Haemosporidian parasites belonging to Haemoproteus cause avian diseases, however, vectors remain unidentified for the majority of described species. We used the laboratory-reared biting midges Culicoides nubeculosus to determine if the sporogonic development of three widespread Haemoproteus parasites completes in this insect. The midges were reared and fed on one common blackbird, white wagtail and thrush nightingale naturally infected with Haemoproteus minutus, Haemoproteus motacillae and Haemoproteus attenuatus, respectively. The engorged females were dissected in order to follow their sporogonic development. Microscopic examination was used to identify sporogonic stages. Bayesian phylogeny based on partial cytochrome b gene was constructed in order to determine phylogenetic relationships among Culicoides species-transmitted haemoproteids. All three parasites completed sporogony. Phylogenetic analysis placed Culicoides species transmitted haemoproteids in one well-supported clade, proving that such analysis readily indicates groups of dipteran insects transmitting avian haemoproteids. Available data show that 11 species of Culicoides have been proved to support complete sporogony of 18 species of avian haemoproteids. The majority of Culicoides species can act as vectors for many Haemoproteus parasites, indicating the low specificity of these parasites to biting midges, whose are globally distributed. This calls for control of haemoproteid infections during geographical translocation of infected birds.

The immunoprotective potential of Babesia divergens antigens released in supernatants of in vitro cultures of the parasite is generally known. Among a number of parasite molecules, a 37 kDa protein has been found in the supernatants of Babesia divergens cultures. In this report the cloning and biochemical characterization of this protein, called Bd37, are describedThe sequence data reported herein have been deposited in EMBL under Accession No. AJ422214. . In addition, the processing of the protein was studied in vitro. Results suggest that Bd37 is encoded by a single copy gene. Bd37 appears to be a merozoite-associated molecule attached to the surface by a glycosylphosphatidylinositol moiety containing a palmitate residue attached to the inositol ring. In addition, it is demonstrated that both extremities of the protein are linked by a disulphide bond. Results further indicate that a soluble, hydrophilic form of Bd37 can be released from the merozoite surface by GPI-specific phospholipase D. The potential role the Bd37 protein and the GPI anchor are discussed.

We have previously shown that oral treatment with the leaf extract of the plant Kalanchoe pinnata (Kp) significantly decreases the lesion size and the parasite load in BALB/c mice infected with Leishmania amazonensis. Here we report on the mode of action of Kp, particularly on the induction of nitric oxide (NO) production by macrophages. We observed that Kp has no direct inhibitory activity on extracellular promastigotes, but effectively decreases the intracellular amastigote growth in a dose-related fashion. A 58% reduction in amastigote growth induced by 500 μg/ml Kp was associated with a 6-fold increase in the production of NO by the macrophages. IFN-γ synergistically enhanced the NO-stimulating effect of Kp in culture. Co-treatment with the inducible NO synthase enzyme inhibitor l-NG-monomethyl-arginine abolished the antileishmanial effect of Kp in vitro and in L. amazonensis-infected BALB/c mice. These results indicate that the protective effect of Kp in leishmaniasis may not be due to a direct effect on the parasite itself but rather to activation of the reactive nitrogen intermediates pathway of macrophages.

Nosema ceranae is a widespread honeybee parasite, considered to be one of the pathogens involved in the colony losses phenomenon. To date, little is known about its intraspecific genetic variability. The few studies on N. ceranae variation have focused on the subunits of ribosomal DNA, which are not ideal for this purpose and have limited resolution. Here we characterized three single copy loci (Actin, Hsp70 and RPB1) in three N. ceranae isolates from Hungary and Hawaii. Our results provide evidence of unexpectedly high levels of intraspecific polymorphism, the coexistence of a wide variety of haplotypes within each bee colony, and the occurrence of genetic recombination in RPB1. Most haplotypes are not shared across isolates and derive from a few frequent haplotypes by a reduced number of singletons (mutations that appear usually just once in the sample), which suggest that they have a fairly recent origin. Overall, our data indicate that this pathogen has experienced a recent population expansion. The presence of multiple haplotypes within individual isolates could be explained by the existence of different strains of N. ceranae infecting honeybee colonies in the field which complicates, and must not be overlooked, further analysis of host–parasite interactions.

