Earlier schemes of classification of the family Heterophyidae have been based in large part on such features as shape of body, presence of oral spines, number and position of testes, and distribution of vitellaria (Witenberg, 1929; Ciurea, 1933; Mueller & Van Cleave, 1932). Price (1940a) was the first to make extensive use of features of the ventrogenital complex (ventral sucker, gonotyl, genital pore, terminal male duct) and excretory bladder, and produced the first reasonable classification of both the family Heterophyidae and the superfamily Opisthorchioidea. In despite of the obvious significance of the rationale of Price's approach, later authors (Morozov, 1952, 1955; Yamaguti, 1958) have largely ignored the ventrogenital complex and recently discovered life-history data, and have used much the same sorts of features as earlier authors.

Proliferative kidney disease (PKD) is a parasitic infection of salmonid fish characterized by an apparently abnormal immune response to the presence of the myxozoan parasite, Tetracapsuloides bryosalmonae. In order to examine the nature of the immune response at the molecular level, the expression of a range of immune regulatory genes, including cytokines and cyclooxygenase (COX)-2 was examined in naive unexposed fish and in naive fish exposed to parasite-infected water at three points during the course of a natural outbreak of PKD. Since fish with advanced PKD pathology generally exhibit increased susceptibility to secondary infections which is typical of stress/cortisol-mediated immune suppression, a further aim of this work was to examine in vitro the influence of the glucocorticoid cortisol on the bacterial lipopolysaccharide (LPS)-induced expression of the trout cytokine genes studied. Two weeks after the initial sampling, naive exposed fish showed a specific profile of up-regulated tumor necrosis factor (TNF)-α2, COX-2 and, to a lesser extent, transforming growth factor (TGF)-β1 expression. As the disease pathology increased, TNF-α2 and COX-2 expression returned to normal levels. Stress levels of cortisol suppressed the LPS inducibility of pro-inflammatory cytokine genes, although TGF-β1 and TNF-α2 appeared to be refractory. These data demonstrate that specific immune responses at the molecular level are affected during PKD infection, with the cortisol suppression of cytokine expression in vitro providing a possible link to PKD-mediated cytokine down-regulation and immune suppression.

RNA interference (RNAi) on parasitic nematodes has been described as successful and useful for the identification of novel drug and vaccine candidates. In this study we have evaluated this technology on the cattle parasite Ostertagia ostertagi. Eight different genes were targeted in L1 and L3 O. ostertagi larvae, by electroporation and soaking in dsRNA respectively. Down-regulation of target transcript levels was evaluated by semi-quantitative reverse transcriptase (RT) PCR. In L3 larvae, variable decreases in mRNA levels were observed for 5 genes, ranging from a complete knock down (tropomyosin, β-tubulin) to a minor decrease (ATPsynthase, superoxide dismutase, polyprotein allergen). However, repeated experiments indicated that effects were sometimes difficult to reproduce. RNAi for ubiquitin, a transthyretin-like protein and a 17 kDa excretion secretion (ES) protein never resulted in a knock down of the transcript. The mRNA levels of 7 non-target genes showed no difference between larvae soaked in C. elegans control dsRNA versus O. ostertagi tropomyosin dsRNA, supporting that the observed reductions are specific for the target gene. Electroporation of L1 larvae proved to be less effective. Reductions in mRNA levels were only noticed for 2 genes and were not reproducible. In conclusion, the results indicate that the RNAi pathway is probably present in O. ostertagi but that the current RNAi techniques can not be used as a reliable screening method.

Schistosomiasis remains a serious world-wide public health problem with a still unfulfilled need for routine cost-effective methods of diagnosis. Such methods are required not only for people in endemic areas, but increasingly for tourists who may have become infected during visits to such places. This article reviews the wide range of immunoassays and antigenic preparations that have been shown to have potential for diagnosis of schistosomiasis by the indirect method of antibody detection. Antigens in native form derived from cercariae, adult worms and eggs are considered, as well as schistosome antigens produced by recombinant DNA technology and the schistosome cross-reactive antigen, keyhole limpet haemocyanin (KLH). Respective advantages and disadvantages of antibody detection, circulating antigen detection and parasitological methods of diagnosis are analysed. It is suggested that due to the relative insensitivity of both parasitology and antigen detection, antibody detection methods could find increasing use in situations of low infection intensity.

