Associations between social rank, immunodepression and resistance to Babesia microti infection within single-sex groups of male house mice suggest rank-dependent suites of response involving different hormonal and immune changes in relation to aggressive behaviour and group size prior to infection. Reduced resistance among high-ranking males was associated with increased serum testosterone and corticosterone concentration and reduced serum immunoglobulin, but was independent of group size. Among low-ranking males, hormonal changes were not associated with resistance to B. microti but changes in corticosterone concentration and measures of immunodepression increased with group size and aggressive behaviour. The results concur with earlier findings suggesting differences between high- and low-ranking mice in their physiological responses to social experience and consequently reduced resistance to B. microti infection among high-ranking individuals.

An experiment was conducted to determine the rate and maximum percentage of sporulation of Eimeria acervulina oocysts at various environmental conditions relating to temperature (21 versus 33 °C) and relative humidity (RH) (40 versus 80%). Measurements were made during 44 h after excretion of oocysts in 3 substrates: dry litter, clammy litter and pure faeces respectively. Maximum sporulation percentage in both dry (22·6%) and clammy litter (19·5%) was higher (P < 0·005) than in pure faeces (11·6%). Neither temperature nor RH had a significant influence on percentage of oocysts that sporulated. Under these simulated practical conditions approximately 25% of all oocysts sporulated, whereas sporulation under optimal conditions (29 °C, aeration, 2% K2Cr2O7) showed a higher (68%) sporulation ability of oocysts. At 33 °C sporulation proceeded at a faster pace than at 21 °C (P < 0·005). With respect to RH and substrate, once sporulation started, the rate of increase to maximum percentage was not different. Time of onset of sporulation was influenced by temperature (P < 0·0001) and RH (P < 0·001). Time of onset occurred 15 h later at 21 °C compared with 33 °C and 5 h later at 40% RH compared with 80%. Also, an interaction effect (P < 0·01) was found with effect of RH being stronger at 21 °C compared with 33 °C. It was concluded that the most important aspect in the epidemiology of E. acervulina during a flock cycle is the time of onset of sporulation with temperature being the most important factor.

A simulation model for the population dynamics of Eimeria acervulina infection in broilers is presented. The model describes the development of the numbers of parasites in the various life-stages during the growing period of broilers and the empty house period between grow-outs. The model includes assumptions with respect to development of immunity to E. acervulina infection and effects of application of anticoccidial drugs. The model consists of a set of difference equations that are solved numerically at 1 h intervals. Under constant conditions, an equilibrium level was reached after a few grow-outs during which infection always peaked around the 21st day in the growing period. Within a growing period, infection peaked earlier (later) than the 21st day in case initial numbers of sporulated oocysts were higher (lower) than the equilibrium number.

A simulation model for effects of Eimeria acervulina infection on technical and economic characteristics in broiler production is presented. The model describes development over time of the growth depression, feed intake reduction, and decrease in feed efficiency associated with infection. The model also shows a phase of compensatory growth during which earlier negative effects are counterbalanced, at least partly. Major assumptions made were: infection with E. acervulina occurs in each flock; production is affected in each flock; compensatory growth takes place because immunity develops and cell regeneration occurs. The results show that the pattern of development of the production characteristics during a flock cycle depends on the initial contamination level. Both a high and low initial contamination level results in a lower average daily gain, a worse feed to gain ratio, and a reduced net revenue compared to an intermediate contamination level.

We evaluated the quantitation of two schistosome circulating antigens in serum and urine as a tool for the assessment of the efficacy of praziquantel dosage regimens (40 versus 60 mg/kgbw). In addition we compared the efficacy of two different brands of praziquantel (Biltricide® and Distocide®), given at the same dosage (40 mg/kgbw). Thirty five Egyptian hospitalized schistosomiasis mansoni patients participated in this study. Thirteen patients (Group 1) received 60 mg/kgbw Biltricide®, administered in 3 oral doses of 20 mg in one day; 22 individuals (Group 2) were treated with 40 mg/kgbw (12 Biltricide®, 10 Distocide®), given in one oral dose. Circulating anodic antigen (CAA) and circulating cathodic antigen (CCA) were quantitated by monoclonal antibody-based ELISA's before, and 1, 3 and 6 weeks after chemotherapy. Before treatment, all patients were positive for at least one of the circulating antigen assays. Three to six weeks after treatment significantly more patients were found to be negative in Group 1 compared to Group 2 (X2 = 7·13, P = 0·008, n = 35). Also the levels of CCA and CAA in serum and of CCA in urine were found to be significantly higher in Group 2 (Mann- Whitney U < 85, P < 0·05, n = 35). These results were confirmed by parasitological data. No differences were found between treatment with Biltricide® or Distocide®. Our results indicate that praziquantel treatment of schistosomiasis with 60 mg/kgbw divided in 3 doses results in a higher cure rate compared to 40 mg/kgbw as a single dose, and provide further evidence for the use of the CAA and CCA assays as a sensitive method to monitor the efficacy of chemotherapy, particularly when circulating antigen assays are combined by parallel testing.

