Pentastomids are arthropod parasites which attain maturity in the respiratory tracts of vertebrates. All pentastomids exhibit an obvious sexual dimorphism in that fully mature females are invariably much bigger than males. Unusually though, copulation, which happens only once in the lifetime of females, occurs when both sexes are approximately the same size and the uterus of the female is undeveloped. This is because sperm is stored in spermathecae which, being positioned at the junction of the oviduct and uterus, become more remote from the vagina as the uterus elongates during development. The provision of spermathecae for sperm storage allows oocytes, shed continuously from the ovary, to be fertilized from time to time. What renders this process quite remarkable, and possibly unique, is the extreme length of the patent period. In one species this can extend up to an estimated 10 years and is associated with an egg production of about 106 eggs/female/year. Spermathecal structure provides some clues as to how prolonged sperm storage and the continuous fertilization of oocytes is accomplished. The upper region of the access duct is a narrow, chitinized tube which tapers to as little as 3 μm diameter at the point of entry into the spermatheca. The chitinous (?) lining of the spermatheca is relatively impermeable and apparently functions to totally isolate sperm from external influences: stored sperm are arranged in bundles and whorls and remain totally quiescent and inactive during the storage period. The entire spermatheca is invested by muscle fibres, the contraction of which will express small numbers of sperms through the narrow access duct. These are then reactivated by secretions from the female reproductive tract: the extreme narrowness of the spermathecal duct provides the fine adjustment of the system. The narrowness of the duct also creates problems during the process of insemination, since sperm discharged directly into the female reproductive tract would be unlikely to find the spermatheca through such a structure. Accordingly, the male cirrus is much modified to directly penetrate the spermathecal duct during copulation. Sperms, stored in the male seminal vesicle, are apparently activated prior to sperm transfer, and swim along the cirrus to be guided directly into the spermathecal lumen. Some unusual variations on the normal pentastomid life-cycle are discussed.

Peritoneal hydatids of Echinococcus granulosus equinus of 9 months standing in BALB/c mice occurred as free cysts or cysts within cyst masses. Both showed wide variation in size and in host tissue response, and all had a well-developed laminated layer separating the host tissue response from the germinal layer. In the smallest cyst-mass cysts the host tissue response was present as remnants of the initial cellular attack involving eosinophils. Slightly larger cyst-mass cysts possessed a primary macrophage invasion which phagocytosed the remnants of the initial attack and also, though to little effect, the laminated layer material. In the largest cyst-mass cysts a second macrophage invasion, of monocyte origin, had commenced and transformation stages of these cells to macrophages were observed. No fibroblasts surrounded individual cyst-mass cysts but they were present around the cyst mass, encapsulating it and possibly preventing further host cell invasion. Thus, the host tissue response around individual cyst-mass cysts remained ‘preserved’ at an early stage such as existed at the time of encapsulation. Small free cysts showed a primary macrophage invasion and transformation stages of cells of a secondary infiltration of peritoneal origin. Peripheral to the macrophages were fibroblasts demonstrating limited fibrinogenesis, and each cyst was surrounded by a layer of mesothelial cells. Large free cysts, also delimited by a mesothelial layer, possessed peripheral connective tissue, a deep fibrous layer and a monolayer of very compressed macrophages lying adjacent to the laminated layer. It is emphasized that an understanding of the host tissue response in cysts of different sizes and from different locations is an essential pre-requisite for the design of experimental studies.

The technique of tail amputation is utilized as a method for interrupting the migration process of Schistosoma mansoni schistosomula from the skin of Fischer rats infected by exposure of the tail to cercariae. The yields of schistosomula recovered from the lungs at different times post-infection are compared, using rats with or without tail amputation. Residence times of schistosomula in skin and lungs, as well as their transit time and efficiency of migration between these sites, are estimated. At least one-third of the infecting cercariae migrate from skin to lung in rats. Amputation of the tail on days 4 or 5 post-infection isolates a definable number of schistosomula in the lung and their migration to the portal circulation can be followed. The kinetics of this migration in rats and mice is compared and a significant difference is revealed.

