Both asexual and sexual development of Plasmodium berghei was synchronized without chemical intervention using in vitro culture techniques. Combined in vivo and in vitro experiments were performed on the relationship between age, morphology and maturity of gametocytes. Schizogony took 22–23 h in the experiments. At 26 h post-invasion (p.i) the first males became capable of exflagellation. By 20 h p.i. the first gametocytes were recognizable in Giemsa-stained smears but the sex was hardly distinguishable until maturity (26 h p.i.). Survival time of gametocytes was estimated at 26 h in vitro (half-life 13 h) and the same survival time was suggested for gametocytes in vivo. Schizonts of P. berghei apparently disappeared from the peripheral circulation upon maturity, rupturing almost immediately. Mature schizonts in vitro persisted up to 48 h p.i. in non-agitated cultures. No evidence was collected for sequestration of any sub-population of gametocytes.

The morphology of Kronborgia pugettensis sp. nov., an endoparasite from the haemocoele of the caridean shrimp Heptacarpus kincaidi is described from specimens collected in the San Juan Island region of the western coast of North America. Members of this species are unisexual as are other members of the genus, but only females were found. An ootype and a type of accessory reproductive gland not known from other Kronborgia spp, are described, but they are probably present in other members of the genus. The life-history is similar to that of other Kronborgia spp. After the female reaches maturity, it emerges from the host and secretes a tubiform cocoon around itself and deposits its egg capsules within it. Embryogenesis lasts about 4 months. A free-swimming larva locates the new host, attaches to its exterior, and secretes a cyst around itself. It then penetrates through the exoskeleton to reach the haemocoele. K. pugettensis is easily distinguished from other members of the genus by the shape of the cocoon, type of host, and geographic distribution. This is the second species of fecampiid described from North America, but several undescribed species are known to exist from their cocoons.

Unisexual females, (80–100 days post-infection) were cultured in vitro and exposed to mature male worms from either mixed or single-sex infections. A comparison was made of the uptake of [3H]thymidine (used as a marker for DNA synthesis) by isolated unisexual females or by those females exposed to males derived from either mixed or unisexual infections during in vitro culture. All males were observed to be paired with females by approximately 18 h post-culture. At 24 h post-culture females paired with males from mixed infections had an elevated uptake of [3H]thymidine compared to females that had not paired and uptake was also significantly different (P ≥ 0–001) from females exposed to males from unisexual infections. By 48 h there was no significant difference between uptake in females paired with either type of male. The frequency of uptake in the paired females was approximately double that in unstimulated females while non-paired females in the presence of males had an increase of between 35 nd 45%. In a second experiment the effect of depriving a mature egg-laying female of the paired male was investigated. These females had a continual decrease in uptake of [3H]thymidine compared to mature females maintained with male partners. The uptake of [3H]thymidine was used as a marker for DNA synthesis and the effect of the male worm on DNA synthesis in the unisexual female is discussed in view of the above results.

This ultrastructural study has shown that there is a layer of dense flocculent material on the surface of juvenile Fasciola hepatica incubated in vitro with specific bovine antiserum. This material corresponds to the complexes of secreted glycocalyx and bovine antibody previously characterized by fluorescence microscopy. Bovine eosinophils attach closely to those regions of the parasite's surface that are free of flocculent precipitates. This close attachment is followed by degranulation of the eosinophils into the narrow zone between the cells and the parasite. Only in these regions is damage, in the form of vacuolation of the tegument, seen within the juvenile F. hepatica. It is concluded that the inability of bovine eosinophils to kill juvenile F. hepatica in the presence of specific antiserum results from the presence of a protective layer, consisting of antigen/antibody complexes, on the parasite's surface.

