An evaluation has been made of the biological and epidemiological parameters that determine the basic reproductive rates of Taenia hydatigena and T. ovis. These host-parasite systems are characterized by (i) no overcrowding in either host; (ii) no parasite-induced mortality of either host; (iii) no density-dependent constraint in the definitive host, but a strong, rapidly mobilized, short-acting immunity in the intermediate host and (iv) egg production which, in the natural environment, is high enough to prevent superinfection. It is considered that tapeworms with these characteristics are more stable to fluctuations in environmental conditions and to control measures such as dog dosing, than species which have a low egg production and infectivity such as Echinococcus granulosus. Reciprocal immunity exists between T. hydatigena and T. ovis in sheep. Exposure to T. hydatigena suppresses infection by T. ovis, but not by E. granulosus. This has important epidemiological consequences where these parasites co-exist.

Molecular characterization of the Plasmodium falciparum genome has led to identification of polymorphic loci and the mechanisms generating genetic diversity in this parasite. This information has resulted in the development of molecular methods to type parasite diversity in the field. Consequently, we are now in a position to describe the population genetics and dynamics of P. falciparum. The limited number of field studies that have been conducted to date have revealed an extraordinary degree of genetic diversity in natural parasite populations. Heterozygous recombination which occurs during meiosis appears to be one mechanism for generating genetic diversity. The rate at which such recombination occurs in natural parasite populations defines the genetic structure of the parasite population and can influence the ability of the parasite to respond to selection pressure. The high frequency of single genotype infections and the female-biased gametocyte sex ratios found in hyperendemic malaria areas suggest that self-fertilization occurs frequently. Population- wide surveys of allele frequencies in endemic areas have, however, shown no evidence of linkage disequilibrium and are consistent with a panmictic population structure. We argue that these studies have only sampled symptomatic infections, within which rare or recombinant genotypes may be disproportionately represented. They also take no account of the spatial structure of P. falciparum populations. Systematic investigations of the amount of heterozygosity in small areas as part of population-wide surveys are required to define the genetic structure of P. falciparum populations. Population dynamic studies which consider genetic heterogeneity of P. falciparum have shown fluctuations of different serotypes in space and time. The host immune response appears to play an important role in generating these dynamics. Integrated field and laboratory studies, which consider the interaction between population genetics and dynamics, will be necessary to describe the population biology of P. falciparum.

Confocal scanning laser microscopy has been employed with immunocytochemical techniques to map the distribution of serotoninergic and peptidergic components in the nervous system of the monogenean gill-parasite, Diclidophora merlangi; results are compared with the distribution of cholinergic components, following histochemical staining for cholinesterase activity. While all three neurochemical elements are present in the central and peripheral nervous systems, the cholinergic and peptidergic systems dominate the CNS, whereas the PNS has a majority of serotoninergic nerve fibres. The cholinergic and peptidergic neuronal pathways overlap extensively in staining patterns, suggesting possible co-localization of acetylcholine and neuropeptides. Within the peptidergic nervous system, immunoreactivity to the pancreatic polypeptide family of peptides and FMRFamide were the most prevalent.

Metastrongyloids of cats are emerging pathogens that may cause fatal broncho-pulmonary disease. Infestation of definitive hosts occurs after ingestion of intermediate or paratenic hosts. Among metastrongyloids of cats, Troglostrongylus brevior and Troglostrongylus subcrenatus (Strongylida: Crenosomatidae) have recently been described as agents of severe broncho-pulmonary disease. Here, we provide, for the first time, observational evidence suggesting the direct transmission of T. brevior from queen cat to suckling kittens. This new knowledge will have a significant impact on current scientific information of this parasite and shed new light into the biology and epidemiology of metastrongyloid nematodes.

Serum taken from mice 12–15 weeks after a primary infection of Sehistosoma mansoni transfers to normal recipients a partial resistance to subsequent schistosome challenge. The transfer of immunity is evident not oniy in the reduced recovery of mature parasites from the liver, but also in the diminished numbers of invading schistosomula recovered from the lungs. Since the recipients of immune serum exhibit a level of resistance equivalent on average to 47 % of that found in actively immunized animals, the results suggest that humoral factors play a major role in the effector mechanism of schistosome immunity.

