The monogenean Gyrodactylus salaris Malmberg, 1957 is an economically important parasite on Atlantic salmon whereas the morphologically very similar G. thymalli Žitňan, 1960 on grayling is considered harmless. Even molecular markers cannot unambiguously discriminate both species. The nuclear internal transcribed spacer (ITS) sequences are identical in both species, and although mitochondrial cytochrome oxidase I (COI) sequences show substantial variation, no support for monophyly of either species is found. Analysis of nucleotide sequences of the intergenic spacer (IGS) have, however, been interpreted as support for 2 species. Here, IGS and COI sequences from 81 G. salaris and G. thymalli specimens of 39 populations across the species' distribution range were determined. Mitochondrial diversity was not reflected in the nuclear marker. Since various 23 bp IGS repeat types usually differ by just one nucleotide and sequences primarily differ in the number and order of repeat types, alignments may be biased and arbitrary, impeding meaningful phylogenetic analyses. The hypothesis that parasites on rainbow trout represent hybrids of both species is rejected. The presence or absence of particular repeat types is not considered informative. We interpret the IGS data as support for G. salaris and G. thymalli being a single species.

Parasites often induce behavioural changes in their host. However, it is not necessarily easy to determine whether these changes are representative of an adaptation of the parasite (parasite manipulation), an adaptive response of the host or a side-effect of infection. In a solitary parasitoid of Drosophila larvae (Leptopilina boulardi), viral particles (LbFV) modify the host acceptance behaviour of infected females by increasing their tendency to superparasitize. This behavioural alteration allows for the horizontal transmission of the virus within superparasitized Drosophila larvae. To add support for or against the ‘manipulation hypothesis’, we investigated whether other behavioural components of the parasitoid are affected by viral infection, and whether other forms of horizontal transmission exist. Neither the ability of females to locate host kairomones nor their daily rhythm of locomotor activity was affected by viral infection. However, infected females showed a lower rate of locomotor activity, suggesting a physiological cost of infection. The searching paths of females were also unaffected. Males from infected and uninfected lines showed the same ability to locate females'sexual pheromones. Moreover, alternative modes of horizontal transmission (through food consumption and/or contact with the same Drosophila larvae) did not lead to viral contamination of the parasitoid. The overall specificity of behavioural alteration and of viral horizontal transmission is consistent with the hypothesis that the virus manipulates the behaviour of the parasitoid.

Vertically transmitted parasites may have positive, neutral or negative effects on host fitness, and are also predicted to exhibit sex-specific virulence to increase the proportion or fitness of the transmitting sex. We investigated these predictions in a study on the survival and sex ratio of offspring of the amphipod Gammarus roeseli from females infected by the vertically transmitted microsporidia Nosema granulosis. We found, to our knowledge, the first evidence for a positive relationship between N. granulosis infection and host survival. Infection was associated with sex ratio distortion, not by male-killing, but probably by parasite-induced feminization of putative G. roeseli males. This microsporidia also feminizes another amphipod host, Gammarus duebeni, which is phylogenetically and biogeographically distant from G. roeseli. Our study suggests that the reproductive system of gammarids is easily exploited by these vertically-transmitted parasites, although the effects of infections on host fitness may depend on specific host-parasite species interactions.

