The localization and distribution of the serotoninergic and peptidergic elements of the nervous system of larval Schistosoma mansoni have been investigated using an indirect immunofluorescence technique in conjunction with confocal scanning laser microscopy (CSLM). A range of antisera was used, raised to the biogenic amine, 5-hydroxytryptamine (5-HT, or serotonin), the vertebrate peptides pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY) and to the native invertebrate peptide, FMRFamide; all these antisera were shown previously to be immunopositive in the adult worm. No immunoreactivity to 5-HT was detected in any of the larval stages, but both miracidia and cercariae were consistently immunoreactive to all 4 peptides. The peptidergic nervous system of the miracidium is relatively simple, taking the form of a central neural mass with associated paired anterior and posterior nerve tracts. The cercarial peptidergic nervous system comprises a central commissure joining paired anterior ganglia, from which emanate paired dorsal and ventral nerve tracts, which terminate at the body/tail junction. The excretory bladder region of the tail is also immunoreactive for the 4 peptides, and a fine pair of nerve tracts extends the length of the tail shaft. Immunoreactive nerve cell bodies are also evident in the midbody region of the intrasprocystic cercariae, these same structures being immunoreactive for the neuronal marker, neurone-specific enolase (NSE). The organization of the larval peptidergic nervous system is compared to that of the cholinergic nervous system and contrasted with the peptidergic system in the adult worm. The absence of immunoreactivity to 5-HT is discussed in relation to the proposed development of the aminergic nervous system upon establishment in the definitive host.

Objective.Rhipicephalus sanguineus is the most widespread tick in the world and a vector of many pathogens infecting dogs and humans. To date, there have been no investigations on the biology of R. sanguineus under natural Mediterranean climatic conditions. Methods. The biology of R. sanguineus was studied under laboratory and natural climatic conditions in southern Italy. Several biological parameters were compared in different seasons, and their correlation with climatic data was assessed. Results. The life cycle of R. sanguineus lasted for a mean of 101·4 and 116·2 days under laboratory and environmental conditions respectively. Reproductive parameters of wild-collected females kept in the environment in May were significantly different from first-generation females maintained in September–October, and the correlation between reproductive parameters and climate data varied according to season. Conclusion. These results indicate that the Mediterranean climate affects the biology of R. sanguineus, which compensates its losses during the initial phases of its cycle in spring with high feeding and moulting rates of larvae and nymphs during summer as well as with high egg production during autumn. These results advance our knowledge of the biology of R. sanguineus and will be useful for the understanding of the eco-epidemiology of tick-borne diseases that this tick transmits.

The study aims were to quantify the survivorship of Anguillicola crassus in eels in seawater and investigate transmission in estuarine (50% seawater) and marine (100% seawater) simulated conditions. Most A. crassus were able to survive and reproduce in 50% and 100% seawater eels for up to 6 months and therefore could survive during the spawning migration of eels to the Sargasso Sea. Up to 10% of the parasite metapopulation were damaged after long-term exposure to 50% or 100% seawater. Transmission of A. crassus was completed in 50% and 100% seawater in the laboratory, although infection levels in 100% seawater hosts were always lower. Transmission in estuaries is ecologically possible since the copepod Eurytemora affinis is susceptible to infection and is the dominant autochthonous species in most tidal estuaries in the Northern hemisphere. Transmission at sea is unlikely due to lack of suitable intermediate hosts. The data support the suggestion that dissemination of A. crassus has been facilitated by natural movement of eels in coastal waters. It is possible that A. crassus could impair the success of the eel spawning migration to the Sargasso Sea, although there is no direct evidence to support this.

In recent years, Trichomonas gallinae emerged as the causative agent of an infectious disease of passerine birds in Europe leading to epidemic mortality of especially greenfinches Chloris chloris and chaffinches Fringilla coelebs. After the appearance of finch trichomonosis in the UK and Fennoscandia, the disease spread to Central Europe. Finch trichomonosis first reached Austria and Slovenia in 2012. In the present study the genetic heterogeneity of T. gallinae isolates from incidents in Austria and Slovenia were investigated and compared with British isolates. For this purpose comparative sequence analyses of the four genomic loci ITS1-5.8S-ITS2, 18S rRNA, rpb1 and Fe-hydrogenase were performed. The results corroborate that one clonal T. gallinae strain caused the emerging infectious disease within passerine birds and that the disease is continuing to spread in Europe. The same clonal strain was also found in a columbid bird from Austria. Additionally, the present study demonstrates clearly the importance of multi-locus sequence typing for discrimination of circulating T. gallinae strains.