We estimate the incidence rate, serological conversion rate and basic case reproduction number (R0) of Leishmania infantum from a cohort study of 126 domestic dogs exposed to natural infection rates over 2 years on Marajó Island, Pará State, Brazil. The analysis includes new methods for (1) determining the number of seropositives in cross-sectional serological data, (2) identifying seroconversions in longitudinal studies, based on both the number of antibody units and their rate of change through time, (3) estimating incidence and serological pre-patent periods and (4) calculating R0 for a potentially fatal, vector-borne disease under seasonal transmission. Longitudinal and cross-sectional serological (ELISA) analyses gave similar estimates of the proportion of dogs positive. However, longitudinal analysis allowed the calculation of pre-patent periods, and hence the more accurate estimation of incidence: an infection–conversion model fitted by maximum likelihood to serological data yielded seasonally varying per capita incidence rates with a mean of 8·66×10−3/day (mean time to infection 115 days, 95% C.L. 107–126 days), and a median pre-patent period of 94 (95% C.L. 82–111) days. These results were used in conjunction with theory and dog demographic data to estimate the basic reproduction number, R0, as 5·9 (95% C.L. 4·4–7·4). R0 is a determinant of the scale of the leishmaniasis control problem, and we comment on the options for control.

This study reports the distribution of parasite genotypes for the trypanosome Crithidia bombi across individual units (the colonies) in host populations of a social insect, the bumble bee Bombus terrestris. A number of microsatellite primers were developed and several of them were found to be polymorphic in our samples. Furthermore, a simple algorithm was used to identify the likely multi-locus genotypes present in multiply infected host individuals. The results demonstrated a remarkably high degree of genetic diversity among infections. A first sample from 1997 could only use a low resolution with 2 loci and showed a total of 11 different genotypes of C. bombi from 12 colonies. The sample from 2000 was analysed at 6 polymorphic loci and contained data from 8 colonies that were infected by 27 different C. bombi genotypes. Roughly 16% of all individual bees but half of all colonies (2000 sample) were infected with more than 1 genotype. The infections in the different colonies were also genetically distinct from each other, and the parasite population as a whole was in linkage disequilibrium and deviated from Hardy–Weinberg expectations. The highly structured and genetically diversified population of C. bombi is likely to result from strong genotypic host–parasite interactions.

The complete nucleotide sequence of the mitochondrial (mt) DNA molecule of the liverfluke, Fasciola hepatica (phylum Platyhelminthes, class Trematoda, family Fasciolidae), was determined. It comprises 14462 bp, contains 12 protein-encoding, 2 ribosomal and 22 transfer RNA genes, and is the second complete flatworm (and the first trematode) mitochondrial sequence to be described in detail. All of the genes are transcribed from the same strand. Of the genes typically found in mitochondrial genomes of eumetazoans, only atp8 is absent. The nad4L and nad4 genes overlap by 40 nt. Most intergenic sequences are very short. Two larger non-coding regions are present. The longer one (817 nt) is located between trnG and cox3 and consists of 8 identical tandem repeats of 85 nt, rich in G and C, followed by 1 imperfect repeat. The shorter non-coding region (187 nt) exhibits no special features and is separated from the longer region by trnG. The gene arrangement resembles that of some other trematodes including the eastern Asian Schistosoma species (and cyclophyllidean cestode species) but it is strikingly different from that of the African schistosomes, represented by Schistosoma mansoni. The genetic code is as inferred previously for flatworms. Transfer RNA genes range in length from 58 to 70 nt, their products producing characteristic ‘clover leaf’ structures, except for tRNA(SUCN) and tRNA(SAGN) lacking the DHU arm.

