CBA mice deprived of their T-cells by means of thymectomy and administration of rabbit anti-mouse thymocyte serum before a primary infection of Schistosoma mansoni, failed to resist re-infection to the same extent as immunologically intact controls. However, little difference was made to the degree of resistance to re-infection acquired by thymectomized S. mansoni-infected mice when the anti-thymocyte serum was administered after the primary infection and just before the challenge. Hydrocortisone acetate, when administered to primarily infected mice just before reinfection appeared to reduce the degree of acquired resistance, but the drug also had a schistosomicidal effect on the challenge-derived organisms. An attempt to specifically suppress the acquisition of resistance against S. mansoni by administering serum from homologously infected, resistant donors before and during the course of a primary infection failed, and partial depletion of complement activity with cobra venom factor that was administered just before challenge also failed to affect resistance. Serum obtained from mice with chronic S. mansoni infections and injected intradermally at the site of subsequent administration of a homologous percutaneous challenge infection failed to markedly inhibit maturation of the challenge in the recipient. Recipients of 3 ml of serum from heavily infected donors were also not significantly more resistant to challenge than untreated controls. However, isolated skin from heavily infected mice allowed fewer cercariae to penetrate and emerge transformed as schistosomula than did isolated skin from uninfected controls.

In an attempt to determine the mechanisms of resistance to schistosome infection operative in bulinid snails, an histological study was carried out on batches of a refractory strain of Bulinus truncatus and two highly susceptible strains of B. africanus killed at various intervals of time, ranging from 7 h to over 3 months, after exposure to infection with Schistosoma mattheei miracidia.

It was found that only a small fraction (less than 1%) of the miracidia penetrated B. truncatus, and that they penetrated only a few of the snails exposed. Much larger proportions (up to 26%) of the miracidia penetrated B. africanus and they penetrated nearly all the snails exposed.

In the susceptible B. africanus only miracidia which settled in the loose tissues of the head-foot proceeded to develop: those settling in dense tissue failed to develop and degenerated within about 2 days.

A strong infiltration reaction of amoebocytes rapidly destroyed the few miracidia that successfully penetrated B. truncatus. In contrast, B. africanus did not respond to the presence of miracidia in its tissues, nor to mother or to daughter sporocysts before the onset of cercarial shedding: it reacted only to cercariae.

It is concluded (a) that the combination of an effective surface barrier and an active defence mechanism due to amoebocytes were responsible for complete resistance to S. mattheei infection in B. truncatus (b) that successful establishment of S. mattheei infection in B. africanus depended on the failure of the snail to recognize the schistosome miracidia and sporocysts as foreign material.

I am indebted to Professor George S. Nelson for advice and encouragement in this work which was carried out at the London School of Hygiene and Tropical Medicine. I am most grateful to Dr R. J. Pitchford, Dr C. A. Wright and Dr F. Arfaa for providing material for schistosome and snail cultures and to the East African Community and the British Ministry of Overseas Development for financial support.

1. The anatomy of the adult miracidium of Parorchis acanthus is described.

2. Development of the miracidium and of the succeeding larval stages has been followed.

3. Cleavage of the fertilized egg gives rise to an ectodermal and a propagatory cell. The former gives rise to the soma, and the latter, after contributing a few small cells to the body of the miracidium, becomes the germ cell.

4. This germ cell develops in the same way into the first generation redia while still in the body of the miracidium.

5. The fully formed miracidium contains a single fully formed redia when it penetrates the intermediate molluscan host.

6. In the intermediate host the miracidium disintegrates and second generation rediae are formed from the germ cells in the body cavity of the parent redia.

7. The germ cells in the body cavity of the daughter redia give rise to cercariae by a similar process of embryonic development to that found in preceding generations.

8. The germ cells in the cercariae become differentiated into the adult genitalia.

9. No evidence has been found to show that rediae or cercariae are derived from cells in the wall of the redia.

10. No ovary, maturation process, parthenogenesis, or fertilization takes place in any of the larval stages, but germinal lineage is apparent throughout the life cycle.

1. The osmotic pressure of the medium in which the nematodes were cultured has been shown to be equivalent to that of about 30 mM. NaCl.

2. The effects of immersion in a saline solution isotonic with the above, in glass-distilled water, and in hypertonic saline solutions, have been described. Particular attention was paid to changes in the length of the body, in the appearance and behaviour of the alimentary canal, and in the movement of the body.

