Molecular nature of virulence in Entamoeba histolytica

For many years virulence of pathogenic Entamoeba histolytica has been attributed to the capacity of the parasite to destroy tissues through the expression and/or secretion of various molecules. Such view is supported mainly by in vitro experimentation, whereas data obtained by using animal models of the disease have clearly demonstrated that the host's inflammatory response is primarily responsible for tissue damage. This review analyzes the content and/or activity of some of the presumed toxic amebic molecules present in amebic strains with different degrees of virulence compared to various parasite in vitro functions that are supposed to correlate with in vivo virulence. The analysis suggests that amebic virulence is primarily determined by the parasite's capacity to adapt and survive the aerobic conditions present in animal tissues. This initial episode in the host–parasite relationship is an absolute requirement for the further development of tissue lesions, which result from the concerted action of many molecules derived from both, the host and the parasite.     

Is the population genetic structure of complex life cycle parasites determined by the geographic range of the most motile host?

Due to their particular way of life, dispersal of parasites is often mediated by their host's biology. Dispersal distance is relevant for parasites because high degree of dispersal leads to high gene flow, which counters the rate of parasite local adaptation in the host populations. Parasites with complex life cycles need to exploit sequentially more than one host species to complete their life cycle. Most trematode parasites have such complex life cycles involving invertebrate and vertebrate hosts. The spatial scales of invertebrate and vertebrate host populations are often different, which may decrease the probability that the parasite cycles locally in the intermediate host population. We used neutral microsatellite markers to determine genetic structure in Diplostomum pseudospathaceum parasites collected from local populations of freshwater snails (Lymnaea stagnalis). D. pseudospathaceum is a trematode that has two intermediate hosts (snail and fish) and a highly motile definitive host (bird). We found that the parasite population infecting the local snail populations showed no genetic structure over a large geographic range (over 300 km). We also did not detect evidence for isolation by distance in the parasite. We conclude that dispersal in the motile definitive host is likely to prevent emergence of local population genetic structure in the parasite. Our results suggest that parasite dispersal in the definitive host may limit local cycling of the parasites in the intermediate host populations.     

To cool or not to cool? Intestinal coccidians disrupt the behavioral hypothermia of lizards in response to tick infestation

It is generally accepted that parasites exert negative effects on their hosts and that natural selection favors specific host responses that mitigate this impact. It is also known that some components of the host immune system often co-evolve with parasite antigens resulting in a host-parasite arms race. In addition to immunological components of the anti-parasitic response, host behavioral responses are also important in this arms race and natural selection may favor avoidance strategies that preclude contact with parasites, or shifts in the host’s thermoregulatory strategy to combat active infections (e.g., behavioral fever). Ticks are widespread parasites with direct and indirect costs on their vertebrate hosts. Their saliva provokes hemolysis in the blood of their hosts and can transmit a plethora of tick-borne pathogens. We enquired whether tick infestation by Ixodes pacificus can provoke a thermoregulatory response in Sceloporus occidentalis. For this, we compared the thermoregulatory behavior of tick-infested lizards against tick-infested lizards co-infected with two different species of coccidians (Lankesterella occidentalis and Acroeimeria sceloporis). After this, lizards were kept in individual terraria with a basking spot and fed ad libitum. We found that tick-infested lizards sought cooler temperatures in proportion to their tick load, and this response was independent of the co-infection status by L. occidentalis. This was consistent in April and June (when tick loads were significantly lower) and suggests a conservative strategy to save energy which might have been selected to overcome tick infestations during phenological peaks of this parasite. However, this behavior was not observed in lizards co-infected with A. sceloporis, suggesting that co-infection with this intestinal parasite prompt lizards to be active. Cost of tick infestation was confirmed because housed lizards lost weight at a constant ratio to initial tick load, independently of other infections. The broader implications of these findings are discussed in the context of climate change.     