The fate of irradiated, immunizing cercariae and challenge schistosomula was investigated in mice using a quantitative, histological technique which would appear to be more efficient in estimating parasite numbers in skin and lungs than does the tissue mincing and incubation recovery technique used previously by other workers. There was evidence for slight retardation of irradiated (20 krad.) parasite migration in skin, but death of schistosomula in the skin appeared negligible. It was observed that the majority of irradiated parasites remained in the lungs until at least day 21 after infection, and that most schistosomula observed at this time were dead. In mice immunized with irradiated cercariae there was no evidence of attrition of the challenge infection in the skin. The migration profile of challenge parasites in the lungs of immunized animals was not significantly different from that of normal parasites in the lungs of naive animals. Dead challenge parasites were observed in the lungs but their numbers were not considered great enough to account for the final attrition measured by perfusion. It is suggested that the remainder of the attrition of challenge schistosomula may occur in the systemic and/or hepatic portal circulation.

Both asexual and sexual development of Plasmodium berghei was synchronized without chemical intervention using in vitro culture techniques. Combined in vivo and in vitro experiments were performed on the relationship between age, morphology and maturity of gametocytes. Schizogony took 22–23 h in the experiments. At 26 h post-invasion (p.i) the first males became capable of exflagellation. By 20 h p.i. the first gametocytes were recognizable in Giemsa-stained smears but the sex was hardly distinguishable until maturity (26 h p.i.). Survival time of gametocytes was estimated at 26 h in vitro (half-life 13 h) and the same survival time was suggested for gametocytes in vivo. Schizonts of P. berghei apparently disappeared from the peripheral circulation upon maturity, rupturing almost immediately. Mature schizonts in vitro persisted up to 48 h p.i. in non-agitated cultures. No evidence was collected for sequestration of any sub-population of gametocytes.

The paper examines the possible contributions to be made by Geographic Information Systems (GIS) to studies on human and animal trypanosomiasis in Africa. The epidemiological characteristics of trypanosomiasis are reviewed in the light of the formula for the basic reproductive rate or number of vector-borne diseases. The paper then describes how important biological characteristics of the vectors of trypanosomiasis in West Africa may be monitored using data from the NOAA series of meteorological satellites. This will lead to an understanding of the spatial distribution of both vectors and disease. An alternative, statistical approach to understanding the spatial distribution of tsetse, based on linear discriminant analysis, is illustrated with the example of Glossina morsitans in Zimbabwe, Kenya and Tanzania. In the case of Zimbabwe, a single climatic variable, the maximum of the mean monthly temperature, correctly predicts the pre-rinderpest distribution of tsetse over 82% of the country; additional climatic and vegetation variables do not improve considerably on this figure. In the cases of Kenya and Tanzania, however, another variable, the maximum of the mean monthly Normalized Difference Vegetation Index, is the single most important variable, giving correct predictions over 69 % of the area; the other climatic and vegetation variables improve this to 82 % overall. Such statistical analyses can guide field work towards the correct biological interpretation of the distributional limits of vectors and may also be used to make predictions about the impact of global change on vector ranges. Examples are given of the areas of Zimbabwe which would become climatically suitable for tsetse given mean temperature increases of 1, 2 and 3 °Centigrade. Five possible causes for sleeping sickness outbreaks are given, illustrated by the analysis of field data or from the output of mathematical models. One cause is abiotic (variation in rainfall), three are biotic (variation in vectorial potential, host immunity, or parasite virulence) and one is historical (the impact of explorers, colonizers and dictators). The implications for disease monitoring, in order to anticipate sleeping sickness outbreaks, are briefly discussed. It is concluded that present data are inadequate to distinguish between these hypotheses. The idea that sleeping sickness outbreaks are periodic (i.e. cyclical) is only barely supported by hard data. Hence it is even difficult to conclude whether the major cause of sleeping sickness outbreaks is biotic (which, in model situations, tends to produce cyclical epidemics) or abiotic. The conclusions emphasize that until we understand more about the variation in space and time of tsetse and trypanosomiasis distribution and abundance we shall not be in a position to benefit from the advances made by GIS. The potential is there, however, to re-introduce the spatial and temporal elements into epidemiological studies that are currently often neglected.

