Observations on the larvae of sheep trichostrongyles on hill pastures showed:

1. A reduction in larval populations occurred when the ‘moving-flock’ type management was used. The large proportion of eggs passed in faeces is deposited on a part of the pasture where little or no grazing occurs.

2. Local concentrations of larvae occurred on the grazed portions of a pasture when grazing was uneven. Thus, the apparent low rate of stocking on hill pastures does not give a true picture of the potential dangers of parasitic disease. Estimates of pasture contamination by Taylor's method of sampling were deceptively low.

3. The number of larvae on grass blades was lowest when the grass was wet with dew. Sampling of pastures between 12 noon and 5 p.m g.m.t. gave maximal and most constant estimates.

4. There was an increase in the number of larvae on a pasture during the warmer months of the year. While low temperatures prevent or delay hatching, the increase in population was not entirely due to the increased rate of development when temperature was high. The increase in numbers was partly due to an increase in the number of adult worms in the sheep hosts.

5. Removal of sheep for 3 weeks reduced the number of larvae on a pasture. This reduction was not apparent, however, until after the first 12 days. After 3 weeks the pasture population was reduced by 55 %, while after 4 weeks (i.e. 7 days after the return of the sheep) the reduction was 90%. Two weeks later there was a small increase in the larval population, but this was again reduced with the onset of colder conditions.

Ixodid female ticks take one comparatively large bloodmeal which they convert to a single large egg mass and then they die. To examine the outcome of interrupted feeding, equal numbers of male and female Rhipicephalus appendiculatus adult ticks were fed on guinea pigs (host 1) for either 2, 4, or 6 days, or to engorgement (8 days). All of the fully engorged (D8) females laid a single large egg mass (80–160 mg/tick), while 85% of the day 6-fed (D6) female ticks (n=20) each laid a small egg mass (6·1 mg/tick). None of the females that had fed for 2 or 4 days oviposited. Ninety percent (n=20) of the day 2-fed (D2) females survived for 4 weeks after their feeding was interrupted, whereas 65% (n=20) of the day 4-fed (D4) females survived. All of the surviving partially fed female ticks (D2 and D4) attached to a second guinea pig (host 2) and attained engorged body weights that were not significantly different from those of the control females (P<0·05). Female ticks that engorged following interrupted feeding layed egg masses comparable to the controls, indicating that engorgement on host 2 was successful. The salivary gland protein profile of female ticks changed constantly during feeding. However, when feeding was interrupted, the protein expression pattern switched back to that of the non-parasitic state, presumably to enable the partially fed ticks to survive and reattach on the new host. This observation indicates that female ixodid ticks have a natural ability to survive and re-establish successful feeding on a new host if the first attempt at feeding is unsuccessful. Such an interrupted feeding mechanism supports the hypothesis that partially engorged ticks may play a role in tick-borne pathogen transmission.

The effect of tumour necrosis factor-α on malaria-infected mice was studied. C57B1/6J mice infected with Plasmodium berghei K173 exhibited an increased sensitivity to exogenous TNF. Injection of 15 μg TNF was lethal to some of the animals when given 5–7 days after infection, while when given later on in the infection (i.e. days 8–10) amounts as low as 2·5 μg TNF appeared to be lethal in all mice. The pathology in infected mice treated with TNF resembled that found in the brains of infected mice dying with cerebral malaria. Infected mice treated with TNF, however, also developed severe pathological changes in other organs. On the contrary, treatment with sublethal amounts of TNF (1·0 μg or less) given on days 8 and 9 after infection, protected mice against the development of cerebral malaria. In addition, infected mice exhibited an enhanced sensitivity for treatment with lipopolysaccharide (LPS). Sublethal amounts of LPS, however, did not prevent mortality as in TNF-treated mice (LPS-treated mice died at about the same time as infected mice that developed cerebral malaria), but no cerebral haemorrhages were found in the majority of LPS treated, infected animals. Treatment with dexamethasone during infection protected mice against the development of cerebral malaria, but did not suppress their increased sensitivity to exogenous TNF. Treatment of mice with liposome-encapsulated dichloromethylene diphosphonate (lip-Cl2MDP), used to eliminate macrophages (an important source of TNF), prevented the development of cerebral malaria, but only when given before day 5 of infection. Mice protected by treatment with lip Cl2MDP, however, remained sensitive for LPS on the eighth day of infection.