3. The results obtained have been discussed, and the following tentative conclusions reached:

(a) The main aqueous exchanges occur through the cuticle and protoplasm covering the periphery of the body, and are controlled by the protoplasm rather than by the cuticle. The cuticle thus does not act as aja impermeable outer covering which insulates the animal from its environment.

(b) The species normally lives in an environment which is hypotonic to the body fluids, and maintains its internal osmotic pressure by means of an active method of osmotic regulation. The alimentary canal is probably partially, but not wholly, responsible for this regulation.

(c) Swelling occurs in distilled water because the removal of water in the initial stages is not rapid enough to cope with the extra inflow. Later the excess water is removed.

(d) In concentrated solutions, water is removed from the body, and the internal osmotic pressure may also be increased to some extent by the penetration of osmotically active substances to the body, through the cuticle and hypodermis of the body wall. Penetration through the hypodermis is slower than through the cuticle.

(e) During recovery in concentrated solutions, there is a slow removal of osmotically active material from the body. The hypertrophy of the lateral and post-anal organs, which sometimes occurs under these conditions, may be connected with this removal.

Investigations were carried out on the influence of the local environment on the preparasitic stages of B. phlebotomum and the following observations were made.

1. The infective larvae are positively thermo-tropic and positively phototropic. These observations confirm those of Schwartz (1924). The larvae are not negatively geotropic and do not climb grass, but remain in the dung, gaining access to the host by adherence of the dung to the skin.

2. Lack of air, such as would occur in tightly packed faeces; immersion of the faeces in water; temperatures below 10° C.; and lack of moisture, are all inhibitory to development. Infective larvae are resistant to at least 5 days‘ direct exposure to an atmosphere of relative humidity 75 at a temperature of 25° C.; in dry faeces they are resistant to 14 days‘ exposure to the same atmosphere.

3. In Northern Nigeria desiccation is the most important inhibitory factor in development. The extreme dryness of the pastures in the dry season prevents development of the larvae and adherence of dung to the skin. Penetration of the skin of the host probably occurs only in the rainy season, maximal penetration probably occurring just after the rains are regularly established.

4. The female lays 600 eggs in 12 hr. in the early rains, but the output of eggs by the females varies considerably throughout the year. It is greatest in the early rains and lowest in the later months of the dry season.

5. The female hookworm burden of nomadic cattle fluctuates throughout the year. The maximum burden occurs in the later months of the dry season.

6. Three factors, all probably bound up with a single factor, i.e. moisture, seem to influence the degree of pasture infectivity. (a) Inhibition of larval development; (b) failure of faeces to adhere to the skin; (c) fluctuation in the egg-laying rate of the female hookworms. The fluctuating pasture infectivity results in a fluctuating hookworm burden.

7. The period of maximum hookworm burden in nomadic cattle more or less coincides with the period of incidence of a disease in Stock Farm cattle, associated with hookworm infestation.

In the discussion an attempt is made to correlate these observations both in accounting for the seasonal incidence of ‘hookworm disease’ and in suggesting methods of control.

This paper was written during the tenure of a Cooper Centenary Fellowship for which the author is indebted to the Council of the Veterinary Educational Trust. The writer's thanks are also due to his laboratory assistant in Nigeria, Mr R. A. O. Shonekan.

Environmental sex determination in the mermithid nematode Romanomermis culicivorax is examined in the context of parasite reproductive success and population regulation. Experimental results show that the sex ratio of the nematode within its mosquito host (Culex quinquefasciatus) is dependent on parasite density. Sex ratios are biased to females at low parasite burdens and to males at high parasite burdens. Low temperature further enhances female-biased ratios. The net effect of density-dependent sex determination on parasite and host population growth is shown to be critically dependent on the frequency distribution of parasite numbers/host. Mermithid parasite distributions within natural host populations show low degrees of aggregation relative to other helminth species. The population regulation of the parasite is examined with respect to environmental sex determination and parasite-induced host mortalities by means of simple mathematical models of the dynamics of parasite transmission via its life-cycle. The significance of regulatory constraints on population growth are discussed in relation to the use of mermithids as biological control agents of insect pests or disease vectors.