Of the four species of Bulinus found on Madagascar, three species: B. obtusispira, B. liratus and B. bavayi are endemic while the fourth, B. forskalii, is probably a recent introduction from the African mainland. The evolutionary relationships of these species with Bulinus species from Africa were studied by phylogenetic analysis of DNA sequence variation at two mitochondrial loci: cytochrome oxidase subunit I (COI) and large ribosomal subunit (LSU) or 16S. The observed levels of nucleotide divergence within Bulinus were substantial but may underestimate the true levels as there was evidence of ‘saturation' of transitional substitutions at both loci. A putative secondary structure model for the sequenced segment of the 16S was developed. Subsequent phylogenetic analysis using transversional changes only for both loci, showed that there were contrasting levels of divergence within the four species groups. B. obtusispira was consistently placed within the B. africanus group, appearing ancestral to this group and was closest to the basal node within Bulinus. Together with B. bavayi, the two species appear to have been isolated on Madagascar for a long time, contrasting with both B. liratus and B. forskalii that appear more recent colonisers; however, estimate of exact times of divergence is problematic. A PCR-RFLP assay was developed to enable identification and discrimination of B. obtusispira and B. liratus using discriminatory variation within the COI. To enable population genetic analysis within B. obtusispira, microsatellite markers were developed using an enrichment method and 8 primer pairs are reported. Laboratory infection experiments using Madasgacan S. haematobium from the Mahabo area showed that certain populations of B. obtusispira, B. liratus and B. bavayi were compatible.

Certain species of the protozoan genus Besnoitia cause clinical disease in livestock and wildlife. In the present paper a new species, Besnoitia neotomofelis is described from the southern planes woodrat (Neotoma micropus). The parasite was detected by bioassay of woodrat tissues in outbred Swiss Webster mice in an attempt to isolate Toxoplasma gondii. Initially, the organism was misdiagnosed as T. gondii because it was highly pathogenic for mice and its tachyzoites resembled T. gondii tachyzoites. Further studies revealed that it differed structurally and biologically from T. gondii. Tachyzoites were successfully cultivated and maintained in vitro in bovine monocytes and African green monkey kidney cells, and in vivo in mice. Non-dividing, uninucleate tachyzoites were approximately 1×5 μm in size. Longitudinally-cut bradyzoites in tissue sections measured 1·5–1·6×7·7–9·3 μm. Tissue cysts were microscopic, up to 210 μm long, and were infective orally to mice. Cats fed tissue cysts shed unsporulated 13×14 μm sized oocysts. All mice inoculated with B. neotomofelis died of acute besnoitiosis, irrespective of the dose, and Norwegian rats became infected but remained asymptomatic. Entero-epithelial stages (schizonts, gamonts) were found in cats fed tissue cysts. Large (up to 40×50 μm) first-generation schizonts developed in the lamina propria of the small intestine of cats. A second generation of small sized (8 μm) schizonts containing 4–8 merozoites was detected in enterocytes of the small intestine. Gamonts and oocysts were seen in goblet cells of the small intestinal epithelium. Tachyzoites were present in mesenteric lymph nodes of cats. Phylogenetic analysis indicated that B. neotomofelis was related to other Besnoitia species from rodents, rabbits, and opossums. Besnoitia neotomofelis is distinct from the 3 other species of Besnoitia, B. wallacei, B. darlingi and B. oryctofelisi that utilize cats as a definitive host.

This cross-sectional study determined the prevalence and distribution of schistosome and soil-transmitted helminth (STH) infections among 1,308 children aged 10–18 years in 34 primary schools in 8 informal urban settlements in Kisumu City, western Kenya. Stool samples were collected and examined for eggs of Schistosoma mansoni and STH (Hookworms, Ascaris lumbricoides and Trichuris trichiura) using the Kato-Katz technique. Haematuria was used as a proxy indicator of urinary schistosomiasis. Schools and water bodies were mapped using a geographical information system. Overall, 34% of children were infected with one or more helminth species whereas 16·2% of children were infected with one or more STH species. Schools in closest proximity to Lake Victoria and River Nyamasaria had the highest S. mansoni prevalence while schools with STH were more homogenously distributed. Mean school prevalence of S. mansoni infection was 21% (range=0–69·7%), S. haematobium 3·6% (range=0–12%), hookworms 6·1% (range=0–20%), A. lumbricoides 4·9% (range=0–18·4%), and T. trichiura 7·7% (range=0–18·6%). Helminth-related morbidities were not associated with infection. Our study demonstrates that schistosomiasis and STH are important health priorities among schools in informal settlements of Kisumu City, and highlights the need for routine deworming in similar settings.

