The pulmonary and portal vasculature of naïve mice of the 129/Ola and CBA/Ca strains has been studied by means of the vasculature casting technique. This involves injection of pigmented vinylite resin into the arterial and venous systems, followed by digestion of the tissues with KOH. The peripheral vessels of the arterial and portal systems of CBA/Ca mice were numerous and highly branched. In contrast, casts prepared from 70–80% of naïve 129/Ola mice showed dramatic reductions in the number and extent of the peripheral vessels. In addition, such vessels appeared severely truncated. The remaining 20–30% of naïve 129/Ola mice yielded lung and liver casts that were indistinguishable from the CBA/Ca casts. Casts prepared from 129/Ola mice infected 6 weeks previously with Schistosoma mansoni cercariae showed the same segregation; faecal smears, together with observations of presence or absence of gross pathology in such mice confirmed that the vascular changes correlated with the ‘non-permissive trait’. We propose that such alterations facilitate the reportedly abnormal migration of schistosomes from the liver to the lungs in ‘non-permissive’ 129/Ola mice.

The excretions-secretions (E-S) of Acanthocheilonema viteae consist mainly of one product, molecular weight 62 kDa. This molecule is synthesized during the vertebrate phase of the parasite life-cycle and is first detectable in the E-S of L4 parasites. It is cross-reactive with E-S of human filarial parasites as a consequence of possessing a phosphorylcholine (PC) moiety. The 62 kDa molecule has been employed as a model for the study of the origin and fate of filarial E-S. Immunohistological analysis has shown the molecule to be located predominantly in the parasite gut. Transplantation of adult female [S] methionine pulsed worms into uninfected jirds resulted in the radio-labelled secreted 62 kDa antigen being detected in the bloodstream within 4 h by SDS–PAGE/immunoprecipitation analysis. The systemic half-life of the molecule as estimated by clearance of injected, purified I-labelled material was measured in naive and infected jird hosts.

The DNA of a parasite is the ultimate blueprint of that parasite, the one characteristic which normally remains unchanged during every stage of the life-cycle. All the DNA sequence in the egg of a species of parasite are also in the larvae and adults of the same species. The same DNA is present in the parasite whether it is in a free-living stage, in an invertebrate vector or in a vertebrate host such as man. The molecular basis for DNA diagnosis is to allow labelled single-stranded species or strain-specific DNA sequences, selected from well-characterized reference species, to find and hybridize with homologous DNA from, or in, the unknown isolates of parasites. DNA probes are now available for most vector borne parasitic diseases. Parasitological identification problems are mostly concerned with distinguishing closely related strains or subspecies, for example detecting Taenia solium eggs as opposed to T. saginata eggs, or finding which of the 15 man-infecting subspecies of Leishmania is present in a single cutaneous lesion, the commonest clinical sign of the disease, or in a sandfly. For efficient hybridization by the present methods there has to be enough of a particular sequence present in a parasite's genome to make a feasible target. Therefore, DNA probes for parasites have been selected from repetitive, reiterated or multicopy DNA with intrinsic extensive sequence variation. DNA, which is free of coding restraint, can evolve rapidly to give differences between species, so that introns, ribosome gene spacers, variant genes, pseudo-genes and non-conserved DNA have all been used for DNA diagnosis. The major problems of sequence selection have been greatly aided by the use of recombinant DNA methods, which have the added advantage of economical production of DNA probes. The unique characteristics of kinetoplast mini-circle DNA in Leishmania has allowed the selection of a complex species, subspecies, strain and even isolate-specific DNA probes. These have been used successfully for Southern filter endonuclease fragment DNA identification, for dot-blot recognition of less than 200 parasites and non-radioactive detection of DNA sequence homology by ‘in situ’ hybridization and light microscopy in a single Leishmania cell. The adaptation of the forensic human genetic fingerprinting technique has allowed identification of L. braziliensis DNA in human biopsy material, even the presence of a vast excess of human DNA. Fingerprinting with ribosomal spacer-derived recombinant DNA probes has been used to discriminate Echinococcus species. The identification of Taenia species has been accomplished using probes from a genomic library of size-selected DNA fragments, and synthetic oligonucleotides are available for Onchocerca subspecies detection. The new technologies combining repeated genomic sequence probes with pulse field separation of the chromosomes of parasites, has opened up new avenues of research. Double probes for the simultaneous identification of the insect vector and the carried parasite have recently been reported. Lastly, the polymerase chain reaction technique has given us the opportunity of amplifying even single copy genes in one parasite to give sufficient DNA for positive identification. DNA diagnosis in parasitology is now on a par with the DNA diagnosis of viruses and human genetic disorders. The last ten years have seen many exciting developments in DNA diagnosis; the next years should see DNA diagnosis routinely used in epidemiological and clinical situations in countries where parasitic diseases are a major public health problem.

