Characteristics of four species of Eimeria isolated from Apodemus sylvaticus are described. From oocyst morphology, three are identified as E. apionodes, E. hungaryensis and E. uptoni, the fourth being unnamed. Cloning demonstrated that oocysts of E. hungaryensis were polymorphic. Previous literature relating to the taxonomy is discussed and discrepancies outlined. Species isolated from A. sylvaticus were not transmissible to Clethrionomys glareolus, but both E. hungaryensis and E. apionodes have been passaged through immunosuppressed laboratory mice, the former species more than 16 times. In both A. sylvaticus and immunosuppressed laboratory mice, endogenous development of E. hungaryensis occurred mainly in enterocytes near the tips of the villi in the first half of the small intestine, with a few parasites in the rest of the small intestine and into the large intestine. The pre-patent period was 2 days in both hosts but oocyst output was higher in the natural hosts. E. apionodes parasitized enterocytes on the sides or at the base of the villi, mainly in the last 90% of the small intestine with a few parasites in the large intestine. The pre-patent period was 7 days. Parasites, probably E. hungaryensis and E. apionodes, were also isolated from A. flavicollis.

Host selection among humans by Anopheles punctulatus was studied in an area of Papua New Guinea endemic for malaria and filariasis. Blood films were made from the stomach contents of freshly engorged mosquitoes found resting on the walls of houses in which the parasite status of the occupants was known. Engorgement rates on humans were non-random but could not be consistently related to the parasite status of individuals in the houses for either malaria or filaria. In some households, anophelines preferentially fed on parasitaemic individuals while in other households aparasitaemic individuals were significantly more often selected. This finding is believed to reflect the fact that malaria and filarial infections in this endemic area are predominantly asymptomatic. There were no significant differences in axillary temperatures between malaria or microfilariae positive or negative individuals.

Sex ratio theory usually predicts an equilibrium sex ratio and equal proportions of males and females in a population, including the progenitors of the reproductive cells of protozoans. This proposal was tested with three species of malarial parasites of lizards, Plasmodium mexicanum of the western fence lizard, and P. agamae and P. giganteum of the African rainbow lizard, using single samples from naturally infected lizards, repeated samples from free-ranging lizards (P. mexicanum only), and repeated samples from laboratory maintained animals. Macrogametocytes were usually more abundant than microgametocytes, and were slightly larger, revealing a typically greater investment of resources by the progenitors of female reproductive cells. However, the proportion of microgametocytes varied among the three species and among infections within each species of Plasmodium. The sex ratio of gametocytes often remained constant within infections followed over time even if the absolute number of gametocytes was changing. However, the equilibrium sex ratio of gametocytes varied among those infections that had an unchanging microgametocyte proportion. Thus, although an equilibrium sex ratio apparently occurs for most infections, there appears to be no characteristic proportion of microgametocytes for any of the species. Potential explanations for this conflict with theory are presented.

The biosynthesis of phospholipids is extensive in Plasmodium knowlesi-infected simian erythrocytes due to the synthesis of membranes by this single-cell eukaryote in a host erythrocyte devoid of any pathway for lipid biosynthesis. In the present paper, we show that the incorporation of [3H]glycerol, which reflects de novo biosynthesis, is better studied at 300 μM−1 mM than at the trace doses, since this non-physiological precursor does not modify the amount of phosphatidylcholine biosynthesis from [3H]choline. Time-course incorporation of radioactive glycerol, oleate, lysophosphatidylcholine, choline, and inositol in RPMI 1640 medium containing nutrients for lipid synthesis showed that the optimum incubation time for phospholipid studies is 60–90 min, after which radioactive incorporation slows considerably. On the other hand, studies with [14C]serine revealed that incubation for 2–3 h is necessary for isotopic labelling of phosphatidylcholine via phosphatidylserine decarboxylation and phosphatidylethanolamine N-methylation. Incorporation of the various fatty acids into individual lipids was related to the molecular species composing each of them. Studies with [14C palmitoyl] lysophosphatidylcholine showed a very fast intracellular release of radioactive fatty acids, which indicates a potent lysophospholipase activity. Taken together, these data define the indispensable conditions for an experimental system suitable for further studies.