A double clone of Trypanosoma platydactyli Catouillard, 1909, derived from a single trypomastigote from the blood of the Moorish gecko, Tarentola mauritanica, was grown in vitro. Morphogenesis of the parasites led to a stable population of promastigotes which were identical, in general morphology, ultrastructure and the electrophoretic mobility of 8 enzymes, to those cultures previously considered to be Leishmania tarentolae Wenyon, 1921. These cultures included the strain isolated from the type locality in Algeria by Parrot (1949) and later used as a laboratory model for the genus Leishmania. T. platydactyli and L. tarenlolae are synonymized and the present status of saurian Leishmania parasites is discussed.

With the exception of gudgeon (Gobio gobio) infection of cyprinid fish with the plerocercoid of Ligula intestinalis is associated with a host-tissue response. The nature and specificity of this response has been investigated using transplantation techniques. In roach there is an intense cellular response to implanted Sepharose, Spurr's resin and L. intestinalis, irrespective of donor host species. However, tegument damage only occurs in gudgeon-Ligula and cultured ro&ch-Ligula which has been exposed to gudgeon-plasma. L1 and L2 cell types predominate within the cellular response. Once the host leucocytes have breached the tegument, they migrate over the basal lamina and penetrate into the sub-tegumental region. In gudgeon an intense cellular attack occurs against Sepharose, Spurr's resin, ro&ch-Ligula and cultured gudgeon-Ligula which had been implanted directly or exposed to roach plasma. Tegument damage only occurs in the latter. In contrast, neither direct implantation of gudgeon-Ligula nor cultured roach-Ligula which has been exposed to gudgeon plasma evokes a cellular response. It is suggested that in these two instances, as in natural infections of gudgeon, protective host proteins may be absorbed onto the surface of the parasite.

Extracts and essential oils from plants are important natural sources of pesticides. These compounds are considered an alternative to control ectoparasites of veterinary importance. Schinus molle, an endemic species of Brazil, produces a high level of essential oil and several other compounds. The aim of this work was to determinate the chemical composition of extracts and essential oils of S. molle and further to evaluate the activity against eggs and adults of Ctenocephalides felis felis, a predominant flea that infests dogs and cats in Brazil. In an in vitro assay, the non-polar (n-hexane) extract showed 100% efficacy (800 µg cm−2; LD50 = 524·80 µg cm−2) at 24 and 48 h. Its major compound was lupenone (50·25%). Essential oils from fruits and leaves were evaluated, and had 100% efficacy against adult fleas at 800 µg cm−2 (LD50 = 353·95 µg cm−2) and at 50 µg cm−2 (LD50 = 12·02 µg cm−2), respectively. On the other hand, the essential oil from fruits and leaves was not active against flea eggs. This is the first study that reports the insecticidal effects of essential oils and extracts obtained from Schinus molle against Ctenocephalides felis felis.

The ciliated protozoan Cryptocaryon irritans, a parasite of seawater fishes, was found to express an antigen that elicits antibodies in rabbits and tiger puffer (Takifugu ruburipes). Serum from rabbits and fish immunized with theronts had agglutination/immobilization activity against theronts in vitro; fish serum antibody levels (measured by enzyme-linked immunosorbent assays: ELISA) correlated with this activity. Anti-theront antibody levels in fish were significantly higher in the immunized group as compared with control fish at 2 weeks after booster immunization (injection of bovine serum albumin; Student's t-test, P<0·01). Biochemical analyses indicated that a Triton X-114-soluble 32 kDa theront integral membrane protein may be the agglutination/immobilization antigen. Indirect immunofluorescence staining of theronts suggested that this 32 kDa antigen was expressed on the surface of cilia. The full-length 32 kDa antigen cDNA contained 1147 basepairs, encoding a 328-amino acid protein including hydrophobic N- and C-termini. As with Tetrahymena and Paramecium spp., TAA and TAG appear to be used as glutamine codons in the 32 kDa antigen gene.