Raillietiella gigliolii is a cephalobaenid pentastomid which inhabits the lungs of the South American worm-lizard Amphisbaena alba. The host is a facultative inquiline of nests of the leaf-cutting ant Atta cephalotes where it feeds occasionally (and possibly by accident) on ants but more often on beetles and their larvae which are themselves inquilines of ant nests. Ants store exhausted leaf-substrate in special underground chambers which serve as refuse dumps and it is here that larvae of the three-horned rhinoceros beetle Coetosis biloba feed: these larvae are also known to be prey items of A. alba. From observations of captive colonies of Atta, we have demonstrated that pentastomid-egg contaminated faeces of A. alba, introduced into the colony, are rapidly cut up and thrown onto the refuse dump, where, under natural circumstances they will be eaten by Coelosis. These larvae have an unusual and highly specialized gut physiology and parasite eggs will develop to an infective stage within the haemocoel in 70–96 days. Cockroaches are refractory to infection. Ants are the vital link in transmission since they literally deliver eggs to the Coelosis larvae. The strong trophic links which exist between the various components of the life-cycle offset a low fecundity of 100 eggs/female parasite/day but nonetheless maintain a high prevalence (86%) of infection.

Lung flukes, Paragonimus ohirai, from Kinosaki, Japan, characterized by 3 electrophoretic variants, FF, FS and SS, of glutamic-oxaloacetic transaminase (GOT: EC 2.6.1.1) were crossed in the laboratory. In the case of a cross between FS and SS, a parent, FS, produced 9 SS and 16 FS clones in the offspring, numbers not significantly different from that expected (1:1) from Mendelian inheritance. From the other cross FS × FF, 20 clones originating from the respective 20 miracidia were obtained from a parent FS. The phenotype segregation ratio was 6 FF and 14 FS, which is not significantly different from Mendelian inheritance expectation. Only 3 clones were obtained from the other parent FF, but these segregated in a ratio of 2 FS: 1 FF. These breeding data indicate that the GOT isozymes of P. ohirai are controlled by 2 codominant alleles, GotF and Gots at a single locus whose products aggregate randomly, forming a dimer.

An enzyme analysis of diploid and triploid Paragonimus westermani was conducted using starch gel electrophoresis. In total, 16 enzymes, probably encoded by 18 loci, were studied for 3 populations of the diploid form sampled from 2 localities, and 4 populations of the triploid form from 4 localities. Comparison of the enzymes of the triploid and the diploid digeneans showed 5 different patterns; diaphorase (EC 1.6.2.2), glutamic-oxaloacetic transaminase (EC 2.6.1.1), hexokinase (EC 2.7.1.1), leucylglycylglycine aminopeptidase (EC 3.4.1.3), and phosphoglucomutase (EC 2.7.5.1). On the basis of the numbers of bands and their patterns, all individuals of the triploid are probably heterozygous at each of these 5 loci and homozygous at the remaining 13 loci. The occurrence of fixed heterozygotes found in triploid populations cannot be easily explained by only a single mutation. It is suggested that the variability may have been introduced by hybridization with a different sub-species or a closely related species and may, thus, have been maintained since the time of the origin of triploids.

A total of 65 enzymes in the 8 European pathotypes of the potato cyst nematodes Globodera pallida and G. rostochiensis were examined by isoelectric focusing (IEF) for possible polymorphic variants which may aid rapid pathotype recognition. Quantitative variation was seen with glucose oxidase (EC 1.1.3.4), with high levels of this enzyme present in G. rostochiensis Ro3. Mannose phosphate isomerase (EC 5.3.1.8) also varied between the pathotypes with G. rostochiensis Ro4 and Ro5 differing from the other pathotypes in the relative absorbance of some bands. Phosphoglucomutase was examined on a narrow-range pH gel and additional differences to those described previously were detected.

Curie-point pyrolysis gas-liquid chromatography (PGLC) was used to detect gross chemical differences between the 8 European pathotypes of the potato cyst nematodes Globodera rostochiensis and G. pallida. No qualitative differences were seen with either flame ionization or flame photometric detection systems, although quantitative differences between pathotypes were detected. Cluster analysis of these differences failed to group the pathotypes into species, but separation of some economically important pathotypes was achieved. The potential of PGLC for nematode identification is discussed.

