Surgical bursectomy or thymectomy at day old had little effect on primary or subsequent infections with E. brunetti or E. maxima.

Whole body irradiation of day-old chickens with 700 or 750 R did not affect the development of immunity to these Eimeria spp., but in some experiments it affected oocyst production during initial infections.

In one of four trials, birds surgically bursectomized and irradiated produced twice as many E. brunetti oocysts during an initial infection as the irradiated only controls; in the remaining three trials, oocyst production in the operated birds was either similar to or slightly greater than that in the controls. Combined bursectomy and irradiation did not affect immunity to re-infection.

Combined thymectomy and irradiation had little effect on primary infections (increased oocyst output was found in only one of six trials) and no effect on subsequent immunity; mean body weights and circulating lymphocyte numbers were decreased but tuberculin sensitivity was reduced in only a small proportion of the birds, indicating that the cell mediated immune responses of the majority were not affected.

Small (0.75 mg) daily doses of cortisone acetate given to irradiated birds or irradiated thymectomized birds, did not affect resistance to reinfection with E. brunetti.

We wish to thank Dr B. M. Freeman for surgical bursectomy; Mr E. A. King, Department of Radiotherapeutics, University of Cambridge, for irradiation and members of the technical staff of the Parasitology Department for their willing and skilled assistance.

Heat shock proteins (HSPs) were documented for the first time in both somatic extracts and excretory/secretory (ES) products of the infective-stage larvae of Trichinella spiralis and T. pseudospiralis. Larvae recovered from muscles of infected mice were heat shocked at 37, 40, 43 and 45 °C in RPMI 1640 medium containing L-[35S]methionine. Somatic extracts and ES products of heat-shocked worms were then analysed by SDS-PAGE, autoradiography and laser densitometry. Prominent bands of HSPs were observed at 43 °C which is the optimal heat shock temperature. The major HSPs in somatic extracts of T. spiralis were 20, 47, 50, 70, 80 and 86 kDa. When the temperature was increased from 37 to 43 °C, the greatest increase in absorbance was observed in HSPs 70 and 86. In vitro translation of mRNA in a nuclease-treated rabbit reticulocyte lysate system showed an increase in the synthesis of the 80 kDa protein. This suggests that the production of HSP 80 is regulated at the transcriptional level. The major HSPs in the ES products were 11, 45, 53 and 64 kDa. In T. pseudospiralis, the major HSPs in the somatic extracts were 20, 26, 31, 50, 53, 70, 80 and 86 kDa, and in the ES products, 11, 35, 37, 41 and 64 kDa.

We used selection to test for a genetic component to the pattern of emergence of infective juveniles from the host cadaver in the entomopathogenic nematode, Steinernema glaseri (Steiner), and whether other traits would respond to selection on this major and complex life-history character. We selected for early (‘ fast’) and late (‘ slow’) emerging lines by perpetuating nematodes that emerged on the first and after the seventh day of emergence respectively. After 12 cycles of selection, the pattern for the slow line but not the fast line differed significantly from the base population. Cumulative emergence for the slow line was less than the base population from Days 4 to 14 of the 18-day emergence period. The maximum difference occurred on the fourth day when 72·6% of emergence was complete for the base population but only 55·4% for the slow line. Decreases in infective juvenile size over the emergence period were consistent with the change in emergence pattern, but variation in sex ratios was not. No differences in infectivity were found. These results indicate that the emergence pattern has a genetic component, and that genetic variability for this trait occurs in natural populations. Furthermore, the asymmetric response to selection suggests that our field population is under strong selection for a highly skewed early emergence.

The ultrastructural changes accompanying oocyte differentiation in the ovaries of the monogeneans, Diclidophora merlangi, Diplozoon paradoxum and Calicotyle kröyeri have been described. In each case, oogenesis in the ovary proceeds as far as meiotic prophase in the primary oocyte. A three-stage sequence of development based on oocyte morphology is proposed: (1) Oogonia and early, immature primary oocytes are typically undifferentiated, with chromatin-laden nuclei occupying most of the cell volume. The cytoplasm contains small clumps of mitochrondria and unattached ribosomal aggregates. There is evidence of mitosis and, in later stages, meiotic prophase is indicated by the appearance of nuclear synaptonemal complexes. (2) Maturing primary oocytes are characterized by increased nucleolar volume associated with the production of RNA for export to the cytoplasm. An organized GER and Golgi apparatus are established and involved in the synthesis and packaging of membrane-limited cortical granules. Annulate lamellae and nucleolus-like bodies appear in the cytoplasm and, with development, the cells increase in size and, peripherally, become interdigitated. (3) Mature primary oocytes represent a resting phase when cellular activity is minimal. Golgi disappear and the ER fragments or becomes reduced in dimensions. Mitochondria and free ribo-somes are numerous and cortical granules move to the cell periphery. The cells separate and, when mature, are released from the ovary. There are minor species differences in oocyte ultrastructure and development.