Experiments were carried out to determine the susceptibility of mammalian cells to infection with different species of Theileria in vitro. Sporozoites of Theileria parva (parva), Theileria parva (lawrencei) and Theileria taurotragi were isolated from Rhipicephalus appendiculatus ticks by grinding infected ticks in medium, filtering the suspension and concentrating by centrifugation. The sporozoites were used in attempts to infect in vitro peripheral blood leucocytes harvested from 16 different mammalian species which included 12 species of Bovidae from 6 different sub-families. The technique was shown to be both sensitive and reproducible. The sporozoites of T. parva (parva) infected and transformed cells from 2 species of the sub-family Bovinae, the two cattle types and African buffalo. Theileria parva (lawrencei) infected and transformed cells from the two cattle types, African buffalo and Defassa waterbuck. Theileria taurotragi sporozoites infected in vitro cells from 11 different species of Bovidae which were members of 6 sub-families; Bovinae, Tragelaphinae, Reduncinae, Alcelaphinae, Antilopinae and Caprinae. Transformed lymphoblastoid cell lines were established from 7 of the species infected. Sporozoite attachment and infection was not observed with non-susceptible bovid host cells, nor were any of the non-bovid leucocytes infected by the parasites. The host range observed in this study corresponded to the known host range in vivo.

The high energy costs of egg-shell production in many helminths suggests that this structure plays an important role in their biology. The mechanical and chemical resistance of the egg-shell and the barrier it provides to the entry and loss of material are important in the survival of the free-living stages within the egg. The presence of egg filaments may increase the availability of infective stages and the operculum may be important in the infective process.

A 2 hectare paddock on the Muguga Estate, Kiambu District of Kenya (altitude 2100 m) free of Theileria parva-iniected ticks was seeded by applying Rhipicephalus appendiculatus nymphs to cattle infected with Theileria parva (Kiambu 4). It was estimated that over 50000 engorged nymphs fell onto the pasture from 4 cattle with high parasitaemias during the cold season (June). Samples of these ticks were placed in plastic cylinders under the vegetation in the paddock and sexual stages of T. parva were detected in gut lumen smears in these samples up to 21 days alter repletion. Zygotes were first observed to transform into kinetes on day 55 and parasites were first detected in salivary glands of adults on day 64. Moulting of the nymphs started on day 60 after repletion and was completed by day 87. Cattle introduced into the paddock showed their first infestation with adult R. appendiculatus on day 64 after repletion of the nymphs and the infestation level gradually increased. On day 76 after repletion, 17% of a sample of adult ticks infesting cattle showed salivary gland infections with T. parva and this increased to over 70% by day 150. From day 360 onwards, a decrease in both the percentage of ticks infected and the number of salivary gland acini infected/tick was noted. In addition, T. parva infections within the salivary glands required a longer period of feeding in the older ticks before they developed into sporozoites. Cycling of Theileria through ticks was prevented by the removal of female ticks before they completed repletion. Lethal challenge levels of T. parva for cattle existed in the paddock up to day 547 after repletion, after which introduced cattle showed no infection or only a sub-lethal infection. Cattle introduced into the paddock on day 808 after repletion showed no evidence of T. parva infection. The tick infestation of introduced cattle decreased markedly from day 368 and infestation had virtually ceased by day 808

The feeding behaviour of Glossina morsitans morsitans infected with Trypanosoma vivax, T. congolense or T. brucei was studied and compared with that of uninfected control tsetse. The following parameters were measured. (1) Mean number of feeds taken in 20 consecutive days; (2) mean rate of blood meal intake in μg/sec of the total days fed; (3) mean weight of blood meal intake/day of the total days fed; (4) mean total blood meal engorgement in 20 days; (5) mean number of probes/day of the days fed; (6) mean frequency of the multiple probe feeds and (7) mean number of pre-feeding probes of the multiple probe feeds. Evidence was found to suggest that the rosettes of T. vivax and T. congolense in the proboscis, and colonies of T. brucei parasites in the salivary glands, did not significantly alter the feeding behaviour of the vector.