Between December 2007 and March 2009, small mammals were captured in 6 Atlantic Forest patches in Brazil. We assessed tick-host associations and whether they differ among forest strata, sites, seasons, and host age classes or between sexes. Moreover, we assessed the exposure of animals to Rickettsia spp. In total, 432 animals were captured and 808 ticks were found on 32·9% of them. Significant differences were found among host species, collection sites, and forest strata; microhabitat preference was a strong risk factor for tick infestation. The highest tick density rates were recorded in forest fragments settled in rural areas; 91·3% of the ticks were collected from animals trapped in these forest fragments. A high prevalence (68·8%) of antibodies to Rickettsia spp. was detected among animals. This study suggests that disturbed Atlantic Forest fragments provide an environment for ticks and small mammals, which are highly exposed to rickettsiae. It also indicates that forest patches settled in rural areas are usually associated with higher small mammal diversity as well as with higher tick density rates.

Penetration and maturation of Schistosoma mansoni were compared in four strains of mice under controlled conditions, with the C3H as the reference strain. Differences were observed in (1) average numbers of cercariae penetrating, (2) percentage of individual mice penetrated by at least 90% of the available cercariae, (3) worm burdens, (4) maturation of penetrants, and (5) consistency of mice with reference to these characteristics. Criteria 2, 4 and 5 have not been evaluated in this way before.

Best average penetration (47%), greatest consistency, and highest percentage of samples with at least 90% of the cercariae penetrating were in mice of the C3H strain (85%). Of the other strains the Swiss albino strain most resembled strain C3H. Beige and hairless mice were comparatively resistant to penetration since only 60 and 30% respectively of these hosts had at least 90% of the cercariae penetrating. Hairless mice gave the best average worm recovery (23%) and parasite maturation (53%) and beige the poorest (15 and 34%). Mice of the Swiss and beige strains were highly variable in terms of parasite maturation.

The establishment of reproducible differences in infections of hosts at the sub-specific level provides basic information for investigation of factors which influence penetration and maturation and which are thus involved in host specificity and innate resistance in closely related animals.

The authors are grateful to Dr Margaret Deringer, National Cancer Institute, National Institutes of Health, for the hairless mice and to Mildred Walters, Naval Medical Research Institute, for assistance in the production of cercariae.

Although there is a growing recognition that the transfer of diseases between humans and non-human primates can be of great significance for conservation biology, there have been only a few studies focusing on parasites. In this study, saddleback (Saguinus fuscicollis) and moustached tamarin (Saguinus mystax) from the rainforest of the Peruvian lowlands were used as models to determine helminth parasite associations between canopy-dwelling primate species and a nearby human settlement. The human population showed high prevalences of infestation with a number of nematodes, including Ascaris lumbricoides (88·9%), Trichuris trichiura (37%) and hookworms (55·6%). However, the ova of these geohelminths were not detectable in tamarin faeces. Thus, no direct parasite transfer from humans to non-human primates could be documented. However, tamarin groups with more frequent contact to humans and their facilities had significantly higher prevalences and egg output of Prosthenorchis elegans, an important primate pathogen, than a forest group. In contrast, a cestode was significantly more common with more egg output in sylvatic than in human-associated groups. Human alteration of the habitat is likely to play a major role in determining the occurrence, prevalence and intensity of helminth infestation of wild non-human primates.

Since the first description of Marteilia refringens (Paramyxea) in flat oysters Ostrea edulis in 1968 in the Aber Wrach, Brittany (France), the life-cycle of this parasite has remained unknown. However, recent studies, conducted in the ‘claire’ system, have proposed the planktonic copepod Acartia grani as a potential intermediate host for the parasite. Nevertheless, experimental transmission of the parasite through the copepod has failed. Recent studies in this field have reported the presence of the parasite in zooplankton from the bays of the Delta de l'Ebre, a more complex and natural estuarine environment than that of the claire. As a result, 2 new Marteilia host species were proposed: the copepods Oithona sp. (Cyclopoida) and an indeterminate Harpaticoida. Consequently, the objective of the present work was to study the dynamics of Marteilia in the zooplankton community from one of the bays, Alfacs Bay, as well as the dynamics of the parasite in cultivated mussels during 1 complete year. Six different zooplankton taxa appeared to be parasitized by M. refringens, including copepods (3 Calanoida, Acartia discaudata, A. clausi and A. italica; 1 Cyclopoida, Oithona sp.; and 1 Harpacticoida, Euterpina acutifrons), and larval stages of decapod crustaceans (zoea larvae of Brachyura, probably Portumnus sp.). These taxa are thus proposed as new subjects for study, since they could be intermediate hosts in the infection process of mussels by Marteilia.

