Sera from rabbits vaccinated with irradiated cercariae mediated cell (P388D1 or mouse peritoneal macrophage) adherence to lung-stage schistosomula (LS) but such antibody-mediated cell adherence was short-lived in contrast to cell adherence to mechanically transformed schistosomula (MS). Thus LS lost 50% of their adherent cells within 3–6 h in culture and up to 90% by 24 h, whereas adherence to MS was undiminished during this time. Rapid loss of adherent cells was unique to schistosomula that had developed to the lung stage because schistosomula recovered from the skin up to 3 days post-infection did not exhibit the rapid cell loss shown by 3-day LS. To determine whether cell loss was caused by loss of surface antigenicity during culture LS were cultured on their own for up to 24 h and at various intervals samples of schistosomula were tested for antigenicity by addition of immune serum and cells. Levels of adherence to both MS and LS were maintained throughout the incubation period. When antibody-opsonized schistosomula were washed and indicator cells added at progressive intervals, persistence of adherence was again demonstrated, showing that antibody binding to LS had not promoted surface antigen loss or degradation of bound antibody. It was then shown, by adding fresh macrophages to cultures up to 24 h old that LS which had lost their adherent cells nevertheless retained bound antibody, and comparison of adherence of ‘used’ and ‘fresh’ cells to MS and LS showed that the cytoadherence properties of macrophages were not significantly reduced during their culture with LS from which cells had been lost. Cell loss was shown to be dependent upon protein synthesis by LS since cell loss was significantly reduced when the schistosomula were pre-incubated in puromycin or actinomycin D. Transient cell adherence to LS may comprise an important immune evasion stratagem by schistosomula.

The effects of exposure to infective larvae on the antibody response to a cocktail of specific recombinant antigens of Onchocerca volvulus and to a worm extract were evaluated by comparing the responses of individuals from a vector controlled area with those from an area of continuing transmission by ELISA. Individuals from the vector controlled areas were found to have reduced responses to both antigen preparations. Amicrofilerdermic (mf–) individuals from the area of vector control exhibited significantly lower total and subclass IgG responses to the worm extract. In contrast, the responses to the cocktail of specific recombinants were significantly reduced in individuals from the area of vector control who were still microfilerdermia positive (mf+). The distribution of IgG subclass specific responses was similar to both antigen preparations, both dominated by the IgG4 and IgG1 subclasses. IgG1 responses to the worm extract remained elevated in the vector controlled individuals but IgG4 was significantly reduced in the mf – individuals. Both subclasses reflected the total IgG response to the cocktail of recombinants and were significantly reduced in individuals from the vector controlled area, when compared to individuals from the hyperendemic area. IgG1 responses to the cocktail of recombinants are significantly lower than IgG4 in all individuals and virtually absent in individuals from the vector-controlled area. Measuring total IgG and IgG4 is more sensitive than IgG1 in detecting infection, 100 or 97% respectively, but they remain elevated in the individuals from the vector controlled areas even after 8–10 years interruption of transmission. These results have important implications for the serological monitoring of control programmes in individuals who have previously been infected.

Two mathematical models are developed that investigate the transmission dynamics of Theileria parva by the ixodid tickRhipicephalus appendiculatus to cattle in endemically stable areas. A method of estimating the rate of infection to cattle ofT. parva at the endemically stable state is given. Empirical estimates of ail the parameters in the model are available. The degree to which animals that have recovered from theileriosis (the ‘carrier’ state) are able to transmit the infection to tick nymphs or larvae is a crucial determinant of the dynamics of infection in a herd. Two control methods influencing the transmission of infection are considered – infection and treatment immunization and the reduction in tick feeding by acaricide application. The impact of each method on the transmission of infection is evaluated. Future developments and the data required to predict the dynamics of T. parva infections in cattle and ticks are discussed.

The characterization of four Trypanosoma vivax isolates from Colombia in South America showed that although minor phenotypic differences existed between them, these parasites are antigenically related and belong to a single serodeme. Characterization by isoenzyme assay, karyotyping and DNA probe analysis, showed the Colombian isolates to be more similar to the West African than to Kenyan T. vivax. There was, however, little serological cross-reactivity between South American and African groups of T. vivax. Although the T. vivax isolates from Colombia were pathogenic for dairy calves which showed the typical sign of progressive emaciation, these parasites failed to infect mice or tsetseand could not be cultivated as bloodstream forms in vitro.This study represents initial attempts to establish the phenotypic and serological diversity amongst T. vivax isolates from South America.