A qualitative and quantitative analysis was made of the release of surface-associated molecules from developing Dirofilaria immitis infective-stage larvae (L3). D. immitis L3s were labelled with 125I using an Iodogen catalysed reaction and either maintained in culture or placed in chambers that were implanted into Lewis rats. The larvae released 10–20% of the labelled material each day during the first 4 days of in vitro and in vivo development. The loss of surface-labelled peptides from developing larvae corresponded with an increase in the amount of trichloroacetic acid-precipitable radioactivity found in the culture medium. SDS—PAGE analysis of the labelled material showed that the same 35 and 6 kDa components found in larval extracts were shed into culture medium by the developing parasites. Metabolic labelling studies and experiments in which larvae were labelled after different times in culture indicated that, once released, the surfaceassociated molecules were not replaced, and that this net loss of surface peptides resulted in a reduction in the antigenic potential of the cuticular surface. Antibodies from both immunized rabbits and naturally infected dogs immunoprecipitated the 35 kDa component. In contrast, the 6 kDa molecule was not recognized by the antibodies in any of the sera tested. Shedding of surface peptides and reducing surface antigenicity may represent mechanisms by which D. immitis infective-stage larvae evade immune attack.

The spontaneous activity of the somatic muscle of the amphibian lung fluke, Haplometra cylindracea has been recorded in vitro, using an isometric force transducer system. Normal movement consisted of a continuous series of regular contractions which were maintained for over 4 h. Acetylcholine (ACh) inhibited motility and at a concentration of 1 χ 10–3 M induced a flaccid paralysis. A similar response occurred with the cholinomimetic drugs, carbachol and nicotine, although in these instances the inhibition was less easily reversed by washing with frog Ringer. The inhibitory effect of ACh was blocked by d-tubocurarine but not by atropine.

The integrity of the hepatic portal vasculature was examined, relative to the resistance to Schistosoma mansoni observed in 68°0 of 129/Ola mice. The passage of microspheres to the lungs, following their injection via the superior mesenteric vein, indicated the presence of shunts in the majority of both naive and infected mice. There was a negative association between shunting of microspheres to the lungs and paucity of liver worms at 28/35 days post-infection. Schistosomula accumulated in the livers of resistant mice at a slower rate than in susceptible animals, and after day 21 relocated to the lungs. Many lung schistosomula injected via the superior mesenteric passed immediately to the lungs; the shunts thus greatly reduce the probability of trapping in the liver. Some parasites migrated back from the lungs, successfully lodged in the liver and began to feed on blood. Latex infusion demonstrated the location of large intrahepatic connections between the portal and hepatic veins. We suggest that as these liver worms grow, migrating upstream into progressively larger vessels, they reach the connections, pass out of the hepatic portal system, and relocate to the lungs. The presence of the natural shunts thus accounts for the resistant status of the mice.

Phase-interference microscopic examination of the infective, post-infective 3rd-stage and 4th-stage larvae of Dirofilaria immitis has identified a single cell body in each of the paired lateral amphidial nerves which undergoes characteristic morphological change during the development from 3rd- to 4th-stage larvae. Acetaldehyde—fuchsin staining of worm sections revealed fuchsinophilic material in the precise location of the amphidial nerve-cell bodies observed by phaseinterference microscopy. This material was found in 70% of infective larvae recovered from mosquitoes and in 100% of larvae recovered from micropore chambers 24 h after implantation into BALB/C mice. In pre-moult larvae recovered at 42 h (Experiment 1) and at 48 h (Experiment 2) fuchsinophilic material was demonstrable, but no staining was observed in those larvae in which separation of the 3rd- and 4th-stage cuticles had occurred. No such material was observed in 4thstage larvae recovered from chambers after 74 h, and in these larvae the amphidial nerve-cell bodies were not discernible. These cytological observations are consistent with a cycle of elaboration and release of neurosecretion associated with moulting in D. immitis.

