The first step in studying the epidemiology of a disease is the accurate identification of the pathogen. Traditional reliance on morphological identification has given way to the use of molecular methods for the detection and identification of pathogens, greatly improving our understanding of epidemiology. For the African tsetse-transmitted trypanosomes, the growth of PCR methods for identification of trypanosomes has led to increased appreciation of trypanosome genetic diversity and discovery of hitherto unknown trypanosome species, as well as greater knowledge about the number and type of trypanosome infections circulating in mammalian hosts and vectors. Sequence data and phylogenetic analysis have provided quantitative information on the relatedness of different trypanosome species and allowed the new trypanosome genotypes discovered through the use of species identification methods in the field to be accurately placed in the phylogenetic tree.

According to the Red Queen hypothesis for sex, cross-fertilization should be positively associated with the probability of exposure (risk) to virulent parasites. Unfortunately, risk is difficult to measure in the wild, and prevalence of infection is often substituted for risk. Here I suggest that prevalence of infection may not generally suffice as a surrogate for risk, since the Red Queen model can make opposite predictions depending on the distribution of risk in the wild. Specifically, the results of a matching-alleles model suggest that asexual populations should be more infected than sexual populations, when (1) the variance in risk among populations is small, and (2) the mean risk of exposure to parasites is near the point where selection switches to favouring sex over asex. If, however, the variance in risk among populations is large, sexual reproduction should be positively associated with the prevalence of infection. In addition, the coefficient of variation for reproductive mode should increase sharply at the switch point. In light of these results, I re-evaluated data from two studies on the distribution of males in 95 populations of a freshwater snail (Potamopyrgus antipodarum). Populations of these snails are often mixtures of sexual and asexual individuals, and the frequency of males is correlated with the frequency of sexual females in the population. The results show a large, highly skewed variance among populations for prevalence of infection by larval trematodes. The results also show a positive, significant relationship between prevalence of infection and the frequency of males, with a sharp increase in the coefficient of variation at intermediate prevalence. In addition, experimental studies suggest that some of the necessary conditions of the Red Queen hypothesis are also met in this system. Specifically, the most common trematode infecting these snails is (1) adapted to infecting local host populations of the snail, and is (2) more infective to clones that were common in the recent past. It is too early to know if the parasite theory is sufficient to explain the widespread distribution of sex. I suggest that the theory is not sufficient, but that parasites in combination with mutation accumulation in clonal lines may explain the maintenance of sex in species that occasionally produce apomictic mutants.

We investigated the diversity of Bartonella in Apodemus agrarius, an important rodent of peri-domestic habitats, which has spread into Europe in the past 1000 years. Spleen samples of 344 A. agrarius from Eastern Slovakia were screened for the presence of Bartonella spp. using 16S–23S rRNA internal transcribed spacer region and bacteria were detected in 9% of rodents. Based on sequencing of three housekeeping genes (gltA, rpoB and groEL) Bartonella genotypes were ascribed to the species typical for mice and voles: B. grahamii, B. taylorii and B. birtlesii. However, the study also confirmed presence of genotypes belonging to the B. clarridgeiae/B. rochalimae clade, and the B. elizabethae/B. tribocorum clade, which are not commonly found in woodland rodents. In addition, a potential recombination event between these two genotypes was noted, which highlights an important role of A. agrarius in shaping Bartonella diversity and evolution.

1. We have discovered that Trypanblau and Trypanrot are highly efficient remedies in the treatment of canine piroplasmosis, no drug or mode of treatment having hitherto been found to exert any appreciable effect upon this very fatal disease.

2. The drugs exert a direct and observable effect upon the parasites (a) by causing the pyriform parasites to quickly disappear, and (b) in most cases, by causing the total disappearance of the parasites from microscopic observation in the peripheral blood.

3. The disappearance of the parasites from the blood is usually temporary, the parasites reappearing in small numbers after an interval of 9 to 12 days, but the dogs, as a rule, show no symptoms and gradually progress toward recovery. In our experience there is but slight loss of weight in treated animals, this being in marked contrast to what is usually observed in dogs which recover naturally. In the two dogs which died of a relapse the parasites reappeared after an interval of 4 to 5 days.

4. The experiments were conducted upon 21 dogs of all ages and of a variety of breeds. Many of the dogs were highly susceptible puppies. Twelve dogs were subjected to curative treatment, one dog received preventive treatment, and eight dogs served as controls.

5. Twelve dogs received treatment as follows:

(a) Two dogs were treated with Trypanrot and survived 111 days and 20 days respectively. In the latter case the dog was treated in an advanced stage of the disease, the parasites disappeared, and death appeared to be due to the after-effects of the malady.