The presence or absence of parasites within host populations is the result of a complex of factors, both biotic and abiotic. This study uses a non-parametric classification tree approach to evaluate the relative importance of key abiotic and biotic drivers controlling the presence/absence of parasites with complex life cycles in a sentinel, the common killifish Fundulus heteroclitus. Parasite communities were classified from 480 individuals representing 15 fish from 4 distinct marsh sites in each of 4 consecutive seasons between 2006 and 2007. Abiotic parameters were recorded at continuous water monitoring stations located at each of the 4 sites. Classification trees identified the presence of benthic invertebrate species (Gammarus sp. and Littorina sp.) as the most important variables in determining parasite presence: secondary splitters were dominated by abiotic variables including conductance, pH and temperature. Seventy percent of hosts were successfully classified into the correct category (infected/uninfected) based on only these criteria. The presence of competent definitive hosts was not considered to be an important explanatory variable. These data suggest that the most important determinant of the presence of these parasite populations in the common killifish is the availability of diverse communities of benthic invertebrates.

Schistosome parasites, despite being multicellular organisms several millimetres in length, can survive in the bloodstream of mammalian hosts for decades. The remarkable and complex adaptation exemplified in the host–parasite relationship in schistosomiasis may include not only immune evasion by the parasite, but also immune exploitation. While the developmental and adult stages of the parasite are by and large invisible to the immune response, the parasite egg induces a granulomatous reaction which not only protects the host from a diffusible parasite toxin, but also is required for normal transmission of parasite eggs from the host to the external environment. Other possible mechanisms of immune exploitation by schistosomes are discussed including skewing of cytokine responses, effects of cytokines on worm fecundity, exploitation of endothelial cell adherence, and induction of IgE.

Plasmodium falciparum displays a large and remarkable variety of heat shock protein 40 family members (PfHsp40s). The majority of the PfHsp40s are poorly characterized, and although the functions of some of them have been suggested, their exact mechanism of action is still elusive and their interacting partners and client proteins are unknown. The P. falciparum heat shock protein 70 family members (PfHsp70s) have been more extensively characterized than the PfHsp40s, with certain members shown to function as molecular chaperones. However, little is known about the PfHsp70-PfHsp40 chaperone partnerships. There is mounting evidence that these chaperones are important not only in protein homoeostasis and cytoprotection, but also in protein trafficking across the parasitophorous vacuole (PV) and into the infected erythrocyte. We propose that certain members of these chaperone families work together to maintain exported proteins in an unfolded state until they reach their final destination. In this review, we critically evaluate what is known and not known about PfHsp40s and PfHsp70s.

Previous studies on infection of Trypanosoma cruzi in the Poço das Antas Biological Reserve population of wild free-ranging Leontopithecus rosalia have shown the presence of genotype T. cruzi II, associated in Brazil with human disease. Herein, this study has been extended, the infection being evaluated in L. rosalia of 3 different tamarin populations, inhabiting distinct forest areas located in the same Atlantic Coastal Rainforest. Edentata, Marsupialia, Rodentia and Chiroptera were examined exclusively in the Poço das Antas Biological Reserve. Excluding Chiroptera, T. cruzi infection was found in all orders. Biochemical and molecular characterization demonstrated that golden lion tamarins maintained stable infections by T. cruzi II. The isolates from the other mammals corresponded to T. cruzi I, suggesting independent transmission cycles occurring among the sylvatic mammals inside Poço das Antas Biological Reserve. Significant differences in the infection patterns presented by the 3 populations of wild and captive-born golden lion tamarins were noticed. In Poço das Antas a considerably higher number of positive haemocultures from tamarins with positive serological titres was observed in comparison to those obtained from other areas. The implications for conservation and public health of an active sylvatic cycle in the Atlantic Coastal Rainforest of Rio de Janeiro are discussed.

