A nutritional characteristic of trypanosomatid protozoa is that in vitro they need a haem-compound as a growth factor, which is supplied as haemoglobin, haematin or haemin. Because haemin and related porphyrins are an important source of oxidative stress in biological systems, the effect of haemin on growth, protein content and the antioxidant defence system in Trypanosoma cruzi was evaluated. We have observed that, in epimastigotes grown under different haemin concentrations in the culture medium (0–30 mg/l), 5 mg/l was the haemin concentration yielding optimum growth. Above 15 mg/l there was a clear decrease in growth rate, producing the epimastigote to amastigote transformation. Such morphological change was observed together with a marked injury of the enzymatic machinery of the parasite, leading to diminished protein synthesis as well as lower activity of the antioxidant enzymes (superoxide dismutase, ascorbate peroxidase and trypanothione reductase), reduced total thiol content and a marked increase in the HaemOx-1 activity and expression. The current work demonstrates that there is a correlation between higher haemin concentrations in the culture medium and oxidative damage in the cells. Under these conditions induction of HaemOx-1 would indicate the important role of this enzyme as an antioxidant defence response in Trypanosoma cruzi.

In the first part of this paper fifteen species were described, eight of which were from birds. In the present part an additional fifteen are dealt with.

Natural infection with the cephalobid nematode, Daubaylia potomaca, is reported in Helisoma antrosum pericarinatum ( = H. anceps pericarinatum) collected in the vicinity of the University of Michigan Biological Station on Douglas Lake, Cheboygan, Michigan.

The life-cycle of D. potomaca, which appears to be unique, has been determined and reproduced experimentally utilizing Australorbis glabratus and other snails as hosts. The cycle involves escape from the host by gravid females, invasion by the females of new hosts and the reproduction therein of generation after generation of parasites, and the eventual emergence of female worms from dying hosts to the external aquatic environment. The parasite does not lead a saprozoic existence, and no dauer larvae were seen. Except for the newly emerged female, no stage of D. potomaca is ‘free-living’, and there is no evidence that transmission can occur except through the agency of these females. However, parasitic females removed at any time from their hosts by dissection are fully capable of infecting new hosts under experimental conditions.

Dissection of A. glabratus infected with known numbers of female D. potomaca disclosed that the gravid nematodes deposit about two eggs per day in the tissues, that larvae may be found after 5 days of infection, and that the first ‘new’ generation of adult males and females appears within the host 11 or more days after infection. Continued reproduction in the host may give rise to very large nematode populations. A. glabratus exposed to five female D. potomaca usually became infected and all infected snails were killed within 44 days. Death of infected snails was preceded, in nearly every case, by the emergence of female D. potomaca from their hosts. It is suggested that this phenomenon reflects an adaptive response of the worms to changes in the internal milieu of the snail.

Attempts to infect certain physid and lymnaeid snails with D. potomaca failed, and varying degrees of success were achieved in attempts to infect several planorbids. Of the latter, the most suitable hosts appeared to be A. glabratus (Puerto Rico) and Helisoma sp. (Michigan); the least suitable host was Helisoma caribaeum (Puerto Rico and St Croix), in which an intense tissue reaction killed all nematode eggs before they could hatch. Other susceptible planorbids (in declining order of apparent suitability as hosts) were Helisoma trivolvis ? macrostomum (Canada), Bulinus truncatus (Egypt) and Planorbarius corneus (Massachusetts).

Some of the problems associated with the possible use of D. potomaca as a means of biological control for A. glabratus are discussed.

It is a pleasure to thank the persons whose names appear in the text for aid or counsel graciously extended. I am also grateful to Dr George L. Graham (University of Pennsylvania) for certain citations in the literature, to Dr M. R. Tripp (now at the University of Delaware) for receiving the shipments of snails in Boston, and to Dr A. H. Stockard, Director of the University of Michigan Biological Station, for facilitating the field work.

Ivermectin is a promising drug for the treatment of lymphatic filariasis. A meta-analysis of trials investigating the effects of a single treatment suggested a dose-dependent effect on the production of microfilariae (mf) by adult Wuchereria bancrofti parasites. A mathematical model that describes the parasite dynamics in the human host and the impact of ivermectin treatment is presented and its outputs compared with these trials. The calculated trend in mf density after treatment appears to be particularly sensitive to the assumption about the mean life-span of mf. Adopting 0.5–2 years as a range of plausible values for this mf life-span, the model is used to estimate the impact of treatment on the parasite. It is found that irrespective of dosage, ivermectin eliminates 100% of the blood mf from a patient. Furthermore, at a dosage level of 400 μg/kg a single treatment irreversibly reduces the mf production of the adult parasites by at least 65%. For a dosage of 200 μg/kg this reduction is at least 35%. No such effect can be concluded from the results of trials using lower dosages.