Two control dogs died respectively on the 9th and 7th day after they were inoculated.

(b) Ten dogs were treated with Trypanblau and seven made a good recovery. Three of these dogs are still alive, having been respectively under observation for periods of 90, 83 and 65 days. Four of the recovered dogs died respectively of distemper on the 42nd and 43rd day, of severe mange on the 52nd day and of distemper and mange on the 69th day. We failed to cure the disease in 3 out of 10 cases treated: two small wormy ill-nourished puppies died from a relapse (on the 13th and 15th day) after they appeared to be on the road to recovery; one dog, treated only 3 hours prior to death, and when 50% of its corpuscles were infected, succumbed to the disease, but even in this case a visible effect was exerted upon the parasites by the drug.

Of the 7 control dogs, 6 died of piroplasmosis within 7–13 days after inoculation with virulent blood, and the remaining dog died of piroplasmosis on the 36th day after inoculation.

6. After the parasites disappear under the influence of the drug, the temperature usually falls to normal, but slight rises of temperature may at times occur, together with the subsequent reappearance of the parasites.

7. When the parasites reappear after treatment they occur in small numbers and can usually be detected only at the edge of the film. In such cases the parasites tend to occur in higher multiples [(PPPP) or more] within the individual corpuscles. The same appearance has been observed in one of the control dogs, which died of chronic piroplasmosis on the 36th day.

8. In the two puppies 10 and 11, in which death followed upon a relapse, the parasites reappeared after a shorter interval (4 to 5 days) than in any of the other dogs. The blood counts made during the fatal relapse are remarkable as showing a high percentage of dividing forms (D); that rapid invasion of fresh corpuscles was proceeding is also shown by the high percentage of single pyriform parasites (P) which occurred in the corpuscles.

9. One experiment is reported (Dog 13) in which Trypanblau was injected 24 hours after the animal was inoculated, with the result that no parasites appeared in the dog's blood up to the 65th day, and the dog remained perfectly well. The control dog died of piroplasmosis on the 7th day. (This experiment has since been successfully repeated.)

Further experiments of this character will be shortly reported upon.

10. Arsacetin and Soamin exert no curative effect upon canine piroplasmosis.

The Regent Honeyeater (Xanthomyza phyrigia) is an endangered Australian bird species. Breeding populations have been established at Australian zoos in support of re-introduction programmes. This species is the host of a new species of Isospora (Apicomplexa). Oocysts are spherical, 25·8 (22·5–28·75) by 23·8 (20–26·25) μm with a colourless to pale yellow smooth wall undergoing rapid exogenous sporulation, 90% sporulated oocysts in 8 h at 20°C. Each oocyst contains 1 polar granule. Sporocysts are ovoid, 18·67 (17–19) by 9·49 (9–10) μm with a flat Stieda body and spherical substieda body devoid of a hyaline body. The asexual stages and sexual phase is within the enterocytes of the duodenum and jejunum. Faeces collected in the morning (AM, n=84) and in the afternoon (PM, n=90) revealed significant diurnal periodicity in oocyst shedding; 21% (18 of 84) of the AM were positive with the mean of 499 oocysts.g−1 compared to the PM with 91% (82 of 90) bird faeces positive with the mean of 129 723 oocysts.g−1. Therefore, parasite checks for these birds should be carried out in the afternoon to obtain an accurate result. The ecological significance of the high parasite burden in captive birds requires further investigation and comparison to the wild counterparts.

Haemosporidian parasites infecting birds show distinct heterogeneity in their distribution among host species. However, despite numerous studies on the prevalence and diversity of parasite communities across species, very little is known on patterns of differences between them. Such data is lacking because up to date the majority of studies explored the patterns of variation in infections in different years, different time of sampling within a year or a breeding cycle, different study sites or was based on a small sample size, all of which may affect the estimates of prevalence and parasite diversity. Here, the prevalence, richness and diversity of haemosporidian parasites from the genera Plasmodium and Haemoproteus were studied in two closely related non-migratory hole-nesting passerines: Great Tits and Blue Tits. Birds were sampled in sympatrically breeding populations during two seasons at the same stage of their breeding cycle – late nestling care. Great Tits were more prevalently infected with Plasmodium and Haemoproteus parasites (97·1 vs 71·2%), harboured a higher proportion of multiple infections (26·2 vs 3·2%) and had a more diverse parasite community (11 vs 5 parasite lineages) than Blue Tits. Observed differences between two host species are discussed with reference to their breeding densities and immunological and behavioural characteristics.