A reproducible and standardized assay for measuring the cytoadherence of knobby Plasmodium falciparum-infected red cells to amelanotic melanoma cells was developed. Adhesion was dependent on temperature, haematocrit, and parasitaemia. Addition of EDTA to the binding medium reduced adhesion. Removal of protease-sensitive molecules on the surface of the infected cell abolished cytoadherence, whereas removal of carbohydrate residues by treatment of cells with neuraminidase or galactosidase promoted adhesion. Calcium, magnesium, fibrinogen or fibronectin in the medium had no effect on adhesion nor was there any enhancement of adhesion by pre-loading infected cells with calcium. Serum was essential for good adhesion. Adhesion was species specific for target cells; human endothelial or amelanotic melanoma cells were suitable target cells whereas bovine cells were not. The amelanotic melanoma cell could be formalin-fixed and still retain its adhesion properties. The binding properties of formalin-fixed amelanotic melanoma cells were not identical to those of endothelial or unfixed target cells.

The transport processes of D-glucose in Plasmodium yoelii-infected mouse erythrocytes were investigated using 2-deoxy-D-glucose (2DOG), a non-metabolizable analogue of D-glucose. Infected cells showed an increase in the uptake of 2DOG compared to uninfected controls, and an effect which was more prominent in cells with mature-stage parasites. Kinetic studies measuring the initial rates of 2DOG uptake revealed two components in infected cells with late trophozoite and schizont-stage parasites: a simple diffusion system and a carrier (transporter)-mediated system. The transporter was common for D-glucose and 2DOG and had a kinetic constant indicating a high affinity for 2DOG (the Km = 0·18 mM and the Vmax = 0·61 mmol/1010 cells/min), as compared to the constant of the mouse erythrocyte carrier (the Km = 10 mM and the Vmax = 1·8 mmol/1010 cells/min). Determination of the distribution of [3H]2DOG in infected cells and experiments with metabolic inhibitors indicated that the simple diffusion system localizes in the membrane of host cells and the transporter in the parasite plasma membrane. The parasite glucose transporter was much less sensitive to cytochalasin B than that of the host cells and the uptake of 2DOG via the transporter was dependent on energy. Based on these findings, the following features emerge: D-glucose first gains access to the cytosol of infected erythrocytes via the simple diffusion system, which appears after infection by the parasite, and an active uptake against the concentration gradient takes place at the parasite plasma membrane via the parasite glucose transporter in an energy dependent manner. Finally, an energy transduction mechanism for the transport of glucose across the parasite plasma membrane is discussed.

In this study, the advantage of the use of chloroquine (CQ) containing liposomes (lipCQ) over free CQ in the chemotherapy of murine malaria (Plasmodium berghei) was demonstrated. The maximum permissible dose per intraperitoneal injection was 0·8 and 10 mg for CQ and lipCQ, respectively. An increase in therapeutic and prophylactic efficacy of lipCQ in comparison with free CQ at a 0·8 mg CQ dose level was found. It was possible to obtain 100% efficacy (injection at day 5 after infection; parasitaemia 4–8%) with one single intraperitoneal injection of 6 mg lipCQ. Moreover, the ability to increase the doses of CQ per injection after liposome encapsulation allowed successful treatment of infections with CQ-resistant Plasmodium berghei which could not be cured by a 7-day course with the maximum tolerable dose of free CQ of 0·8 mg/mouse/day.

Blastocrithidia triatomae parasitizes vectors of Chagas' disease and is very difficult to cultivate in conventional media. However, co-cultivation with a cell line of its host Triatoma infestans (TI-32; in Schneider's Drosophila medium supplemented with 20% foetal calf serum and 10% tryptose phosphate broth) led to vigorous growth at 24 or 28 °C without an adaptation phase. More than 60 primary cultures were initiated successfully without any failures. Subcultures could be started immediately at weekly intervals. The doubling time was similar in both the primary cultures (41 h) and the 33rd subculture (39 h). The importance of the reduviid cells for B. triatomae became clear after removal of the insect cells, when multiplication of epimastigotes stopped and mainly cysts were formed. Cysts produced in vitro were infective for reduviids. Scanning electron microscopy showed that B. triatomae attached to the host cells, inserted its flagellum into them and destroyed them.

A study of Leptomonas lygaei in Lygaeus pandurus is described. Flagellates with promastigote configuration were found in the midgut, ileum and rectum. Cysts and encysting stages (straphangers) were found in the rectum either attached to the flagella of promastigotes or free in the lumen. In the posterior midgut, the crypts were frequently filled with flagellates, but no attachment to host epithelium was observed. A close association between the flagellates and extracellular membrane layers was observed. In the rectum the flagellates frequently attached by hemidesmosomes to the cuticle of the gland cells and less frequently to the rest of the rectal wall. Interflagellar desmosomes between the expanded sheaths of flagella of adjacent parasites were also observed. Straphanger cysts attached to parental flagella by the formation of zonular desmosomes. Differences were apparent between the organelles of the encysting stages and the parental flagellates. In mature cysts, the cellular organelles were unrecognizable. A comparison between the cysts and flagellates of L. lygaei and Blastocrithidia familiaris is made.