We conducted the first comprehensive study on the spatiotemporal structure of trematode communities in the large-mouthed valve snail, Valvata macrostoma. A total of 1103 snails were examined monthly between May and October 2007 from Lake Konnevesi, Central Finland, from a shallow (1–2 m deep) and an offshore site (5–6 m deep), located ca. 50–70 m apart. Snails were infected by 10 trematode species. The species composition and prevalence were strikingly different between the sites with high species diversity in the shallow site (all 10 species; total prevalence of sporocysts/rediae 12·1%, metacercariae 55·4%) compared to the deeper site (3 species; prevalence 15·0% and 1·9%, respectively). This difference persisted throughout our study and is probably related to the spatial distribution of bird definitive hosts, whereas the seasonal parasite dynamics are likely to be affected by changes in the age-structure of the snail population. The probability of sporocyst infections increased with snail size, but no such trend was observed in redial or metacercarial infections which decreased with host size. Our results show that generally well-described spatiotemporal differences in trematode infection of molluscs can emerge in very narrow spatial and temporal scales, which emphasizes the importance of these factors in community studies.

Among the potential effects of parasitism on host condition, the ‘increased host abilities’ hypothesis is a counterintuitive pattern which might be predicted in complex-life-cycle parasites. In the case of trophic transmission, a parasite increasing its intermediate host's performance facing non-host predators improves its probability of transmission to an adequate, definitive host. In the present study, we investigated the cost of infection with the acanthocephalan Polymorphus minutus on the locomotor/escape performance of its intermediate host, the crustacean Gammarus roeseli. This parasite alters the behaviour of its intermediate host making it more vulnerable to predation by avian definitive hosts. We assessed the swimming speeds of gammarids using a stressful treatment and their escape abilities under predation pressure. Despite the encystment of P. minutus in the abdomen of its intermediate host, infected amphipods had significantly higher swimming speeds than uninfected ones (increases of up to 35%). Furthermore, when interacting with the non-host crustacean predator Dikerogammarus villosus, the highest escape speeds and greatest distances covered by invertebrates were observed for parasitized animals. The altered behaviour observed among the manipulated invertebrates supported the ‘increased host abilities’ hypothesis, which has until now remained untested experimentally. The tactic of increasing the ability of infected intermediate hosts to evade potential predation attempts by non-host species is discussed.

The germ-cell cycle and method of reproduction in the daughter sporocysts of the bucephalid Cercaria bucephalopsis haimeana (Lacaze-Duthiers, 1854) and the gymnophallid Cercaria dichotoma Lebour, 1911 (non Müller) from the edible cockle, Cardium edule L., have been investigated by the examination of stained sections. A review is given of the principal interpretative theories which attempt to explain the nature of the reproduction and the homology of the germinal sacs of the Digenea. It is suggested that the life-cycle includes a cyclic alternation of homologous generations. Of these, the miracidium/mother-sporocyst and the daughter germinal-sac generations usually reproduce by apomictic parthenogenesis but polyembryony may also occur secondarily in some species.

We are very grateful to Dr Gwendolen Rees and Professor E. W. Knight-Jones for considerable help with the manuscript.

Four monoclonal antibodies, raised against macroschizonts of Theileria parva, were studied to characterize their antigen binding specificity. The indirect fluorescent antibody test showed that the antigen(s) recognized were present in the macroschizont stage of the parasite life-cycle but not in piroplasm, kinete or sporozoite stages. Immunoblot analysis of macroschizont stage antigens suggested that all four antibodies recognized the same antigen. This was a molecule which varied in molecular mass between different parasite stocks, ranging from 68000 to 95000 Da. The antigen was localized by immunoelectron microscopy to the surface of the intracellular macroschizonts.

Life-cycle stages of the microsporidia Nosema bombycis, the pathogen causing silkworm pebrine, were separated and purified by an improved method of Percoll-gradient centrifugation. Soluble protein fractions of late sporoblasts (spore precursor cells) and mature spores were analysed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Protein spots were recovered from gels and analysed by mass spectrometry. The most abundant differential protein spot was identified by database search to be a hypothetical spore wall protein. Using immunoelectron microscopy, we demonstrated that HSWP5 is localized to the exospore of mature spores and renamed it as spore wall protein 5 (NbSWP5). Further spore phagocytosis assays indicated that NbSWP5 can protect spores from phagocytic uptake by cultured insect cells. This spore wall protein may function both for structural integrity and in modulating host cell invasion.