We describe a new freshwater myxosporean species Ceratomyxa gracillima n. sp. from the gall bladder of the Amazonian catfish Brachyplatystoma rousseauxii; the first myxozoan recorded in this host. The new Ceratomyxa was described on the basis of its host, myxospore morphometry, ssrDNA and internal transcribed spacer region (ITS-1) sequences. Infected fish were sampled from geographically distant localities: the Tapajós River, Pará State, the Amazon River, Amapá State and the Solimões River, Amazonas State. Immature and mature plasmodia were slender, tapered at both ends, and exhibited vermiform motility. The ribosomal sequences from parasite isolates from the three localities were identical, and distinct from all other Ceratomyxa sequences. No population-level genetic variation was observed, even in the typically more variable ITS-1 region. This absence of genetic variation in widely separated parasite samples suggests high gene flow as a result of panmixia in the parasite populations. Maximum likelihood and maximum parsimony analyses placed C. gracillima n. sp. sister to Ceratomyxa vermiformis in a subclade together with Ceratomyxa brasiliensis and Ceratomyxa amazonensis, all of which have Amazonian hosts. This subclade, together with other Ceratomyxa from freshwater hosts, formed an apparently early diverging lineage. The Amazonian freshwater Ceratomyxa species may represent a radiation that originated during marine incursions into the Amazon basin that introduced an ancestral lineage in the late Oligocene or early Miocene.

These studies were aimed at giving a clearer idea of the source of the antigens responsible for the induction of resistance by larval schistosomes. Firstly, the resistance induced by radiation-attenuated (20 krad.) infections was compared with that induced by the larval phase of unirradiated infections. The superiority of the irradiated larvae was demonstrated by infecting mice with 500 female cercariae which were either unirradiated or irradiated, and challenging them 3 weeks later with male cercariae. At perfusion, when primary and challenge worms could be distinguished by sex, significantly greater resistance was demonstrated in the groups vaccinated with irradiated larvae (56 cf. 31% and 48 cf. 26% in two experiments). Secondly, in order to establish how long immunizing larvae need to survive to stimulate optimum resistance, mice were either treated with schistosomulicidal drugs or had the site of infection excised at various intervals post-vaccination. Treatment or excision at up to 24 h did not result in significant resistance to challenge (6–16%). Similar treatments at 1 week did not prevent significant resistance developing (26–37%) but the levels of resistance were significantly lower than those manifest by vaccinated, untreated mice (51–71%). It is concluded that the immunizing larvae need to persist for between 1 and 2 weeks in order to stimulate optimum protection and that antigens released during protracted survival and/or death of the larvae in either the skin or the lungs are probably key factors in the induction of resistance.

In sub-Saharan Africa, over 22 million people are estimated to be co-infected with both helminths and HIV-1. Several studies have suggested that de-worming individuals with HIV-1 may delay HIV-1 disease progression, and that the benefit of de-worming may vary by individual helminth species. We conducted a systematic review and meta-analysis of the published literature to determine the effect of treatment of individual helminth infections on markers of HIV-1 progression (CD4 count and HIV viral load). There was a trend towards an association between treatment for Schistosoma mansoni and a decrease in HIV viral load (Weighted mean difference (WMD)=−0·10; 95% Confidence interval (CI): −0·24, 0·03), although this association was not seen for Ascaris lumbricoides, hookworm or Trichuris trichiura. Treatment of A. lumbricoides, S. mansoni, hookworm or T. trichiura was not associated with a change in CD4 count. While pooled data from randomized trials suggested clinical benefit of de-worming for individual helminth species, these effects decreased when observational data were included in the pooled analysis. While further trials are needed to confirm the role of anthelmintic treatment in HIV-1 co-infected individuals, providing anthelmintics to individuals with HIV-1 may be a safe, inexpensive and practical intervention to slow progression of HIV-1.

The acanthocephalan parasite Acanthocephalus dirus induces a colour change in the intermediate host, the aquatic isopod Caecidotea intermedius, which increases transmission to definitive hosts (creek chub, sunfish). We examined the potential for conflict to occur between infective (cystacanth) and non-infective (acanthor, acanthella) stages of A. dirus over the level of colour modification that should be induced when these stages share a host. Using a field survey, we showed that host sharing by infective and non-infective stages was relatively common and that infective and non-infective stages differed in their effects on colour modification. Non-infective stages induced a colour change over 40% of the body, whereas infective stages induced a colour change over 80%. Thus, conflict could occur between stages over the level of modification that should be induced. We then showed that mixed-stage infections induced a colour change in the host that was consistent with the level of modification induced by the infective stage. We discuss the potential significance of these results to patterns of host modification and their effects on stage-related survival in nature.