Intestinal schistosomiasis continues to be a major public health problem in sub-Saharan Africa, and is endemic in communities around Lake Victoria. Interest is growing in the molecular evolution and population genetic structure of Schistosoma mansoni and we describe a detailed analysis of the molecular epidemiology and phylogeography of S. mansoni from Lake Victoria. In total, 388 cytochrome oxidase 1 (COI) sequences were obtained from 25 sites along the Ugandan, Tanzanian and Kenyan shorelines of Lake Victoria, and 122 unique barcodes were identified; 9 corresponded to previously discovered barcodes from Lakes Victoria and Albert. A subset of the data, composed of COI sequences from miracidia from 10 individual children, was used for population genetics analyses; these results were corroborated by microsatellite analysis of 4 isolates of lab-passaged adult worms. Overall, 12 barcodes were found to be shared across all 3 countries, whereas the majority occurred singly and were locally restricted. The population genetics analyses were in agreement in revealing high diversity at the level of the human host and negligible population structuring by location. The lack of correlation between genetic distance and geographical distance in these data may be attributed to the confounding influence of high intra-individual diversity as well as human migration between communities.

The most important function of any parasite is to secure transmission to new hosts. The gametocyte, the stage which has become developmentally committed to the sexual cycle, provides a critical link in the transmission of Plasmodium falciparum from the human host to the anopheline mosquito vector. It is therefore imperative that our determination to understand the biology of the gametocyte is greater than the technical obstacles which have resulted in the gametocyte being left very much out of the limelight by the intensive investigation of the asexual bloodstream parasite. Here we explore the areas of gametocyte biology which by nature of their relevance to control and pathology as well as basic biology, are the subjects of investigation in our laboratory. We also point out areas in need of particular attention.

Serum-derived fatty acids are essential for the intraerythrocytic proliferation of Plasmodium falciparum in humans. We previously reported that only limited combinations of fatty acids can support long-term parasite culture, and palmitic acid (C16:0)/oleic acid (C18:1, n-9), palmitic acid (C16:0)/vaccenic acid (C18:1, n-7), or stearic acid (C18:0) are required in these combinations, implying that these fatty acids are key molecules for intraerythrocytic parasite growth (Mi-Ichi et al.2006). Here, we analysed profiles of parasitaemia changes as well as morphologies during the erythrocytic cycle and confirmed the importance of C16:0 and C18:1, n-9. We also provide evidence that C18:1, n-9 but not other C18 monoenoic or dienoic acids maintain the synchronicity of parasite development in serum-free medium when paired with C16:0, resulting in maintained exponential growth. Thus, C18:1, n-9 is indispensable for the intraerythrocytic proliferation of P. falciparum.

The short-lived infective copepodid stages of the copepod Salmincola edwardsii, ectoparasitic on brook trout, Salvelinus fontinalis, are under strong selective pressure to evolve efficient host-finding mechanisms. In laboratory experiments, we quantified the effects of visual and mechanical stimulation on the activity of the copepodids. We found that shadows passing above the copepodids and shock waves passing through the water generated marked increases in the rate and length of upward swimming bursts, which resulted in more time spent suspended in the water column and greater distances travelled. However, we found no strong evidence for a relationship between the magnitude of the copepodids' response and the intensity of stimulation. Although these responses are not host-specific, they allow the parasites to conserve their limited energy stores by increasing their host-finding activity only when potential hosts are nearby.