The development of the male reproductive system in Gyrodactylus gasterostei has been followed using parasites of known age maintained on isolated hosts. The penis develops shortly after the parasite has given birth for the first time (at an age of 24–30 h at 13°C) and the first active spermatozoa appear after 40–50 h. Spermatogenesis occurs more rapidly in G. gasterostei than in any other parasitic flatworm (including those from warm-blooded hosts) in which it has been measured, and the onset of male maturity is further hastened by a reduction in the number of pre-spermatogenic germ cell divisions. Spermatogonia have a diploid chromosome complement of 12, and spermatocytes undergo meiosis to produce haploid spermatozoa. No evidence of aneuploidy in spermatozoa was obtained. Although the development of haploid spermatozoa suggests that sexual reproduction can occur, production of embryos by isolated flukes which lack a mature male system indicates that other means of reproduction may also be employed.

Heterogeneity in susceptibility of guppies (Poecilia reticulata) to infection with Gyrodactylus bullatarudis was examined by comparing the establishment, reproduction and survival of the parasite on 4 inbred strains of the fish host following low and high levels of exposure to infection. The 4 strains fell into 2 broad categories of innately ‘resistant’ and ‘susceptible’ fish on the basis of strain differences in (a) the rates of growth and decay of the parasite populations, (b) the proportional occurrence of susceptible and resistant fish within each strain, (c) the average duration of primary infection, and (d) the duration of resistance to reinfection following recovery from a primary exposure. The results suggest genetic control of resistance/susceptibility traits and experiments with hybrids tentatively suggest dominance of resistance over susceptibility. The potential of the guppy – Gyrodactylus laboratory system for the investigation of the role of genetic factors in the study of host–parasite interactions is discussed.

Media are described which stimulate the eggs of Notocotylus attenuatus to hatch in vitro. Hatching involves opening of the operculum and emergence of the opercular cord to form a tube measuring approximately 75 μm in length along which the sporocyst passes. The sporocyst is then released from the end of the tube. It is suggested that in vivo the opercular cord serves to inject the sporocyst through the snail host's gut wall into the haemocoel.

The effect of the protostrongylid nematode, Elaphostrongylus rangiferi, on the fecundity and survival of an intermediate snail host, Arianta arbustorum, was studied. Over an experimental period of 12 weeks, infected snails had a significantly lower egg production than uninfected control snails. No significant difference in survival was found between infected and uninfected adult snails. Infected juvenile snails showed a consistent decrease in survival with increasing mean density of parasites. A linear relationship was found between snail instantaneous death rate over a 28–day interval and the mean number of parasites/snail. When calculated for 4–day intervals, the instantaneous death rate was highest between 8 and 12 days post–infection. It is concluded that the most pathogenic phase of this parasite is during the first moult.

The abundance of Profilicollis botulus was monitored in eider ducks for 3 years. No significant difference in P. botulus abundance was found between eiders taken alive and those collected dead from the shore line, or between adult males and adult females. Adult male eiders had a seasonal pattern of infection. Incubating female eiders lost their infection in almost all cases, suggesting that a seasonal pattern of infection must also occur in these birds. Eiders in their first winter had P. botulus abundances which were approximately 10 times that of adult eiders, but by the following summer the abundance had declined to that found in adults. Data are compared with previous studies undertaken on the Ythan estuary. Captive eiders were used to determine the developmental rate of female P. botulus and the percentage of administered cystacanths which established in the intestine. There was no evidence to suggest that male and female P. botulus have different life–spans.

Succinate dehydrogenase and fumarate reductase activities from a participate fraction of 4 isolates of the supposed ‘species’ of Trichinella, T. spiralis, T. pseudospiralis, T. nativa and T. nelsoni have been determined using spectrophotometric methods. The ‘in vitro’ effect of the anthelmintics thiabendazole, mebendazole and ivermectin on succinate dehydrogenase and fumarate reductase activities from the above isolates is also described. The significance of the results is discussed in the context of the controversy that surrounds speciation of the genus Trichinella.