Normal human serum (HS) contains trypanolytic activity and agglutinins to Trypanosoma equiperdum, while such activities are not found in sera from a range of animals susceptible to infection. HS given to T. equiperdum-infected mice caused a rapid decrease in the number of circulating trypanosomes and protection from lethal infection. Trypanolytic activity of human serum was found to be associated, after DEAE chromatography and Sephadex G-200 gel filtration, with the fraction containing 19S antibodies. Immunofluorescence assays confirmed a binding of human IgM and C1q complement component onto the surface of T. equiperdum. Anti-T. equiperdum activity of HS was specifically directed to T. equiperdum surface components and not to some mouse serum components adsorbed on parasites during the growth in the host, because HS adsorbed in vivo in CD-1 mice retained full protective and agglutinating properties. Trypanocidal activity appears in human serum about the 7th month after birth and persists until late in life. On the contrary, human purified high-density lipoprotein had no significant in vitro or in vivo trypanocidal activity. In conclusion, strong natural anti-T. equiperdum activity in human serum was mainly mediated by natural antibodies of the IgM class. The presence of natural IgM active against T. equiperdum in HS could represent one of the natural mechanisms of resistance of refractory hosts against trypanosome infections. This phenomenon provides further evidence that host specificity of trypanosomes may be partly conditioned by the presence of natural antibodies.

Schistosoma mansoni cercariae in water were shown to possess a largely aerobic energy metabolism, the Krebs cycle being the main terminal of carbohydrate breakdown. A metabolic transition towards a more anaerobic breakdown of carbo-hydrate could be achieved by incubation conditions which also stimulated biological transformation. Incubation of cercariae in a simple salt medium containing 5 mM glucose induced such a metabolic transition: beside carbon dioxide large amounts of lactate and pyruvate were excreted. The results indicate that the production of pyruvate was coupled to electron transfer in the respiratory chain. Some aspects of this unusual pyruvate production are discussed. The observed change in the end-product pattern of carbohydrate breakdown is very rapid: most of the switch occurred within 2 h. Our results show that the metabolic transition was triggered by the biological transformation itself, or by the same event that induces the biological transformation. The metabolic and the biological changes proceeded synchronously.

125I-Schistosoma mansoni schistosomulum surface antigens were immunoprecipitated with human antibodies from individual Egyptian patients diagnosed as being either acutely or chronically infected with S. mansoni. Both sets of patients were found to have IgG antibodies in their sera capable of immunoprecipitating the major Mr > 200, 38 and 32K antigens. However, the immunoprecipitation of the Mr 200K antigen was found to constitute a significantly greater proportion of the total precipitate achieved with acute sera than with chronic sera. The Mr 38 and 32K antigens were more variably precipitated by the acute sera than the chronic sera but the proportion of the total precipitation that these two antigens constituted was not found to be significantly different between the two sets of sera. Immunoprecipitation with pooled antibodies absorbed with egg and adult worm homogenates which had been treated to remove either carbohydrate or polypeptide epitopes demonstrated that the Mr > 200K antigen was the principal target of egg-cross-reactive anti-carbohydrate antibody amongst the antigens detected. The Mr 38 and 32K antigens were found to be precipitated by antibodies to protease-sensitive and periodate-insensitive polypeptide epitopes. These results are consistent with egg-cross-reactive anti-carbohydrate IgG antibody making a greater contribution to schistosomulum surface recognition in acute infection than in chronic infection. Indeed the presence of a higher level of egg-cross-reactive and anti-carbohydrate antibody directed against schistosomulum surface epitopes in an acute serum pool than in a chronic serum pool was confirmed by measurement of antibody binding to whole schistosomula.