An examination with light and electron microscopes has been made of the cell type which is considered by some authors to be a sporozoan parasite, Rhabdospora thelohani, and by others to be a tissue cell of fishes and amphibians.

The cell type was found in the minnow Phoxinus phoxinus in many tissue sites, including olfactory and pharygneal epithelia, kidney, eyes and brain. In some specimens it could not be found in any tissue.

The cell has a thick fibrous wall, a basal nucleus, an apical mitochondrial zone, a Golgi body, and from 8 to 50 characteristic refractile rodlets.

The rodlets give a negative reaction to the Feulgen test for nuclear material, but are positive to the periodic acid–Schiff stain, a test for polysaccharide. Glycogen does not appear to be present in the rodlets.

Under certain circumstances the rods rupture the cell wall and may protrude in the apical region of the cell.

The structures described are discussed with the evidence for and against a parasitic interpretation of the cell. It is concluded that Rhabdospora is a protozoan parasite, whose exact functional and taxonomic status is as yet unknown.

I am grateful to Professor G. E. H. Foxon and to Professor P. C. C. Garnham for their encouragement, advice, and helpful criticism of the manuscript. I am further indebted to the National Spastics Society for the use of the electron microscope housed in Guy&s Hospital Medical School, and to Mr I. D. Bradbrook for his assistance with the histological techniques.

1. Extracts made from whole nematode worms, Litomosoides carinii, and from Ascaris ‘heads’ and body wall tissue, contain a substance similar to acetylcholine.

2. In the case of extracts from Litomosoides whole worms, this substance was shown to be probably acetylcholine itself.

3. Tissue from the anterior end of Ascaris (including the nerve ring), contains about 15 times as much acetylcholine as the body wall preparations; i.e. 0·39μg./g. as compared with 0·025μg/g. of wet weight.

4. There appeared to be rather more acetylcholine present in the Litomosoides males than in the females; 0·92μg./g. as compared with 0·63μg./g. of wet weight.

5. The microfilariae of the filariid nematode of the dog, Dirofilaria repens, contain as much as 2·4;μg./g. of acetylcholine. In this respect it resembles some other motile parasites of the blood, such as trypanosomes.

A contingency table approach was used to explore the influence of location, host species and time on the genetic composition of a Trypanosoma brucei population in Lambwe Valley, Kenya. Significant differences in zymodeme frequencies were noticed over comparatively short geographical distances suggesting that transmission of T. brucei is somewhat localized. A significant association was observed between zymodeme and the mammalian host from which T. brucei was derived. The association was consistent in different localities in Lambwe valley and remained stable for at least 32 months. These observations indicate that zymodemes are adapted to different host species and that genetic exchange has not disrupted host associations over this time-scale. A major change in the composition of the T. brucei population during a sleeping sickness outbreak in 1980 was confirmed. But while new zymodemes emerged, a decline in overall diversity was noted during times of high sleeping sickness incidence. The results can be explained by selection of T. brucei zymodemes for particular transmission cycles. Although it is not necessary to invoke genetic exchange, sex may help T. brucei to adapt to changes in selection pressures. Such a hypothesis helps to explain why T. brucei appears largely clonal in the short term, even though population studies indicate that sex is responsible for much genetic diversity in the long term. It also explains why neighbouring populations of T. brucei are composed of a different range of zymodemes formed from the same alleles. Such a view implies that genetic exchange has an important role in the microevolution of T. brucei populations.

The life-history of I. pacificus when fed on guinea-pigs under laboratory conditions is described. Feeding of the female proceeds for 10–11 days. Between 120 and 191 mg of concentrated food is found in the female's gut on detaching from this host, and this is directly related to the unfed body weight of the tick.

The egg-laying pattern, in general, shows an initial peak, after which the number of eggs tails off, the whole process lasting approximately 33–40 days. The number of eggs laid by females varies between 790 and 1300. The preoviposition period is about 11–16 days.

Larvae hatch in about 53–55 days, and their feeding time ranges from 4 to 9 days, although in the present experiments larvae emerging from early laid eggs took about 10 days to complete engorgements.