Strong neutral genetic differentiation in a host,but not in its parasite

The genetic diversity and population structure of a parasite with a complex life cycle generally depends on the dispersal by its most motile host. Given that high gene flow is assumed to hinder local adaptation, this can impose significant constraints on a parasite's potential to adapt to local environmental conditions, intermediate host populations, and ultimately to host-parasite coevolution. Here, we aimed to examine the population genetic basis for local host-parasite interactions between the eye fluke Diplostomum lineage 6, a digenean trematode with a multi-host life cycle (including a snail, a fish, and a bird) and its second intermediate host, the three-spined stickleback Gasterosteus aculeatus L. We developed the first microsatellite primers for D. lineage 6 and used them together with published stickleback markers to analyse host and parasite population structures in 19 freshwater lakes, which differ in their local environmental characteristics regarding water chemistry and Diplostomum abundance. Our analyses suggest that one parasite population successfully infects a range of genetically differentiated stickleback populations. The lack of neutral genetic differentiation in D. lineage 6, which could be attributed to the motility of the parasite's definitive host as well as its life cycle characteristics, makes local host-parasite co-adaptations seem more likely on a larger geographical scale than among the lakes of our study site. Our study provides a suitable background for future studies in this system and the first microsatellite primers for a widespread fish parasite.     

The impact of tick-borne pathogen infection in Indian bovines is determined by host type but not the genotype of Theileria annulata

Tick-borne pathogens (TBP) are a major source of production loss and a welfare concern in livestock across the globe. Consequently, there is a trade-off between keeping animals that are tolerant to TBP infection, but are less productive than more susceptible breeds. Theileria annulata is a major TBP of bovines, with different host types (i.e. exotic and native cattle breeds, and buffalo) displaying demonstrable differences in clinical susceptibility to infection. However, the extent to which these differences are driven by genetic/physiological differences between hosts, or by different parasite populations/genotypes preferentially establishing infection in different host breeds and species is unclear. In this study, three different bovine host types in India were blood sampled to test for the presence of various TBP, including Theileria annulata, to determine whether native cattle (Bos indicus breeds), crossbreed cattle (Bos taurus x Bos indicus breeds) or water buffalo (Bubalus bubalis) differ in the physiological consequences of infection. Population genetic analyses of T. annulata isolated from the three different host types was also performed, using a panel of mini- and micro-satellite markers, to test for sub-structuring of the parasite population among host types. We discovered that compared to other host types, “carrier” crossbreed cattle showed a higher level of haematological pathology when infected with T. annulata. Despite this finding, we found no evidence for differences in the genotypes of T. annulata infecting different host types, although buffalo appeared to harbour fewer mixed parasite genotype infections, indicating they are not the major reservoir of parasite diversity. The apparent tolerance/resistance of native breed cattle and buffalo to the impacts of T. annulata infection is thus most likely to be driven by host genotype, rather than differences in the parasite population. Our results suggest that an improved understanding of the genetic factors that underpin disease resistance could help to ameliorate future economic loss due to TBP or tropical theileriosis.     

Species Identification of Birds Nasal Trichobilharzia in Sari,North of Iran

Background

Cercarial dermatitis is known as an endemic parasitic disease in North of Iran, a hypersensitive skin reaction to the penetration of nonhuman schistosome larvae into human skin. In recent studies in this region, final and intermediate hosts were recognized and Trichobilharzia was identified as the main causative agent of cercarial dermatitis in this region, but to date the parasite species haven''t been identified. Therefore this study was performed to species identification of nasal Trichobilharzia in infected birds for the first time.

Methods

A total of 45 Anas clypeata birds identified as final host, were collected from Sari in North of Iran and infected nasal tissues analyzed using molecular techniques. Genomic DNA was isolated by phenol/chloroform extraction method and ITS region of rDNA were amplified with specific primers its5Trem and its4Trem, then sequenced area were compared with existing records in GenBank.

Results

Twelve samples were infected with Trichobilharzia and results of PCR reaction indicated that all of them belonged to T. regenti. The sequence alignment of present work isolates and those deposited in GenBank showed differences in nucleotide sequences of repeat region in ITS1.

Conclusion

Trichobilharzia regenti is the most frequent parasite of Anatid birds in North of Iran. This corresponds to the distribution of this parasite along the flyway of migratory birds, which annually migrate from Siberia and northern countries of Caspian Sea to wintering areas in southern regions of it.     

Recent developments in the field of Chagas' disease

Chagas'' disease is an endemic human trypanosomiasis produced by Trypanosoma cruzi, which is usually transmitted by blood-sucking triatomid bugs. This article reports recent developments in research on Chagas'' disease and outlines some of the basic investigations that have immediate practical application or that contribute to our knowledge of the parasite and its relationship with the vertebrate host.     

