Cyphocarax gilbert (Szidat, L., 1948) is a fish commonly found in coastal drainage of eastern Brazil. This fish is sometimes caught with signs of infection by the crustacean Riggia paranensis, a haematophagous parasite. A remarkable feature of infected fish is that they lack gonads. In this paper we have analysed the frequency of parasitism, the gonadal development of non-infected fish and the profile of plasma proteins in both infected and non-infected specimens. Two reproductive periods/year were observed, beginning in February and August. On average, 40% of fish were infected, in the Itabapoana River (Brazil). Sex-specific proteins were identified by electrophoresis. SDS-PAGE analysis demonstrated that a 143 kDa female-specific glycolipoprotein (FSP) is a calcium-binding phosphoprotein. FSP was isolated through ultracentrifugation and SDS-PAGE analysis showed that the native protein is composed of three polypeptides of 143, 100 and 70 kDa. Both FSP and a 33 kDa male-specific protein (MSP) are absent from infected fish plasma. FSP levels in female plasma changes with the developmental stage of gonads. Altogether these data suggest that the FSP corresponds to fish vitellogenin. Furthermore, the absence of the above-mentioned proteins in infected fish suggests that R. paranensis might interfere with the regular hormonal process of fish vitellogenesis.

1. The functional anatomy of the ducts associated with Mehlis's gland in the echinostome Artyfechinostomum mehrai (Faruqui) recovered from a girl has been described.

2. The mechanism and histochemistry of egg-shell formation are described and discussed.

Appreciation is extended to Dr V. S. Raghunathan, M.D., Physician and Superintendent, and Dr T. Sreenivasan, M.D., of King George Hospital, Visakhapatnam, for their kind co-operation and the gift of specimens of A. mehrai. My thanks are due to Professor P. N. Ganapati for facilities and encouragement.

A method has been devised to show that malaria parasites can be detected serologically in infected blood with a high degree of sensitivity. Using a murine malaria model, parasites were demonstrated in a solid-phase radio-immunoassay which measured antibody-binding inhibition. Lysed red blood cells (r.b.c.) were incubated with labelled specific antibody and were then reacted in antigen-coated tubes. The degree of inhibition of antibody binding in the tubes correlated with the level of parasitaemia in the test blood. Using homologous antisera the test detected infection at a level of 1 parasite/million r.b.c. The specificity of the method was shown by comparison of antibody-binding inhibition in normal and infected r.b.c. and in r.b.c. from non-infected mice with induced reticulocytosis. The sensitivity was shown in vitro in tests of serially diluted blood of high parasitaemia and in vivo for the detection of early infection. The presence of antibody in the test blood did not significantly affect the sensitivity of parasite detection.

Self/non-self-discrimination by vertebrate immune systems is based on the recognition of the presence of peptides in proteins of a parasite that are not contained in the proteins of a host. Therefore, a reduction of the number of ‘words’ in its own peptide vocabulary could be an efficient evolutionary strategy of parasites for escaping recognition. Here, we compared peptide vocabularies of 30 endoparasitic and 17 free-living unicellular organisms and also eight multicellular parasitic and 16 multicellular free-living organisms. We found that both unicellular and multicellular parasites used a significantly lower number of different pentapeptides than free-living controls. Impoverished pentapeptide vocabularies in parasites were observed across all five clades that contain both the parasitic and free-living species. The effect of parasitism on a number of peptides used in an organism's proteins is larger than effects of all other studied factors, including the size of a proteome, the number of encoded proteins, etc. This decrease of pentapeptide diversity was partly compensated for by an increased number of hexapeptides. Our results support the hypothesis of parasitism-associated reduction of peptide vocabulary and suggest that T-cell receptors mostly recognize the five amino acids-long part of peptides that are presented in the groove of major histocompatibility complex molecules.

Infection with Plasmodium vivax was established in splenectomized Callithrix jacchus marmosets by inoculation of parasitized blood from Aotus trivirgatus carrying the Vietnam Palo-Alto line of P. vivax. Subsequent blood passage through intact marmosets resulted in higher peak parasitaemias (about 1% of red cells infected) and the loss of stainable Schüffner's dots in infected cells. Primary infections with the adapted line were patent for 74 days or more, and induced both a substantial antibody response, as determined by indirect fluorescence, and some lymphocytosis, but no marked anaemia. Marmosets which had recovered from their primary infection (or in which it was drug-cured) suffered abbreviated patency with low-grade parasitaemia on re-infection.