Here we discuss proteomic analyses of whole cell preparations of the mosquito stages of malaria parasite development (i.e. gametocytes, microgamete, ookinete, oocyst and sporozoite) of Plasmodium berghei. We also include critiques of the proteomes of two cell fractions from the purified ookinete, namely the micronemes and cell surface. Whereas we summarise key biological interpretations of the data, we also try to identify key methodological constraints we have met, only some of which we were able to resolve. Recognising the need to translate the potential of current genome sequencing into functional understanding, we report our efforts to develop more powerful combinations of methods for the in silico prediction of protein function and location. We have applied this analysis to the proteome of the male gamete, a cell whose very simple structural organisation facilitated interpretation of data. Some of the in silico predictions made have now been supported by ongoing protein tagging and genetic knockout studies. We hope this discussion may assist future studies.

Despite metabarcoding being widely used to analyse bacterial community composition, its application in parasitological research remains limited. What interest there has been has focused on previously intractable research settings where traditional methods are inappropriate, for example, in longitudinal studies and studies involving endangered species. In settings such as these, non-invasive sampling combined with metabarcoding can provide a fast and accurate assessment of component communities. In this paper we review the use of metabarcoding in the study of helminth communities in wild mammals, outlining the necessary procedures from sample collection to statistical analysis. We highlight the limitations of the metabarcoding approach and speculate on what type of parasitological study would benefit from such methods in the future.

During the acute phase of Trypanosoma cruzi infection, strong haematological and immune system alterations are observed. The parasite expresses trans-sialidase, a virulence factor responsible for the sialylation of its surface glycoconjugates. This enzyme is also shed to the bloodstream where it is associated with immune system alterations triggered during the infection. During experimental and human infections, the host elicits antibodies able to neutralize the enzyme activity that would be responsible for restricting systemic trans-sialidase to the early steps of the infection, when major immune alterations are induced. The actual relevance of these antibodies was tested by passive transference of monoclonal neutralizing antibodies in acute infection models displaying extreme sensitivity to the infection. Mice were inoculated with virulent parasite strains that induce high parasitaemia, early mortality and strong immune tissue abnormalities. The trans-sialidase-neutralizing antibodies were able to preserve B cell areas both in ganglia and spleen as well as the thymus architecture even in these extreme models. Although no differences between control and treated mice regarding animal survival were found, a major role for the humoral response in controlling the damage of the immune system induced by a systemically distributed virulence factor was defined in an infection with a eukaryotic pathogen.

A study has been carried out to investigate whether the action of triclabendazole (TCBZ) against Fasciola hepatica is altered by inhibition of drug metabolism. The cytochrome P450 (CYP P450) system was inhibited using piperonyl butoxide (PB). The Oberon TCBZ-resistant and Cullompton TCBZ-susceptible isolates were used for these experiments. The CYP P450 system was inhibited by a 2 h pre-incubation in PB (100 μm). Flukes were then incubated for a further 22 h in NCTC medium containing either PB; PB+nicotinamide adenine dinucleotide phosphate (NADPH) (1 nm); PB+NADPH+TCBZ (15 μg/ml); or PB+NADPH+TCBZ.SO (15 μg/ml). Morphological changes resulting from drug treatment and following metabolic inhibition were assessed using scanning electron microscopy. After treatment with either TCBZ or TCBZ.SO alone, there was greater disruption to the TCBZ-susceptible than the resistant isolate. However, co-incubation with PB and TCBZ/TCBZ.SO lead to more severe surface changes to the TCBZ-resistant Oberon isolate than with each drug on its own. With the TCBZ-susceptible Cullompton isolate, there was limited potentiation of drug action, and only with TCBZ.SO. The results support the concept of altered drug metabolism in TCBZ-resistant flukes and this process may play a role in the development of drug resistance.

A PCR procedure for the detection of Opisthorchis viverrini in experimentally infected bithynid snails and cyprinoid fishes was developed. This procedure was based on primers designed from a pOV-A6 specific probe sequence giving a 330 base-pair product. The detection was accomplished in host tissue homogenates to which a single cercaria or metacercaria was introduced. PCR can detect as little as a single cercaria artificially inoculated in a snail or a single metacercaria artificially inoculated in a fish sample. The method gave a 100% positivity rate for all infected snails or fishes. The method did not yield a 330 base-pair amplified product with other digenean fluke DNAs such as Haplorchis taichui, Centrocestus spp., Echinostoma malayanum, Fasciola gigantica, animal schistosomes, Paragonimus heterotremus or Haplorchoides spp. The assay has great potential for application in epidemiological surveys of both snail and fish intermediate hosts as well as for investigation of foodborne parasites in freshwater fishes.