Estimating novel potential drug targets of Plasmodium falciparum by analysing the metabolic network of knock-out strains in silico

Malaria is one of the world's most common and serious diseases causing death of about 3 million people each year. Its most severe occurrence is caused by the protozoan Plasmodium falciparum. Biomedical research could enable treating the disease by effectively and specifically targeting essential enzymes of this parasite. However, the parasite has developed resistance to existing drugs making it indispensable to discover new drugs. We have established a simple computational tool which analyses the topology of the metabolic network of P. falciparum to identify essential enzymes as possible drug targets. We investigated the essentiality of a reaction in the metabolic network by deleting (knocking-out) such a reaction in silico. The algorithm selected neighbouring compounds of the investigated reaction that had to be produced by alternative biochemical pathways. Using breadth first searches, we tested qualitatively if these products could be generated by reactions that serve as potential deviations of the metabolic flux. With this we identified 70 essential reactions. Our results were compared with a comprehensive list of 38 targets of approved malaria drugs. When combining our approach with an in silico analysis performed recently [Yeh, I., Hanekamp, T., Tsoka, S., Karp, P.D., Altman, R.B., 2004. Computational analysis of Plasmodium falciparum metabolism: organizing genomic information to facilitate drug discovery. Genome Res. 14, 917–924] we could improve the precision of the prediction results. Finally we present a refined list of 22 new potential candidate targets for P. falciparum, half of which have reasonable evidence to be valid targets against micro-organisms and cancer.     

Molecular epidemiology,evolution, and phylogeny of Entamoeba spp.

Entamoeba histolytica is a protozoan parasite and the causative agent of amoebiasis in humans. The estimations of the worldwide burden of amoebiasis by the WHO indicated that approximately 500 million people were infected with the parasite and 10% of these individuals had invasive amoebiasis. However, our understanding of the disease burden and epidemiology of human amebiasis has undergone dramatic changes over the last two decades based on molecular analyses. The development of Entamoeba genomics has also provided some interesting and valuable information on the evolution and population structure of this parasite. In addition, the use of a number of molecular markers has greatly expanded our understanding of Entamoeba host range and genetic diversity. In this review, we re-assessed Entamoeba prevalence and species in humans, non-human primates, other animals, and the environment in the context of molecular data. Some issues regarding the evolution and phylogeny of different Entamoeba species lineages are also discussed.     

Modulation of immune response in experimental Chagas disease

Trypanosoma cruzi (T. cruzi), the etiological agent of Chagas disease, affects nearly 18 million people in Latin America and 90 million are at risk of infection. The parasite presents two stages of medical importance in the host, the amastigote, intracellular replicating form, and the extracellular trypomastigote, the infective form. Thus infection by T. cruzi induces a complex immune response that involves effectors and regulatory mechanisms. That is why control of the infection requires a strong humoral and cellular immune response; hence, the outcome of host-parasite interaction in the early stages of infection is extremely important. A critical event during this period of the infection is innate immune response, in which the macrophage’s role is vital. Thus, after being phagocytized, the parasite is able to develop intracellularly; however, during later periods, these cells induce its elimination by means of toxic metabolites. In turn, as the infection progresses, adaptive immune response mechanisms are triggered through the TH1 and TH2 responses. Finally, T. cruzi, like other protozoa such as Leishmania and Toxoplasma, have numerous evasive mechanisms to the immune response that make it possible to spread around the host. In our Laboratory we have developed a vaccination model in mice with Trypanosoma rangeli, nonpathogenic to humans, which modulates the immune response to infection by T. cruzi, thus protecting them. Vaccinated animals showed an important innate response (modulation of NO and other metabolites, cytokines, activation of macrophages), a strong adaptive cellular response and significant increase in specific antibodies. The modulation caused early elimination of the parasites, low parasitaemia, the absence of histological lesions and high survival rates. Even though progress has been made in the knowledge of some of these mechanisms, new studies must be conducted which could target further prophylactic and therapeutic trials against T. cruzi infection.     

Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3 (RMS-3)