An experiment using lysimeters suggested that the eggs of Taenia saginata and Ascaris lumbricoides survive for only a short time when applied to pasture in sewage sludge. However, a subsequent experiment which followed the survival of eggs throughout the soil profile demonstrated that some T. saginata eggs could still be found at 200 days on the soil surface, and that survival increased down the profile. Rainfall is shown to be able to wash eggs into the soil where they may be afforded protection from radiation and desiccation; this may have little epidemiological significance.

The sequential, quantitative histological analysis of adult Hyalomma anatolicum anatolicum feeding sites on rabbits showed that the cellular reactions at tick feeding sites change in character and magnitude with time after attachment, and differ considerably from primary infestation to tertiary infestation. Neutrophils (62–68%) followed by mononuclear cells (22–24%) were the major component of the cellular infiltrate throughout primary infestation. Eosinophils accounted for 10% of the cellular infiltrate at 24 h after attachment. Their proportions declined thereafter, forming 9% of the infiltrate at 72 h and 5% at 144 h. Basophil infiltration in most of the feeding lesions was noticed by 24 h, but always in low numbers. Despite a slight increase in absolute numbers the proportion of basophils fell from 3% at 24 h to 2– at 144 h. In contrast to primary infestation the cellular reactions on tertiary infestations were characterized by a significant increase in the numbers of basophils and eosinophils and marked degranulation of mast cells and basophils as early as 24 h after attachment. As the feeding advanced there was an increase in the mean proportions of basophils from 4% to 9% and mononuclear cells from 22% to 38%, and a decrease in the proportion of eosinophils from 21% to 9%. However, neutrophils (42–53%) remained the major component of the cellular infiltrate. There was no significant difference in the nature and sequence of cellular events in the feeding sites of male and female ticks, although the lesions produced by feeding males were 5–10 fold smaller than those of females. Mast cells, basophils and eosinophils appeared to be the major host cells involved in the resistance response, manifested by protracted feeding, reduced engorgement weight and poor egg laying. The mechanisms by which these cells might interact to mediate resistance are suggested and discussed.

According to the World Health Organization the global estimate of malaria is over 200 million infections, the majority of which are caused by the most life-threatening species, Plasmodium falciparum (Report of the Steering Committees of the Scientific Working Groups on Malaria, World Health Organization, June 1983). The causative agent of the disease, the malarial parasite, requires two hosts: a blood-sucking mosquito and a blood-containing vertebrate. Commonly, infection of the vertebrate begins when an infected mosquito bites a suitable vertebrate and injects minute sporozoites into the bloodstream. Within 30 mm the introduced sporozoites leave the bloodstream and enter parenchymal cells of the liver (mammals) or endothelial cells (birds). In these sites the parasite undergoes asexual multiplication (= exo-erythrocytic schizogony) producing daughter progeny called merozoites. The exo-erythrocytic merozoites are released from the tissues into the circulation where they invade red blood cells. Within an erythrocyte the merozoite undergoes asexual multiplication (= erythrocytic schizogony) producing a substantial number of merozoites. The erythrocyte lyses, merozoites are released, and invasion of another erythrocyte may then take place. The synchronous rupture of the red cell and merozoite release is marked by the periodic fever–chill cycles so characteristic of the malarial infection. Some merozoites continue to reinvade other erythrocytes and multiply by asexual means, whereas others enter erythrocytes and differentiate into sexual stages, male or female gametocytes. When a suitable mosquito feeds on an infected vertebrate gametocytes are ingested and the sexual cycle of development is initiated. In the mosquito stomach the gametocytes transform into gametes, fertilization takes place, the resultant worm-like zygote penetrates the cells of the mosquito gut and comes to lie on the outer surface of the stomach. Here each zygote forms a cyst-like body, the oocyst, within which thousands of sporozoites are produced by asexual multiplication. When the swollen oocysts burst, sporozoites are freed and these make their way to the salivary gland. At the next blood feeding the mosquito injects the infective sporozoites and the life-cycle is completed.