The effect of Sanguinicola inermis on serum antibody and complement activity in Cyprinus carpio was assessed using an ELISA and haemolytic assays. Possible immune evasion strategies were assessed using immunodetection of host proteins on the surface of the parasite. Carp acclimatized to 20 or 25 °C were infected by exposure to 500 cercariae or injected intraperitoneally with 150 cercariae, and serum monitored over a 63-day period. In cercariae-injected carp, irrespective of time and temperature, a significant increase occurred in complement activity being greatest at 25 °C. In addition, fish exposed to the cercariae of S. inermis and maintained at 20 °C the level of complement activity was significantly higher after 5 weeks compared to controls. At 20 °C intraperitoneal injections of parasites increased serum antibody levels which peaked after 7 days. In contrast, at 25 °C, antibody levels were maintained over 63 days. Exposure of fish to infection did not appear to stimulate antibody production. Immunofluorescence studies revealed ‘host-like’ molecules on the surface of the cercarial body exposed to carp serum and adult flukes obtained directly from the fish or cultured for 24 h in L15 medium. The possible role of ‘host-like’ molecules in immune evasion is discussed and the response at different temperatures is related to infection dynamics.

The growth and development of Dipylidium caninum, Hymenolepis citelli, H. diminuta, H. microstoma, and H. nana were examined in Ctenocephalides felis, and the Hymenolepis species further examined in the reference host Tribolium confusum. This is the first recorded infection of C. felis with these four species of Hymenolepis and also of a flea with H. citelli.

Growth and development of the four species of Hymenolepis were comparable in C. felis and T. confusum. Observations relative to scolex formation indicated that perhaps only one basic sequence occurs in the genus Hymenolepis.

Growth and metamorphosis of the flea host had no important effects upon Hymenolepis development. However, presumably owing to greater crowding in C. felis than in T. confusum, an increased amount of retarded development was noted in cestodes from fleas. In contrast, the growth and metamorphosis of the flea host did affect the development of D. caninum. Little growth occurred in larval fleas but with the onset of pupation an increased growth rate was initiated, the cestode reaching almost its maximum size prior to the emergence of the adult flea. Cysticercoid differentiation and maturation mostly occurred within the adult.

Sera from swine, rabbit, and dog, that had never been exposed to Toxoplasma gondii, demonstrated significant killing of T. gondii tachyzoites in vitro, while cat serum did not. Swine and rabbit sera contained natural IgM antibody against the tachyzoites, and the classical complement pathway was activated by the binding of natural IgM antibody to the tachyzoites, leading to lysis. Anti-T. gondii antibodies, induced in swine or cat infected with T. gondii, had no killing effect by themselves but killed the tachyzoites in the presence of swine complement. However, the anti-T. gondii antibodies of swine or cat demonstrated a very low killing effect in the presence of cat complement. This suggests that T. gondii tachyzoites have an evasion mechanism to prevent lysis which is specific for cat complement.

The arbitrary genus Phytomonas includes a biologically diverse group of kinetoplastids that live in a wide variety of plant environments. To understand better the subdivisions within the phytomonads and the variability within groups, the exon, intron and non-transcribed spacer sequences of the spliced leader RNA gene were compared among isolates of the phloem-restricted members. A total of 29 isolates associated with disease in coconut, oil palm and red ginger (Alpinia purpurata, Zingibreaceae) were examined, all originating from plantations in South America and the Caribbean over a 12-year period. Analysis of non-transcribed spacer sequences revealed 2 main groups, I and II; group II could be further subdivided into 2 subgroups, IIa and IIb. Three classes of spliced leader (SL) RNA gene were seen, with SLI corresponding to group I, SLIIa to group IIa, and SLIIb to group IIb. Two isolates showed some characteristics of both major groups. Group-specific oligonucleotide probes for hybridization studies were tested, and a multiplex amplification scheme was devised to allow direct differentiation between the 2 major groups of phloem-restricted Phytomonas. These results provide tools for diagnostic and molecular epidemiology of plant trypanosomes that are pathogenic for commercially important flowers and palms.