The elongation factor-1 alpha (EF-1α) is a highly conserved ubiquitous protein that is involved in translation and is desirable for use in phylogenetic studies on Blastocystis, an enigmatic intestinal parasite with a contentious taxonomic position. In the present study, a PCR product (BEα) that codes for a major part of the coding region of the EF-1α protein was amplified. Genome walking experiments together with cloning were implemented to elucidate the 5′ and 3′ ends of the EF-1α gene of the human isolate, Blastocystis hominis C. The genomic organization and the potential transcription factor binding sites of the 5′ end of B. hominis C EF-1α were identified. A comparative study on the deduced amino acid sequences of BEα of 13 Blastocystis isolates from various hosts was done to evaluate the phylogenetic relationship among the speciesThe nucleotide sequence data reported in this paper are available in the GenBank database with the accession numbers as shown in Table 1.. A phylogenetic reconstruction analysis with other eukaryotic EF-1α sequences was carried out to trace the phylogenetic position of Blastocystis among eukaryotic organisms.

Samples of the snail Velacumantus australis from an area of 10 yards in diameter, were examined to determine if the incidence of larval trematodes varied with the density of the snails. The incidence in susceptible snails from areas where the density was less than 20 snails/sq. ft. was found to be significantly greater than in snails from areas where the density was higher than this. Further analysis showed that the variation in incidence, with density, was due almost entirely to the juvenile part of the population.

This investigation compared the effect of two common types of diet on worm burdens and faecal egg counts of pigs inoculated with Ascaris suum and Oesophagostomum dentatum. The diets were both considered nutritionally sufficient for pigs of this age, and gave high and comparable growth rates. Pigs were inoculated with both worm species at 14 weeks of age and slaughtered 3 or 12 weeks later. At 3 weeks, A. suum larvae were numerous in the small intestine of pigs fed ground barley plus protein supplement, but were not detected from pigs fed a commercial full-constituent pelleted feed. At 12 weeks, however, average adult worm burdens were low and similar in both groups. At both slaughter times, O. dentatum worm burdens were significantly higher in pigs fed barley plus protein than in pigs given commercial feed, particularly for female worms, and the commercial diet led to worms being located further along the large intestine, and to reduced worm fecundity. The chemical composition of large intestinal contents differed significantly between pigs fed the two diets.

Cercaria patialensis Soparkar, 1924, emerging from the snail Melanoides tuberculatus (Müller) has been encountered at Waltair. The cercariae begin to emerge from the snail at dusk and continue to do so during the period of darkness.

Stages resembling the adult but without the development of eggs have been encountered in nature on the fresh water fishes Panchax panchax (Ham. & Buch.) and Esomus danricus (Bleeker). These stages have also been raised experimentally by exposing P. panchax, E. danricus and fingerlings of Catla catla Ham. & Buch. to the cercariae.

The cercaria is referable to the genus Transversotrema Witenberg, 1944.

It is suggested that there may not be a metacercarial stage in the life-cycle of Transversotrema spp.

Young white mice were infected with different doses of eggs or cysticercoids of Hymenolepis nana and the subsequent faecal egg output was measured over a 3-day period. This was then correlated with worm numbers, determined after killing the hosts. The longest worm in each infection was measured and it was shown that crowding decreased the linear dimensions, number of eggs per proglottis and rate of proglottis production of these worms, but that their shape was not affected.

The relation between total egg output and worm numbers was complex. In both egg-derived and cysticercoid-derived infections the egg production rose to a peak, fell and then levelled off to a constant rate with increasing worm numbers. Cysticercoid-derived worms were larger than egg-derived worms, but produced approximately the same number of eggs per unit of biomass. The differences in the egg output curves for the two types of infection were interpreted as being due to their size difference; this in turn may be a consequence of the host immunological response.