Female Dermacentor andersoni were induced to feed on defibrinated bovine blood through fixed mouse skin membranes. Their feeding behaviour was recorded electronically, the tick being incorporated into the circuit and acting as a variable resistor during the periods of sucking, salivation and rest. Recordings of the feeding behaviour of these ticks were similar to those of ticks feeding on rabbits. Known concentrations of histamine, serotonin, dopamine, prostaglandin E1 and prostaglandin F2α were added singly or in combination to the feeding medium of ticks attached to the mouse skin membrane. Addition of both histamine and serotonin was followed by a significant reduction in the amplitude or recordings associated with sucking and salivation. The concentration of mediators producing this effect was less than 10 mM, a concentration comparable to that of histamine found locally in the skin of tick-resistant guinea-pigs. Comparisons are made between the effects produced by these mediators in ticks and the effects of serotonin and other amines known to produce modulation of complex behaviour patterns in other invertebrates.

Theileria annulata was transmitted transtadially by an Israeli strain of Hyalomma excavatum from larvae to nymphs and from nymphs to adults. However, the adults developing from larvae which engorged on infected calves and fed during the nymphal stage on non-infected calves, rabbits or gerbils (Meriones tristrami), did not transmit theileriosis. Suspensions of infected unfed nymphs or adults did not produce theileriosis but both nymphs and adults of the same batch became infective to cattle after 2 days of feeding. Suspensions of salivary glands or of whole ticks derived from infected adults that remained attached to rabbits for more than 7 days, were not infective to cattle.

Previous studies on the spermatogenesis and oogenesis of digeneans, monogeneans and cestodes are reviewed, including those in which isotopic labelling techniques have been utilized in order to determine the temporal duration of spermatogenic development stages. Similar labelling experiments have also provided information on the development and movement of oocytes in the female reproductive tract as well as indicating patterns of cell division and development in vitelline glands. The literature relating to the inseminative behaviour of parasitic platyhelminths is considered from a number of viewpoints including the patterns of cross-and self-insemination that occur in hermaphroditic digeneans, the mating behaviour of dioecious schistosomes, spermatophore production and insemination among monogeneans and the inseminative behaviour of polyzoic cestodes. The implications of selfing among parasitic platyhelminths are considered with respect to possible genetically deleterious consequences. In addition, consideration is given to the mechanisms underlying the fact that some species require cross-insemination or at least the presence of another worm in order to undergo normal growth and sexual development. Finally, experimental studies are reviewed which shed light on the strength of the barriers against hybridization that exist between well-defined species of parasitic platyhelminths.

The numbers, distribution in the small intestine, sexual development and growth (dry weight) of 5-week-old Moniliformis dubius (Acanthocephala) were investigated experimentally in adult, female CFHB rats fed on theoretically isoenergetic diets containing known amounts of fructose in combination with either maize-oil fatty acids or maize oil and two concentrations of casein. There was no obvious development of M. dubius when there was no fructose in the host's diet. In contrast, estimated consumption by the host of as little as about 2 g of fructose during the 5-week infection period was accompanied by marked sexual dimorphism and weight gain in most of the M. dubius present. The dry weights of M. dubius of both sexes were positively correlated with fructose concentrations ranging from 0 to 2·5 % (w/w) in the diets containing fatty acids. Significant, but not substantial, increases in M. dubius dry weight were observed as the dietary fructose concentration was raised to 12 % (w/w). Similar trends were observed when the fructose was offered to the infected rats with maize oil, but in general, fructose added to the fatty-acid based diets supported most M. dubius growth. Differences in the distribution pattern of the worms in rats fed on the fatty-acid or maize-oil based diets were observed and their possible significance is discussed.

Previous observations, which showed that treatment of mice with hydrocortisone acetate around the time of their infection with Schistosoma mansoni reduced the mature worm burden, have been confirmed. The number of eggs produced by the surviving worms during early patency was also significantly reduced. Cyclophosphamide and another steroid immunosuppressant, betamethasone, also caused a reduction in fecundity of S. mansoni when given at the time of infection, as did T-cell deprivation of the mice by adult thymectomy and injection of anti-thymocyte serum 1 month before infection. There was no effect of these three treatments on worm numbers. In contrast, injection of mice with anti-thymocyte serum at the time of infection marginally increased the size of the mature worm burden. The deleterious effects of hydrocortisone acetate on S. mansoni worm numbers and fecundity were only apparent if the steroid was given within 1 week of infection. Indomethacin, a compound which inhibits tissue inflammatory reactions by inhibiting prostaglandin synthesis, and which therefore mimics one of the actions of corticosteroids, had no effect on S. mansoni worm maturation and fecundity.