Variation in the immunogenicity of 3 isolates of Trichinella spiralis was assessed by the parameters of adult worm recovery, mast cell, eosinophil and antibody responses in mice of defined response phenotype. The levels of the protective, inflammatory and immune responses induced by infection differed between the isolates. Isolates showed considerable variation in the capacity to elicit mast cell and eosinophil responses. All induced increases in parasite-specific antibody, levels of total (IgGAM) antibody and of IgM and IgG isotypes rose steadily after infection, but there were significant differences in levels of response. The IgGAM response was correlated with the number of worms present, i.e. the greatest response was seen in low responder (C57BL/10) mice infected with the longest-surviving isolates. All isolates elicited specific IgG1 and IgG2a antibodies after infection, although, again, there were isolate-specific differences in the levels and kinetics of response. Levels of these isotypes were always higher, although not significantly so, in high-responder NIH mice. Low-responder mice showed higher IgE serum levels than high-responder mice after infection, one isolate giving much higher IgE values than the other two.

Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogeneous parasites infective to livestock and other mammalian hosts; three different genotypes of this parasite have been described previously. Restriction enzyme fragment length polymorphisms (RFLPs) in both kinetoplast DNA minicircle and nuclear DNA sequences, and randomly amplified polymorphic deoxyribonucleic acid (RAPD) patterns have been used here to demonstrate the existence of another type of T. (N.) congolense that is genotypically distinct from those that have so far been characterized at the molecular level. A highly repetitive, tandemly arranged DNA sequence and oligonucleotide primers, for use in polymerase chain reaction (PCR) amplification are described, which can be used for specific identification of the trypanosome and its distinction from others within the Nannomonas subgenus.

Three morphologically distinct types of muscle fibres isolated from the platyhelminth Schistosoma mansoni have been studied with whole cell current- and voltage-clamp techniques. Fibres showed a marked time-dependent decrease in membrane resistance in response to depolarizing current injections. Voltage-clamp experiments revealed the presence of two distinct voltage-gated outward currents. The most prominent current is a slowly activating, slowly and incompletely inactivating potassium current similar to delayed rectifier currents which have been described in a variety of cell types from a variety of organisms. Also present is a faster activating, quickly and completely inactivating potassium current that shares functional characteristics with ‘A’-currents. All three of the cell types studied possess a delayed rectifier current, but only two of the three types have ‘A’-currents. Though depolarization with high K+ leads to contraction of the dispersed fibres, no voltage-gated inward currents could be detected by whole cell voltage-clamp under any of our conditions.

The effect of experimental non-host snail (NHS) communities on the infection of Lymnaea truncatula with Paramphistomum daubneyi and Fasciola hepatica miracidia was studied. The results obtained indicated that NHS had variable effects on basic population parameters of host infection. Specifically, NHS were able to produce aggregation of sporocyst number/snail in the experimental host populations. Proposed hypotheses for decoy effect mechanisms failed to explain completely the experimental results. A new mechanism is suggested, based on a distance response of miracidia to snail chemo-attractants, to explain effects of NHS on miracidial host finding behaviour. It is assumed that miracidia have evolved the ability to discriminate among individual snail chemical attractants in a community already at a distance. This attribute of miracidial behaviour allows NHS to induce variations in the effectiveness of individual host localization by miracidia. The resulting aggregation of successful infections/host is assumed to be the specific component of snail decoy effect at the community level. It was also established that NHS affected trematode reproduction in the host inducing a compensatory increase in the intramolluscan population. Intraspecific competition among developmental stages is suggested to be the mechanism of this compensatory reaction.