We are disappointed with Ostfeld's parochial commentary (Ostfeld, 2013) because we hoped we had addressed the dilution theory debate from a wider perspective, taking into account studies beyond those originating in New York State (Randolph & Dobson, 2012). We emphasized the inherent variability in the effects of biodiversity on the risk of zoonotic disease, noting explicitly that Ostfeld's own work allowed for positive, neutral and negative outcomes depending on the precise circumstances and biological interactions. The wider the range of studies, the greater the evidence for neutral and positive outcomes (i.e. biodiversity may have no effect or exacerbate infection risk) as the literature bias is gradually overcome. Publication bias, however, may still persist and be identified by appropriate meta-analyses (Salkeld et al., 2013).

Monensin (Mon) is an anticoccidial polyether ionophore widely used to control coccidiosis. The extensive use of polyether ionophores on poultry farms resulted in widespread resistance, but the underlying resistance mechanisms are unknown in detail. For analysing the mode of action by which resistance against polyether ionophores is obtained, we induced in vitro Mon resistance in Toxoplasma gondii-RH strain (MonR-RH) and compared it with the sensitive parental strain (Sen-RH). The proteome assessment of MonR-RH and Sen-RH strains was obtained after isotopic labelling using stable isotope labelling by amino acid in cell culture. Relative proteomic quantification between resistant and sensitive strains was performed using liquid chromatography-mass spectrometry/mass spectrometry. Overall, 1024 proteins were quantified and 52 proteins of them were regulated. The bioinformatic analysis revealed regulation of cytoskeletal and transmembrane proteins being involved in transport mechanisms, metal ion-binding and invasion. During invasion, actin and microneme protein 8 (MIC8) are seem to be important for conoid extrusion and forming moving junction with host cells, respectively. Actin was significantly upregulated, while MIC8 was downregulated, which indicate an invasion reduction in the resistant strain. Resistance against Mon is not a simple process but it involves reduced invasion and egress activity of T. gondii tachyzoites while intracellular replication is enhanced.

An infection approach was adopted for examining consequential heat shock (HS) or stress response in brain and spleen tissues from Wistar rats. Stress in this system was due to interactions with the infecting helminth, Trichinella spiralis, or its body-dwelling larval stages, or products thereof. It was argued that in the infection model used, elements effecting stress in the brain would differ from those in the spleen. HS responses were measured by quantitation of 4 levels of HS proteins (HSP25, HSP60, HSP70 and HSP90) with time, in infected and uninfected rat tissues using an assay depending on immunoblotting specifically to detect the separate HSPs and image analysis to measure HSP content. In brain and spleen tissue from uninfected rats, a continuous expression of the above HSPs was observed at levels which hardly varied throughout the experiment. In contrast, HSP25, HSP60 and HSP70 levels in infected rat tissues varied and apparently depended on ‘infection cycle’-related events. Thus, an enhanced expression of HSP25 and HSP60 and of these plus HSP70 was observed at certain, yet different, time-points during infection in rat spleen and rat brain, respectively. Interestingly, HSP90 expression in spleen tissue from infected rats versus controls, was significantly reduced throughout the experiment suggesting some important (as yet undefined) role for HSP90 in the infection cycle. These studies seem to have provided evidence for the occurrence of soluble factors causing altered HSP expression at both sides of the blood-brain barrier in rats with a primary T. spiralis infection.

Sialic acid on the red cell surface plays a major role in invasion by the malaria parasite Plasmodium falciparum. The NeuAc(α2,3) Gal motif on the O-linked tetrasaccharides of the red cell glycophorins is a recognition site for the parasite erythrocyte-binding antigen (EBA-175). Consequently, the interaction of P. falciparum and the red cell might share homology with that of the influenza virus. The cellular interactions of P. falciparum were examined for their sensitivity to 4-guanidino-2,3-didehydro-D-N-acetyl neuraminic acid (4-guanidino Neu5Ac2en), a potent inhibitor of influenza virus sialidase. Parasite invasion and subsequent development was unaffected by the sialidase inhibitor. The inhibitor did not affect rosette formation of parasite-infected erythrocytes with uninfected cells nor their cytoadherence to C32 melanoma cells. Furthermore, we were unable to confirm the presence of a previously reported parasite sialidase using sensitive fluorometric or haemagglutination assays, neither was any malarial trans-sialidase identified. We conclude that P. falciparum possesses neither sialidase nor trans-sialidase activity and that an inhibitor of influenza virus sialidase has no effect on important cellular interactions of this parasite.