Large malaria epidemics in the East African highlands during the mid and late 1990s kindled a stream of research on the role that global warming might have on malaria transmission. Most of the inferences using temporal information have been derived from a malaria incidence time series from Kericho. Here, we report a detailed analysis of 5 monthly time series, between 15 and 41 years long, from West Kenya encompassing an altitudinal gradient along Lake Victoria basin. We found decreasing, but heterogeneous, malaria trends since the late 1980s at low altitudes (<1600 m), and the early 2000s at high altitudes (>1600 m). Regime shifts were present in 3 of the series and were synchronous in the 2 time series from high altitudes. At low altitude, regime shifts were associated with a shift from increasing to decreasing malaria transmission, as well as a decrease in variability. At higher altitudes, regime shifts reflected an increase in malaria transmission variability. The heterogeneity in malaria trends probably reflects the multitude of factors that can drive malaria transmission and highlights the need for both spatially and temporally fine-grained data to make sound inferences about the impacts of climate change and control/elimination interventions on malaria transmission.

Ascariasis is an important infection in humans (Ascaris lumbricoides) and pigs (Ascaris suum) and individuals appear to be predisposed to either heavy or light worm burdens. These extremes of susceptibility and resistance are represented in a mouse model by 2 strains of mice, CBA mice showing high resistance to infection and C57BL/6 which are highly susceptible, as reflected in worm burdens in the lungs 6–7 days after infection. In an attempt to identify the point at which the difference between these 2 strains is first manifested, we quantified worm burdens at key stages during infection leading up to the pulmonary stage of development. Thus mice were inoculated with fully embryonated A. suum eggs and larval burdens were enumerated in the large intestine and rectum, liver and lungs of the 2 strains at 6 h post-inoculation (p.i.) and on each of days 1–8 p.i. inclusively. A higher percentage of the total inoculum was recovered from the intestine/rectum of C57BL/6j mice in contrast to CBA/Ca mice at 6 h p.i. Larvae were recovered from the intestinal contents and also whilst actively migrating through the large intestinal wall. The number of larvae recovered was significantly reduced in CBA/Ca mice in contrast to C57BL/6j mice between the phase of migration from the liver and arrival in the lungs. The combined results of the inoculation of mice with corticosteroids and the examination of the change in profile and number of leukocytes present in bronchoalveolar lavage fluid suggested that the pulmonary inflammatory immune response was not prominently involved in primary protection of mice to A. suum infection in the latter days of infection in the lungs. The susceptible C57BL/6j mice produced a BAL response almost twice as intense as that of resistant CBA/Ca mice with stronger neutrophil, lymphocyte and eosinophil but not macrophage components, suggesting that the difference in worm burdens between the strains was generated earlier in the course of infection. These results were further corroborated by a histological examination of the lung tissues which showed that the passage of the larval stages of A. suum through the mouse lungs was associated with a marked inflammatory response in both strains. Again, C57BL/6j mice exhibited increased inflammation relative to CBA/Ca mice. Hence some hepatic/post-hepatic factor that varies between the 2 strains, but exerts its effect before the lung phase plays a critical role in determining the success of larvae through the host tissues. The possible sites of this host defence are reviewed.

Interference microscopy has been used to study the rate at which water enters and is lost from larval and adult stages of Ditylenchus dipsaci and D. myceliophagus. It is shown that the cuticle of the dry nematode, irrespective of stage, is completely permeable to water. In contrast, marked intra– and interspecific differences were found in the rate at which stages dry at 0 and 50% relative humidity. All stages of D. myceliophagus dry very rapidly: in this species, death, as a consequence of severe desiccation, appears to be linked to a lack of any marked ability to control water loss. For D. dipsaci, those stages known to survive desiccation will appear to be the slower dryers. In general, the “order of performance” in slowing the rate of drying is the same as the order previously found for survival of desiccation. Both the 4th– and, to a lesser extent, the 3rd–stage larvae of D. dipsaci show evidence of an intrinsic ability to control water loss.

An analysis of the drying curve for 4th–stage larvae of D. dipsaci supports the indications of interference photographs that the rate of drying is slowed by a decrease in permeability of the drying cuticle.

Vacuum desiccation studies further demonstrate the remarkable ability of the 4th–stage larva of D. dipsaci to control its rate of drying. The survival of 4th– and 3rd–stage larvae of D. dipsaci is diminished by very rapid drying, and the survival of other stages of this species and larval and adult stages of D. myceliophagus is precluded. The decrease in survival of the 4th–stage larvae of D. dipsaci is shown to be associated with a more rapid rate of water loss.