Aerobic resistance of Trichomonas vaginalis to metronidazole was induced in vitro by anaerobic cultivation of drug-susceptible trichomonads with low concentrations of the drug (2–3 μg/ml) for 50 days. Minimal lethal concentrations (MLC) for metronidazole of the resistant derivatives were high in aerobic susceptibility assays (MLC = 216–261.5 μg/ml) but low in anaerobic assays (MLC = 4.2–6.3 μg/ml), surpassing MLC values of their parent strain approximately 50-fold and 3-fold under aerobiosis and anaerobiosis, respectively. Sensitivity to metronidazole under anaerobic conditions and activity of the hydrogenosomal enzyme pyruvate: ferredoxin oxidoreductase indicated that the resistance was of the aerobic type. Dependence of the resistance manifestation on O2 was further confirmed by susceptibility assays in vitro performed in defined gas mixtures of different oxygen content (1–20%). Five percent concentration of O2 proved to be the threshold required for resistance demonstration and the MLC values further increased with increasing O2 concentrations. The in vitro-induced resistance was also demonstrated in vivo by subcutaneous mouse assay. The dose of metronidazole needed to cure 50% of infected mice (DC50) was 223 mg/kg × 3 for resistant derivative MR-3a but 6.6 mg/kg × 3 only for its drug-susceptible parent strain. The metronidazole – resistant strains developed in this study correspond by their properties to drug-resistant T. vaginalis strains isolated from patients refractory to treatment, and promise to be a useful tool in the study of 5-nitroimidazole aerobic resistance.

As there is little evidence of pollution affecting the health of fish and shellfish on a global scale, this paper attempts to put into perspective the pollution/fish disease relationship by reviewing examples of studies and reports in the historic and current literature. Although there is no dispute that pollution can affect the health of aquatic organisms under laboratory conditions and may be responsible for the decline of populations of such animals in some inland waters and some estuaries, most of the evidence for pollution causing or increasing disease in fish in open waters is circumstantial. Historical data proves that almost all fish and shellfish diseases known today have been described since the end of the last century. However, it is also known that water pollution, especially in inland waters, has for the past 400-500 years been the result of urbanization and industrialization. This has resulted in some major rivers becoming devoid of or deficient in fish stocks. The concern that pollution may influence the health status of fish and shellfish stocks has increased over the past 20 years. Initial attention was paid to epidermal diseases, including fin-rot in demersal fish, and protozoan diseases in molluscs in the heavily polluted bays and estuaries in North America. As the interest in this subject spread, it became political, and often controversial, especially amongst the North Sea countries. The disagreements have largely been settled amongst scientists because international bodies, such as the International Council for Exploration of the Sea (ICES), established workshops to investigate sampling methods and disease-reporting techniques. Recommendations from those workshops have contributed to some form of standardization for field work and the subject, although largely subjective, has some objective approaches which are described. As there are variable, interacting biological and physical influences in the aquatic environment, it is difficult to establish the background prevalences of diseases in populations offish and shellfish. Examples of the influences of climatic changes are presented, and these show that short-term catastrophes can be directly related. However, a more long-term problem is water acidification resulting largely from anthropogenic activities. In parts of Scandinavia this has, and is, leading to decimation offish stocks in inland waters. In general, diseases in fish and shellfish are very localized, but there is concern amongst scientists that certain cancers, especially liver tumours, occurring in demersal fish inhabiting polluted estuarine and coastal waters, are related to the release of chemicals, e.g. hydrocarbons, pesticides and heavy metals. This subject is discussed in detail, with examples of the author's own studies in North Sea fish. It is concluded that cancers in fish are of extremely low prevalence, and only present in a very few species, and then only in the oldest animals. Though changes in disease pattern may well be an indication of adverse environmental effects, further research is necessary for conclusive evidence.