Clinical immunity to Plasmodium falciparum malaria develops after repeated exposure to the parasite. At least 2 P. falciparum variant antigens encoded by multicopy gene families (var and rif) are targets of this adaptive antibody-mediated immunity. A third multigene family of variant antigens comprises the stevor genes. Here, 4 different stevor sequences were selected for cloning and expression in Escherichia coli and His6–tagged fusion proteins were used for assessing the development of immunity. In a cross-sectional analysis of clinically immune adults living in a malaria endemic area in Ghana, high levels of anti-STEVOR IgG antibody titres were determined in ELISA. A cross-sectional study of 90 nine-month-old Ghanaian infants using 1 recombinant STEVOR showed that the antibody responses correlated positively with the number of parasitaemia episodes. In a longitudinal investigation of 17 immunologically naïve 9-month-old infants, 3 different patterns of anti-STEVOR antibody responses could be distinguished (high, transient and low). Children with high anti-STEVOR-antibody levels exhibited an elevated risk for developing parasitaemia episodes. Overall, a protective effect could not be attributed to antibodies against the STEVOR proteins chosen for the study presented here.

The rate of expulsion of penetration gland material was determined from photomicrographs of Hymenolepis diminuta hexacanths stained with neutral red dye at intervals after hatching in vitro. Identical secretory rates were recorded for hexacanths incubated in either Tyrode's solution or Tyrode's solution plus an extract of the midgut of Tenebrio molitor. In both media the reduction in stained material observed in the glands after 135 min incubations was equivalent to that observed for hexacanths that had had opportunity to penetrate in vivo during the same time period. These results were interpreted as indicating that the in vivo secretory pattern was similar to that observed in vitro, and that chemical stimuli from the tissue extracts were not required to initiate secretion. Microdensitometric readings also demonstrated a quantitative decrease in dye within the glands as incubation time increased. Ultrastructural examination of the glands showed that their secretory inclusions were exported via ducts to the epithelial cytoplasm where they accumulated in discrete blebs enclosed by the surface membrane. These inclusion-filled, membrane-bounded blebs were apparently formed and released continuously until, after approximately 2 h, only a few inclusions remained in the penetration glands.

The capsule, which surrounds the acanthocephalan parasite Moniliformis dubius in its intermediate host, the cockroach, Periplaneta americana, has been studied in the electron microscope. Some observations were also made with the light microscope.

The capsule surrounding the developing parasite (acanthella) taken from the haemocoel of its host consists largely of closely packed vesicles, which are empty of electron-dense contents.

The vesicles arise from the surface membrane of the insect's haemocytes, which surround the parasite. Long tubular protrusions arise from the haemocyte's surface and, on separating, swell to form rounded vesicles.

The capsule, which surrounds the later resting stage (cystacanth), has apparently been stretched by the growth of the parasite after haemocytic activity has declined. An amorphous, electron–dense component is also evident at this stage.

We are grateful to Mrs A. Baker and Miss R. Jantunen for their technical assistance and to Dr A. C. S. Crossley for his comments on the work.

Indirect immunofluorescent studies have been used to localize a glycoprotein (gp32) in the amphids of 2nd-stage juveniles (J2) of Meloidogyne incognita parasitic in the roots of tomato plants, male Meloidogyne javanica and some young adult female M. incognita. Immuno-electron microscopic studies showed an apparent absence of gp32 in the amphids of mature adult female M. incognita. Chemoattraction experiments demonstrated that the binding of gp32 antibodies to the amphidial secretions of infective J2 M. incognita partly impaired the ability of the nematodes to locate tomato roots.