Moulting of larvae to nymphs at 90% R.H. took 37–38 days, and nymphs fed for 7–11 days, with most becoming replete on the ninth.

Very few females emerged from these nymphs, and those that did emerged in just over 30 days. The reasons for this are not clear. No diapause appeared to be necessary to complete the life-cycle, and with due allowance for hardening of the mouthparts the life-history of one generation was about 7 months. Females died within 3 or 4 months if hosts were not available.

From the literature and from hitherto unpublished records some 55 vertebrates have been implicated as hosts to I. pacificus; these hosts include a wide range of mammals, six species of birds and possibly nine species of lizards.

The species has been listed from British Columbia and from five western states of U.S.A. The need for more intensive ecological work on this species is emphasized.

We are indebted to Glen M. Kohls, J. D. Gregson and D. E. Johnstone for permission to use unpublished records from their respective institutions. One of us (D.R.A) is also indebted to the Wellcome Trustees for financial assistance to visit Canada during the preliminary stages of the work.

Trypanosomes are consistently present in the milk of lactating mice with primary phase T. cruzi infections (Peru strain). The infection is transmitted to the young, although rarely, mainly due to the difficulty of establishing infection via the oral route. Lactating females which had recovered from the primary phase of T. cruzi infection consistently transferred antibody to their young during suckling. The antibody in the serum of the young and milk of the mother was detectable by the indirect fluorescent antibody technique. In the case of the Peru strain the presence of antibody transferred to the young in this way correlated with very good protection against T. cruzi. Antibody transferred to the young was no longer detectable 6 weeks after birth. The indirect fluorescent antibody technique using bloodform antigen proved specific and highly sensitive throughout, giving prolonged and comparably high titres in mice infected with chronic and acute strains of T. cruzi.

The life-cycle of canine Isospora bigemina was studied in dogs, cattle, cats and mice. Under experimental conditions dogs served as both definitive and intermediate hosts. Unsporulated oocysts (11 × 12 μm) were shed in the faeces and sporulated outside the host within 12 h at 30–37 °C and 36 to 48 h at 23 °C. Sporulated oocysts measured 12 × 13 μm and contained 2 sporocysts which in turn contained 4 sporozoites. Sporocysts averaged 6 × 9 μm and sporozoites averaged 2 × 6 μm. Although no stages were found microscopically in tissues or in faeces of 23 dogs orally inoculated with sporulated oocysts, the oocysts were infectious because 5 of 14 dogs that ingested tissues of dogs fed oocysts, shed oocysts. Prepatent periods were 7 to 15 days; patency was 1–3 days. No dogs became ill. In the naturally infected dog and one experimentally infected dog, schizonts were found in epithelial cells, distal to the host cell nucleus at the tips of villi throughout the small intestine. Schizonts were 5–7 μm and contained 3–12 merozoites. In the naturally infected dog, gametes were found in the same location. Male gametocytes were 6–8 μm and contained 6–12 microgametes, and female gametes were 7–8 μm.

Sporulated I. bigemina oocysts from a naturally infected dog were not infectious to cattle, cats or mice. Structurally identical oocysts were shed by 2 dogs after ingesting hearts and diaphragm from naturally infected cattle; these oocysts were also not infectious to cattle. Although dogs acted as both intermediate and definitive hosts under experimentation, this is not likely to occur in nature. A canine-bovine-canine cycle would appear to be the natural mode of infection but I. bigemina was not infectious to cattle under experimental conditions.

Adult Oesophagostomum columbianum populations were larger in sheep fed low-protein diets than in adequately fed animals. Diet did not influence the numbers of larva which became established in sheep. Sheep fed high-protein diets eliminated more worms and were more immunologically competent than poorly fed animals. More encapsulated larvae, showing arrested development, were found in adequately fed sheep than in those fed low-protein diets. Adult O. columbianum produced eggs at an earlier time after infection in poorly fed sheep than worms in well fed sheep. More eggs/female worm were produced in sheep on a low-protein diet compared with the number of eggs produced in well fed hosts over the last week of the infection. The effects of immunity on the behaviour of the worm in both host diet groups is discussed.