The epidemiology of trypanosome infections of Glossina pallidipes was studied at a riverine site in the Zambezi Valley, Zimbabwe for a period of 13 months. Over 9000 flies were captured using a single trap. These flies were dissected, screened for trypanosome infection, sexed, and aged using both wing fray and (for females) ovarian category indices. Midgut infections were identified to species using recently developed DNA probes. The overall prevalence of mature infections was 5·5%, comprising 3·1% Trypanosoma vivax-type and 2·4% T. congolense-type (which included very low prevalences of T. brucei, T. simiae and another Nannomonas species). The prevalence of infection increased with age. For T. vivax-type infections in flies aged by ovarian category this relationship could be described by a simple ‘catalytic’ model assuming a constant per capita rate of infection. For T. congolense-type infections this model tended to over-estimate prevalence in older age classes, implying that the rate of infection decreases with age, and/or that infected flies have higher mortality rates, and/or that a significant fraction of the population is resistant to infection. Prevalences of infection also varied between months. This variation was more marked for T. vivax-type infections and was negatively correlated with both temperature and rainfall. The shape of the age-prevalence relationship, however, did not vary significantly between months. These observations are not fully explained by variation in the age-structure of the tsetse population and are consistent with temporal variation in the rate of infection (rather than in the trypanosome developmental period or in effects of infection on fly mortality). Possible causes of this variation are discussed.

Stratospheric ozone depletion threatens to increase exposure to ultraviolet (UV) radiation which is known to be a factor in a number of diseases. There is little doubt that cumulative exposure to UV radiation is important in the aetiology of non-melanoma skin cancers. Evidence is also strong for a link with cutaneous malignant melanoma, although here it appears to be intermittent intense exposure that is most damaging. More controversial is the view that exposure to solar radiation is a significant factor in ocular damage, particularly in the formation of cataracts. Earlier studies pointing to such an effect have been criticized and alternative aetiological hypotheses have been proposed. However, other studies do show an effect of UV exposure on cortical cataract. Concern is also growing that UV may be capable of activating viruses and have immunological effects that might exacerbate infectious disease. Very worrying is the possibility that UV exposure can activate the human immunodeficiency virus which might accelerate the onset of AIDS. Any such health effects that have been observed in human populations are the result of exposure to existing, naturally occurring levels of UV radiation. There is, therefore, great concern about the possible exacerbation of these impacts as a result of increased exposure to UV radiation associated with stratospheric ozone depletion. However, any assessment of the nature and scale of such impacts on human health has to deal with several major problems and these are the focus of this paper. There are uncertainties about recent trends in stratospheric ozone and problems in the prediction of future changes. Following on from this are the difficulties of estimating what effects these changes will have on UV flux at ground level in populated areas. Further problems arise in the prediction of changes in biologically significant doses to humans which might be affected by changes in behaviour as well as by changes in the environment. Finally, the limitations of existing epidemiological knowledge of the effects of UV exposure are a constraint on our ability to predict what the health effects of any changed UV doses might be.

Teneral Glossina morsitans centralis were fed on the flanks of African buffalo, N'Dama or Boran cattle infected with Trypanosoma vivax IL 2337. The infected tsetse were maintained on goats and on day 25 after the infected feed, the surviving tsetse were dissected to determine the infection rates. The mean mature infection rates (% ± S.E.) in the tsetse fed on buffalo, N'Dama and Boran cattle were 34·3 ± 9·9, 33·7 ± 13·4 and 58·9 ± 7·1, respectively. Logistic regression analysis indicated that infection rates in the labrum and hypopharynx of the tsetse were significantly lower when fed on the infected buffalo or N'Dama than Boran cattle. Similarly, the risk of infection was significantly lower in male than female tsetse. When teneral G. m. centralis, G. pallidipes, G. p. gambiensis, G. brevipalpis and G. longipennis were fed simultaneously on either the buffalo cow, the N'Dama bull or the Boran steer infected with T. vivax IL 2337, the mature infection rates were higher in the two morsitans group than the two fusca group tsetse, whilst G. p. gambiensis was relatively refractory to the infection, irrespective of the host species on which they fed. Logistic regression analysis indicated that the infection rates in the labrum and hypopharynx were significantly different amongst the five tsetse species for each of the three infected host animals. Nevertheless, the trypanotolerant African buffalo and N'Dama cattle may serve as reservoirs of T. vivax infection as can trypanosusceptible Boran cattle.