Population outbreaks of house mice (Mus domesticus) occur periodically in the wheatlands of southeastern Australia. This paper uses mathematical models to assist in the evaluation of the potential of a nematode, Capillaria hepatica, as a biological control agent to reduce the severity of these ‘plagues’. C. hepatica is unique amongst helminths of mammals in that its eggs are released only upon the death of an infected host. The major goal of the modelling in this paper is to determine the impact of this feature on the population dynamics of the host-parasite interaction. Simple differential equation models are used to examine the general properties of the system and determine which population parameters are most crucial to the outcome of the interaction. These models are supplemented by age-structured models which investigate the initial behaviour of the system after introduction of the parasite. The necessity of host death for transmission is a strongly destabilizing factor, suggesting that C. hepatica cannot regulate most populations stably in the absence of strong resource limitation, although it has the potential to depress mouse populations below infection-free levels. Although C. hepatica influences mouse fecundity at lower burdens than it affects mortality, the age-structured models show that parasite-induced host death cannot be neglected. Because transmission requires host death, the parasite life-cycle operates on a time-scale similar to that of the hosts, and introduction of the parasite as early as possible in the development period of an outbreak will therefore be necessary to achieve substantial reductions in plague intensity.

The parameters of species richness, abundance and diversity of the intestinal helminth infracommunities of a sample of 269 Common gulls, Larus canus, were examined over one season at Agdenes, Norway. The communities developed rapidly in unfledged gulls and the diversity parameters were of similar magnitude in immature and adult birds. All parameters were low in the early part of the season, reached maxima in June or July and fluctuated erratically or declined thereafter. Comparison of summary parameters with those from other species of gulls indicated that helminth community diversity in Common gulls is fairly typical of gulls in general. In gulls as a group, helminth communities show common features of high species richness, low abundance, few or no core species and a number of species that occur rarely and in low numbers. Expectations of particularly high parasite community diversity were not fully realised because species lists and diversity of the component community are not a very good basis for predicting diversity at the infracommunity level.

Distribution of Leptorhynchoides thecatus (Acanthocephala: Rhadinorhynchidae) among the pyloric caeca and the relationship between site and rate of maturation were studied in laboratory infectons of 10, 25 and 40 cystacanths fed to green sunfish, Lepomis cyanellus. After 1 week fish fed at each intensity had significantly different numbers of worms. By the 3rd week post-infection, parasites disappeared from the anterior portion of the intestine. At this time the mean numbers of worms recovered from 25 and 40-cystacanth infections were not significantly different. At the end of the 1st week, the area where caeca join the alimentary tract (between caecal area) and caeca numbered 6 and 7 contained significantly more worms than the other sites. By the 3rd week post-infection only caecum 7 contained significantly more worms, and at 5 weeks there was no significant difference between the number of worms present in any caecum or the between caecal area. Initially worms in the more intense infections matured more slowly, but by the 3rd week post-infection there was no significant difference in the states of maturity. The rate of maturation was not related to the site occupied.

The results are presented of an experimental study of the population biology of chronic Trichuris muris (Nematoda) infection in cortisone-treated CBA/Ca mice. Attention is focused upon both the validity of the common use of faecal egg counts to demonstrate density dependence in helminth fecundity, and the identification of other possible density-dependent mechanisms that may regulate worm numbers in chronic trichuriasis. The results show that faecal egg counts, although demonstrating high daily variation, are not an artefact of host faecal output but a significant density-dependent function of worm burden. This finding contrasts with the observations on Heligmosomoides polygyrus infection in outbred MF1 mice, but accords with similar studies in a wide variety of host - helminth systems. Worm establishment in the murine host is found to be a density related function of infection dose. This is attributed to the probable existence of a physical gut-carrying capacity in the murine host for T. muris. Worm distribution in the gut is also shown to be density dependent, with worms being displaced from the caecum to the colon at increasing intensities of infection. The sex ratio of the adult parasites, however, is found to be both unitary and independent of worm burden. Evidence for a significant density-dependent decline in female T. muris growth or size is presented. The results also show a significant positive association between female T. muris weight and per capita fecundity. These findings indicate that the stunted growth of individual worms at high parasite densities may be a potential mechanism underlying density dependence in helminth fecundity.

The time-course of low and high intensity primary infections with Heligmosomoides polygyrus was monitored in SJL and SWR mice, both of which usually expel worms within 7 weeks of larval administration. Worm expulsion in these strains was not dependent on the intensity of infection, with low and high intensity worm burdens being lost within the same period of time. The ability to expel worms rapidly was inherited in a dominant manner in F1 offspring of SJL or SWR mice mated with C57Bl10 mice; the latter being a strain in which no loss of worms was evident within 10 weeks of infection. However, neither (SJL × C57Bl10)F1 nor (SWR × C57Bl10)F1 mice expelled worms as rapidly as the parental SJL and SWR strains. (SWR × B10G)F1 [H-2q] mice eliminated worms faster than (SWR × C57Bl10)F1 [H-2bq], suggesting that the b haplotype had a moderating influence on the expulsion process. In fact (SWR × B10G)F1 mice showed a significant reduction in worm burdens by week 4 but by weeks 6–8 the rate of worm loss had slowed considerably. In contrast, SJL and SWR mice, whilst initiating rejection slightly later, (after week 4) expelled all worms within the following 2 weeks. Thus two distinct patterns of response were observed among the fast responder strains as exemplified by SWR and SJL mice on the one hand and (SWR × B10G)F1 on the other. Our results support the hypothesis that the course of a primary infection with H. polygyrus is influenced by multiple host gene loci, some of which are encoded within the MHC. SJL and SWR mice probably have similar if not identical gene combinations at loci which determine a fast responder phenotype, distinguishing them from the other mouse strains which have been studied.

