We have examined metabolic rate, lipid and carbohydrate of female Aedes aegypti during 10 days following a malaria-infected bloodmeal. In parallel, we determined bloodmeal size, portions retained and diuresed, and subsequent fecundity. We found that mosquitoes obtained identical masses of blood when feeding on an infected or control host. However, infected mosquitoes lost more mass during diuresis and retained a smaller mass. Infection led to a significant reduction in fecundity, the extent of which could not be explained by the difference in post-diuresis bloodmeal mass alone. We found no differences in lipid or carbohydrate content between infected and control mosquitoes during the 10 days post-infection, although infected mosquitoes had a lower body mass than controls. Metabolic rates were not different between groups, except during blood digestion, where the metabolic rate was lower in infected mosquitoes. These results suggest that infection by malaria does not lead to an increase in metabolic rate during the phases of midgut invasion and sporogony. However, infection does have a measurable effect on fecundity and subsequent body mass of the infected females.

We monitored temporal changes in the magnitude of altered host behaviour in minnows (Pimephales promelas) experimentally infected with metacercariae of a brain-encysting trematode (Ornithodiplostomum ptychocheilus). This parasite develops and then encysts in a region of the brain that mediates the optomotor response (OMR), an innate behaviour that links visual stimuli with motor performance. The OMR of infected and uninfected minnows was evaluated between 0 and 10 weeks post-infection (p.i.), an interval spanning the development period of metacercariae to infectivity in birds. Trials involved monitoring the time an individual minnow spent following a spinning drum that had been painted with alternating black and white stripes. At 2 and 4 weeks p.i., infected minnows followed the drum 40% less often than controls. Differences between controls and infected fish declined thereafter, and were undetectable by 10 weeks p.i. Both control and infected fish habituated equally rapidly to the spinning drum. However, the difference in performance between controls and infected fish was 29% for experienced fish and 48% for fish that had never experienced the drum. Because maximum parasite-induced reduction in OMR coincided with the period of maximum parasite development, the behavioural effects are most likely due to unavoidable pathology in the brain associated with developing larvae.

Evolutionary and closer structural relationships are demonstrated by phylogenetic analysis, peptide prediction and molecular modelling between Solanum tuberosum apyrase, Schistosoma mansoni SmATPase 2 and Leishmania braziliensis NDPase. Specific protein domains are suggested to be potentially involved in the immune response, and also seem to be conserved during host and parasite co-evolution. Significant IgG antibody reactivity was observed in sera from patients with American cutaneous leishmaniasis (ACL) and schistosomiasis using potato apyrase as antigen in ELISA. S. mansoni adult worm or egg, L. braziliensis promastigote (Lb) and Trypanosoma cruzi epimastigote (EPI) have ATP diphosphohydrolases, and antigenic preparations of them were evaluated. In ACL patients, IgG seropositivity was about 43% and 90% for Lb and potato apyrase, respectively, while IgM was lower (<19%) for both. In schistosomiasis patients IgM (>40%) or IgG (100%) seropositivity for both soluble egg (SEA) and adult worm (SWAP) antigens was higher than that found for potato apyrase (IgM=10%; IgG=39%). In Chagas disease, IgG seropositivity for EPI and potato apyrase was 97% and 17%, respectively, while the IgM was low (3%) for both antigens. The study of the conserved domains from both parasite proteins and potato apyrase could lead to the development of new drug targets or molecular markers.

The genus Clinostomum Leidy, 1856 (Digenea: Clinostomidae) has been reported in all ecozones of the world and a clear separation between the species of the ‘Old World’ and ‘New World’ has been recognized based on molecular studies. Recent works on Afrotropical species include redescriptions of C. cutaneum and C. phalacrocoracis, while C. tilapiae has yet to be studied using modern taxonomic approaches. In the present research, morphological redescription of C. tilapiae metacercariae from a new host, Synodontis batensoda sampled at Anambra River Basin, Nigeria, together with molecular analysis of nuclear internal transcribed spacer rDNA and cytochrome c oxidase 1 mtDNA are reported. We also provide morphological and molecular data from four further putative species of Clinostomum (morphotypes 1–4) from different areas of Africa, as well as the first report of C. phalacrocoracis in South Africa.