There was a greater cellular proliferation in the intestines of infected adequately fed sheep than in infected animals on low-protein diets. These changes were most pronounced in the large intestine where the adult parasites were found. The macrophage-lymphocyte series of cells underwent hyperplasia in well fed animals but these changes were reduced, particularly among the plasma cells, in sheep fed low-protein diets. Increased mucin and mast cell counts were observed in sheep on high-protein, but not in hosts on low-protein diets: the intestinal populations of eosinophil and globule leucocytes were also reduced in poorly fed sheep.

The relationship of these various cellular reactions and their effect on the protective immunity of sheep to O. columbianum is discussed. It was concluded that the increased susceptibility of protein deprived sheep to O. columbianum infections was associated with malfunctions of the innate immunity of the gut, involving decreased peristalsis and failure of the mucin cell response, and with reduction of the adaptive immune response which was reflected by impoverished lymphocyte and plasma cell reactions, and possibly with poor cooperation between sensitized lymphocytes, antibodies and the mast cell-granulocyte effector mechanisms in protective immunity.

Infection of ewes by Toxoplasma gondii may induce abortions, thus it has consequences for sheep production. Colima, Western State of Mexico, has favourable climatic conditions for transmission and both domestic and wild cats live there. The aim of this study was to determine the frequency of specific antibodies in sheep from the coast, a mountain, and a hill of Colima. Serum samples from 351 sheep were tested by a previously standardized indirect ELISA. The frequency of infection was estimated and the farm location and flock size, as well as the animals' age and sex were analysed as risk factors for toxoplasmosis. The frequency of antibodies depended on the altitude, being higher at sea level than at 1200 metres above sea level (OR=3·77, 95% CI=1·79–7·94, P<0·0001), and the size of the flock, being higher in the large ones (OR=2·23, 95% CI=1·35–3·71, P=0·002). Older animals were more frequently positive and with a stronger response than young ones (OR=1·77, 95% CI=1·07–2·93, P=0·016). No differences were observed between male and female sheep. In conclusion, toxoplasmosis is present in sheep of Colima with variations related to altitude, flock size and age.

Duodenal mucosae of uninfected lambs and lambs inoculated at least 16 days earlier with 85000–140000 Trichostrongylus colubriformis larvae were examined with the scanning electron microscope. Normal duodenum had tall spatulate villi with surface folds upon which goblet cells and a regular pattern of hexagonal enterocytes were seen. Micro villi on normal enterocytes were closely packed and imparted a granular surface texture. In heavily infected areas of gut the villi were atrophic, the mucosa sometimes being composed of irregular masses and ridges, with crypt mouths, often surrounded by collars of cells, opening into the surface. More severely affected mucosae were flat, with protuberant collars of cells surrounding crypt mouths. There were rounded bodies, interpreted as sloughing enterocytes, or inflammatory cells, on the mucosal surface. Apices of enterocytes were domed and microvilli were sparse and irregular. Micro-organisms were numerous on cell surfaces. Nematodes were located in sinuous thin-walled tunnels in the epithelium. The mucosal microtopography is compared with that of coeliac disease of humans, nippostrongylosis in rats and with villus atrophy in pigs.

Cyclospora cayetanensis is a coccidian parasite associated with diarrheal illness. In the USA, foodborne outbreaks of cyclosporiasis have been documented almost every year since the mid-1990s. The typical approach used to identify this parasite in human stools is an examination of acid-fast-stained smears under bright-field microscopy. UV fluorescence microscopy of wet mounts is more sensitive and specific than acid-fast staining but requires a fluorescence microscope with a special filter not commonly available in diagnostic laboratories. In this study, we evaluated a new DNA extraction method based on the Universal Nucleic Acid Extraction (UNEX) buffer and compared the performances of four published real-time polymerase chain reaction (PCR) assays for the specific detection of C. cayetanensis in stool. The UNEX-based method had an improved capability to recover DNA from oocysts compared with the FastDNA stool extraction method. The best-performing real-time PCR assay was a C. cayetanensis-specific TaqMan PCR that targets the 18S ribosomal RNA gene. This new testing algorithm should be useful for detection of C. cayetanensis in human stool samples.