PCR assays have been developed mainly to assist investigations into the epidemiology of Giardia duodenalis, the only species in the Giardia genus having zoonotic potential. However, a reliable identification of all species is of practical importance, particularly when water samples and samples from wild animals are investigated. The aim of the present work was to genotype Giardia species and G. duodenalis assemblages using as a target the region spanning the 5.8S gene and the 2 flanking internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene. Primers were designed to match strongly conserved regions in the 3′ end of the small subunit and in the 5′ end of the large subunit ribosomal genes. The corresponding region (about 310 bp) was amplified from 49 isolates of both human and animal origin, representing all G. duodenalis assemblages as well as G. muris and G. microti. Sequence comparison and phylogenetic analysis showed that G. ardeae, G. muris, G. microti as well as the 7 G. duodenalis assemblages can be easily distinguished. Since the major subgroups within the zoonotic assemblages A and B can be identified by sequence analysis, this assay is also informative for molecular epidemiological studies.

The swallow bug, Oeciacus vicarius Horvath, is of common occurrence in North America, chiefly, if not entirely, upon the cliff swallows, Petrochelidon lunifrons (Say); it has been recorded from the barn swallow, probably in error.

It appears to be really distinct from the European swallow bug, Oeciacus hirundinis (Linnacus).

The habits, structure, and specific characters of the bug do not seem to have been adequately studied hitherto.

This paper deals with the external structure of the bug and of such of its biology as could be worked out under the handicap that its natural host could not be kept in captivity.

The bugs attack man readily, but do not appear to adapt themselves readily to living upon man or in his dwellings.

Extracts and essential oils from plants are important natural sources of pesticides. These compounds are considered an alternative to control ectoparasites of veterinary importance. Schinus molle, an endemic species of Brazil, produces a high level of essential oil and several other compounds. The aim of this work was to determinate the chemical composition of extracts and essential oils of S. molle and further to evaluate the activity against eggs and adults of Ctenocephalides felis felis, a predominant flea that infests dogs and cats in Brazil. In an in vitro assay, the non-polar (n-hexane) extract showed 100% efficacy (800 µg cm−2; LD50 = 524·80 µg cm−2) at 24 and 48 h. Its major compound was lupenone (50·25%). Essential oils from fruits and leaves were evaluated, and had 100% efficacy against adult fleas at 800 µg cm−2 (LD50 = 353·95 µg cm−2) and at 50 µg cm−2 (LD50 = 12·02 µg cm−2), respectively. On the other hand, the essential oil from fruits and leaves was not active against flea eggs. This is the first study that reports the insecticidal effects of essential oils and extracts obtained from Schinus molle against Ctenocephalides felis felis.

A 58 kDa excretory–secretory product (ESP) of Giardia lamblia has been characterized. The ESP was purified over 508-fold by a combination of ammonium sulphate precipitation and sequential chromatography on affinity matrix and a gel filtration column. The homogeneity of the purified protein was established by sodium dodecyl sulphate polyacrylamide gel electrophoresis (Mr, 58 kDa) and analytical isoelectrophoresis (pI 4·75). The purified protein was recognized by the pooled sera of G. lamblia-positive patients as well as an antiserum raised against crude Giardia extract, thus indicating it to be an immunodominant parasite product. The ESP was found to agglutinate rabbit erythrocytes. The haemagglutinating activity of this protein was inhibited strongly by thyroglobulin, fetuin, asialofetuin and monosialoganglioside but not by simple sugars. The purified protein was characterized immunochemically and was found to be heat stable as well as protease sensitive. Lectin-binding studies of the purified ESP and its sensitivity to periodic-acid silver staining as well as to metaperiodate treatment clearly indicated its glycoprotein nature. The major localization site of the ESP was found to be on the surface of the parasite as revealed by flow cytometric analysis. Further, this glycoprotein induced fluid accumulation in ligated rabbit ileal loops and revealed a positive skin permeability reaction in the rabbit.