This paper describes the structure of the microfilarial reservoir of Loa loa in an endemic population of central Cameroon. The possible effects of age and sex on the prevalence and intensity of microfilaraemia have been explored. Logistic analysis showed that the prevalence of microfilaraemia increased significantly with age, reaching 60% in the oldest males. This result suggests that the figure commonly reported, according to which only one third of the infected individuals were microfilaraemic, should be reconsidered; in addition, as part of surveys of loiasis, crude microfilaraemia prevalence values should be replaced by adjusted ones. The intensity of infection did not show any age-specific change. As a result, even if the oldest members of the male population are clearly the most at risk of developing post-ivermectin serious adverse reactions, especially Loa-encephalopathy, the other members of the population are not risk-free. Therefore, in those areas where the African Programme for Onchocerciasis Control is undertaking regular mass distributions of ivermectin for onchocerciasis control, and where loiasis is co-endemic, no subpopulation should be excluded from surveillance and monitoring during community directed treatments with ivermectin.

Myxobolus cerebralis is the causative agent of whirling disease that has significant economical and ecological impacts on trout populations. Although intensive studies have been conducted to understand its effects on and interactions with its fish host, only limited information is available about how and to what extent M. cerebralis affects its oligochaete host, Tubifex tubifex. We investigated the effects of M. cerebralis on survival, growth, reproduction, and feeding activity of T. tubifex. Mature, immature and juvenile worms were exposed to myxospores and their infection prevalence, mortality, sexual development, reproduction and spore production were compared with unexposed worms. The parasite affected neither survival nor growth but inhibited clitellar development and reduced cocoon production by over 80%. Numbers of actinospores released from mature worms were nearly 9-fold higher than that of immature worms. When non-clitellated infected worms were kept at 30°C for 4 days, spore release ceased and they re-developed a clitellum. These results suggest parasite-induced castration. Comparative monitoring of defecation rate revealed that M. cerebralis reduced feeding activity of T. tubifex by approximately 40%. Low energy intake and impaired energetic allocation may be the underlying mechanism behind reduced fecundity of infected T. tubifex.

Brain cysts from three strains of Toxoplasma gondii were purified and analysed by SDS-PAGE and immunoblotting. Western blot analysis revealed the presence of major cyst/bradyzoite antigens at Mr 25000, 36000 and 67000 common to all strains and recognized by many individual sera. There was also evidence of strain variation in cyst antigens with, for example, antigens at Mr 88000, 28000 and 26000 unique to the 17025 strain. There was some overlap in the molecular weight of cyst and tachyzoite antigens, but the major tachyzoite surface antigen (P30) could not be detected in brain cysts.

Two recombinant Toxoplasma gondii polypeptides, H4 and H11, were tested as diagnostic antigens in enzyme-linked immunosorbent assays (ELISAs). The results obtained by ELISAs based on single H4 (H4-ELISA), on single H11 (H11-ELISA) and on a mixture of H4 and H11 (H4/H11-ELISA) were compared with results obtained by an ELISA based on traditional ELISA antigen (TEA-ELISA), an indirect fluorescent antibody test (IFAT), the Sabin-Feldman dye test (SFDT) and a direct agglutination test (DAT). A total of 306 cats from a suburban cat population were tested of which about 45% showed serological evidence of T. gondii infection. Infection rates varied from about 32% for cats kept indoors to about 55% for stray cats. Specificities > 99% were observed for all ELISAs based on the recombinant antigens (H4-ELISA, H11-ELISA and H4/H11-ELISA). The H4/H11-ELISA also reached a sensitivity of 95% which compared very favourably with those observed for the TEA-ELISA (98%) and for the IFAT (94%). Negative and positive predictive values for the H4/H11-ELISA were 96 and < 99%, respectively. Antibody titres measured by the H4/H11-ELISA also correlated well with those measured by the SFDT and the DAT. Hence, the H4/H11-ELISA appears to be a very suitable test for sero-epidemiological studies on T. gondii infections in cats.