The aim of this study was to investigate the intra-litter infection dynamics of Isospora suis under natural conditions, and to study any association between parasite transmission and the contamination level of the farrowing pen by applying different interventions in order to reduce the transmission of I. suis infection within the litter. The study was divided in 2 trials including in total 22 litters (254 piglets). The first trial included 4 litters (where standard procedures practiced routinely on the farm piglets were applied) and the piglets were followed coprologically from farrowing until 2 weeks after weaning. The sows of those litters were also examined at various intervals before and after farrowing. The second trial included the application of 3 different management procedures: (A) standard farm hygiene and management procedures, (B) standard farm hygiene and management procedures+the first piglets found to excrete I. suis oocysts in each pen were removed from the pen, and (C) reduced cleaning. Each procedure was studied in 2 litters. This was replicated 3 times to yield a total of 18 litters. The results suggested that (i) the sow does not play an important role in transmission of I. suis in the farrowing pen; (ii) in natural infections, both the age of the piglet age at onset of oocyst excretion and the oocyst excretion patterns may vary considerably; (iii) the course of oocyst excretion or development of diarrhoea is related to the time of initial infection and (iii) piglets, which are heavy at birth, are more prone to acquire I. suis infection. Moreover, it was demonstrated that cleaning could be an effective means of restricting the spread of the parasite within the litter and thus the development of diarrhoea.

Two experiments were carried out in which calves reared parasite-free were infected with a single dose of 3rd-stage larvae of Cooperia oncophora. In the first experiment the calves received 20000 or 200000 infective larvae and they were autopsied 28 or 56 days after the infection. In the second experiment the doses were the same but the animals were killed 14, 84 or 140 days after infection. If a dose of 20000 was given, clinical signs were never observed, while at a dose level of 200000 the weight gain was less on 56 and 84 days after the infection compared with the low-infected groups or the control animals. After 140 days the differences in weight gain were compensated. Faecal egg output was higher in the 200000 groups only in the first period of patency, thereafter the calves in the 20000 groups produced more. No obvious differences between the two infection levels were observed with regard to the haematological data. In the low-infected groups worm counts were only slightly lower when the results after 28 days were compared with those after 56 days. Also, the worm numbers after 14 days were almost equal to those after 84 days, while at 140 days 1 animal still had the same number, the other one had lost its worm burden. In the high-infected groups the worm loss was much quicker. After 28 days a great part of the population had already been lost. Obviously, at the 200000 level the reaction of the host against the parasite was much stronger. Adult worms were expelled at a higher rate than early 4th-larval stages. In the first experiment worm measurements revealed differences between the length of females, males and spicules of males, these being significantly longer in the low-infected groups. Analysis of the distribution of worms over the small intestine showed that in the low-infected groups worms were mainly restricted to the first 6 metres. In the high-infected groups the worm population was distributed more evenly over the whole small intestine.

Cystic echinococcosis (CE) is a high prevalence zoonosis among the rural population of Tacuarembó (Uruguay). The correlation between serological data and the incidence of risk factors was studied in a survey carried out in 1998 among rural communities where 480 individuals were examined by means of abdominal sonography (local prevalence=0·8%). Serum samples (305) were analysed by ELISA to determine specific IgG against crude antigens from Echinococcus granulosus. A total of 27 individuals exhibiting no detectable changes in abdominal sonographic examination were found to be seropositive (‘ultrasound normal group’). Of these individuals 9 were seroreactive against purified antigen B. A significant degree of correlation was found between seroreactivity and the incidence of some risk factors (CE antecedent in the family, P<0·005 and use of rural water, P<0·0001) among this group. Follow-up of individuals of the ‘ultrasound normal group’ was carried out after 2 years to evaluate the implications of this serological reactivity. No predictive value for cyst development was assessed with complementary image study; in contrast transient antibodies were observed with both crude and purified antigen as approximately 60% of individuals became negative when re-sampled.

