Prototheca richardsi, an unpigmented heterotrophic alga, causes growth inhibition in amphibian larvae and has proved refractory to culture in Vitro. P. richardsi replication is dependent on regular passaging through tadpole digestive systems; uptake of thymidine by free-living Prototheca cells and incorporation into DNA are very low by comparison with leucine uptake and incorporation into protein, but DNA synthesis is detectable in cells isolated from tadpole intestines. DNA replication was elicited 6–8 h after ingestion in protothecans fed to tadpoles and subsequently re-isolated from them, providing that the tadpoles were fed subsequent to the ingestion. It appears that passaging through tadpole intestines provides an essential stimulus to maintaining an active cell division cycle in P. richardsi.

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.

A DNA polymerase activity has been detected and characterized in crude extracts from tachzoites of Toxoplasma gondii. The enzyme has a sedimentation coefficient of 6·4 S, corresponding to an approximate molecular weight of 150000 assuming a globular shape. Like mammalian DNA polymerase α, the DNA polymerase of T. gondii was sensitive to N-ethylmaleimide and inhibited by high ionic strength. However, the enzyme activity was not inhibited by aphidicolin which is an inhibitor of mammalian DNA polymerases α, δ and ε and also cytosine-β-D-arabinofuranoside-5′-triphosphate which is an inhibitor of α polymerase. The activity was inhibited by 2′,3′-dideoxythymidine-5′-triphosphate which is an inhibitor of mammalian DNA polymerase β and γ. Magnesium ions (Mg2+) were absolutely required for activity and its optimal concentration was 6 mM. The optimum potassium (K+) concentration was 50 mM and a higher concentration of K+ markedly inhibited the activity. Activity was optimal at pH 8. Monoclonal antibodies against human DNA polymerase did not bind to DNA polymerase of T. gondii. Thus the T. gondii enzyme differs from the human enzymes and may be a useful target for the design of toxoplasmacidal drugs.

Rickettsia-like organisms (RLO) from tsetse midguts and mosquito cell cultures showed high levels of endochitinase activity. A line of Glossina morsitans morsitans highly susceptible to midgut trypanosome infection and with high incidence of RLO infection showed significantly greater chitinolytic activity than G. austeni which had low RLO incidence and were correspondingly refractory to midgut infection. Midgut infection rates of Trypanosoma brucei rhodesiense in G. m. morsitans showed a dose-related increase when flies were fed N-acetyl-D-glucosamine (GlcNAc) in the infective meal and for 4 subsequent days. A model is proposed for susceptibility to trypanosome infection based on the generation of GlcNAc by RLO endochitinase activity in tsetse pupae inhibiting midgut lectin in teneral flies.

An entire 18S rRNA gene sequence from Schistosoma spindale (1990 bases) and partial 18S rRNA gene sequences from S. haematobium (1950 bases) and S. japonicum (1648 bases) have been determined. Together with the previously published sequence for the S. mansoni 18S rRNA gene, these data encompass the 4 recognized Schistosoma species groups. Although Schistosoma 18S rRNA genes are highly conserved, the sequences permit a preliminary molecular phylogeny to be established for the genus. This identifies S. haematobium and S. spindale as sister taxa in a clade with S. mansoni. S. japonicum does not appear to be closely related to this clade. Much of the observed variation occurs within a ‘hypervariable’ stretch of the gene corresponding to part of the V4 region of 18S rRNA. Despite this variation, the 3 new sequences fit models of 18S rRNA secondary structure predicted from the S. mansoni sequence.

The sequences reported in this paper have been submitted to the EMBL Database under accession numbers Z11976 (S. haematobium), Z11590 (S. japonicum) and Z11979 (S. spindale).

The complete small subunit ribosomal RNA (srRNA) gene of Theileria parva was cloned and sequenced. Two primers were designed which permitted the specific amplification of part of the Theileria srRNA gene from Theileria-infected cell line samples which were predominantly (> 95%) bovine DNA. The sequence of the central (variable) region of the srRNA genes of T. annulata, T. taurotragi, T. mutans and two unidentified parasites referred to as Theileria sp. (buffalo) and Theileria sp. (Marula) were obtained. An alignment of the sequences was generated from which 6 oligonucleotide probes, corresponding to species-specific regions, were designed. These probes were demonstrated to provide unequivocal identification of each of the 6 species either by direct detection of parasite srRNA or by hybridization to amplified parasite srRNA genes. The probes were not able to distinguish buffalo-derived T. parva, the causal agent of Corridor disease, from cattle-derived T. parva, the causal agent of East Coast fever.