The avian schistosomatid Trichobilharzia ocellata plays an important role as causative agent of cercarial dermatitis of humans in Europe. In order to improve the taxonomic knowledge on this parasite, studies of miracidial chemo-orientation as well as experimental infections of different snail species were conducted using strains of T. ocellata and T. franki. Both schistosomes exhibited a high intermediate host specificity. The miracidia clearly preferred the SCW (snail-conditioned water) of the respective natural intermediate host to SCW of other sympatric snail species. T. ocellata proved to be capable of infecting Lymnaea stagnalis and Stagnicola palustris, but could not develop in Radix ovata or R. auricularia. T. franki established an infection in specimens of R. auricularia and R. ovata, but not in L. stagnalis or S. palustris. The results imply that the intermediate host spectrum of T. ocellata is limited to L. stagnalis and S. palustris. Findings of T. ocellata (or Cercaria ocellata) that originated from snails of the genus Radix are likely to have actually belonged to species such as T. franki or T. regenti. The assumption that T. szidati is synonymous to T. ocellata is also discussed.

Unsporulated oocysts were recovered from the allantoic cavity of 5 chicken embryos inoculated with a single sporozoite of Eimeria tenella; some of these oocysts sporulated. Both macrogametes and microgametes are required for the formation of viable oocysts. The results conclusively demonstrate the bisexual nature of the sporozoite, suggest that sexual differentiation is influenced by environmental factors, and show that true clones of Eimeria can be established from individual sporozoites.

The effect of sainfoin (Onobrychis viciifolia) hay consumption on the pathophysiology and local cellular responses of growing lambs during infection with Trichostrongylus colubriformis was investigated. Thirty-two lambs, 16 weeks of age, were allocated to 1 of 4 treatment groups (n=8) that were offered either grass (G) or sainfoin (S) hay while concurrently either infected (+), or not (−) with 12 000 L3 T. colubriformis larvae per week for 6 weeks. Liveweight gains were affected by diet (P=0·002) and reduced by infection (P<0·005). Faecal egg count was reduced in S+ compared to G+from days 35 to 42 (P=0·001); however, total egg output, worm burdens at day 42 and worm fecundity were similar between diets (P>0·05). Feeding sainfoin appeared to enhance immune cell development with tissue eosinophils, mast cells and pan T cells present in greater concentrations in S+ than in G+ animals. However, further studies are required to determine if the enhanced immune cell development is a consequence of a greater nutrient supply or a direct influence of sainfoin metabolites on local inflammatory responses to the gastrointestinal nematode T. colubriformis.

Hosts and parasites interact with each other in a variety of ways, and this diversity of interactions is reflected in the networks they form. To test for differences in interaction patterns of ecto- and endoparasites we analysed subnetworks formed by each kind of parasites and their host fish species in fish–parasite networks for 22 localities. We assessed the proportion of parasite species per host species, the relationship between parasite fauna composition and host taxonomy, connectance, nestedness and modularity of each subnetwork (n = 44). Furthermore, we evaluated the similarity in host species composition among modules in ecto- and endoparasite subnetworks. We found several differences between subnetworks of fish ecto- and endoparasites. The association with a higher number of host species observed among endoparasites resulted in higher connectance and nestedness, and lower values of modularity in their subnetworks than in those of ectoparasites. Taxonomically related host species tended to share ecto- or endoparasites with the same interaction intensity, but the species composition of hosts tended to differ between modules formed by ecto- and endoparasites. Our results suggest that different evolutionary and ecological processes are responsible for organizing the networks formed by ecto- and endoparasites and fish.