We characterized partial sequences of 18S rDNA from sedge warblers infected with a parasite described previously as Hepatozoon kabeeni. Prevalence was 47% in sampled birds. We detected 3 parasite haplotypes in 62 sequenced samples from infected animals. In phylogenetic analyses, 2 of the putative Hepatozoon haplotypes closely resembled Lankesterella minima and L. valsainensis. The third haplotype grouped in a wider clade composed of Caryospora and Eimeria. None of the haplotypes showed resemblance to sequences of Hepatozoon from reptiles and mammals. Molecular detection results were consistent with those from microscopy of stained blood smears, confirming that the primers indeed amplified the parasite sequences. Here we provide evidence that the avian Hepatozoon-like parasites are most likely Lankesterella, supporting the suggestion that the systematic position of avian Hepatozoon-like species needs to be revised.

In total 445 faecal samples were collected from 259 European badgers (Meles meles) in Wytham Woods, Oxfordshire, UK (462080). Microscopical examination revealed infection with 2 species of coccidia Eimeria melis and Isospora melis. From the initial examination of each animal, point prevalence rates of 0·44 and 0·35 were calculated for Eimeria and Isospora respectively. The intensity of infection was significantly greater for Eimeria than Isopora and the distribution of intensities was highly skewed for both species, with a few individuals shedding the majority of oocysts. Incidence and recovery rates for both coccidia species were calculated from longitudinal data collected at 3-monthly intervals from a subset of the adult badger population, and the predicted prevalence rates based on these were similar to the point prevalence rates. This suggests little, if any, parasite-induced mortality in the adult population. In contrast, there was a marked and significant reduction in the point prevalence and intensity of infection with Eimeria from cub to adult badger suggesting a degree of acquired immunity to Eimeria melis on initial exposure and/or that there is significant Eimeria-associated mortality in the cub population. No such relationship was found for Isospora infection. In those adult badgers with co-infections there was a direct relationship between the intensity of Eimeria and Isospora. The taxonomic status of these parasites suggests a heteroxenous life-cycle for I. melis, and direct transmission of E. melis. However, the greater than expected prevalence of co-infection is consistent with a common source of infection, such as communal latrines.

Oocyst wall formation in Eimeria maxima was studied during the macrogamete stage in intestinal cells of the chick and in unsporulated oocysts isolated from faeces. The outer of the 2 membranes bounding the mature macrogamete separated from the surface but remained as a veil surrounding the developing oocyst throughout the whole intracellular process. Wall-forming bodies Type I were initially applied to the limiting membrane of the zygote cytoplasm; a layer of material similar to their contents was then formed around the zygote. As this occurred a new double membrane was formed surrounding the oocyst cytoplasm. The outer wall layer was initially homogenous in appearance but later developed into 2 zones, an outer amorphous region and an inner osmiophilic region. The inner layer of the oocyst wall was formed from the contents of the wallforming bodies Type II which dispersed between the outer wall and the limiting membranes of the oocyst cytoplasm. There was evidence of an additional membrane formed external to the outer wall. The outer membranes were not present around the wall of oocysts passed in the faeces of chicks, but the same wall zonation was evident, although the inner osmiophilic zone of the outer wall layer was markedly thinner in comparison with the same zone seen in the tissues.

Phyllobothrium piriei sp.nov. (Cestoda: Tetraphyllidea) from the intestine of Raja naevus in the North Sea resembles P. dagnallium, P. radioductum and P. britannicum more closely than any other known species of the genus but differs from these mainly in its host, in having distinctly bifid bothridia with faint marginal loculi and about 150 testes in each proglottid. Brief comments are made on the preference shown by the adult for the posterior region of the intestine and on a modification of its bothridium for attachment to up to 12 villi.

This work was supported by generous financial aid from the Natural Environment Research Council. I wish also to record my gratitude to Professor V. C. Wynne-Edwards and Dr P. A. Orkin for reading and suggesting improvements to the manuscript and to the Director and staff Marine Laboratory, Aberdeen, in particular Dr B. B. Rae and Mr Stuart Pirie, after whom this species has been named, for providing live rays. Alasdair McVivar of this department, who has now begun an independent study of the intestine of Raja naevus as a habitat for helminth parasites, kindly provided information for Fig. 9.