Susceptibility to oxygen and properties relative to oxygen metabolism were compared in metronidazole-resistant and susceptible strains of Trichomonas vaginalis. The study involved clinical isolates displaying the aerobic type of resistance, as well as resistant strains developed in vitro, both with aerobic (MR-3) and anaerobic (MR-5, MR-100) resistance. Elevated sensitivity to oxygen of the resistant clinical isolates was observed. Progressive increase of susceptibility to oxygen also accompanied in vitro development of resistance. No correlation was found between the activity of NADH oxidase and aerobic resistance, while the in vitro derivative with fully developed anaerobic resistance (MR-100) showed about 50% decrease of NADH oxidase activity. The superoxide dismutase (SOD) activity was elevated in both resistant clinical isolates and in in vitro-developed resistant strains. The changes in levels of ferredoxin were insufficient to support ferredoxin deficiency as a cause of aerobic metronidazole resistance. Western blot analysis and electron paramagnetic resonance spectroscopy of purified hydrogenosomes showed that ferredoxin is expressed in aerobically resistant strains and has intact iron-sulfur clusters. Down-regulation of ferredoxin was demonstrated only in the late phase of development of the anaerobic resistance (MR-100). The results support a link between aerobic resistance and defective oxygen scavenging. The increased levels of intracellular oxygen, beneficial to resistant parasites when they interact with the drug, may have adverse effects on their fitness as shown by their increased sensitivity to oxidative stress.

The surface morphology of schistosomula extracted from the skin, lungs and hepatic portal system (hps) of mice was investigated from Days 0 to 18 post-infection. Skin schistosomula and newly arrived schistosomula from the hps were of similar dimensions but were morphologically distinct. Lung schistosomula were considerably elongated with an estimated 53% increase in surface area compared to skin schistosomula. The pitted tegument of lung schistosomula was formed into ridges and troughs. These were compressed together in contracted individuals recovered from the hps on Day 10. The annular ridges were cross-linked by longitudinal septae which possibly prevent further elongation of the body. A regression of the spines between the mouth and the ventral sucker was observed in Day 2 skin schistosomula. In lung schistosomula only the spines at the anterior and posterior of the body remained. New spines were formed after the schistosomula reached the hps. It is suggested that the spines in the mid-region of the body are selectively disassembled and that their loss facilitates migration along the lumina of capillaries with the residual spines acting as anterior and posterior anchors. The mouth opening was enlarged in schistosomula from the hps recovered from Day 10 onwards. Skin schistosomula lost the cercarial apical tegumentary ridges between 24 and 48 h after penetration but a spineless protrusible area remained. After arrival in the hps this area became integrated into the anterior surface as the oral sucker developed around the sub-terminal mouth. The cercarial ciliated papillae were lost on penetration. The migrating schistosomulum had few visible sensory papillae but following arrival in the hps new papillae were observed.

1. Cerfontaine's (1896) description of the egg is confirmed. The eggs are not retained on the gills during development.

2. Maturation of the oocyte takes place after egg formation. The haploid number of chromosomes is 9.

3. The onset of embryonic development is related to the time of egg formation or sperm entry. Cleavage is total and slightly unequal, leading to the formation of a morula in which there are no germ layers. The larval gut is formed by epiboly round a mass of yolk which becomes incorporated in mid-body. Larval organs and a partially ciliated epithelium are differentiated in situ.

4. During embryonic development yolk is broken down and absorbed, some being incorporated in the body.

5. Hatching does not seem to result from any special stimulus. Mechanical pressure by the larva forces off the operculum, but secretions may weaken the suture.