The study of genetic exchange systems and the use of genetic analysis has been relatively limited in parasites leading to considerable gaps in our basic knowledge. This lack of knowledge makes it difficult to draw firm conclusions as to how these systems evolved. An additional problem is also raised by the difficulties in defining evolutionary distances particularly with the unicellular protozoa, using classical ultrastructural and cytological criteria. While these difficulties have by no means been overcome, the use of rapid sequencing techniques applied to the ribosomal genes has allowed measurement of evolutionary distances, and considerable advances in our understanding of the genetic exchange systems in a few parasitic protozoa have recently been made. The conclusions from these recent sets of analyses are reviewed and then examined together in order to discuss the evolution of genetic exchange systems in parasitic protozoa. The evolutionary distances defined by ribosome sequence analysis show that parasites are an extremely divergent group, with distances which, in some cases, are orders of magnitude greater than the distances between mammals and fish; furthermore these studies suggest that the parasitic protozoa or their free-living ancestors are extremely ancient. These findings support the view that parasitism has occurred independently many times and that the parasitic life-style has been adopted by evolutionarily distinct groups. The recent observation of a non-obligatory genetic exchange system in the diploid but evolutionarily ancient kinetoplastid Trypanosoma brucei suggests that diploidy and meiosis are extremely old. The observation, in parasitic protozoa and helminths, that selfing or non-obligatory mating is a common feature suggests that these processes may be strategies to overcome the cost of meiosis. In this context, the question of what selective forces maintain genetic exchange is discussed.

Using a range of infection techniques, the ability of cercariae of the ectoparasitic digenean Transversotrema patialense to infect 24 species of tropical freshwater fish is examined. Of the 24 species (from 11 families) it is possible to infect 21 (from 8 families) under laboratory conditions. Reproductive success (R) for T. patialense from the cercarial infection process to the production of viable eggs by adult flukes is estimated for infections on two fish hosts, the cyprinid Brwhydanio rerio and the characinid Cheirodon sp. in controlled laboratory conditions. To calculate R, separate estimations of parasite infectivity, survival, fecundity and egg viability were made on populations of both host species. The enhanced reproductive success (R) of T. patialense on Cheirodon sp. is the composite resultant of higher infectivity, lower mean expected adult fluke life-span and higher mean fecundity/surviving worm when compared with infections on B. rerio under identical experimental conditions. Different patterns of egg viability of the two hosts do not materially influence differential reproductive success. A variety of formal techniques for handling the interacting components of reproductive success are described. The relationship between these techniques and the concept of basic reproductive rate, R0 (Macdonald, 1957; Anderson, 1982) is assessed.

Antineoplastic cyclophosphamide (CY) significantly reduced the survival and the weight of Hymenolepis microstoma in mice. Corticosteroid dexamethasone (DEX), on the other hand, did not influence the worm survival and the worms in DEX-treated mice became significantly heavier. Both immunosuppressants abolished H. microstoma-induced bile duct hypertrophy and submucous enteritis and lowered the number of mast cells (MC) in these regions. CY appeared to be more effective in depleting the bile duct MC whereas DEX seemed to affect more the duodenal MC population.

During the last 2 decades there have been numerous reports of the emergence of mefloquine resistance in Southeast Asia and nearly 50% resistance is reported in Thailand. A World Health Organization report (2001) considers mefloquine as an important component of ACT (artesunate+mefloquine) which is the first line of treatment for the control of uncomplicated/multi-drug resistant (MDR) Plasmodium falciparum malaria. In view of the emergence of resistance towards this drug, it is proposed to develop new drug combinations to prolong the protective life of this drug. Prior studies have suggested that mefloquine resistance can be overcome by a variety of agents such as ketoconazole, cyproheptadine, penfluridol, Icajine and NP30. The present investigation reports that clarithromycin (CLTR), a new macrolide, being a potent inhibitor of Cyt. P450 3A4, can exert significant resistance reversal action against mefloquine resistance of plasmodia. Experiments were carried out to find out the curative dose of CLTR against multi-drug resistant P. yoelii nigeriensis. Mefloquine (MFQ) and clarithromycin (CLTR) combinations have been used for the treatment of this MDR parasite. Different dose combinations of these two drugs were given to the infected mice on day 0 (prophylactic) and day 1 with established infection (therapeutic) to see the combined effect of these combinations against the MDR malaria infection. With a dose of 32 mg/kg MFQ and 225 mg/kg CLTR, 100% cure was observed, while in single drug groups, treated with MFQ or CLTR, the cure was zero and 40% respectively. Therapeutically, MFQ and CLTR combinations 32+300 mg/kg doses cleared the established parasitaemia on day 10. Single treatment with MFQ or CLTR showed considerable suppression of parasitaemia on day 14 but neither was curative. Follow-up of therapeutically treated mice showed enhanced anti-malarial action as reflected by their 100% clearance of parasitaemia. The present study reveals that CLTR is a useful antibiotic to be used as companion drug with mefloquine in order to overcome mefloquine resistance in plasmodia.