Changes in host phenotype are often attributed to manipulation that enables parasites to complete trophic transmission cycles. We characterized changes in host phenotype in a colonial host–endoparasite system that lacks trophic transmission (the freshwater bryozoan Fredericella sultana and myxozoan parasite Tetracapsuloides bryosalmonae). We show that parasitism exerts opposing phenotypic effects at the colony and module levels. Thus, overt infection (the development of infectious spores in the host body cavity) was linked to a reduction in colony size and growth rate, while colony modules exhibited a form of gigantism. Larger modules may support larger parasite sacs and increase metabolite availability to the parasite. Host metabolic rates were lower in overtly infected relative to uninfected hosts that were not investing in propagule production. This suggests a role for direct resource competition and active parasite manipulation (castration) in driving the expression of the infected phenotype. The malformed offspring (statoblasts) of infected colonies had greatly reduced hatching success. Coupled with the severe reduction in statoblast production this suggests that vertical transmission is rare in overtly infected modules. We show that although the parasite can occasionally infect statoblasts during overt infections, no infections were detected in the surviving mature offspring, suggesting that during overt infections, horizontal transmission incurs a trade-off with vertical transmission.

Molecular phylogeography has revolutionised our ability to infer past biogeographic events from cross-sectional data on current parasite populations. In ecological parasitology, this approach has been used to address fundamental questions concerning host-parasite co-evolution and geographic patterns of spread, and has raised many technical issues and problems of interpretation. For applied parasitologists, the added complexity inherent in adding population genetic structure to perceived parasite distributions can sometimes seem to cloud rather than clarify approaches to control. In this paper, we use case studies firstly to illustrate the potential extent of cryptic diversity in parasite and parasitoid populations, secondly to consider how anthropogenic influences including movement of domestic animals affect the geographic distribution and host associations of parasite genotypes, and thirdly to explore the applied relevance of these processes to parasites of socio-economic importance. The contribution of phylogeographic approaches to deeper understanding of parasite biology in these cases is assessed. Thus, molecular data on the emerging parasites Angiostrongylus vasorum in dogs and wild canids, and the myiasis-causing flies Lucilia spp. in sheep and Cochliomyia hominovorax in humans, lead to clear implications for control efforts to limit global spread. Broader applications of molecular phylogeography to understanding parasite distributions in an era of rapid global change are also discussed.

The immune response to leishmaniasis is complex, and the result of infection depends on both the genetic composition of the Leishmania species and the immunity of the host. Clinical and experimental evidence suggest that the activation of B cells leads to exacerbation of visceral leishmaniasis. However, the role of B-1 cells (a subtype of B lymphocytes) in the pathogenesis of experimental visceral leishmaniasis has not yet been elucidated. In this study, we investigated the importance of B-1 cells in experimental infection with Leishmania. (L.) chagasi. Our results showed that BALB/XID mice (X-linked immunodeficient mice which are genetically deficient in B-1 cells) infected with L. (L.) chagasi for 45 days had a significant reduction in parasite load in the spleen when compared with control mice. Cytokine analysis showed that the BALB/XID mice had lower amounts of IL-10 in their sera compared with control group. In addition, the transfer of B-1 cells from wild type mice into IL-10KO animals led to an increase in susceptibility to L. (L.) chagasi infection in the IL-10KO mice, suggesting that the IL-10 produced by these cells is important in experimental infection. Our results suggest that B-1 cells may play an important role in susceptibility to L. (L.) chagasi.

There has been a recent shift in the literature on mosquito/Plasmodium interactions with an increasingly large number of theoretical and experimental studies focusing on their population biology and evolutionary processes. Ecological immunology of mosquito-malaria interactions – the study of the mechanisms and function of mosquito immune responses to Plasmodium in their ecological and evolutionary context – is particularly important for our understanding of malaria transmission and how to control it. Indeed, describing the processes that create and maintain variation in mosquito immune responses and parasite virulence in natural populations may be as important to this endeavor as describing the immune responses themselves. For historical reasons, Ecological Immunology still largely relies on studies based on non-natural model systems. There are many reasons why current research should favour studies conducted closer to the field and more realistic experimental systems whenever possible. As a result, a number of researchers have raised concerns over the use of artificial host-parasite associations to generate inferences about population-level processes. Here I discuss and review several lines of evidence that, I believe, best illustrate and summarize the limitations of inferences generated using non-natural model systems.

An ultrastructural study of microfilariae of Wuchereria bancrofti was performed after in vitro treatment with diethylcarbamazine. One of the first morphological alterations produced by treatment with 5, 10 or 50 μg/ml of DEC was the loss of microfilarial sheaths. Drastic effects of DEC were only detected when microfilariae were exsheathed. Microfilariae treated with 5, 10 or 50 μg/ml DEC showed severely affected organelles, formation of several vacuoles mainly in the hypodermis, and cytolysis. Some microfilariae showed extreme cellular disorganization with abundance of electron-dense degenerating organelles, numerous large vacuoles, and nuclear condensation. Lamellar bodies probably related to an assembly of endoplasmic reticulum membranes were observed in some damaged microfilariae. Thus, in vitro treatment with concentrations of DEC similar to therapeutic conditions had direct effect by causing a loss of the microfilarial sheath of W. bancrofti with subsequent damage of organelles and apoptosis.