The DNA from a number of free-living and parasitic nematode species was examined to determine the genomic number and distribution of DNA sequences encoding two evolutionary conserved proteins; the major sperm protein (MSP) and nematode actin. Ascaris and Caenorhabditis MSP cDNA sequences and Ascaris genomic actin sequences were used to probe Southern blots of Eco RI and Hin d III digested nematode DNA. The number of MSP genes varied widely between the 1 MSP gene in Ascaris and the 60 MSP genes in Caenorhabditis. Filarial nematodes appeared to have 1—4 MSP genes while the plant and insect parasitic species showed from 5–12 MSP-hybridizing restriction fragments. Mammalian intestinal parasites showed between 1 and 13 bands hybridizing with the MSP probes. Blots probed to estimate the number of actin genes showed that, with the exception of Ascaris which contains more than 20 germ line sequences that encode actin, all of the nematodes tested had between 3 and 9 bands that hybridized to the Ascaris genomic actin probe. The possible use of highly conserved sequences such as MSP and actin to differentiate between nematode species in diagnostic and taxonomic studies is discussed.

The invasion and establishment of Heterodera glycines has been studied for 0–120 h after addition to host roots using a synchronized infection system. A high degree of synchrony in development was noted, with juvenile nematodes reaching their feeding sites adjacent to the root endodermis and discharging their subventral pharyngeal glands by 24 h post-invasion (p.i.). The moult to the 3rd-stage juvenile occurred approximately 89 h after entry to the root. Using an enzyme-linked immunosorbent assay with monoclonal antibodies (MAbs) whose tissue specificities had already been defined by immuno-cytochemistry, it was found that MAbs recognizing intestinal lipid droplets and granules showed a major increase in reactivity 18–24 h p.i. before declining by 39 h p.i. In contrast, the reactivity of two MAbs recognizing adult cuticle increased significantly only after 89 h p.i., following the second moult. MAbs to the two subventral pharyngeal gland cells showed a variety of patterns of changing reactivity during the experiment. One increased within 24 h of invasion before declining whereas a second fell without any initial rise. Two further MAbs specific for the subventral glands initially declined but showed a secondary increase in reactivity after 72 h and could be detected in adult females. This pattern was also seen in a MAb specific for the dorsal pharyngeal gland cell. These changes are discussed in the context of events taking place following invasion, with particular reference to the initiation and maintenance of a feeding syncytium by the developing nematode.

Maturity, mass and sex were determined for individual Raphidascaris acus from northern pike, Esox lucius from Dauphin Lake, Manitoba. Length, width and total fecundity of female worms; seasonal changes in worm numbers, growth and maturity; and inequalities in mass and fecundity were quantified for 8 sample periods between April and September. Recruitment of R. acus is most rapid in early spring and continues throughout summer. An accumulation of larvae over winter leads to a peak in the number of gravid worms in spring, while increased feeding by post-spawning pike on yellow perch, Perca fluviatilis, leads to lower levels of gravid worms throughout summer and early autumn. Gravid worms found in spring versus summer were similar with respect to all measured variables and do not appear to be from genetically distinct populations. The strength of the correlation between fecundity and mass for worms was variable between pike and between sample periods, and only weakly correlated when the data were pooled. Much of this variation could be attributed to continued growth of gravid worms after maturation. Worms found in June were smaller, less fecund and less variable and inequalities in mass and fecundity measured using Gini coefficients (GCs) and Lorenz curves (LCs) were most similar at this time. For all other sample periods inequality in fecundity was greater than the corresponding value for mass. Similarity between GCs based on mass and fecundity was maximum when the coefficient of variation (C.V.) for mass was greater than the corresponding C.V. for fecundity. These observations suggest that one must be cautious when applying GCs and LCs to the analysis of parasite populations.