6. The free-swimming larva is gyrodactyloid with a winged haptor bearing four pairs of lateral, one pair of postero-lateral and one pair of median hooks. Four pairs of flame cells are present and one pair of longitudinal, lateral, excretory ducts, which open antero-laterally. The larva swims by ciliary action and does not respond to jarring, light and shade stimuli, or, when vigorous, to the presence of the excised gill of the host. Tired larvae are doubtfully attracted by host gill tissue. Larval haptorial type is thought to be determined phylogenetically rather than as a result of adaptation to the type of host tissue to which the larva attaches itself.

7. The larva attaches itself to the host's gill and sloughs off the ciliated coat. Growth follows with the development of the adult clamps in pairs, starting posteriorly, each pair replacing a pair of lateral, larval hooks. Primordia of the reproductive organs are laid down in the larval stage, but are only finally elaborated when the adult form has been attained. Maturity is not reached at a definite size and the ovary and testes mature simultaneously.

8. Feeding is at least partly on the host's blood.

9. The longest time which may elapse between formation and laying of an egg is about 5 hr. The average time taken for embryonic development is 18·3 days at 14·25° C., but temperature affects the developmental rate. Ciliated larvae survive about 24 hr. in the absence of a host. The second larval stage is reached between 5 and 13 days after hatching, the third larval stage after more than 38 days. The immature stage is thought to be reached within 3 months and the adult stage within 6 months.

10. Reproduction occurs throughout the year. The animal is functionally hermaphrodite at all times once maturity is reached.

11. A certain degree of osmo-regulation is possible in adults, but this does not extend to ciliated larvae.

12. Adults rest along a gill filament on its inner face and do not move about. Larvae are more ready to move.

13. The average number of flukes per host is 2·24. The 1st and 2nd pairs of gills carry three times as many flukes as the 3rd and 4th pairs.

14. The host suffers no apparent harm from the presence of the flukes.

15. Premunition of Gadus virens against Diclidophora denticulata does not occur. Age resistance is probably developed to the extent that larvae cannot establish themselves on older fish and is probably due to mechanical difficulties and changed habits of the host. The ability of this fluke to parasitize species of Gadus does not extend to G. callarias.

16. D. denticulata is recorded for the first time from St Andrews Bay and the Firth of Forth.

Among flatworms with parasitic and commensal modes of existence, parthenogenesis and asexual multiplication appear to be largely confined to the Digenea and Cestoda, the only parasitic platyhelminths that routinely utilize indirect life-cycles. Parthenogenesis is apparently restricted to a minority of adult digeneans and cestodes inhabiting their final hosts, and a survey is made of the particular modes of parthenogenesis (i.e. apomictic, automictic and generative) which are employed by such adults. Asexual (amictic) multiplication, in the form of fissioning, is demonstrated by young adults of the cyclophyllidean cestode, Mesoces-toides corti, but is otherwise not exhibited by adult cestodes or digeneans, other than in the perplexing phenomenon of proglottid formation in polyzoic tapeworms. Secondary multiplication is of ubiquitous occurrence in digenean life-cycles in the form of the proliferation which takes place within sporocysts and rediae (germinal sacs) located in the first intermediate host. The controversy concerning the nature of this multiplication is reconsidered in the context of recent findings which have centred on cellular aspects. On the basis of present evidence germinal sac multiplication should be regarded as an asexual rather than a parthen-ogenetic process. The cestode asexual multiplication which occurs in intermediate hosts is a function of the metacestode stage of development. Metacestode proliferation is only known from about 20 species and 6 families of polyzoic cestodes with approximately half the described instances occurring in the family Taeniidae. The organization of these proliferative metacestodes, findings concerning their totipotent stem cells and the ontogeny of buds and new scolices are all reviewed. Finally, the capacity for population expansion of multiplicative larval digeneans and metacestodes are compared, while the ecological roles and the genetical consequences of both parthenogenesis and amictic multiplication in the two taxa are also examined.