Nematode membrane preparations contain high amounts of low-affinity specific L-glutamate binding sites. The numbers of these sites were increased in 2 isolates, one field-derived and the other laboratory-derived, of ivermectin-resistant Haemonchus contortus and a field isolate of ivermectin-resistant Telodorsagia circumcincta, when compared to control, drug- sensitive isolates. Specific [3H]ivermectin binding to these membrane preparations showed no differences between ivermectin-sensitive and resistant isolates and the number of ivermectin binding sites was approximately 100-fold less than the number of L-glutamate binding sites. Kinetic analysis of L-glutamate binding suggested the presence of at least 2 classes of binding site. L-Glutamate binding was blocked by ibotenic acid, kynurenic acid and β-hydroxyaspartate, but not by ivermectin, argiopine, kainate, quisqualate or NMDA. Competition assays with ibotenic acid suggested that there were 2 distinct populations of glutamate binding sites and that the site with the lower affinity for ibotenate was upregulated in the ivermectin-resistant nematodes. In the field isolate of resistant H. contortus we found no coding changes in the cDNAs encoding glutamate-gated chloride channel subunits HG2, HG3 and HG4, nor were any changes in channel expression detected using subunit-specific antibodies. The low-affinity binding site is unlikely to be associated with the ivermectin receptor in these nematodes.

Infection parameters of Posthodiplostomum cuticola, a digenean parasite with a complex life-cycle, were investigated in fish (the second intermediate host) from 6 floodplain water bodies over 2 years. A broad range of factors related to abiotic characteristics of localities, density of the first intermediate (planorbid snails) and definitive (wading birds) hosts and fish community structure were tested for their effects on P. cuticola infection in juvenile and adult fish. Characters of the littoral zone and flood duration were found to be important factors for the presence of the first intermediate and definitive hosts. Visitation time of definitive bird hosts was also related to adult fish host density. Localities with P. cuticola infected fish were visited by a higher number of bird species. Infection of P. cuticola in fish and similarities in infection among fish host assemblages were correlated with fish host density and fish species composition. Parasite infection in both adult and juvenile fishes was associated with the slope of the bank and the bottom type, in particular in juvenile fish assemblages with snail host density. We conclude that habitat characteristics, snail host density and fish community structure contribute significantly to P. cuticola infection in fish hosts.

The present study deals with the infection dynamics of Diplostomum spathaceum and with the direct parasite-associated mortality of its experimental fish host, guppies (Lebistes reticulatis) and brook trout (Salvelinus fontinalis). The relationship between exposure density and cercarial penetration of the hosts is essentially linear. There is a significant decrease in the proportion of penetrating cercariae that establish themselves in the fish lens at a high exposure density. The host death rate increases exponentially with parasite burden in both guppies and brook trout.

The fact that the Schistosoma mansoni egg has two ATP diphosphohydrolase (EC 3.6.1.5) isoforms with different net charges and an identical molecular weight of 63000, identified by non-denaturing polyacrylamide gel electrophoresis and immunological cross-reactivity with potato apyrase antibodies, is shown. In soluble egg antigen (SEA), only the isoform with the lower net negative charge was detected and seemed to be the predominant species in this preparation. By confocal fluorescence microscopy, using anti-potato apyrase antibodies, the S. mansoni egg ATP diphosphohydrolase was detected on the external surface of miracidium and in von Lichtenberg's envelope. Intense fluorescence was also seen in the outer side of the egg-shell, entrapped by the surface microspines, suggesting that a soluble isoform is secreted. ATP diphosphohydrolase antigenicity was tested using the vegetable protein as antigen. The purified potato apyrase was recognized in Western blots by antibodies present in sera from experimentally S. mansoni-infected mice. In addition, high levels of IgG anti-ATP diphosphohydrolase antibodies were detected by ELISA in the same sera. This work represents the first demonstration of antigenic properties of S. mansoni ATP diphosphohydrolase and immunological cross-reactivity between potato apyrase and sera from infected individuals.