Changes during development in the elemental contents (Ca, P, K, Mg and S) of male and female Schistosoma mansoni from both mixed and single sex infections have been measured for the first time. In mixed infections, there is a significant difference between the calcium content of male and female worms; female worms having between 2 and 3 times more calcium. In mature female worms from mixed infections, after an initial rise in the calcium content corresponding to the time when vitelline development occurs, the amount of calcium ranged from 0.36 to 0.65 mg/g dry wt, whereas the amounts in similarly aged male worms ranged between 0.19 and 0.32 mg/g dry wt. While the type of infection does not affect the calcium content of male worms, a significant difference in the calcium levels in female worms from mixed and single sex infections was found. The amount of calcium in females from single sex infections ranged from 0.21 to 0.39 mg/g dry wt and this is considerably less than the levels recorded in females from mixed infections. This difference in calcium content between mixed and single sex infections can be related to the differences in degree of development of the vitelline gland in the two types of female worm. The amounts of phosphorus, potassium, magnesium and sulphur in both male and female worms from both single and mixed infections are similar, although females from mixed infections contained slightly more phosphorus than either females from single sex infections or either type of male worm.

Growth of Hymenolepis nana in mice is analysed by identifying designated stages of proglottid maturation in the strobila, and by determining the numbers of proglottids between them and the posterior terminus of the worm, as a function of time after cysticercoid infection. The proglottid maturation rate was fairly constant between stage A (onset of the somatic primordium formation) and stage B (first appearance of sperms in the seminal vesicle), but gradually fell after stage B, during pre-patent development of the worm. The actual rate of proglottization during this period coincided well with that estimated from the number of proglottids between stages A and B on an assumption of a certain maturation rate. This procedure is extended to estimate the rates of proglottid formation and shedding during later development. It is deduced that the absolute rate of proglottization increases until day 12, gradually decreasing afterwards, and that from 2300 to 2800 proglottids are shed by day 20, until which from 4000 to 4500 proglottids have passed stage A. It is shown that the proportionate increase of the gravid region in older worms depends not only on a decrease in the rate of proglottization, but also on the prolonged period during which proglottids stay in the strobila.

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.

The immunogenicity of the secretory exo-antigen (EXA) originating in the stichosome of Trichuris muris has been investigated. High levels of immunity to challenge were produced using EXA (100 μg) by subcutaneous administration. Vaccination with a range of doses of EXA in Freunds' Incomplete Adjuvant (FIA) showed a dose response; 1 μg of EXA reduced the worm burden by 50% and doses of 10 μg or more reduced the worm burden by 80–90% at Day 9 post-infection. Worms recovered from vaccinated mice were significantly smaller than those from controls. A precipitin line identifiable with EXA was shown to immunize mice against T. muris challenge by vaccination. The protective antigen contained in EXA has been partially isolated using gel filtration, ion exchange filtration and isoelectric focusing and has been shown to be a protein of 30000 molecular weight having a pi of 6·8–7·3.

The encapsulated embryos of platyhelminths may be retained and complete their development in utero in a range of circumstances. However, hatching within the parent (the criterion of ovoviviparity) is relatively rare and larvae generally emerge only after deposition. Viviparity is characterized by the nutritional dependency of the unencapsulated larva upon the parent, but in several cases larvae retained within a shell also receive parental nutrients during intra-uterine development. Uptake of exogenous nutrients via shell pores occurs in Schistosoma mansoni but the eggs, which gain all the advantages of intra-uterine retention, are supported by host nutrients.