Gyrdicotylus gallieni is a viviparous gyrodactylid monogenean that infects the buccal cavity and pharynx oi Xenopus laevis laevis. Offspring directly recolonize the same host as their parent and transmission is carried out exclusively by adult worms which are capable of leech-like stepping locomotion. The infrapopulation dynamics (at 20 °C) of this species were studied by experimental exposure to single worms of previously uninfected hosts (which were dissected at various time intervals post-infection). Infrapopulations increased exponentially until 50 days post-infection (p.i.), during which time the intrinsic rate of increase was estimated by regression as 0·065/parasite/day. After this, infrapopulation sizes showed greater variability, with established infections becoming extinct 2–5 months p.i.: elimination was probably due to someform of host reaction, as parasite lineages were maintained for up to 10 months by transfers to successive uninfected hosts.The development of infrapopulations is similar to that reported for other viviparous gyrodactylids parasitic on the bodysurface of teleosts. However, the intrinsic rate of increase was comparatively slow in G. gallieni and the duration of infection in isolated hosts prolonged. Slow-growing infrapopulations may elicit a host response more slowly and extend the period during which dispersal to a new host or host population can occur. This might be important in G. gallieni for which the internal site of infection could limit the transmission rate, as worm migration from the oral cavity or accidental detachment and expulsion is necessary for host-host transfer to occur. Exposure of wild caught X. l. laevis of unknown infectionhistory to 1 (n = 33) or 10 (n = 10) worms produced only 2 established infrapopulations (in both cases hosts initially infected by single worms), compared with establishment of over 60% in naive hosts infected with single worms: this suggests that host resistance may be an important factor in the population dynamics of G. gallieni.

The effect of experimental non-host snail (NHS) communities on the infection of Lymnaea truncatula with Paramphistomum daubneyi and Fasciola hepatica miracidia was studied. The results obtained indicated that NHS had variable effects on basic population parameters of host infection. Specifically, NHS were able to produce aggregation of sporocyst number/snail in the experimental host populations. Proposed hypotheses for decoy effect mechanisms failed to explain completely the experimental results. A new mechanism is suggested, based on a distance response of miracidia to snail chemo-attractants, to explain effects of NHS on miracidial host finding behaviour. It is assumed that miracidia have evolved the ability to discriminate among individual snail chemical attractants in a community already at a distance. This attribute of miracidial behaviour allows NHS to induce variations in the effectiveness of individual host localization by miracidia. The resulting aggregation of successful infections/host is assumed to be the specific component of snail decoy effect at the community level. It was also established that NHS affected trematode reproduction in the host inducing a compensatory increase in the intramolluscan population. Intraspecific competition among developmental stages is suggested to be the mechanism of this compensatory reaction.

A structure called ‘veil’ in Theileria sergenti-, T. buffeli- or T. orientalis-infected bovine erythrocytes was purified from erythrocyte lysates by Percoll density-gradient centrifugation. On electron microscopical examination, the veils consisted of electron-dense material showing a periodicity of striations and were not surrounded by membranes. Analyses of veil proteins by one- and two-dimensional polyacrylamide gel electrophoresis revealed that the veils contain haemoglobin and several other basic proteins of molecular weight ranging from 15·5 to 46 kDa. By comparing the protein patterns of the veil with those of purified piroplasms and uninfected bovine erythrocytes, these basic proteins appeared to be of parasite origin. It would appear likely that the veils formed by intra-erythrocytic precipitation of haemoglobin and proteins excreted by the parasites. Differences in veil constituents were found between T. sergenti, T. buffeli and T. orientalis.

MicroRNAs (miRNAs) are short noncoding RNAs, involved in the regulation of parasite diseases. However, a role of miRNAs in Echinococcus multilocularis infection remains largely unknown. In this study, we first found the expression levels of key genes involved in miRNA biogenesis and function, including Ago2, Xpo5, Tarbp2 and DgcR8, were obviously altered in the macrophage RAW264·7 cells exposed to E. multilocularis metacestodes. Compared with the control, 18 and 32 known miRNAs were found to be differentially expressed (P < 0·05 and fold change >2) in the macrophages exposed to E. multilocularis metacestodes for 6 and 12 h, respectively. Among these, several are known to be involved in regulating cytokine activities and immune responses. Quantitative real-time polymerase chain reaction results showed that the expression of nine selected miRNAs was consistent with the sequencing data at each treatment time points. Moreover, there were statistically significant correlations between the expression levels of miRNAs and their corresponding targeted genes. Our data give us some clues to pinpoint a role of miRNAs in the course of infection and immunity of E. multilocularis.