The localization of the 28 kDa Schistosoma mansoni glutathione S-transferase (Sm28 GST) has been investigated using immunohistochemistry and electron microscopy and the results compared with previously published data. This study confirms the wide distribution of this antigen in the parasite. In male and female worms, Sm28 GST is localized in the tegument, the parenchyma, the oesophageal epithelium and in genital organs. Sm28 GST was clearly detected in germinal and sustentacular cells. The decrease of staining intensity during the differentiation of germinal cells suggests a down-regulated expression of the molecule. At the ultrastructural level, this antigen was abundant in nuclei and less present in the cytoplasm. The marked heterogeneity observed in the staining of individual worms indicates that Sm28 GST seems to be closely associated with the parasite’s metabolism. The results are discussed in relation to the biological and protective functions of the protein.

In 1915 Glaser demonstrated for the first time that a nuclear polyhedrosis of a lepidopterous larva (Lymantria dispar), was induced by an invisible filterable virus isolated from the blood of diseased larvae. Nuclear polyhedroses of the larvae of over a 100 insect species are now known, and those of them which have been investigated have been shown to be due to viruses. In these polyhedroses the nuclei of susceptible cells, hypodermis, tracheae, fat and blood cells, etc., enlarge and become packed with infectious crystalline inclusion bodies, the polyhedra, from which the diseases get their name. The pupae and imagos of these insects are not generally susceptible to polyhedral virus diseases, and so far no nuclear polyhedrosis of imaginal tissues in the larvae has been reported. This has led to the suggestion that imaginal tissues are not susceptible to polyhedral virus diseases.

Acid proteases hydrolysing haemoglobin, first described by Timms & Bueding (1959), were studied to investigate the interspecific and intraspecific distribution in adult parasitic helminths. Acid protease activity was detected in a variety of adult helminths, irrespective of their habitats, namely, parasitic nematodes, Angiostrongylus cantonensis, Dirofilaria immitis, Trichuris muris and Ascaris suum, a trematode, Paramphistomum sp. and cestodes, Diphyllobothrium erinacei and Hymenolepis nana. Much higher specific activity was observed in the blood nematodes, A. cantonensis and D. immitis than in the gastro-intestinal helminths examined. The intraspecific distribution of the acid protease activity within organs was studied in those nematodes from which the organs could be anatomically separated. The highest specific activity was observed in the intestine of A. cantonensis, D. immitis and A. suum. The activity was also found in other organs of these nematodes, namely, the reproductive organs and the body wall, but to a much lesser extent. The pH optima for the hydrolysis of haemoglobin were found to be in the pH range of 3·1 to 4·6. Each of the enzyme solutions from the 7 species of helminths hydrolysed haemoglobin faster than the other proteins examined but some activity on myoglobin, casein, albumin and other proteins was demonstrated. Pepstatin, a potent inhibitor of carboxyl proteases, inhibited the hydrolysis of haemoglobin by the acid proteases of the 7 species of the parasites. Acid protease activity from A. cantonensis and A. suum was not inhibited in the presence of thiol-, serine- and metallo-protease inhibitors at a concentration of 10−6 M. This fact, together with the inhibitory effect of pepstatin, suggests that the enzyme activity is likely to be due to carboxyl protease(s). Acid proteases such as cathepsin D and pepsin were simultaneously used for comparison of some enzymological characteristics. The substrate specificity of the enzyme solutions from the helminths seemed to resemble that of cathepsin D rather than that of pepsin. The effects of pH and pepstatin on the acid protease activity of the helminths were similar to those on pepsin and cathepsin D.

1. A qualitative paper chromatographic analysis of the free amino acids in Rhodnius urine and excreta was made: eleven amino acids or related compounds were detected, cystine and cysteic acid in urine, and histidine, histamine, leucine, taurine, glycine, valine, phenylalanine and alanine in the excretory fractions.

2. In addition to these, three yellow pigment spots were separated from the excreta; one of these is of unknown composition and two are peptides.

3. A possible relationship between certain of the excretory amino acids and haemolymph ones is discussed.

I am greatly indebted to the late Professor P. A. Buxton, C.M.G., F.R.S., for his stimulating criticism during this study, and to the members of the School for very useful discussion, and to Mr Paul Baily for performing the column chromatographic analyses on my behalf. The study forms part of a Thesis for the degree of Ph.D. approved by the University of London, and was financed by the South African Council for Scientific and Industrial Research. I wish to acknowledge their generous financial support.