Although the mosquito vectors of malaria have an effective immune system capable of encapsulating many foreign particles, they rarely encapsulate malaria parasites in natural populations. A possible reason for this apparent paradox is that infection by malaria reduces the capability of the mosquito to mount an effective immune response. To investigate this possibility, we blood-fed Aedes aegypti mosquitoes on an uninfected chicken or on one infected with Plasmodium gallinaceum, and compared the proportions of the infected and uninfected mosquitoes that melanized a negatively charged Sephadex bead injected into the thorax 1, 2 and 4 days after blood-feeding. About 40% of the uninfected mosquitoes, but less than 25% of the infected ones, melanized the bead. The difference between infected and uninfected mosquitoes was most obvious 1 day after infection (at the parasite's ookinete stage), while the difference diminished during the early oocyst stage (2 days after infection) and disappeared at the later oocyst stage (4 days after infection). These results suggest that the parasite can either actively suppress its vector's immune response or that it modifies the blood of its chicken host in a way that reduces the efficacy of the mosquito's immune system. In either case, the reduction of immunocompetence can have important consequences for malaria control, in particular for the current effort being invested into the genetic manipulation of mosquitoes.

The present material of buffalo–dog origin could not be transmitted to the laboratory animals, such as white rats, white mice, rabbits, golden hamsters, guinea-pigs and chicks.

Secondary development took place only in mice.

The strobilar phase was infective to a cat but did not show sexual differentiation even at 30 days, and worms were further observed to be at a degenerative stage.

The buffalo was highly suited to the perpetuation of the parasite, forming larger cysts and massive production of scoleces; nearly 90% of the cysts were fertile.

The strobila had mostly two segments and not more than three.

The number and distribution of testes were more posterior to the genital pore and were also observed posterior to the vitellaria, occasionally.

The position of the genital pore in the gravid as well as mature segments was both anterior and posterior but with an average of 52.8%.

Uterine sacculations were observed to range from 28 to 54.

Echinococcus material of buffalo–dog origin does not appear to fall into a definite ‘subspecies’ pattern. It is suggested that it be merely recognized as a further ‘mutant’, the existence of which can be accounted for on theoretical grounds.

The authors are thankful to the Director, Indian Veterinary Research Institute, and to Dr H. D. Srivastava, Head of Division of Parasitology, I.V.R.I., for providing the necessary laboratory facilities.

1. Standard conditions are described for preparing suspensions of washed Trypanosoma lewisi and T. equiperdum in modified Ringer–phosphate solution.

2. Oxygen consumption was measured with differential manometers, using microflasks containing 2–5 × 107 trypanosomes in 0·9 ml. of reaction mixture. Measurements of oxygen uptake were carried out at 37° C.

3. T. lewisi respired slowly in the absence of substrate for up to 2 hr. The trypanosomes suffered little damage when stored at 5° C. for 24 hr. without substrate. No oxygen uptake was observed with T. equiperdum in the absence of substrate. The trypanosomes were viable after 24 hr. at 5° C. with glucose or glycerol as substrate, but not in the absence of substrate.

4. With glucose as substrate, the rate of oxygen consumption by T. lewisi increased with the age of infection. This change was more marked with glutamine as substrate.

5. With glucosamine as substrate, the oxygen uptake of T. lewisi was at a slightly lower rate than with glucose. The rate of oxygen uptake was still lower with Na l-glutamic acid, asparagine, aspartic acid, casein hydrolysate, yeast extract and Difco Bacto-peptone. Thirteen other amino-acids had no effect on the motility of the trypanosomes.

6. With glycerol as substrate, the oxygen uptake of T. equiperdum was at a slightly lower rate than with glucose. The rate of oxygen uptake was very low with yeast extract, casein hydrolysate and Difco Bacto-peptone. No oxygen uptake or motility was recorded with glutamine, Na l-glutamic acid, glucosamine, asparagine, aspartic acid, dl-alanine, or Na acetate. Thirteen other amino-acids had no effect on the motility of the trypanosomes.

7. Ammonia was liberated from glutamine by adult and reproductive phase T. lewisi.

This experiment was designed to determine the effects of under-dosing on the frequency of benzimidazole resistant allele in the nematode Teladorsagia circumcincta. Fenbendazole (FBZ) was tested at 1/32, 1/16, 1/8 and 1/4 of the recommended dose for sheep (5 mg/kg body weight). The fraction of the susceptible homozygote (SS), susceptible heterozygote (RS) and resistant homozygote (RR) genotypes were compared among FBZ dose groups to evaluate differences between SS and RS genotype selective advantage. Almost all SS genotype worms were eliminated by 1/4 of the FBZ recommended dose, whereas a significant fraction of the RS genotype worms survived treatment. The selective advantage was 4·5 times higher for the RS genotype. This selective advantage was determined at 1/4 of the manufacturer's recommended dose of FBZ. This value should be taken as an indictor of the selective advantage of RS over the SS genotype when lambs are under-dosed. A computer simulation was used to study the putative spread of anthelmintic resistance over a range of RS selective advantages (2, 4·5 and 10-fold), with two average sizes of individual host worm population (20 or 2000 worms/host) and two initial R allele frequencies (0·1% or 1%). In all situations, the lowest selective advantage of the RS genotype over the SS genotype was sufficient to promote the spread of resistance in susceptible populations. When the RS genotype had no selective advantage over the SS genotype, genetic drift almost always led to the loss of the R allele, except in the largest populations (average size = 2000 worms).