Rhipicephalus microplus is an important bovine ectoparasite, widely distributed in tropical and subtropical regions of the world causing large economic losses to the cattle industry. Its success as an ectoparasite is associated with its capacity to disarm the antihemostatic and anti-inflammatory reactions of the host. Serpins are protease inhibitors with an important role in the modulation of host–parasite interactions. The cDNA that encodes for a R. microplus serpin was isolated by RACE and subsequently cloned into the pPICZαA vector. Sequence analysis of the cDNA and predicted amino acid showed that this cDNA has a conserved serpin domain. B- and T-cell epitopes were predicted using bioinformatics tools. The recombinant R. microplus serpin (rRMS-3) was secreted into the culture media of Pichia pastoris after methanol induction at 0.2 mg l^?1. qRT-PCR expression analysis of tissues and life cycle stages demonstrated that RMS-3 was mainly expressed in the salivary glands of female adult ticks. Immunological recognition of the rRMS-3 and predicted B-cell epitopes was tested using tick-resistant and susceptible cattle sera. Only sera from tick-resistant bovines recognized the B-cell epitope AHYNPPPPIEFT (Seq7). The recombinant RMS-3 was expressed in P. pastoris, and ELISA screening also showed higher recognition by tick-resistant bovine sera. The results obtained suggest that RMS-3 is highly and specifically secreted into the bite site of R. microplus feeding on tick-resistant bovines. Capillary feeding of semi-engorged ticks with anti-AHYNPPPPIEFT sheep sera led to an 81.16% reduction in the reproduction capacity of R. microplus. Therefore, it is possible to conclude that R. microplus serpin (RMS-3) has an important role in the host–parasite interaction to overcome the immune responses in resistant cattle.     

Serial studies on the evolution of chloroquine resistance in an area of East Africa receiving intermittent malaria chemosuppression

Serial in vitro and in vivo tests for chloroquine sensitivity of Plasmodium falciparum were carried out from 1979 to 1982 in an area of E. Africa where chemosuppression with chloroquine had been attempted since 1977. Within 1½ years there were signs of a decreasing drug response. Chloroquine resistance was first detected in 1981 and this increased markedly in 1982. Other contributory causes for the rise of parasite rates in children were possibly a decline in the efficiency of the drug distribution system and also immunological factors. Evidence of resistance to pyrimethamine was also found. Observations were made of the heterogeneity of the parasites'' responses with emerging resistance. Implications for the future are discussed.     

Relative reproductive success of co-infecting parasite genotypes under intensified within-host competition

In nature, host individuals are commonly simultaneously infected with more than one genotype of the same parasite species. These co-infecting parasites often interact, which can affect their fitness and shape host–parasite ecology and evolution. Many of such interactions take place through competition for limited host resources. Therefore, variation in ecological factors modifying the host resource level could be important in determining the intensity of competition and the outcome of co-infections. We tested this hypothesis by measuring the relative reproductive success of co-infecting genotypes of the trematode parasite Diplostomum pseudospathaceum in its snail host Lymnaea stagnalis while experimentally manipulating snail resource level using contrasting feeding treatments (ad libitum food supply, no food). We found that food deprivation constrained the overall parasite within-host reproduction as the release of parasite transmission stages (cercariae) was reduced. This indicates intensified competition among the parasite genotypes. The genotypic composition of the released cercariae, however, was not affected by the feeding treatments. This suggests that in this system, the relative reproductive success of co-infecting parasite genotypes, which is an important component determining their fitness, is robust to variation in ecological factors modifying the strength of resource competition.     

Computational and experimental analysis identified 6-diazo-5-oxonorleucine as a potential agent for treating infection by Plasmodium falciparum

Plasmodium falciparum (PF) is the most severe malaria parasite. It is developing resistance quickly to existing drugs making it indispensable to discover new drugs. Effective drugs have been discovered targeting metabolic enzymes of the parasite. In order to predict new drug targets, computational methods can be used employing database information of metabolism. Using this data, we performed recently a computational network analysis of metabolism of PF. We analyzed the topology of the network to find reactions which are sensitive against perturbations, i.e., when a single enzyme is blocked by drugs. We now used a refined network comprising also the host enzymes which led to a refined set of the five targets glutamyl–tRNA (gln) amidotransferase, hydroxyethylthiazole kinase, deoxyribose–phophate aldolase, pseudouridylate synthase, and deoxyhypusine synthase. It was shown elsewhere that glutamyl–tRNA (gln) amidotransferase of other microorganisms can be inhibited by 6-diazo-5-oxonorleucine. Performing a half maximal inhibitory concentration (IC₅₀) assay, we showed, that 6-diazo-5-oxonorleucine is also severely affecting viability of PF in blood plasma of the human host. We confirmed this by an in vivo study observing Plasmodium berghei infected mice.     