When cercariae of Schistosoma mansoni and of S. japonicum were irradiated with various levels of u.v. light at 254 nm, their development to perfusable worms was reduced to below 1% at about 200 μW min cm−2. Cercariae attenuated with about 300 μW min cm−2 induced partial resistance against an homologous challenge infection in mice. No differences were observed between the two schistosome species when the same treatment was given to the cercariae. Thus the same u.v. dose can confer immunizing ability to cercariae of both S. mansoni and S. japonicum.

The maintenance of Theileria parva parva infection in an endemic area of Kenya on the shore of Lake Victoria was studied in the field and laboratory. High prevalences of antibodies against T. parva and T. mutans and intra-erythrocytic piroplasms were detected in local zebu (Bos indicus) cattle. The mean infection rate of Theileria parasites in the tick, Rhipicephalus appendiculatus, in field collections was 1·1 %. Most of the infection was attributed to T. parva parva by application of field ticks to susceptible cattle. Five cattle, all about 1·5 years old, were purchased from local owners and transported to the laboratory. All five had oscillating antibody titres against T. parva and T. mutans and had patent theilerial infections during the subsequent 13 months. Uninfected R. appendiculatus nymphs were applied to cattle at 0, 3, 6, 9 and 13 months after transport to Muguga, and 18 out of 23 batches transmitted T. parva parva infection to cattle when 100 resultant R. appendiculatus adults were applied. Infection rates in the tick batches were usually low, with 1 salivary gland acinus infected/tick. Hence, a frequent carrier state of naturally infected cattle has been demonstrated for T. parva parva for the first time, and it is likely that this carrier state is of great importance in maintenance of T. parva parva infection in the field.

Parasite modification of host behaviour influences a number of critical responses, but little is known about the effects on host spatial abilities. This study examined the effects of infection with the intestinal trichostrongylid nematode, Heligmosomoides polygyrus, on spatial water maze learning by male laboratory mice, Mus musculus. In this task individual mice had to learn the spatial location of a submerged hidden platform using extramaze visual cues. Determinations of spatial performance were made on day 19 post-infection with mice that had been administered either 50 or 200 infective larvae of H. polygyrus. The infected mice displayed over 1 day of testing (6 blocks of 4 trials) significantly poorer acquisition and retention of the water maze task than either sham-infected or control mice, with mice that had received 200 infective larvae displaying significantly poorer spatial performance than individuals receiving 50 larvae. The decrease in spatial learning occurred in the absence of either any symptoms of illness and malaise, or any evident motor, visual and motivational impairments. It is suggested that in this single host system the parasitic infection-induced decrease in spatial learning arises as a side-effect of the host's immunological and neuromodulatory responses and represents a fitness cost of response to infection.

Stage-specific molecular and morphogenic markers were used to follow the kinetics of appearance, number, and position of metacyclic promastigotes developing during the course of L. major infection in a natural vector, Phlebotomus papatasi. Expression of surface lipophosphoglycan (LPG) on transformed promastigotes was delayed until the appearance of nectomonad forms on day 3, and continued to be abundantly expressed by all promastigotes thereafter. An epitope associate with arabinose substitution of LPG side-chain oligosaccharides, identified by its differential expression by metacyclics in vitro, was detected on the surface of a low proportion of midgut promastigotes beginning on day 5, and on up to 60% of promatigotes on days 10 and 15. In contrast 100% of the parasites egested from the mouthparts during forced feeding of 15 day infected flies stained strongly for this epitope. At each time-point, the surface expression of the modified LPG was restricted to morphologically distinguished metacyclic forms. Ultrastructural study of the metacyclic surface revealed an approximate 2-fold increase in the thickness of the surface coat compared to nectomonad forms, suggesting elongation of LPG as occurs during metacyclogenesis in vitro. A metacyclic-associated transcript (MAT-1), another marker identified by its differential expression in vitro, also showed selective expression by promastigotes in the fly, and was used in in situ hybridization studies to demonstrate the positioning of metacyclics in the anterior gut.