By interactions with other microbes, free-living amoebae play a significant role in microbiology, environmental biology, physiology, cellular interactions, ecology and evolution. Here, we discuss astonishing interactions of bacteria and amoebae, in the light of evolution and functional aspects impacting human health. In favourable environmental conditions, the interaction of Acanthamoeba with non-virulent bacteria results in lysis of the bacteria. However, the interaction with weak-virulent bacteria results in a symbiotic relationship or amoebal lysis may occur. The microbial survival of amoebae in harsh environments, ability to interact with bacteria, and their ability to aid transmission to susceptible hosts is of great concern to human, animal and ecosystem health.

Biological invasions expose parasites to new invasive hosts in addition to their local hosts. However, local parasites are often less successful in infecting and exploiting their new hosts. This may have major consequences for the competitive ability of hosts, and finally on the fate of the parasite-host community. In Burgundy (Eastern France), the acanthocephalan parasite, Pomphorhynchus laevis, infects 2 amphipod species living in sympatry: the native Gammarus pulex and the invasive Gammarus roeseli. While P. laevis affects the behaviour and the immunity of G. pulex, G. roeseli seems unaffected by the infection. In this study, we examined in detail the ability of the parasite to affect the immune system and resource storage of both gammarid species. We found that the infection was associated with a general decrease of the prophenoloxidase activity, haemocyte density, resistance to an artificial bacterial infection and level of sugar reserves in G. pulex, but not in G. roeseli. These results demonstrate a differential ability of P. laevis to exploit its local and its invasive gammarid hosts. Potential mechanisms of these differential physiological alterations and their potential consequences on the coexistence of both gammarid species in sympatry are discussed.

The impact of haematozoan infection on host fitness has received substantial attention since Hamilton and Zuk posited that parasites are important drivers of sexual selection. However, short-term studies testing the assumption that these parasites consistently reduce host fitness in the wild have produced contradictory results. To address this complex issue, we conducted a long-term study examining the relationship between naturally occurring infection with Haemoproteus and Plasmodium, and lifetime reproductive success and survival of Mountain White-crowned Sparrows. Specifically, we tested the hypothesis that birds infected with haematozoan parasites have reduced survival (as determined by overwinter return rates) and reproductive success. Contrary to expectation, there was no relationship between Haemoproteus and Plasmodium infection and reproduction or survival in males, nor was there a relationship between Plasmodium infection and reproduction in females. Interestingly, Haemoproteus-infected females had significantly higher overwinter return rates and these females fledged more than twice as many chicks during their lifetimes as did uninfected females. We discuss the impact of parasitic infections on host fitness in light of these findings and suggest that, in the case of less virulent pathogens, investment in excessive immune defence may decrease lifetime reproduction.

Dermanyssus gallinae (De Geer 1778), commonly named the poultry red mite (PRM), is considered to be the most harmful ectoparasite in poultry farms in Europe. This species feeds on the blood of laying hens, but spends most of its time hidden in cracks and crevices around hen nests. To control PRM populations in poultry houses, chemical pesticides are currently used; however, concern is growing regarding the harmful residues found in eggs and hens, along with the increased resistance of mites against several compounds. Alternatives to synthetic compounds are now being explored, including vaccines, biological control, physical control and semiochemical control based on the chemical ecology of PRM. This review focused on the different volatile organic compounds (VOCs) identified from D. gallinae and other mite species that have been discovered to control them. Pheromones (aggregation pheromone, sex pheromone and alarm pheromone) and kairomones promoting attraction behaviour in D. gallinae and other mite species are presented, while VOCs from essential oils and plant extracts with repellent properties are also explored. Finally, devices using VOCs on PRM in the field are described, with devices that have been tested on other Acari species being mentioned as potential directions for the future control of PRM.