Work reported at this meeting has described the exploitation of variation in parasite phenotype in disciplines ranging from molecular taxonomy and drug development, through the understanding of host-parasite interaction, to the evolution of parasite populations and determining the potential efficacy of vaccine programmes.

The prevalence of the caecal threadworm Trichostrongylus tenuis in red grouse in the north of Scotland was high despite low grouse densities. Prevalence, intensity and aggregation of threadworms was higher in old than in young grouse. Infections were long-lasting: populations of adult worms could survive for over 2 years in grouse, with little mortality. Parasite egg output decreased with the age of a worm population, largely as a result of a decrease in the fecundity of ageing female worms. Seasonal variations in worm fecundity were also evident.However, there was no evidence of an intensitydependent decrease of worm fecundity with increasing worm numbers in either captive or wild grouse. The long life and high reproductive capacity of T. tenuis probably contribute to its effective transmission and high prevalence.

Two trypanosome clones, representing East and West African homozygotes at 2 isoenzyme loci (T. b. rhodesiense MHOM/ZM/74/58 [CLONE B] and T. b. brucei MSUS/CI/78/TSW 196 [CLONE A]), were cotransmitted through tsetse flies and the resulting trypanosome populations checked for the presence of non-parental karyotypes by pulsed-field gel electrophoresis. Ten clones isolated from these populations proved to have 5 different recombinant genotypes by analysis of nuclear and kinetoplast DNA (kDNA) polymorphisms. It is inferred that genetic exchange occurred between the 2 trypanosome clones in the fly, as previously reported for 2 other T. brucei spp. clones by Jenni and colleagues. For the most part, the hybrid clones shared many characteristics with both parents and their genotypes were consistent with segregation and reassortment of parental alleles. The least amount of genetic material exchanged was kDNA alone. Regarding the mechanism of genetic exchange, several hybrid clones had identical and unique nuclear DNA polymorphisms, but different kDNA type. Assuming that the same reassortment of nuclear markers is unlikely to occur by chance, these clones most probably arose from a predecessor carrying both types of kDNA.

In the laboratory, yields of infective 3rd-stage larvae of Trichostrongylus tenuis were determined by temperature when moisture was adequate. On the moor, eggs in caecal faeces did not develop to infective larvae during the winter. In summer, development did occur and yields of infective larvae were related to temperature: greatest yields were obtained when mean monthly soil surface temperatures ranged between 76 and 103°C. Rainfall was unimportant, as was desiccation. During summer the mean monthly mortality of infective larvae was 44% but over winter survival was negligible.

The resistance of captive reared red grouse to Trichostrongylus tenuis was measured as the proportion of ingested infective 3rd-stage larvae which failed to develop to adult worms. Individual grouse showed wide, repeatable variations in resistance. Seasonal variations may also have occurred. Red grouse acquired little or no effective immunity to reinfection after challenge with a single dose of infective larvae. In trickle-dosed grouse, however, adult worms weakly inhibited the establishment of incoming larvae in an intensity-dependent fashion. Despite this, the proportion of larvae established in trickle infections was similar to that in single-dose challenges.

The growth of individual proglottides of Hymenolepis nana has been studied by measuring the width and length of the proglottides between days 3 and 8 post-infection in mice. Two sets of measurements were obtained. The first involved proglottides immediately posterior to the point of proglottis formation and proglottides at particular stages of development. For the second set, measurements were made of proglottides at certain positions in the worm, specifically the 20th, 100th and 200th proglottides, as counted from the posterior end of the worm. For proglottides at different stages in development, the width of a particular stage was greater in the later days of infection, while the length was almost constant. The length of newly-formed proglottides varied only slightly with time despite differences in the width and in the 3-fold increase in proglottis production that occurs between days 3 and 8 post-infection. The width of the 20th, 100th and 200th proglottides followed parallel growth curves that coincided with each other after an appropriate shift along the time axis. The more anterior the position of the proglottis, the greater was the width at the time of proglottis formation, and so its growth began later on the common growth curve. Growth in length of the 20th proglottis was at first faster than the 100th and 200th proglottides, but later slowed down to a level comparable with them, and the growth curves for the length of the three proglottides were very similar to each other. The rates of growth in the volume of the three proglottides were also estimated. The results are also discussed in relation to other approaches to the study of tapeworm growth and to factors that may be responsible for the growth patterns observed.