The sensitivities of promastigotes and amastigotes of Leishmania donovani to reagent or glucose oxidase-generated hydrogen peroxide (H2O2) were examined in a phagocyte-free system and compared with direct measurements of loss of H2O2 due to reaction with the parasite. Using a combined fluorescence dye uptake/dye exclusion viability assay in conjunction with motility and transformation data it was shown that log-phase promastigotes harvested from recently transformed cultures were intermediate in their H2O2 sensitivity between amastigotes and log-phase promastigotes harvested from long-term subcultures. It was also observed that, while promastigotes are equally sensitive to either form of H2O2 stress, amastigotes are more resistant to single larger amounts of reagent H2O2 than to equivalent amounts of H2O2 generated over a 1 h period. In each case the respective LD50 values obtained for each form of the parasite under each type of H2O2 stress correlated with saturation of their ability to remove H2O2 from the phagocyte-free system. For both promastigotes and amastigotes there was always a time delay after removal of either form of H2O2 stress before H2O2-mediated damage to membranes became apparent. The results suggest that the differential responses of promastigotes and amastigotes to different forms of H2O2 stress may depend upon different H2O2 scavenging mechanisms examined in more detail in the accompanying paper.

Recent work on the inheritance of susceptibility to malaria suggests that, over what may have been a relatively short evolutionary period, a remarkably diverse series of gene families have been modified in response to the selective drive of this single infection. The phenotype consequences are not confined to the red cell, but involve the immune system, cytokines and many other systems. It seems likely that the mechanisms of variation in genetic susceptibility to other infective agents will reflect at least a similar degree of complexity and, if the selective pressures have been present for longer periods of our evolutionary history, may be even more diverse. This may have important implications for work directed at trying to define susceptibility loci for current infectious and non-infectious diseases.

A mathematical model of the transmission dynamics of Cowdria ruminantium by the ixodid tick Amblyomma hebraeum in the bovine host is developed and used to investigate the epidemiology of heartwater across a range of vector challenge. The processes described are supported by empirical data. The pattern of outcome measures (incidence, case-fatality and proportion of infected hosts) predicted agrees with those described anecdotally from field experience and empirical observation, and demonstrates the concept of endemic stability. The underlying theory is explored and it is shown that endemic stability may be due principally to the protection of calves against disease by either innate or maternally derived factors. The role of vertical infection in the establishment and maintenance of endemic stability is also investigated. Although increasing the vertical infection proportion results in endemic stability occurring at progressively lower levels of tick challenge, the concomitant reduction in incidence and case-fatality predictions across the range of tick challenge means the endemically stable state simultaneously becomes less discernible. Model limitations and future developments are discussed. The essential role of a transmission dynamics model in assessing the impact of new vaccines in conjunction with vector control programmes is highlighted.

Iran is an important endemic focus of cystic hydatid disease (CHD) where several species of intermediate host are commonly infected with Echinococcus granulosus. Isolates of E. granulosus were collected from humans and other animals from different geographical areas of Iran and characterized using both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). The sheep and camel strains/genotypes were shown to occur in Iran. The sheep strain was shown to be the most common genotype of E. granulosus affecting sheep, cattle, goats and occasionally camels. The majority of camels were infected with the camel genotype as were 3 of 33 human cases. This is the first time that cases of CHD in humans have been identified in an area where a transmission cycle for the camel genotype exists. In addition, the camel genotype was found to cause infection in both sheep and cattle. Results also demonstrated that both sheep and camel strains can be readily differentiated on the basis of hook morphology alone.

To determine the prevalence of Cryptosporidium in dairy cattle in Guangdong Province, South China, 1440 fecal samples were collected from 10 farms and screened for Cryptosporidium with PCR. The overall prevalence of Cryptosporidium was 4.38% (63/1440), and the infection rates in preweaned calves, postweaned calves, heifers and adults were 6.4% (19/297), 6.19% (33/533), 1.48% (4/271) and 2.06% (7/339), respectively. Three Cryptosporidium species, Cryptosporidium andersoni (n = 33), Cryptosporidium bovis (n = 22) and Cryptosporidium ryanae (n = 8) were detected by DNA sequence analysis of the 63 positive samples, and C. andersoni was identified as the most common species on the dairy cattle farms. In preweaned calves, C. bovis was the most prevalent species (9/19, 47.4%). In contrast, C. andersoni was the predominant species (19/33, 57.6%) in postweaned calves and the only species found in heifers and adults. The zoonotic species Cryptosporidium parvum was not detected in this study. Twenty-four C. andersoni isolates were successfully classified into three multilocus sequence typing (MLST) subtypes. MLST subtype A4,A4,A4,A1 was the predominant subtype, and MLST subtype A2,A5,A2,A1, previously found in sheep, was detected in cattle for the first time. A linkage disequilibrium analysis showed that the C. andersoni isolates had a clonal genetic population structure. However, further molecular studies are required to better understand the epidemiology of Cryptosporidium in Guangdong.