Numerical taxonomy was used to review isoenzyme variation in isolates of Trypanosoma brucei obtained from cattle, tsetse, humans and wildlife from the Lambwe Valley, Kenya. From isoenzyme information alone, it was possible to classify isolates as to source through the use of linear discriminant functions analysis, with an error rate of only 2% in humans, and 14% over all groups. Differentiation was mostly dependent on patterns in the enzymes ASAT, PEPI, and ICD. Parasites from non-human sources, especially tsetse, were characterized by high isoenzyme diversity, and many unique zymodemes. Observed frequencies of genotypes for ICD, ALAT, and ASAT did not agree with expected frequencies based on random mating of a diploid organism. Deviations were particularly large for tsetse isolates, and were mostly due to a deficiency of one homozygote. Cluster analysis revealed complex relationships among isolates, with patterns evolving through time. Major human zymodemes from the 1970s clustered together with most wildlife isolates from East Africa. This chronic human-wildlife transmission cycle was characterized by ASAT pattern I. Other, minor human zymodemes were associated with a human-cattle transmission cycle characterized by ASAT pattern VII. These original chronic transmission cycles appeared to change in 1980 with the appearance of two new zymodemes in humans. These zymodemes involved changes in ALAT and/or PGM to patterns typical of tsetse and cattle isolates. A resultant epidemic was halted with repeated aerial spraying of endosulfan in 1981. Since then, a variety of new zymodemes of unknown human infectivity have appeared. The origins of these changes are discussed in terms of genetic exchange in tsetse, adaptation to human and cattle transmission cycles, and selection resulting from chronic use of insecticides.

The structure and development of the spermatozoon of Dipetalonema viteae has been studied by means of electron microscopy.Spermatogonia are developed from a syncytium in the terminal region of the reproductive tract. The syncytium grows along the length of the testis as an anucleate rachis, carrying with it the developing germ cells. The gametes become detached from the rachis when they have become secondary spermatocytes. The chromosomes which appear in the primary spermatocytes at the onset of meiosis persist throughout all subsequent stages of development. The nucleus is not reconstructed. Cytophores are produced by the spermatids at the end of the second meiotic division. The spermatid is an elongated cell, but the mature spermatozoon, within the male tract, is amoeboid. There are only minor differences between the sperm found in the male and female tracts. The male gametes contain complex membraneous organelles which are developed from the Golgi bodies and endoplasmic reticulum of the primary spermatocytes. These organelles are suggested to have similar origins and functions to the acrosome of the typical mammalian spermatozoon.

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.

Epidemiological rate parameters of host generalist parasites are difficult to estimate, especially in cases where variation in parasite performance can be attributed to host species. Such cases are likely common for generalist parasites of sympatric grazing mammals. In this study, we combined data from experimental exposures in cattle and sheep and natural infections in elk to compare the recruitment, morphology and reproduction of adult Dicrocoelium dendriticum, a generalist trematode that has emerged in sympatric grazing hosts in Cypress Hills Provincial Park, Alberta. Overall, there were no significant differences in the recruitment of metacercariae and in the pre-patency period of adults in experimentally exposed cattle and sheep. All flukes reached reproductive maturity and the degree of reproductive inequality between individual flukes within each infrapopulation was moderate and approximately equal among the three host species. Neither fluke size nor per capita fecundity was constrained by density dependence. Thus, fitness parameters associated with growth and reproduction were approximately equivalent among at least three species of definitive host, two of which are sympatric on pastures in this Park. The generalist life-history strategy of this trematode, which is known to extend to other stages of its life cycle, has likely contributed to its invasion history outside its native range in Europe.

Strong resistance to reinfection developed in pigs as a result of trickle inoculations with Trichuris suis (250 infective eggs twice weekly for 4 weeks). At challenge (4000 eggs) 8 weeks after the last trickle inoculation, we observed a 96% reduction in the mean number of established challenge worms in immunized (TRI/CHA) compared with non-immunized controls (CON/CHA) (74 v. 1753; P = 0·004). Moreover, the total faecal egg excretion was significantly lower in immunized pigs compared with controls (P = 0·0003). The trickle inoculations depressed pig weight gains over the 8-week course of immunization (P = 0·042). It was observed that a small number of pigs developed a weaker level of resistance, reflecting the importance of genetic determinants. The relevance of these data to the population biology of porcine and human whipworm infections is discussed.