Recombinant Schistosoma mansoni calreticulin (SmCaR) was expressed in Escherichia coli, using the glutathione S-transferase fusion protein, and its Ca2+-binding capacity was determined. Results obtained by a 45Ca2+ overlay technique showed that Ca2+-binding site(s) were present in the recombinant CaR indicating that proper folding of the protein was obtained using this system. An antiserum raised against the recombinant SmCaR showed that the native protein (Sm58) was expressed in all stages of the life-cycle from cercariae to the adult worm and in the egg. However, SmCaR seems to be a developmentally regulated protein whose expression can be used to study the post-transformational differentiation of the schistosomulum. Localization of SmCaR demonstrated that the majority of SmCaR was expressed in the epithelia of the digestive duct and in the genital organs. These results suggest that SmCaR, by regulating the Ca2+ concentration, may play an important role during cell proliferation. Finally the presence of SmCaR in miracidia and in the genital organs suggests that the antibody response directed against this protein could interfere in egg production.

We have previously described a rat mAb directed against a peptide derived from the vif protein of HIV-1 that recognized two Schistosoma mansoni (Sm) antigens with a major band at 65 kDa. Epitope mapping of this mAb using overlapping hexapeptides derived from the vif peptide revealed that the motif recognized was PLPSVT. The screening of a Sm cDNA library led to the identification of two clones, Sm70 and Sm65. The two deduced protein sequences did not share any common structural features apart from the epitope recognized by the mAb (see below), and did not show significant identity to sequences present in the data bases. However, the N terminus of the deduced sequence of the Sm70 protein exhibits a consensus sequence known to be an ATP/GTP binding site. Furthermore, the C terminus of the deduced Sm65 protein sequence was found to contain a conserved hexapeptide with a consensus sequence LPETGE reported to be an important motif of the surface proteins of gram-positive cocci. Both proteins exhibit a peptide sequence (PLRSVT for Sm70 and PVGSVT for Sm65) similar to the epitope recognized by the mAb anti-vif. Western blotting experiments showed that the mAb anti-vif reacted with both proteins. However, only Sm65 was recognized by sera from HIV-1- seropositive individuals, whereas both proteins were recognized by S. mansoni-infected patients.

Theileria-free waterbuck (Kobus defassa) born in captivity were successfully infected with Theileria parva sporozoites derived from ticks infected by feeding on African buffalo (Syncerus caffer). All waterbuck underwent mild infections with the development of sporadic schizont and piroplasm parasitosis when inoculated with sporozoite doses lethal to cattle. A carrier state of T. parva was demonstrated by feeding clean R. appendiculatus nymphs on two of these infected waterbuck. Tick batches from these waterbuck on 2 of 5 occasions transmitted lethal Theileria infections to cattle. In a separate experiment, waterbuck cells were infected and transformed in vitro by T. parva sporozoites derived from buffalo but not by cattle-derived T. parva (Muguga) sporozoites. Waterbuck cells infected in vitro with T. parva isolated from buffalo were inoculated into autologous waterbuck but no infections developed. Theileria parva isolates generated in this study from various sources were characterized using anti-T. parva schizont monoclonal antibodies (MAbs), and it was found that buffalo-derived and waterbuck-passaged isolates had different profiles. Species-specific synthetic oligonucleotide probes, restriction fragment length polymorphism (RFLP) analysis with cloned T. parva DNA probes, and DNA sequence analysis of the p67 sporozoite antigen gene confirmed that the waterbuck-passaged parasite was T. parva. The Tpr repetitive probe hybridization patterns from the waterbuck-passaged parasites were different from the other samples tested. The ribosomal genotype of the waterbuck-passaged T. parva was similar to that of cattle-derived T. parva Muguga. Analyses with both probes and MAbs suggested that a minor parasite population present within the T. parva 7014 buffalo- derived stock had been selected during waterbuck passage. A variable region of the p67 sporozoite antigen gene of the waterbuck-passaged T. parva was similar to that of cattle-derived T. parva stocks and different from that of buffalo- derived parasites. Based on these results, methods were suggested to confirm and quantitate the involvement of waterbuck in the epidemiology of cattle theileriosis.

