ fundamentally from the larger more complex ones of vertebrates. It seems therefore that the special properties of the human brain arise more from the fact that it has ten trillion cellular components than from any unusual properties of the components themselves. By studying invertebrates we can gain insight into what basic functions are performed by the cells and molecules of the nervous system and this will contribute to a more fundamental understanding of what goes on in a system in which the same functions are performed by uncountable numbers of neurones. Invertebrate studies are also important for an entirely different reason; they are interesting and important in their own right. Moreover, a relatively small number of invertebrates are pests; either parasites, vectors of serious parasitic diseases or pests of our agricultural production. It is no accident that most of the methods that are used to control such organisms act on their nervous system. That is because the nervous system is a complex chemical machine which works through a great variety of chemical interactions between a wide diversity of receptors and ligands. Many currently used control methods work because they disrupt these interactions. For this reason I would imagine that the new generation of compounds developed to control invertebrates will depend for their activity on interactions with the nervous system. Since most of the chemical effectors (transmitters, modulators and hormones) in the nervous system are peptides, a number of these newly developed approaches will depend upon a fundamental knowledge of peptidergic systems in parasites. This essay is about peptidergic systems and indicates how we might exploit their vulner-ability to interference.

Infection in mice (BALB/cABom) with the intestinal trematode Echinostoma caproni had a negative influence on pregnancy. The effect of the infection set in immediately following implantation (day 5 of pregnancy) resulting in fewer foetuses being present in infected mice on day 9 than in the controls. Ovulation, fertilization of eggs, and implantation itself were obviously not impaired. The infected mice had significantly lower serum progesterone levels on day 5 of pregnancy than the non-infected controls. It is speculated that the progesterone levels in the infected female mice were too low to secure early post-implantation gestation.

The neuropeptide story began in 1928 with the description by Ernst Scharrer of gland-like nerve cells in the hypothalamus of the minnow, Phoxinus laevis. Because these nerve cells were overwhelmingly specialized for secretory activity, overshadowing other neuronal properties, Scharrer termed them ‘neurosecretory neurons’. What was even more remarkable about the cells was that their products were released into the bloodstream to act as hormones, specifically neurohormones. Neurosecretory cells were identified largely on morphological grounds. That is, they could be stained with special techniques, such as chrome-haematoxylin and paraldehyde-fuchsin, although the techniques are far from specific, staining non-neurosecretory cells as well. However, the basis for the ‘special’ neurosecretory techniques is the demonstration of sulphur-containing proteins – so they are indicative of peptide-producing neurones. An alternative characteristic of neurosecretory cells is the presence of large (> 100 nm), dense-cored vesicles at the electron microscope level; these are the so-called elementary granules of neurosecretion, or ENGs. However, implicit in the concept of neurosecretion is that the prime function of the neurosecretory cell is in endocrine regulation, exerting a hormone-like control over some aspect of the organism's metabolism, by controlling endocrine glands and other effector organs. To satisfy this criterion, evidence had to be obtained of cycles of secretory activity within the cell that could be correlated with a change in the physiological condition of the organism.

Sixteen strains of trypanosomes isolated from the blood of 10 freshwater fish species (all but one from South Bohemia) were characterized by enzyme analysis using dise electrophoresis in 5% polyacrylamide gel. Six enzymes were selected: ADH, LDH, MDH, ME, G6PDH and GPI. The most variable were ADH and LDH, whereas MDH, G6PDH and GPI showed the same pattern in all strains analysed. Cluster analysis indicated a very high degree of similarity among strains, even those isolated from phylogenetically distant hosts. Moreover, no correlation between enzyme pattern and host species was found.

Infection with Schistosoma haematobium was studied in a rural community of approximately 500 persons in eastern Zimbabwe. The overall prevalence of infection, as determined by urine egg counts, was 40·1%, and of heavy infections (≥ 50 eggs/10 ml urine) was 11·0%. The prevalence of both heavy and all infections was highest in the 8 to 10-year-old age class. During 1987–88 data were obtained from 102 individuals on intensity of reinfection 14 weeks after treatment with praziquantel, the efficacy of treatment having been determined after 4 weeks. The water contact made by these individuals during 2-week periods immediately following treatment was recorded. The relative abundance of patent infected intermediate host snails, Bulinus globosus, was also monitored. An index of exposure was developed which weighted each water contact by its duration, the type of activity, the time of day, and the abundance of infected snails at the site used. The relationships between rates of reinfection, rates of exposure, and age were examined. Although only 13 individuals showed positive rates of reinfection, there were statistically significant and independent effects of both exposure and age on reinfection rate. Quantitative estimates of reinfection rates suggested that individuals aged 12 years or less acquired substantially more infection (measured as egg output) than individuals more than 12 years old.