Phenol oxidase of Fasciola gigantica exists both as the soluble form as well as the membrane-bound form. The membrane-bound enzyme is considered to be a tyrosinase type because it is capable of oxidizing mono- and diphenol and is inefficient in oxidizing paraphenols. The soluble enzyme is a laccase type showing more affinity to various diphenols and paraphenols. Membrane-bound enzyme exists as isoenzymes, showing 3 fractions, of which the slow-moving fraction is capable of oxidizing both 4-methyl catechol and catechol, whereas the two remaining fractions are specific to 4-methyl catechol only. Soluble enzyme exists as a single homogeneous form showing affinity to both mono- and diphenols. Inhibition of the enzyme by potassium iodide and mercuric chloride indicates the active tyrosyl and SH groups of the enzyme. Inhibition of the enzyme by sodium diethyl dithiocarbamate and phenyl thiourea indicates that copper is the prosthetic group of the enzyme.

Direct surface I radio-isotope labelling techniques and SDS—PAGE analysis were used to compare the proteins and lentil—lectin adherent glycoproteins of the bovine stage of viable Taenia saginata larvae at three points in their development, the invasive oncospheres, immature (4-week-old) and mature (12 to 16-week-old) cysticerci. Some proteins and glycoproteins were present on all three of the ages of the parasite examined but there were also distinct age-specific proteins and glycoproteins detected on oncospheres and 4-week-old cysticerci and a marked difference between the protein/glycoprotein profiles of the parasite was apparent at these earlier stages of development and the mature cysticerci. The latter were characterized by the presence of high, 160–200 kDa molecular weight, lysine rich, glycoproteins, whereas small 16 and 18 kDa glycoproteins and a reduction-sensitive 23 kDa glycoprotein were first detected on 4-week-old immature cysticerci. Antigenic characterization of the isotope-labelled proteins and glycoproteins by immunoprecipitation against a panel of clinically defined bovine sera combined with SDS–PAGE analysis indicated that relatively few proteins were precipitated by sera from T. saginata-infected cattle as compared to the glycoproteins. However, both protein and glycoprotein antigens of possible protective and/or diagnostic significance were identified from oncospheres and cysticerci.

The development and mortality of the eggs of Ancylostoma duodenale and Necator americanus in distilled water were monitored over a range of temperatures between 15 and 35°C. Egg demography was examined within the context of a fourparameter mathematical model of development and mortality. Over the range of temperatures studied, egg mortality (μ) was an increasing exponential function of temperature (T) measured in degrees Celsius. A single model adequately described the mortality of both species (ln [μ] = 0·041*T − 6·87). The minimum time (τ) to hatching was consistently less for A. duodenale (ln [1/T] = −(0·79+53·05*[l/T]) than N. americanus (ln [1/T] = −(0·99 + 53·05*[l/T])). The hatching rate [σ when t > τ) was an increasing function of time and temperature in both cases, but the precise functional relationship was species specific.

A series of compounds that apparently disrupt hormonally regulated processes in insects have been examined for effects on the viability and microfilarial production of adult Brugia pahangi cultured in vitro. The azasteroids, 25-azacoprostane and 25-azacholestane, inhibited the production of microfilariae at 5 ppm, the former also exhibiting macrofilaricidal activity at this concentration. The brassinosteroids examined inhibited microfilarial production at 5 ppm but did not affect worm viability. Azadirachtin also proved to be a significant inhibitor of microfilarial release without effect on worm motility or viability. Of all the compounds tested, the non-steroidal amines appeared to be the most promising as potential filaricides, several of them proving to be macrofilaricidal at 1 ppm and affecting microfilarial production at even lower concentrations.