The effects of the novel benzimidazole, triclabendazole (TCBZ) (‘Fasinex’, Ciba-Geigy), in its active sulphoxide metabolite form (TCBZ-SX), on the tegumental ultrastructure of Fasciola hepatica were determined in vitro by transmission electron microscopy (TEM), using both intact flukes and tissue-slice material. At a concentration of 15 µ/ml, the tegument of the whole adult fluke showed ultrastructural changes only after prolonged time-periods, with vacuolation at the base of the syncytium and accumulation of T2 secretory bodies in the tegumental cells. At a concentration of 50 µ/ml, with both whole flukes and tissue-slices, the tegument appeared extremely abnormal with accumulation of secretory bodies towards the base of the syncytium. With longer incubation times, the tegument was completely sloughed away and the tegumental cells became synthetically inactive. The tegument of the 3-week-old juvenile became progressively convoluted at the apex, while in the basal regions there was severe vacuolation. In the tegumental cells, there were accumulations of T1 secretory bodies. These results confirm TCBZ as a potent fasciolicide, being very effective in disrupting the fluke tegument. They may go some way to explain the mode of action of this important fasciolicide.

Macroparasites almost invariably exhibit overdispersed distributions of parasites/host, yet the specific causes of such aggregations remain poorly understood. The present study focused on the distribution of the parasitic nematode Howardula aoronymphium among its hosts, several species of mycophagous Drosophila. The distribution of parasites/host is close to random for cohorts of flies of a given host species emerging from single mushrooms. At the level of Howardula populations, overdispersion of parasites among hosts results primarily from variation among subgroups of hosts in their exposure to infective-stage nematodes. The sources of variation identified in this study include Drosophila host species, the site where flies bred, mushroom species within sites, and, most importantly, individual mushrooms within mushroom species at a site. For the mean intensity of parasitism observed in this study, the degree of aggregation is typical of macroparasites in general. Combinations of random distributions with different means, resulting from variation among groups in exposure to parasites, may be a common cause of the overdispersion of macroparasites among hosts.

Acetylcholinesterase (AChE) has been purified from the excretory/secretory (ES) products of Trichostrongylus colubriformis (using edrophonium chloride linked to epoxy-activated Sepharose) with yields of 40–50%. Purity was confirmed by polyacrylamide gel electrophoresis (using silver [protein] and Karnovsky [activity] stains) and measurement of specific AChE activity. Further analysis of the purified fractions by gel filtration and sucrose density gradient techniques revealed the existence of 2 forms of hydrophilic AChE (Mr 189 and 80 kDa). From the data we deduce these to be the globular monomer and dimer, G1 and G2 forms of AChE. Inhibition studies using BW284C51, iso-OMPA and excess substrate, along with substrate specificity studies, show both forms to be true acetylcholinesterases. We are currently assessing the protective immunogenicity of purified AChE in sheep.

A post-embedding immunogold technique has been used to examine the subcellular distribution of immunoreactivities to vertebrate pancreatic polypeptide (PP) and to the invertebrate peptide, FMRFamide within the central nervous system (CNS) of the nematode, Ascaris suum. Gold labelling of peptide was localized exclusively over dense-cored vesicles within nerve cell bodies, nerve axons and nerve terminals of the main ganglia and nerve cords in the CNS. Double-labelling of peptides demonstrated an apparent co-localization of PP and FMRFamide immunoreactivities in the same dense-cored vesicles, although populations of dense-cored vesicles that labelled solely for FMRFamide were also evident. Antigen preabsorption studies indicated little or no cross-reactivity between the two antisera.

The excreted/secreted proteinases of Haplometra cylindracea maintained in vitro, were found to hydrolyse the fluorogenic substrates, Z-ArgArg-NHMec and Z-PheArg-NHMec. This activity was shown to be typically that of cysteine proteinases, as turn-over of both substrates could be blocked by pre-incubation with peptidyl diazomethyl ketones. The biotinylated affinity reagent, biotin-Phe Ala-DMK, used in combination with Z-PheTyr(OBut)-DMK, was employed for the labelling and characterization of these cysteine proteinase activities. Three cathepsin B-like species were detected, with molecular weights of 48, 22–23 and 14 kDa, together with a cathepsin L-like enzyme, with a molecular weight of 55 kDa. The proteinases were also found to have hydrolytic activity towards the substrate, Z-GlyGlyArg-NHMec, which could be blocked by pre-incubation with either of the serine proteinase-selective reagents, Z-ArgP(OPh)2, or biotin-LysP(OPh)2, showing the activity to be trypsin-like. Using the biotinylated affinity label to characterize the trypsin-like enzymes revealed two molecular species with molecular weights of 20 and 24 kDa.