A comparison was made between the early events (a sexual stages) in the life-cycle of Eimeria tenella in specifically immunized and control chickens. Particular attention was paid to the quantitative aspects and to the transport of sporozoites within intra-epithelial lymphocytes (IEL) from the enterocytes of the surface epithelium to the enterocytes of the crypts. There was a moderate decrease in the number of parasites initially seen in the mucosa of the immune birds, suggesting that some of the effects of immunity are exerted before penetration of the surface enterocytes, but the reduction in the numbers of developing parasites was more marked. This latter effect was due, at least partly, to failure of transfer of sporozoites from IELs to crypt enterocytes. These findings are discussed in relation to the efficacy of the immunity induced.

Fourteen 9-month-old zebu calves were immunized with 10000 irradiated Schistosoma bovis schistosomula given in 1–3 intramuscular or subcutaneous doses, and 4 more calves were immunized with 10000 irradiated cercariae administered percutaneously in a single dose. Eight weeks after the beginning of the experiment these calves, together with four non-immunized controls were challenged percutaneously with 10000 normal S. bovis cercariae/calf. Comparative clinical, parasitological, pathological and pathophysiological observations subsequently revealed significant differences between the vaccinated and non-vaccinated calves. The vaccinated calves showed significantly higher growth rates, and a superior body composition as indicated by their lower total body water content. The beneficial effects of vaccination were also shown by significantly lower faecal egg outputs in the vaccinated calves and by their lower tissue egg and adult worm counts. The reduced tissue egg counts were also reflected in the milder histopathological changes seen in the vaccinated calves. The vaccinated calves had significantly higher packed cell and circulating red blood cell volumes than the challenged controls, longer red blood cell half lives, and somewhat lower blood volumes and rates of red blood cell synthesis. No untoward clinical effects that could be attributed to vaccination were recorded. These results indicate that zebu cattle can be effectively protected against S. bovis by vaccination with irradiated organisms. We are now evaluating this type of vaccine in a field trial in an enzootic area in the Sudan.

Two mathematical models are developed that investigate the transmission dynamics of Theileria parva by the ixodid tickRhipicephalus appendiculatus to cattle in endemically stable areas. A method of estimating the rate of infection to cattle ofT. parva at the endemically stable state is given. Empirical estimates of ail the parameters in the model are available. The degree to which animals that have recovered from theileriosis (the ‘carrier’ state) are able to transmit the infection to tick nymphs or larvae is a crucial determinant of the dynamics of infection in a herd. Two control methods influencing the transmission of infection are considered – infection and treatment immunization and the reduction in tick feeding by acaricide application. The impact of each method on the transmission of infection is evaluated. Future developments and the data required to predict the dynamics of T. parva infections in cattle and ticks are discussed.

Through the courtesy of Dr Hans Gadow I have had the opportunity of examining a number of skins of five species of Apteryx in the collection of the Cambridge University Museum. From these I have collected at least three species of Mallophaga, which I find to belong to the genus Rallicola. These are described below, with some discussion of the genus to which they belong, and of the significance of the occurrence of that genus upon Apteryx, from which no Mallophaga have previously been recorded.

Intermittent preventive treatment of infants (IPTi) with sulphadoxine pyrimethamine (SP) is recommended as an additional malaria control intervention in high transmission areas of sub-Saharan Africa, provided its protective efficacy is not compromised by SP resistance. A significant obstacle in implementing SP-IPTi, is in establishing the degree of resistance in an area. Since SP monotherapy is discontinued, no contemporary measures of in vivo efficacy can be made, so the World Health Organisation has recommended a cut-off based upon molecular markers, stating that SP-IPTi should not be implemented when the prevalence of the dhps 540E mutation among infections exceeds 50%. We created a geo-referenced database of SP resistance markers in Africa from published literature. By selecting surveys of malaria infected blood samples conducted since 2004 we have mapped the contemporary prevalence of dhps 540E. Additional maps are freely available in interactive form at http://www.drugresistancemaps.org/ipti/. Eight countries in East Africa are classified as unsuitable for SP-IPTi when data are considered at a national level. Fourteen countries in Central and West Africa were classified as suitable while seven countries had no available contemporary data to guide policy. There are clear deficiencies in molecular surveillance data coverage. We discuss requirements for ongoing surveillance of SP resistance markers in support of the use of SP-IPTi.