The influence of subcutaneous infection with protoscoleces of Echinococcus granulosus on the distribution of lymphocyte subsets in draining lymph nodes has been evaluated by immunocytochemical labelling of lymphocyte surface antigens. These studies reveal marked expansion of paracortical Thy-1 +, Lyt-1 + cells and moderate proliferation of surface immunoglobulin-bearing B-cells immediately after infection. The Lyt-1 +:Lyt-2 + ratio decreases rapidly during the first 21 days post-infection and remains below unity until at least 12 weeks post-infection due to severe depletion of Lyt-1 + cells in draining lymph nodes and a significant increase in the percentage of Lyt-2 + cells. The potential role of these Lyt-1 ∓, 2 + (putative T-suppressor) cells in regulation of the anti-parasite immune response and mediation of generalized immunosuppression is discussed in the light of evidence of inhibition of anti-parasite immunity in infected mice.

The metabolites of 5-hydroxytryptamine (5-HT, serotonin) namely, L-tryptophan, 5-hydroxytryptophan, 5-hydroxyindole acetic acid and 5-hydroxytryptophol were measured in perfused tissue and whole worms from adult female Ascaris suum using reversed-phase liquid chromatography with electrochemical detection. The intracellular levels of each metabolite were quantitated in response to several physiological effectors but only Ltryptophan (TRP) caused dose-dependent changes in these metabolites. Serotonin itself could also be absorbed by perfused A. suum muscle and intestinal tissue. When live A. suum were tied at the anterior and posterior regions to restrict TRP absorption by the intestine, TRP was absorbed through the cuticle and converted into 5-HT by the muscle tissue. In untied live parasites TRP absorption was observed in both muscle and intestinal tissue. Collectively, the data indicated that 5-HT may be either absorbed directly or synthesized de novo from absorbed TRP in the isolated tissue of A. suum. The 5-HT, in the live adult female A. suum, can be synthesized de novo from TRP, or 5-HT can be absorbed from the environment both through the cuticle and by the intestine of living parasites. Data also indicated that there was preferential sequestering of 5-HT and the metabolites of 5-HT in the anterior tissues of the worms.

1. The cytological study of 4–8 hr. old zygotes of seven species of Plasmodium, i.e. P. gallinaceum, P. berghei, P. cynomolgi, P. gonderi, P. inui, P. knowlesi and P. vivax, revealed a uninucleate condition at the resting phase.

2. Ookinetes of these species at 12–24 hr were uninucleate and of the usual dimorphic form.

3. In the present studies post-zygotic meiosis was demonstrated in the early oocysts of the above seven species of Plasmodium. The times of investigation for the beginning and completion of meiosis in the early oocysts of these species of Plasmodium were as follows:

4. Different forms of spireme at prophases were observed in the early oocysts of the above species of Plasmodium. This fact indicated that each species has its own characteristic form of spireme; for instance, ‘bow-shaped’ in P. gallinaceum, beaded, filamentous net-work in P. cynomolgi.

5. The diploid and haploid number of chromosomes were demonstrated step by step, during the first reduction division in the early oocysts of seven species of Plasmodium.

6. The succeeding nuclear divisions following the post-zygotic meiosis were traced in early oocysts of seven species of Plasmodium. The divisions took place mitotically, giving rise to tetra- to multinucleate oocysts. The haploid group of chromosomes in each species was quite distinct during the different mitotic phases found in these oocysts.

7. The results obtained from the cytological study of different meiotic and mitotic phases found in the early oocysts of seven species of Plasmodium show that the number of chromosomes is a specific character, for instance, P. gallinaceum has a complex of two haploid chromosomes, P. cynomolgi, four; P. gonderi, three; P. inui, four; P. knowlesi, P. berghei and P. vivax, two.

An ATP diphosphohydrolase was identified in the plasma membranes isolated from promastigote forms of Leishmania amazonensis. Both ATP and ADP were hydrolysed at similar rates by the enzyme. Other nucleotides such as UTP, GTP and CTP were also degraded, revealing a broad substrate specificity. Adding ATP and ADP simultaneously, the amount of hydrolysis achieved was compatible with the presence of a single enzyme. ATPase activity was not affected by addition of vanadate, ouabain, thapsigargin, dicyclohexylcarbodiimide, oligomycin and bafilomycin A, thus excluding involvement of P-, F- and V-type ATPases. The effects of pH in the range 6·5–8·5 were examined using ATP or p-NPP as substrate. At pH 7·4, the phosphatase activity decreased, and did not show a significant contribution to ATP hydrolysis. In addition, the enzyme was not inhibited by levamisole and ammonium molybdate, excluding alkaline phosphatase and nucleotidase activities, respectively. Sodium azide (5–10 mM) caused inhibition of the ATP and ADP hydrolysis in a dose-dependent manner. Calcium was the best activating metal ion for both ATPase and ADPase activities. Ultrastructural cytochemical microscopy showed ATP diphosphohydrolase on the surface and flagellar pocket of the parasite. We have proposed that L. amazonensis ATP diphosphohydrolase may participate in the salvage pathway of nucleosides.