At present, bovine neosporosis is an important worldwide concern because of its wide geographic distribution and economic impact. Abortion is the main clinical sign of bovine neosporosis in both dairy and beef cattle. Ruminant challenge models are critical to evaluate potential vaccine candidates to help tackle bovine neosporosis and to study pathogenesis and host responses to infection. Several research groups have developed ruminant models of Neospora caninum infection independently of others, resulting in a high degree of variability due to the use of different species of animals, breeds, strains/isolates of N. caninum, doses, routes and times of inoculation. Standardization is greatly needed to advance research in a more collaborative, timely and efficient manner. In the absence of widely accepted international guidelines, this manuscript serves to summarize and discuss the different models and parameters currently in use. Parameters essential for the development of non-pregnant and pregnant ruminant models are outlined and the main knowledge gaps are identified. This information could act as the basis to develop a consensus for international standard guidelines for ruminant models of neosporosis that would be helpful for researchers in this field worldwide.

Ouabain applied to adult male Schistosoma mansoni causes a large, non-reversible tension increase of the parasite's musculature and elimination of spontaneous contractions. The tension increase and the time-course of tension development caused by ouabain are dose dependent with significant effects obtained at 3 × 10−6m. Digoxin and digoxigenin act in a similar manner with a relative potency of ouabain ≃ digoxin > digoxigenin. Lowered temperature as well as substitution of Li+ for Na+ increases muscle tension. The membrane potential recorded from the ventral tegument is also affected by treatments which interact with Na+–K+ transport systems. Ouabain (0·1 mm) causes a rapid depolarization without a significant effect on membrane resistance. The tegument is depolarized by temperatures below 30 °C. The effect of temperature is readily reversible and the temperature sensitivity is eliminated by pretreatment with ouabain. Substitution of Li+ for Na+ also causes a depolarization of the tegument. Tracer experiments show both an increase in Na+ and a decrease in K+ in the parasite within 10 min after treatment with ouabain (1 × 10−5m). Decreasing temperature of the bathing medium also causes an accumulation of Na+ as well as a decrease in the animals. The above results indicate a significant role for active Na+–K+ transport in muscle contraction and in maintenance of the tegumental membrane potential. The data also suggest that the Na+–K+ transport in S. mansoni may be electrogenic.

Parasites can have a wide range of pathophysiological effects on the host. This review describes those associated with some parasites of major importance in man and animals. Haemoprotozoan diseases such as trypanosomiasis and malaria are primarily associated with anaemia. Such anaemias have a complex aetiology involving various mechanisms responsible for red cell destruction as well as possible defects in red cell production. In addition to these haematological effects these diseases are associated with marked disturbances in heart function and the nervous, immune and urinary systems. The other major groups of parasitic diseases are those associated with the gastrointestinal tract. The most advanced studies have been conducted on the pathophysiology of gastrointestinal nematode parasites of sheep and have revealed significant effects on feed intake, gastrointestinal function, and protein and energy metabolism. Similar studies have yet to be conducted in other hosts and parasitic diseases. There is also a need to examine in greater detail the factors which can modulate pathophysiological responses by the host to parasitic infections.

The life cycle of Entobdella soleae, a monogenean skin-parasite of a marine bottom-living flatfish (Solea solea) has been described and related to the habits of the host, which buries itself in the sediment for some time each day and when not buried rarely moves more than a few centimetres from the sea bottom.

The eggs of the parasite are laid by specimens attached to the lower surface of the host and are anchored to sand grains on the sea bottom by an egg stalk bearing sticky droplets. The anchoring device may prevent the eggs from being carried by water currents vertically or laterally out of the region inhabited by soles.

The free-swimming oncomiracidium which hatches from the egg invades the anterior part of the upper surface of the fish, which is the only part exposed when the fish is buried in the sand. After a short period of development on the upper surface the parasite emigrates to the lower surface where sexual maturity is reached.

Adult parasites, which are distributed randomly on the lower surface, are orientated with the adhesive organ upstream with respect to water currents produced by the forward locomotion of the host, but the parasites are capable of moving from place to place on the lower surface of the fish. This movement of the parasite may be connected with the nature of the food.

I wish to thank the Director and Staff of the Plymouth Laboratory and in particular Mr J. E. Green, for invaluable assistance. I am also very grateful to Dr J. Llewellyn for advice and criticism. The work was conducted during the tenure of a Fishery Research Training Grant from the Development Commission.