The population dynamics of Sphaerospora dicentrarchi and S. testicularis, myxosporean parasites of sea bass (Dicentrarchus labrax), was studied in different groups of cultured and wild fish. S. dicentrarchi prevalence was higher for wild fish (100%) than for cultured ones (70.5%). Infection levels increased with host age and there were no significant sex differences in cultured fish. Among parasitized organs, the gall bladder and intestine registered the highest infection levels in wild and cultured fish. A progressive increase in infection levels of S. testicularis was observed in cultured males during the spawning season in four consecutive periods. The evolution of the percentage of the different parasitic stages is pointed out. The infection seems to be related to water temperature and gonadal maturation.

The feeding behaviour of parasitic 3rd-stage larvae (L3) of the hookworms Ancylostoma caninum, A. ceylanicum and Necator americanus was examined. Less than 11% of A. caninum L3 recovered from the small intestines of dogs infected orally were feeding at 4–48 h post-infection (p.i.). and none of the A. ceylanicum L3 recovered from the intestines of orally infected hamsters had resumed feeding. All L4 of both species recovered at 36 and 48 h p.i. had resumed feeding. On the other hand, approximately 16% of the A. ceylanicum L3 recovered from the skin of percutaneously infected hamsters at 18 h were feeding, and the percentage feeding increased to nearly 58% at 44 h p.i. Necator americanus L3 recovered from the skin of percutaneously infected neonatal hamsters resumed feeding at 6–12 h p.i. and reached 90–94% by 18 h. Feeding began to decline at 66 h, and reached 29% at 120 h p.i. This decrease was associated with the migration of larvae from the skin to the lungs. By 192 h p.i. over 95% of the larvae had reached the small intestine, and all had moulted to the L4. The results indicate that parasitic L3 resume feeding in the skin during percutaneous infections, and suggest that feeding by hookworm L3 correlates with the resumption of development.

Reinfection with hookworm (Necator americanus) following chemotherapy was studied over 2 years in a rural village in Madang Province, Papua New Guinea. The prevalence of hookworm infection had returned to pre-treatment levels after 2 years, and the geometric mean hookworm burden had returned to 58 % of the pre-treatment value. The rate of acquisition of adult worms was independent of host age, and was estimated as a geometric mean of 2·9–3·3 worms/host/year (arithmetic mean 7·9–8·9 worms/host/year). There was significant predisposition to hookworm infection; the strength of this predisposition did not vary significantly between age or sex classes.

A control programme of opisthorchiasis was carried out for three years in three villages of northeast Thailand in order to compare the effectiveness of two intervention measures. The intervention measures employed were to give praziquantel treatment to all infected people either once (village I) or twice (village II) per year with the integration of regular health education and sanitation improvement. A control village (village III) received no intervention during the study. Sampling of the population in these villages to estimate prevalence, intensity, reinfection and incidence of infection as well as knowledge of opisthorchiasis and number of latrines was performed at 6 monthly intervals throughout the duration of the study. At the initial assessment, there were no significant differences in the prevalence and intensity of infection of villages I and II. Both values were greatly reduced when assessed 6 months after the initiation of treatment, and the decrease in prevalence and intensity of infection did not differ between villages I and II. The control village (village III), during the first 2 years, exhibited slight variation in the prevalence and intensity, and these decreased considerably in the third year of the study. The reinfection rate per 6 months in villages I and II was nearly equal. The incidence rate per 6 months in the two treated villages was lower than that of the control village. Marked improvement in knowledge of opisthorchiasis, behavioural changes of eating raw fish and increased numbers of latrines was evident in all of the villages during this study. The study showed that the effectiveness of annual drug treatment is similar to that of 6 monthly treatments when combined with regular health education and sanitation improvement.

Only green plants can convert the single carbon units of atmospheric carbon dioxide into the multi-carbon organic molecules on which all forms of life depend. Only green plants can provide the oxygen required by man and other aerobic organisms. In addition to his basic need for preformed organic molecules and oxygen, man also depends on plants to provide him, directly or indirectly, with an array of specific compounds such as vitamins and essential amino acids. Inadequate supplies of these may hinder growth and development or give rise to well defined deficiency diseases. At the present time information concerning the distribution and concentrations of such essential nutrients in plants is largely restricted to those plants that are already used as human foods. Nothing or virtually nothing is known about the chemical composition of approximately 250000 wild and little-used species. Amongst these there may be many that could provide us with cheap and plentiful new sources of essential nutrients that could be of enormous benefit to those suffering not only from full-blown deficiency diseases but also suffering sub-normal health due to partial deficiencies. The destruction of much of the world's wilderness areas has already deprived us of the opportunity to evaluate the contributions that a great many plant species might have made towards the elimination of deficiency diseases.