It has been shown that in intact cercariae of Schistosoma mansoni in water, both head and tail had an identical, aerobic energy metabolism. As long as the environment was water, glucose was mainly degraded to carbon dioxide by both head and tail whether or not these two were still connected to each other. Transfer of intact cercariae into a simple salt medium supplemented with glucose resulted in a very rapid transition towards a more anaerobic energy metabolism: the production of lactate and pyruvate increased, whereas the production of carbon dioxide remained more or less constant. A concomitant rise in temperature to 37°C was not essential for this biochemical transition, but made it more pronounced. Experiments on isolated cercarial bodies and tails in a transforming medium demonstrated that the tails oxidized glucose to carbon dioxide, whereas bodies produced mainly pyruvate and lactate. The results showed that the metabolic transition towards a more anaerobic energy metabolism occurred only in the head and not in the tail of the cercariae. Loss of the tail was shown not to be a pre-requisite for this transition, nor did it by itself trigger a metabolic switch in the resulting cercarial body.

The activity of atovaquone against Plasmodium berghei ANKA during sporogonic development has been examined. Anopheles stephensi mosquitoes were fed on gametocyte infected mice which had been treated 8 h previously with atovaquone or diluent alone. Mosquito midguts were examined for oocysts, and salivary gland infections were estimated using an ELISA for the circumsporozoite protein (CSP). The number of oocysts per midgut fell by at least 97% when mosquitoes were fed on mice dosed with 0·1–10 mg atovaquone/kg body weight. This was paralleled by a decrease in the prevalence of oocyst-infected mosquitoes from 70–90% in controls to 40% or 10% respectively. No oocysts were observed at a dose of 100 mg/kg. CSP ELISA results indicated that mosquitoes fed on atovaquone failed to produce sporozoites. Mosquitoes which fed on gametocytaemic, atovaquone-treated mice (0·1–100 mg/kg) did not transmit malaria to naive mice. These results demonstrate that atovaquone has a highly potent inhibitory activity against the mosquito stages of P. berghei.

Parvicapsula minibicornis is a myxozoan parasite implicated in mortalities of both juvenile and pre-spawning adult salmon in the Pacific Northwest of North America. Disease severity and presentation varies between salmon species and geographical localities. To better characterize population structure of the parasite, we sought genetic markers in the P. minibicornis ribosomal RNA gene. We compared samples from California with the type specimen from British Columbia, identified sequence variations, and then sequenced 197 samples from fish, river water and the parasite's polychaete worm host. Although DNA sequences of the parasite were >98·9% similar, there was enough variation to define 15 genotypes. All genotypes were detected in fish samples, although not in all species. A single genotype only was found in sockeye and pink salmon in the Fraser River Basin, but was not detected in sockeye from the adjacent Columbia River Basin. All coho salmon, irrespective of river basin, were infected with a unique mix of 2 genotypes. These data indicated that the P. minibicornis population exhibited strong signals of structuring by both geography and salmonid host species. Particular genotypes may correlate with disease differences seen in salmon populations in the Pacific Northwest.

A study was made of some of the factors which affect the growth of Plasmodium knowlesi in blood diluted in the Harvard medium. Some slight enhancement of growth and re-invasion of new host cells was obtained when P. knowlesi was incubated in low oxygen tensions. Some growth but no re-invasion was obtained under anaerobic conditions. High oxygen tensions were also inhibitory.

The lipids present in the plasma are important for the growth of P. knowlesi in vitro. Stearic acid and cholesterol are required by the parasite but do not account for all the growth promoting properties of plasma. A requirement for vitamin B12 was also demonstrated.

I should like to thank Dr F. Hawking and Dr J. Williamson for their advice and for reading through the manuscript, Mr T. J. Scott-Finnigan for technical assistance, Dr K. Mervyn (Glaxo Research Ltd.) for samples of vitamin B12 analogues and the Hachmeister Company (Pittsburgh) for a sample of TEM 4T. This work received financial assistance from the World Health Organization.