Intra-uterine larval development avoids the hazards of development in the external environment and eliminates the time delay between oviposition and infection. Deposition of immediately infective offspring may be concentrated in time and space to exploit periods of host vulnerability. The control and precision of transmission is illustrated by examples in which the opportunity for invasion is restricted because of either host behaviour or environmental instability. This strategy has been an important factor in the evolution of polystomatid monogeneans, and its effectiveness is demonstrated by comparison of the life-cycles of Polystoma integerrimum and Pseudodiplorchis americanus. Ovoviviparity also increases reproductive potential in some polystomatids by extending the period of multiplication and by increasing established populations through internal re-infection. In Eupolystoma alluaudi, the capacity for ovoviviparity is programmed into larval development and this regulates population growth within individual hosts.

After demonstrating that rats were capable of discriminating between the same diet treated with either flavour 1 or flavour 2 and that the 2 flavours of diet were equipreferred, an experiment was carried out to see whether learned taste aversion might play a role in the reduction of food intake that is commonly observed during the course of a parasitic infection. The results showed that rats, given a subcutaneous inoculation of approximately 6000 third-stage larvae of Nippostrongylus brasiliensis (Nematoda) while feeding on a diet containing flavour 2, strongly preferred to eat diet containing flavour 1 when given simultaneous choice conditions. Uninfected rats showed no preference and ate equal amounts of both flavoured diets. This effect is interpreted as the first experimental demonstration of learned taste aversion using a eukaryotic parasite as the inductive agent.

In this study, we have developed a reliable radioimmunoassay (RIA) for Ascaris suum protein (Asp) and have measured the serum concentration of Asp in control subjects and in patients with helminthiasis and a variety of gastrointestinal diseases. The cross-reactivities of Asp antibody with other helminth worm antigens were also measured by RIA. Ascaris lumbricoides contained approximately the same amount of Asp antigenic substance as did Ascaris suum. Toxocara canis had a high concentration of a substance partially cross-reactive with Asp. Small amounts of substance cross-reacting with Asp were also exhibited by Anisakis larvae, Schistosoma japonicum and Taenia saginata. However, no cross-reactivities of Fasciola hepatica and Paragonimus westermani with Asp were observed in this study. Almost all the serum levels of Asp from normal control subjects were below the sensitivity of this RIA (10 ng/ml). High concentrations of Asp were observed in sera from patients with ascariasis (64.5±18.8 ng/ml: mean ± S.E.), anisakiasis (75.2±28.0 ng/ml), toxocariasis (78.4±31.3 ng/ml) and schistosomiasis japonica (14.3±3.4 ng/ml). Asp levels in sera from patients with other helminthiasis were not significantly high. From these findings, it was suggested that RIA for Asp may not be specific for diagnosis of ascariasis, but might be a suitable screening method for patients with some kinds of helminthiasis. Interestingly, high concentrations of Asp were also observed in sera from patients with ulcerative colitis (24.8±2.5 ng/ml) and Crohn's disease (19.8±5.2 ng/ml), but Asp levels from 7 other gastrointestinal diseases seemed not to be significantly high. These findings remain unexplained but may derive from cross-reactivities between helminthic and gut mucosal antigens.

Prior to scolex-retraction, the tegumentary syncytial cytoplasm of the presumptive rostellar region of Hymenolepis diminuta (from Tenebrio molitor kept at 26 °C) is indistinguishable from that of the rest of the cysticercoid. At 1 day after scolex-retraction differentiation of the rostellum has commenced, the tegumentary cytoplasm containing a small number of membrane-bound, ovoid, electron-dense granules which are absent from all other tegumentary regions of the metacestode. By 3 days after scolex-withdrawal there is a substantial increase in the number of ovoid granules within the rostellar tegumentary syncytium and the Golgi systems of the rostellar cytons are highly secretory. At this stage, although the cyst wall tissues are not fully developed, the metacestode is infective. This early and rapid development of the rostellar region presumably enables the ‘adult’ condition to be readily attained. In older metacestodes there is a progressive accumulation of ovoid granules within the rostellar tegumentary cytoplasm, accompanied by a decrease within the rostellar cytons. At 23 days following scolex-retraction the rostellar syncytium has the appearance of that of the adult tapeworm. The rostellar cytons also produce tegumentary discs and vesicles and are therefore regarded as homologous to the other tegumentary cytons of the metacestode.