Spatial,temporal and host factors structure the Ceratomyxa shasta (Myxozoa) population in the Klamath River basin

The myxozoan parasite Ceratomyxa shasta is a virulent pathogen of salmonid fish in the Klamath River, Oregon/California, USA. We previously defined four principal genotypes of the parasite (O, I, II, III) based on a trinucleotide repeat (ATC)_0–3 in Internal Transcribed Spacer region 1 sequences. Genotypes occur in sympatry and show marked host preference: I in Chinook salmon (Oncorhynchus tschawytscha) and II in non-native rainbow trout (O. mykiss). In the present study, we sequenced the parasite from river water samples collected in May, June and September at three localities below, above and between the Klamath's five dams. We also sampled adult and juvenile coho salmon (O. kisutch), steelhead trout (O. mykiss, anadromous form) and native redband rainbow trout (O. mykiss, freshwater form) and additional Chinook salmon and non-native rainbow trout. We found that the C. shasta population was highly structured spatially, temporally and with respect to fish host species. Genotype O was present in water throughout the basin but detected almost exclusively in steelhead and native rainbow trout. Genotype I was in water only below the dams and detected only in Chinook salmon. Genotype II was detected in coho salmon below the dams, and in non-native rainbow trout exposed both above and below the dams. The same genotypes were detected in adult and juvenile fish of the same species. These findings have major implications for the design of effective surveillance and control programs for this economically and ecologically important fish parasite.     

Occurrence and distribution of parasites in relation to food components of grey gurnard,Eutrigla gurnardus (L.) (Teleostei: Scorpaeniformes), off the Shetland Islands

The research was conducted on the parasite fauna and food composition of Eutrigla gurnardus caught as by-catch in commercial catches of the Atlantic cod Gadus morhua near the Shetland Islands. Thirteen species and two genera of pathogens were identified, including six species and one genus recorded for the first time in this host. Copepoda — Euphausiacea dominated in the stomach contents (they are also the intermediate hosts for most of the parasites found), while Gadidae dominated among the fish. A checklist of E. gurnardus parasites is included.     

Antigenic variation and immunity to Plasmodium knowlesi: antibodies which induce antigenic variation and antibodies which destroy parasites

2 types of antibody response specific for intrastrain antigenic variants of P. knowlesi can be detected in the serum of M. mulatta. One induces a change in the parasite serotype, the other is parasiticidal. The rate of synthesis of both types of antibodies to new variants increases as the host develops protective immunity and their rate of synthesis relative to one another also changes. Both these effects are possibly important in determining the level of parasitaemia.     

Protection induced by Leishmania Major antigens and the imiquimod adjuvant encapsulated on liposomes in experimental cutaneous leishmaniasis

There is a need for new, effective, and less expensive and toxic treatment for Leishmaniasis. It seems that the use of a suitable adjuvant and a delivery system is effective in inducing immune reactions for protection. Liposomes can be applied as immunoadjuvants to trigger immune reactions to different antigens. The adjuvant effects of imiquimod using DSPC liposomes containing SLA (soluble Leishmania antigens) were studied on the type and intensity of the produced immune reaction to the challenge of Leishmania major in BALB/c mice. Liposomes were produced by the lipid film procedure. BALB/C mice were immunized subcutaneously, three times at 2-week intervals and with various formulations. Lesion development and the parasite burden in the spleens and feet after the challenge with Leishmania major, Th1 cytokine (IFN-γ), and the IgG isotype titration were assessed to evaluate the induced immune reaction and the protection level.The group of mice immunized with Liposome DSPC +Imiquimod +SLA revealed less severe footpad swelling, being significantly different (P < .05) from other groups. A higher level of IgG2a and IFN-γ secretion was observed in the mice immunized with Liposome DSPC +Imiquimod +SLA than the control group. These observations imply that the DSPC liposome containing imiquimod induces the Th1 immune response that is protective against the challenge of Leishmania major.     

Skin sensitivity reactions in Nigerian children with bronchial asthma

Skin sensitivity reactions to a variety of antigens in 290 unselected urban Nigerian children suffering from bronchial asthma of varying severity, are reported. The percentages of positive skin reactions to the first four antigens were as follows: Ascaris (25%), Dermatophagoides pteronyssinus (21%), house dust (12%) and feathers (12%). The sizes of reaction were generally smaller than those reported in European and American children. While the relatively high sensitivity to Ascaris was probably fortuitous, further studies are needed to evaluate the actual role, if any, of the parasite in bronchial asthma affecting the African child. The relatively low rate of positive skin reactions indicate that, at present, skin sensitivity tests are of limited value in the identification of aetiological factors in asthma affecting Nigerian children. The development of local materials for skin testing may however enhance the usefulness of this investigation in future.