The distributions of parasitic diseases are determined by complex factors, including many that are distributed in space. A variety of statistical methods are now readily accessible to researchers providing opportunities for describing and ultimately understanding and predicting spatial distributions. This review provides an overview of the spatial statistical methods available to parasitologists, ecologists and epidemiologists and discusses how such methods have yielded new insights into the ecology and epidemiology of infection and disease. The review is structured according to the three major branches of spatial statistics: continuous spatial variation; discrete spatial variation; and spatial point processes.

This review focuses on some of the more ground-shifting advances of recent decades, particularly those at the molecular and cellular level that illuminate mechanisms underpinning the natural ecology of tick-host-pathogen interactions and the consequent epidemiology of zoonotic infections in humans. Knowledge of components of tick saliva, now recognized as the central pillar in the tick's ability to complete its blood meal and the pathogen's differential ability to use particular hosts for transmission, has burgeoned with new molecular techniques. Functional studies have linked a few of them to saliva-assisted transmission of non-systemic infections between co-feeding ticks, the quantitative key to persistent cycles of the most significant tick-borne pathogen in Europe. Human activities, however, may be equally important in determining dynamic patterns of infection incidence in humans.

This study sought to assess the effect of giardiasis on growth of young children. In Salvador, northeast Brazil, 597 children initially aged 6 to 45 months were followed for a year in 1998/9, measured anthropometrically thrice, every 6 months, and monitored for diarrhoea prevalence twice weekly. Stool samples were collected and examined during the second round of anthropometry, and infected children were treated 39 days later, on average (S.D. 20 days). For each 6-month interval, the gains in z-scores of infected and uninfected children were compared, after adjustment for potential confounding factors, including longitudinal prevalence of diarrhoea. No significant difference was found for the first interval but in the second, the gain in adjusted height-for-age z-score was 0·09 less in infected than uninfected children, equivalent to a difference in height gain of 0·5 cm. The shortfall in growth was greater in children who remained free of diarrhoea, and was significantly correlated with the proportion of the second interval during which the child had remained untreated. We conclude that Giardia can impede child growth even when asymptomatic, presumably through malabsorption. This finding challenges the view that young children found to have asymptomatic giardiasis in developing countries should not be treated.

Coccidiosis, a serious disease resulting from infection with parasitic protozoa of the genus Eimeria, causes significant economic losses to the poultry industry, where intensive rearing facilitates transmission of infectious oocysts via the fecal/oral route. Current control relies primarily on prophylactic drugs in feed but, whilst cost effective, the rise of drug resistance and public demands for residue-free meat has encouraged development of alternative control strategies. Chickens that recover from infection with Eimeria develop solid immunity that is directed against the early asexual stages of the parasite life cycle. This has allowed development of a number of vaccines that utilize deliberate infection with controlled doses of virulent oocysts or reproductively attenuated lines of Eimeria. The latter are immunogenic but non-pathogenic. The realization that both prophylactic drugs and attenuated vaccines control but do not eradicate infection with Eimeria encouraged development of a vaccine based upon maternal immunity. Laying hens exposed to Eimeria are able to transfer protective antibodies to hatchlings via egg yolks and these antibodies have been used to identify parasite proteins that are conserved across the genus. When delivered maternally, these provide an economical means of preventing coccidiosis, offering immediate protection to newly hatched chicks.