A descriptive epidemiological survey was undertaken of the ascarid nematode Toxocara canis in 521 red foxes (vulpes, vulpes) during the period January 1986 to July 1990. Age–prevalence and age–intensity profiles show that worm are significantly higher in cubs than in subadult or adult foxes and higher in subadult than in adult foxes. variations in worm burdens occur, with the highest prevalences and intensities being found during the spring, when are born, and in the summer months. Prevalences and intensities then decrease during the autumn and winter months both subadult and adult foxes, but, during this period, prevalences are significantly higher in male than in female Variations in worm burdens in the fox population are likely to be related to the reproductive cycle of the fox, with proportion of cubs becoming infected in utero. The role of the fox in the transmission of T. canis in the urban environment is discussed.

The use of a selective schedule of tests to identify a viable population of isolated adult Onchocerca volvulus (Nematoda: Filarioidea) has been investigated in a large worm population. The study was initiated to develop methodology appropriate to test new candidate macrofilaricides for their in vitro activity against O. volvulus. After removal from the host the viability of isolated intact parasites was estimated by assessing the motility indices of male worms, and the colorimetric quantification of the reduction of the bioreducible tetrazolium reagent XTT and lactate output by female worms. Additionally the motility of whole females and the movement of inner organs of female worms were scored quantitatively. These response parameters were used to sort the adult worms into viability groups at the start of the in vitro culture. The adult worms were then observed for 6 days and viability was assessed regularly during the culture period. At the end of the culture period, the reduction of the water-insoluble tetrazolium reagent MTT was used to determine the formazan formed by the entire male and female worms. The response parameters used at the start of the culture proved to be highly predictive for detecting viable and non-viable adult worms. In the group of worms selected as ‘viable’ around 70% kept their motility and metabolic activity at a high level until the end of the culture compared to the initial level. In contrast, none of the female worms and only 13% of the male worms categorized as ‘poorly viable’ demonstrated a motility index or metabolic level at the end of the culture period that was comparable to that of the worms in the ‘viable’ groups. For female worms the lactate output correlated significantly with weight whereas no correlation was seen between MTT reduction and weight.

Unrelated and initially unfamiliar male CFLP mice, maintained for different periods in groups of 6, differed in both their rate of clearance of Babesia microti and the time taken to reach peak parasitaemia in relation to their aggressive behaviour within groups prior to infection. Males maintained in groups for shorter periods and showing more aggression within their group were slower to clear infection and males showing more marked external evidence of aggressive interaction reached a peak of parasitaemia sooner. Serum IgG and corticosterone analyses were consistent with increased aggression causing stress-induced immunodepression but relationships with aggression and social status were not simple. Males showing more aggression tended to enter their groups with higher levels of corticosterone and, to a lesser extent, reduced levels of IgG compared with other mice. The results thus suggest that increased susceptibility to disease may be a cost to males aggressively maintaining high social status.

Comparisons were made between the protein concentrations and protein profiles of uninfected Gammarus pulex and those infected with cystacanths of Pomphorhynchus laevis. Comparisons of the extractable haemolymph volumes from uninfected and infected G. pulex, when related to host tissue weight, showed that there was no difference in the host tissue/haemolymph volume ratio between infected and uninfected individuals. SDS-PAGE of haemolymph samples resolved 36 bands of protein, the major components being band 6(19 kDa) and 21(33 kDa). Densitometric analysis of haemolymph samples showed that infected gammarids had total protein concentrations elevated by 84%. The (88 kDa) subunit was identified as the monomer of the respiratory protein haemocyanin. This band was significantly elevated in infected individuals (one-way ANOVA P < 0.001); increasing from 30 to 45% of the total haemolymph proteins.

Trichostrongylus tenuis eggs were counted in faeces from individually marked wild red grouse for 8 years. Egg counts varied seasonally and annually. In some years, a sudden increase in mid-April was consistent with delayed maturation of larvae which had overwintered in the birds in a hypobiotic state. A more gradual increase in summer was probably due to uninterrupted maturation of larvae ingested then. Despite 30-fold year-to-year variation in mean egg counts, relative differences in egg counts among known individuals within years tended to persist across years. Rainfall in previous summers explained much of the year-to-year variation in egg counts, probably because parasite recruitment was greatest during wet summers. Grouse density was only weakly related to worm egg counts. The data were not consistent with the hypothesis that the cyclic-type population fluctuation in red grouse numbers observed at the time of this study was caused by the parasites.

Hatching of eggs of Anguillicola crassus was studied at temperatures between 5 and 30°C. Hatching occurred between 10 and 30°C and the incubation time was inversely related to temperature. Survival, spontaneous and excitatory rate of activity of 2nd-stage larvae of A. crassus were also affected by temperature. The percentage of larvae eaten by Paracyclops fimbriatus was not always influenced by the activity of the larvae. Penetration efficiency into the haemocoel of the intermediate host was highest for freshly hatched larvae and declined gradually with age. Completely non-active larvae were still able to penetrate, although penetration efficiency was lower.