We evaluated the quantitation of two schistosome circulating antigens in serum and urine as a tool for the assessment of the efficacy of praziquantel dosage regimens (40 versus 60 mg/kgbw). In addition we compared the efficacy of two different brands of praziquantel (Biltricide® and Distocide®), given at the same dosage (40 mg/kgbw). Thirty five Egyptian hospitalized schistosomiasis mansoni patients participated in this study. Thirteen patients (Group 1) received 60 mg/kgbw Biltricide®, administered in 3 oral doses of 20 mg in one day; 22 individuals (Group 2) were treated with 40 mg/kgbw (12 Biltricide®, 10 Distocide®), given in one oral dose. Circulating anodic antigen (CAA) and circulating cathodic antigen (CCA) were quantitated by monoclonal antibody-based ELISA's before, and 1, 3 and 6 weeks after chemotherapy. Before treatment, all patients were positive for at least one of the circulating antigen assays. Three to six weeks after treatment significantly more patients were found to be negative in Group 1 compared to Group 2 (X2 = 7·13, P = 0·008, n = 35). Also the levels of CCA and CAA in serum and of CCA in urine were found to be significantly higher in Group 2 (Mann- Whitney U < 85, P < 0·05, n = 35). These results were confirmed by parasitological data. No differences were found between treatment with Biltricide® or Distocide®. Our results indicate that praziquantel treatment of schistosomiasis with 60 mg/kgbw divided in 3 doses results in a higher cure rate compared to 40 mg/kgbw as a single dose, and provide further evidence for the use of the CAA and CCA assays as a sensitive method to monitor the efficacy of chemotherapy, particularly when circulating antigen assays are combined by parallel testing.

Changes in the response of black mollies Poecilia latipinna to infection with Ichthyophthirius multifiliis subsequent to single I. multifiliis infections were examined experimentally. Incomplete resistance to infection was established, the degree of which did not depend on the intensity of the initial infection. Resistance was maintained for longer periods, however, by those fish with higher initial levels of infection. Previous experience of infection by their hosts had a significant, though small, effect on the time for which trophozoites remained on the fish. The influence of these results on the population dynamics of the Ichthyophthirius-fish interaction is examined with the aid of simple mathematical models.

Four common blood parasites belonging to the genera Babesia, Hepatozoon, Trypanosoma and Grahamella were detected in sympatric populations of Clethrionomys glareolus and Apodemus sylvaticus. Seasonal changes in the abundance of hosts and parasites were monitored in a longitudinal field survey for 18 months. All parasites in both hosts showed similar seasonal patterns with most transmission occurring in summer and autumn and ceasing in winter. Amongst aged subsets of each host population, Hepatozoon and Grahamella spp. were primarily parasites of young animals whereas Babesia and Trypanosoma spp. generally infected older hosts. Data from the aged subsets were also used to estimate values for the basic reproductive rate, R0, for each parasite. In C. glareolus, R0 = 2·0 for B. microti, 2·6 for H. erhardovae, 1·7 for T. evotomys and 2·8 for Grahamella sp. In A. sylvaticusR0 = 2·2 for Grahamella sp. These estimates are probably representative only for summer and autumn of each year.

Young cercariae were covered with a thin nucleated primitive epithelium which was lost when the true tegument appeared beneath it. The tegument was at first similar to the primitive epithelium in that it was a nucleated, cytoplasmic syncytium but it could be distinguished from the primitive epithelium by its thickened outer membrane and by the layer of interstitial material beneath its basal membrane. The tegumental nuclei became pycnotic and disappeared and, at about the same time, nucleated subtegumental cells became continuous with the tegument. Two types of small, dense bodies were formed in the subtegumental cells and passed into the tegument. Later in development the tegument became filled with dense, granular material and spines were formed. The fully developed cercarial tegument was covered with a surface coat of fibrous material which was specifically attached to the outer membrane of the tegument.

The observations described in this paper formed part of a thesis approved by the University of London for the Degree of Ph.D.

Common bird species were screened during May and June 2007 on Barbados for haemosporidian parasites (Haemosporida) of the genera Haemoproteus and Plasmodium to determine whether the low parasite diversity reported in previous studies might have reflected limited sampling. PCR screening and DNA sequencing revealed a single predominant lineage of Haemoproteus identified as H. coatneyi. Sixty-two out of 257 birds were infected with Haemoproteus spp. on Barbados in 2007. Fifty-nine of the infections were identified as H. coatneyi (lineage HC), the only lineage recovered in the previous study in 1993. Two of the infections recovered from the bananaquit (Coereba flaveola) were identified as Haemoproteus spp. (lineage HD), which is the prevalent haemosporidian parasite in C. flaveola on Grenada. We discuss the possibility of infrequent colonization events and absence of vectors as explanations for Barbados's low avian haemosporidian diversity. In our study, the parasites were absent from the southeast of the island, whereas they were abundant in several host species in the northwest. Accordingly, environmental and host population genetic differences were also investigated between the areas with and without parasites. No host genetic differences were found between the parasite-free and the parasite-afflicted regions. However, the parasite-free region is slightly warmer and drier, and it supports less vegetation than the parasite-afflicted region. The influence that this harsher environment may have on vector survival is discussed.