For the majority of intestinal parasites, real-time PCR-based diagnosis outperforms microscopy. However, the data for Trichuris trichiura have been less convincing and most comparative studies have been performed in populations with low prevalence. This study aims to improve detection of T. trichuria DNA in human stool by evaluating four sample preparation methods. Faecal samples (n = 60) were collected at Flores island, Indonesia and examined by microscopy. Aliquots were taken and a bead-beating procedure was used both on directly frozen stool and on material preserved with 96% ethanol. PCR on frozen samples showed 40% to be positive for T. trichiura, compared with 45% positive by microscopy. The percentage positive increased when using ethanol preservation (45·0%), bead-beating (51·7%) and a combination (55·0%) and all three methods showed significantly higher DNA loads. The various procedures had a less pronounced effect on the PCR results of nine other parasite targets tested. Most prevalent were Ascaris lumbricoides (≈60%), Necator americanus (≈60%), Dientamoeba fragilis (≈50%) and Giardia lamblia (≈12%). To validate the practicality of the procedure, bead-beating was applied in a population-based survey testing 910 stool samples. Findings confirmed bead-beating before DNA extraction to be a highly efficient procedure for the detection of T. trichiura DNA in stool.

A culture chamber fitted with a polycarbonate sieve has been used to isolate Plasmodium knowlesi merozoites as they are released from schizonts. A 3 μm pore-size sieve allows passage of normal erythrocytes and red cells containing rings and trophozoites and can be used to concentrate schizonts from a mixed cell population. A 2 μm pore-size sieve retains normal and parasitized cells and provides uncontaminated merozoites in high yield (5 × 1010 merozoites per ml schizonts). Merozoite viability diminishes rapidly during 30 min after isolation. These preparations should prove valuable for studies of the biochemical, physiological and antigenic properties of this transient extracellular phase of the malaria parasite.

Schistosomes are parasites of considerable medical and veterinary importance and, therefore, all aspects of their biology have been intensively studied. In contrast, our knowledge of species of the largest genus, Trichobilharzia, is insufficient. Because morphological characterization of Trichobilharzia species provides a limited number of criteria for species determination, molecular data are required. In the present paper, we sequenced internal transcribed spacers ITS1 and ITS2, and 5·8S ribosomal RNA (rRNA) genes of 3 European Trichobilharzia species (T. regenti, T. szidati and T. franki). We showed that ITS1 and ITS2 sequences can be used in species identification. Repetitive elements were found in ITS1 of all 3 Trichobilharzia species; their number and length varied depending on the species. Phylogenetic analysis showed that the visceral T. franki is more related to the nasal T. regenti, than to the visceral T. szidati. The newly designed primer, which is specific for T. regenti, might be used as a tool for diagnosis of this potential pathogen.

The morphology and genetic characterization of a new species of trypanosome infecting koalas (Phascolarctos cinereus) are described. Morphological analysis of bloodstream forms and phylogenetic analysis at the 18S rDNA and gGAPDH loci demonstrated this trypanosome species to be genetically distinct and most similar to Trypanosoma bennetti, an avian trypanosome with a genetic distance of 0·9% at the 18S rDNA and 10·7% at the gGAPDH locus. The trypanosome was detected by 18S rDNA PCR in the blood samples of 26 out of 68 (38·2%) koalas studied. The aetiological role of trypanosomes in koala disease is currently poorly defined, although infection with these parasites has been associated with severe clinical signs in a number of koalas. Based on biological and genetic characterization data, this trypanosome species infecting koalas is proposed to be a new species Trypanosome irwini n. sp.

Infections with Haemonchus contortus are a major constraint on ruminant health world-wide. Young lambs are very sensitive to Haemonchus infection. Older lambs and sheep acquire immunity after a continuous or seasonal exposure to the parasite. The mechanisms underlying immunity are still not completely understood. Antibodies, in particular local IgA and IgE, certainly play a role. The role of IgG is less clear. Lymphocyte proliferation responses seem to correlate to immunity. Sheep that have high antigen-induced lymphocyte responses have a low susceptibility to infection. Furthermore, several studies have demonstrated that immunity against H. contortus is associated with mastocytosis and hypersensitivity reactions. More recently, increasing attention is being paid to the role of cytokines (interleukins and γ- interferon) in the activation of specific defence mechanisms. Reverse transcriptase–polymerase chain reaction (RT–PCR) assays to study cytokine mRNA expression have become available. The inability of young lambs to mount a significant Th2 response, which is normally characterized by high IgE levels, mastocytosis and eosinophilia, may account for the phenomenon of unresponsiveness in these animals.