Many plants used as human foods contain compounds that are toxic to man. If intake is sufficiently high, these toxins may cause disease. Breeding programmes designed to eliminate toxins from crops species or reduce their concentrations to acceptable levels depend on genetic variability within the species or the possibility of producing hybrids with the desired characteristics. The motor neurone disease, lathyrism, which affects populations in the Indian sub-continent, Africa and China is caused by a toxin in the seeds of Lathyrus sativus. Surveys of cultivated plant populations have shown great variability in toxin levels and such genetic differences make it possible to select and breed toxin-low varieties. The existence of toxin-free species within the same genus has led to research aimed at producing toxin-free hybrids suitable for agricultural use. Approaches, designed to reduce or eliminate diet-related diseases, depend on the maintenance of the greatest possible diversity among both wild and cultivated plant populations. Such diversity is under threat.

Some 250 plant species are used as sources of drugs in western medicine. Most of these drugs are obtained from plants whose therapeutic value was recognized long before the compounds were isolated. In the developing countries of the world, it is estimated that 25000 plant species may be used in medicinal preparations. Few of these plants have been studied systematically to determine whether their reputations are justified and if so, the nature of the drugs they contain. If only 0.1 % were to yield useful drugs, they could make a major contribution to human health and welfare. The plants and the indigenous populations who understand their uses are both disappearing and it is a matter of great regret that much that could be of value has been and will be lost.

The excreted/secreted proteinases of adult and juvenile Fasciola hepatica maintained in vitro were found to hydrolyse the fluorogenic substrates Cbz-Phe-Arg- and Cbz-Arg-Arg-NHMec. This activity was demonstrated to have a classical cysteine proteinase inhibitor profile, with turn-over of both substrates being blocked by pre-incubation with E64 and peptidyl diazomethanes. The Cbz-Arg-Arg-NHMec hydrolysing activity of the mature fluke exhibited an alkaline stability not characteristic of its mammalian lysosomal counterparts. Further, the biotinylated affinity reagents biotin-Phe-Ala- CHN2 and biotin-Phe-Cys(SBzyl)-CHN2 were used to label and characterize these cysteine proteinases in terms of apparent molecular weight and subsite specificity. Adult fluke media were found to contain four species of molecular weights 66, 58, 50 and 25–26 kDa; juvenile media contained three species of molecular weights 66, 54 and 25–26 kDa. The major 25–26 kDa cysteine proteinase common to both stages was shown to have a subsite specificity similar to that of mammalian cathepsin B.

Adults of the porocephalid pentastomid Porocephalus crotali infect the lung of rattlesnake definitive hosts and larvae develop in rat intermediate hosts. In the latter, nymphs encyst within a variety of tissue sites (commonly abdominal fat bodies and lungs) and each becomes the focus of an eosinophilic granuloma. From an early stage in infections, granulomas become increasingly infiltrated by mast cells which, using conventional histology and paired immunofluorescence against mast cell proteases, appear to be exclusively of the mucosal phenotype. Mucosal mast cells are concentrated along the dorsal region of the parasite and in a plug of tissue containing degenerating cuticles within independent granulomas, which is located between its head and tail. ELISAs against the rat mast cell proteases I and II (RMCP I and II), extracted from abdominal fat, lung, spleen, liver and kidney granulomas at various intervals post-infection, reveal a substantially elevated concentration of RMCP II in all lesions. In fat, concentrations increase up to about 100 days post-infection, at which time moulting ceases and inflammatory responses subside. RMCP II was scarcely detectable in matched control tissues. Unlike infections with certain nematode parasites, where enteric mucosal mast cells secrete RMCP II systemically, concentrations of RMCP II in the serum of infected rats were significantly reduced when compared with age-matched uninfected controls. These results confirm that P. crotali can selectively recruit mucosal mast cells to a variety of tissue sites, most of which are non-mucosal. Possible mechanisms are discussed.