The physiological effects of selected classical transmitters and FMRFamide-related peptides (FaRPs) on dispersed muscle fibres from the marine turbellarian, Procerodes littoralis have been examined. Confocal scanning laser microscopy coupled with fluorescein isothiocyanate (FITC) or tetramethylrhodamine (TRITC)-labelled phalloidin revealed a highly developed body wall muscle system with circular, longitudinal and diagonal layers of muscle fibres. Dispersed muscle fibres contracted when depolarized by exposure to extracellular media with elevated K+ (15–100 mM) in a concentration-dependent manner, with a maximal response of 87% achieved at [ges ] 75 mM. 5-Hydroxytryptamine (5-HT) induced concentration-dependent muscle contraction between 0·01 and 1000 μM, with 10 μM producing a near maximal contraction response of 75%. Acetylcholine (ACh) had less pronounced excitatory effects (0·01–1000 μM), inducing contraction of only 32% of the fibres at 100 μM. The flatworm FMRFamide-related peptides (FaRPs), GYIRFamide, YIRFamide and GNFFRFamide each had concentration-dependent myocontractile effects, indicating the occurrence of at least 1 FaRP receptor on P. littoralis muscle fibres. At 10 μM peptide, GNFFRFamide induced contractions in < 40% of the muscle fibres examined, whereas YIRFamide and GYIRFamide induced contraction in 70 and 75%of muscle fibres, respectively. The order of potency of the peptides was: GYIRFamide > YIRFamide > GNFFRFamide. Pre-incubation of the muscle fibres in 5 μM 5-HT significantly reduced the responses to GYIRFamide, YIRFamide and 5-HT, while the responses to high K+ remained unaltered. Muscle fibres pre-incubated in GYIRFamide (0·1 μM) were also less responsive to 5-HT but not to ACh and high-K+. The GYIRFamide analogue, GYIRdFamide, did not induce muscle contraction (0·01–100 μM) per se, but when co-applied with the myoactive peptides GYIRFamide, YIRFamide or GNFFRFamide, it significantly blocked their ability to elicit contractions. This suggests that the peptides tested may act via a common muscle-based neuropeptide receptor. GYIRdFamide did not alter the contractile effects of high K+, 5-HT or ACh. Collectively, these results indicate that FaRPs, 5-HT and ACh all have the potential to cause muscle contraction in flatworms and that 5-HT and FaRPs alter muscle sensitivity to each other, but do not influence the ability of flatworm muscle fibres to contract.

This study analyses the different populations of Trypanosoma brucei spp. which may coexist within the midgut of wild tsetse flies (Stevens et al. 1994). Cloned trypanosome populations characterized by multilocus enzyme electrophoresis (MLEE) were further analysed by the random amplified polymorphic DNA (RAPD) technique, allowing detection of genetic variation at a finer level than that possible by MLEE. Genetic distance matrices derived from the results of each of the two biochemical methods were calculated and compared using a computer program based on the method of Mantel (1967). The observed correlation was used to investigate the degree of linkage disequilibrium (LD) in the data, association between unrelated polymorphic markers providing a measure of the departure from panmixia. The potential of each biochemical method to detect linkage was evaluated by an extended Mantel test. The MLEE/RAPD correlation test evidenced significant LD within the population, suggesting a predominantly clonal method of reproduction for these West African trypanosomes. Analysis of RAPD data by the extended Mantel test also showed significant LD, while the results with MLEE data were less conclusive, providing an indication of the relative potential of the two techniques to detect fine genetic variation.

Asexual stages and mature gametocytes of Plasmodium falciparum have been produced in a 500 ml suspension culture system containing 6–10% red cells. Medium change was automated and was accomplished using a tangential flow filtration unit. The rate of multiplication of the parasite (isolate NF 54) was consistently high when compared with static cultures and ranged from 6·4 × to 15·3 × per cycle in 8 experiments. Mature gametocytes were present in up to 2% of red cells in 14-day-old cultures. Only minor modifications would be required to further scale-up the volume of the culture.

The tegumental surface of Schistosoma mansoni sporocysts is the site of both nutritive and immunological interactions with haemolymph cells and plasma of Biomphalaria glabrata, the schistosome intermediate host. Within minutes of being placed in host plasma, sporocysts acquire plasma antigens, and within 3 h host plasma antigens are present on the surface at near steady state. Though a wide variety of peptides is acquired, there is selection. Furthermore, some differences occur in the peptides acquired from the plasma of susceptible and resistant strains of snail. Acquired antigens are rapidly processed, and are predominantly undetectable in tegumental extracts after a few hours. In contrast, rabbit antibody on sporocysts remains in situ for at least 48 h, so under some conditions there is stable expression of certain tegumental antigenic determinants.These data, obtained using antibodies to snail plasma antigens and to sporocyst tegumental antigens, are discussed in the light of current ideas on the cellular and molecular basis of susceptibility and resistance in this host#parasite system.

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3–Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Sα rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5·8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Sα rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.