In a 3-year study, we examined landscape features (aspect, slope, sun exposure, canopy cover, type of ground cover, and nearest water source) that were potentially related to prevalence of infection with Plasmodium mexicanum in fence lizards (Sceloporus occidentalis) within a 4.5 ha study area in northern California, USA. Logistic regression analysis showed that ground cover type was the primary mediator of the probability of P. mexicanum infection. Infected lizards were captured more often in rock and/or leaf litter locations than in grassy ones. In another experiment, the study area was divided into 9 sites (0.07–0.33 ha), and infection prevalence was calculated for each. Three sites with high (>30%) infection prevalence had significantly more rocky outcrops and leaf litter than those with low (<20%) or moderate (20–30%) infection prevalence (N = 3 sites each). We conclude that lizard site selection may influence the probability of exposure to infected vectors and thus the likelihood of P. mexicanum infection. We also demonstrate that studies at different spatial scales may be required to understand fully the relationship between landscape features and parasite distribution.

With the increased availability of multilocus sequence data, the lack of concordance of gene trees estimated for independent loci has focused attention on both the biological processes producing the discord and the methodologies used to estimate phylogenetic relationships. What has emerged is a suite of new analytical tools for phylogenetic inference – species tree approaches. In contrast to traditional phylogenetic methods that are stymied by the idiosyncrasies of gene trees, approaches for estimating species trees explicitly take into account the cause of discord among loci and, in the process, provides a direct estimate of phylogenetic history (i.e. the history of species divergence, not divergence of specific loci). We illustrate the utility of species tree estimates with an analysis of a diverse group of feather mites, the pinnatus species group (genus Proctophyllodes). Discord among four sequenced nuclear loci is consistent with theoretical expectations, given the short time separating speciation events (as evident by short internodes relative to terminal branch lengths in the trees). Nevertheless, many of the relationships are well resolved in a Bayesian estimate of the species tree; the analysis also highlights ambiguous aspects of the phylogeny that require additional loci. The broad utility of species tree approaches is discussed, and specifically, their application to groups with high speciation rates – a history of diversification with particular prevalence in host/parasite systems where species interactions can drive rapid diversification.

Female Tenebrio molitor infected with metacestodes of Hymenolepis diminuta exhibit elevated concentrations of female-specific proteins in their haemolymph and the origin of these has been investigated. Following a 4 h in vitro incubation with [14C]leucine, fat bodies from non-infected females secreted 13 times more protein than those from females 12 days post-infection. A comparison of the uptake in vivo of radio-isotope labelled amino acids by ovaries from non-infected and infected beetles of various ages revealed no differences; however, a 51·5% decrease in protein sequestration was detected in females 12 days post-infection. Electrophoresis of homogenates of radio-isotope labelled ovaries demonstrated that the majority of label was associated with vitellin sub-units. It is suggested that the decrease in vitellogenin sequestration associated with infection results in an increase in the haemolymph concentration of these proteins despite a concomitant reduction in their secretion by fat bodies. Both fat body synthesis and ovarian sequestration are under juvenile hormone control and it is proposed that metacestodes of H. diminuta may cause a reduction in the concentration of this hormone in the intermediate host.

1. Liver lesions were easily induced in hamsters with two strains of E. histolytica but not with six others.

2. Serial liver passage increased the invasiveness to the rat's caecum of one strain which had become attenuated by in vitro cultivation.

3. The invasiveness of one of the liver passage substrains was lost a second time after cultivation in vitro and a second series of liver passages restored some degree of virulence.

4. Serial liver passage did not increase the invasiveness of two avirulent strains.

5. The ease of production of amoebic liver lesions in hamsters is not correlated with invasiveness in the rat's caecum.

Parasites are more fecund than free-living relatives. The traditional explanation of this is that parasites have to compensate for massive mortality in the transmission phase of their life cycles, but there are neo-Darwinian problems with this interpretation. Similarly, parasites invest more resources in reproduction than free-living relatives but often live longer as adults, and yet negative correlations are expected between fecundity and longevity. These patterns and paradoxes are discussed within the context of a general life-cycle theory. The theory is also used to address questions concerning the influence of age-specific mortality on life-cycle patterns, the trade-off between gamete size and numbers, and the relative merits of gametic and non-gametic reproduction. Wherever possible, the theory is related to facts about parasites.