A population of three-spined sticklebacks, Gasterosteus aculealus, from a pond in Somerset was sampled about every three weeks for 19 months, a total of 1863 fish being collected. Three species of parasites were found in the fish: Schistocephalus solidus, Diplostomum gasterostei and Echinorhynchus clavula.

Seasonal changes in the incidence and intensity of the three parasitic infections are described. In general the infections increased in summer and autumn and decreased in winter and spring, although in Schistocephalus the differences in the level of infection between the two years were greater than between seasons.

The largest increases in the Schistocephalus and Echinorhynchus infections occurred in August and September and in the Diplostomum infection in May and June. These increases could be related to the feeding activity of the sticklebacks and to the abundance and level of infection of the Cyclops, Asellus and Limnaea pereger populations.

Reduction in the sizes of the infections was by predation and death of heavily infected fish in the case of Schistocephalus and Diplostomum and mainly by death of individual worms in Echinorhynchus.

These mechanisms are shown to be adapted to the life cycles of the parasites.

I am most grateful to Dr H. D. Crofton for his help and encouragement during this investigation and to Dr R. A. Avery for useful discussions and advice. This work was supported by a Science Research Council research studentship and a NATO studentship.

The Anaplasma species are important globally distributed tick-transmitted bacteria of veterinary and public health importance. These pathogens, cause anaplasmosis in domestic and wild animal species including humans. Rhipicephalus, Ixodes, Dermacentor and Amblyomma genera of ticks are the important vectors of Anaplasma. Acute anaplasmosis is usually diagnosed upon blood smear examination followed by antibodies and nucleic acid detection. All age groups are susceptible but prevalence increases with age. Serological cross-reactivity is one of the important issues among Anaplasma species. They co-exist and concurrent infections occur in animals and ticks in same geographic area. These are closely related bacteria and share various common attributes which should be considered while developing vaccines and diagnostic assays. Movement of susceptible animals from non-endemic to endemic regions is the major risk factor of bovine/ovine anaplasmosis and tick-borne fever. Tetracyclines are currently available drugs for clearance of infection and treatment in humans and animals. Worldwide vaccine is not yet available. Identification, elimination of reservoirs, vector control (chemical and biological), endemic stability, habitat modification, rearing of tick resistant breeds, chemotherapy and tick vaccination are major control measures of animal anaplasmosis. Identification of reservoirs and minimizing the high-risk tick exposure activities are important control strategies for human granulocytic anaplasmosis.

Based on morphological and genetic characteristics, we describe a new species of Hepatozoon in the European wild cat (Felis silvestris silvestris), herein named Hepatozoon silvestris sp. nov. The study also provides the first data on the occurrence of H. felis in this wild felid. Hepatozoon meronts were observed in multiple cross-sections of different organs of four (44%) cats. Additionally, extracellular forms, resembling mature gamonts of Hepatozoon, were found in the spleen and myocardium of two cats. Furthermore, tissues of six animals (67%) were positive by PCR. Hepatozoon felis was identified infecting one cat (11%), whereas the 18S rRNA sequences of the remaining five cats (56%) were identical, but distinct from the sequences of H. felis. Phylogenetic analyses revealed that those sequences form a highly supported clade distant from other Hepatozoon spp. Future studies should include domestic cats from the areas where the wild cats positive for H. silvestris sp. nov. were found, in order to investigate their potential role to serve as intermediate hosts of this newly described species. Identification of its definitive host(s) and experimental transmission studies are required for elucidating the full life cycle of this parasite and the possible alternative routes of its transmission.