Large numbers of Theileria parva sporozoites were separated from Rhipicephalus appendiculatus adult ticks by filtration and were then concentrated by centrifugation. The sporozoites were incubated at 37 °C with leucocytes from 6 cattle of Bos indicus and B. taurus types. Giemsastained smears and living preparations under interference contrast microscopy were used to follow the course of the infection of the leucocytes with sporozoites. Sporozoites were seen to attach rapidly to about 25% the leucocytes which they penetrated. After penetration by the sporozoites the morphology of the cells changed to show an increase in cytoplasm and an enlargement of the Golgi apparatus, with which the parasite appeared to become associated. The early intracellular or preschizont stages resembled Babesia parasites. From day 3, the parasite showed the typical morphology of the macroschizont of T. parva. Multiple infections were frequent and up to 8 schizonts were observed arranged around the Golgi apparatus. Multiply infected cells did not survive in culture but some of the cells infected with a single parasite divided to produce 2 infected daughter cells and infected lymphoblastoid cell lines were established in all 21 attempts.

Anthelmintic resistance is a serious problem in veterinary medicine and appears to be developing in some helminths of importance to human health. Anthelmintic drugs remain the principal means of control of helminth infections in animals and humans and the continued dependence on these pharmaceuticals will continue to impose selection pressure for resistance development. Our ability to detect anthelmintic resistance before control breaks down and to monitor the spread of anthelmintic resistance is quite limited. We are currently dependent on biological methods which are not sufficiently sensitive to detect low levels of drug resistance and are particularly difficult to perform on helminth parasites of humans. There is a serious need for new molecular markers for detecting and monitoring for anthelmintic resistance. The problem of anthelmintic resistance is already very serious in nematode parasites of livestock. In addition, there should be great concern about possible anthelmintic resistance development and the lack of tools and efforts for monitoring it as part of the major worldwide programmes to control helminth parasites in people. An international Consortium has been formed to develop Anthelmintic Resistance Single nucleotide polymorphism markers (CARS). Discussions within the Consortium have addressed the need for such markers, the current state of knowledge about them, possible mechanisms of anthelmintic resistance and approaches and constraints to the development of markers. Summaries of the state of the art in these areas are presented in a series of papers in this Special Issue of Parasitology.

In Strongyloides ratti-infected rats, 2 peaks of egg excretion were observed; a large one with maximum egg production on days 7–8 of infection and a small more inconspicuous one around day 25. The second peak, which had been ignored in most studies, was produced by adults in the caecum and the colon. The adults were larger in length and had more embryonated eggs in the uterus compared with adults in the small intestine at day 25 post-infection. It is suggested that parasitic adults once expelled from the small intestine resettle and recover in the large intestine. Filter paper faecal culture carried out for 9 days at different days post-infection revealed that the total number of infective larvae that developed during the second peak was twice the number that developed during the first peak, despite the fact that total egg output during the second peak was less than one twentieth of the first peak. The results suggest that the small second peak was as important as the first one in the transmission of S. ratti.

The behaviour of mice exposed to graded doses of Trichinella spiralis larvae was monitored in an open-field situation. Ambulatory and exploratory activity declined significantly as the infections progressed, whereas the time spent inactive increased concomitantly. The advent of these behavioural changes coincided with the expected release of larvae by the adult worms and showed no clear signs of abating as late as 90 days post-inoculation. The levels of activity were negatively correlated with the inoculating dose. Such behavioural changes may render the host more vulnerable to predation and thus enhance parasite transmission.

The pig is a natural host of Schistosoma japonicum and a useful animal model of human disease. In the present study mRNA levels of Th1 (IFN-γ) and Th2 (IL-10 and IL-4) cytokines were assessed by RT-PCR within tissues from infected pigs. Twelve Danish crossbred pigs were infected by intramuscular injection or orally with 1000 cercariae. Six other pigs served as non-infected controls. Liver and intestinal tissues were collected 10 weeks post-infection, and analysed for their relative levels of cytokine mRNA. Infected pigs developed a Th2 response as characterized by the increased level of mRNA encoding for IL-4 and IL-10 in their large intestine (caecum and colon). In contrast, levels of IFN-γ did not differ between control and infected animals although variation between animals was observed. When comparing the immune response of orally and intramuscularly infected animals, we found that orally infected pigs produced higher IL-4 and IL-10 levels in their caecum and colon respectively. This stronger Th2 response correlated with a previously reported delay in maturation of infection following oral infection. The cytokine expression levels in tissue samples taken from lesion sites and in nearby areas, without obvious lesions, were then compared. Subsequent to an oral infection, the Th2 type cytokine production was higher in the lesion sites of the liver. In conclusion, this study is the first demonstration of IL-4 and IL-10 cytokine response in pig tissues during S. japonicum infection.

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.