1. The reactions, under field conditions, of Lucilia sericata (Mg.) to various substances placed on sheep are described.

2. Attempts to produce either attraction or oviposition by placing cystine or cysteine hydrochloride in the living fleece gave negative results.

3. Several organic sulphur compounds (see Table 1) of a type which might arise from the breakdown of cystine have been tested. All showed some power of attraction for L. sericata. Ethyl mercaptan and dimethyl disulphide were the most powerful as attractants, but none of these substances was able to induce oviposition.

4. Tests with ammonium compounds and carbon dioxide have shown that, at the concentrations used, ammonia acted as an attractant. For oviposition to occur carbon dioxide had to be present.

5. Hydrogen sulphide was detected as a constituent of the fleece atmosphere of certain sheep. This substance, whilst having no attractive powers when used alone on sheep, increased the attractiveness of organic sulphur compounds.

6. Comparative tests with.ammonium carbonate-indole and ammonium carbonate-ethyl mer-captan mixtures have shown marked variations in the relative powers of these preparations to induce oviposition.

7. A distinction is drawn between stimuli which attract L. sericata to sheep and those which induce oviposition. Ammonia and various organic sulphur compounds enhanced attraction, but both ammonia and carbon dioxide were necessary for oviposition. It is suggested that some of these materials may be produced under natural conditions from the break-down of fleece keratin or by the bacterial decomposition of sweat and similar products in the fleece.

East Coast fever (ECF) is a fatal disease of cattle caused by the protozoan parasite Theileria parva. The development of a subunit vaccine, based on the sporozoite-specific surface antigen p67, has been hampered by difficulties in achieving high-level expression of recombinant p67 in a near-authentic form. Therefore two sets of recombinant baculovirus vectors were constructed. The first set, encoding various regions of p67, produced low levels of the corresponding p67 domains in High Five™ cells, despite the presence of large amounts of p67 RNA. The second, consisting of p67 domains fused to the carboxy-terminus of GFP expressed significantly higher levels of p67 protein. The GFP[ratio ]p67 fusion proteins were recognized by a sporozoite-neutralizing monoclonal antibody (TpM12) raised against native p67 whereas non-fused full length p67 expressed in insect cells was not recognized. GFP-tagging therefore, appeared to enhance the stability of p67 and to conserve its folding. The high-level expression of p67 domains in a more authentic form is an important step towards the development of an effective subunit vaccine against ECF.

The guinea-pig has been reassessed as a potential laboratory host for Schistosoma mansoni. Twenty-six per cent of an infective cercarial population survive to maturity in this rodent and there are no gross fluctuations in worm burden subsequent to pairing of male and female parasites. Five day, 4-week and 6-week-old schistosomes grown in guinea-pigs have topographical features that are identical to those exhibited by similarly aged mouse worms, but different from those exhibited by rat worms. Schistosome eggs are never detected in the faeces of infected guinea-pigs, but they can be observed in the pulmonary, hepatic and intestinal tissues. Only 55% of the eggs that can be recovered from the intestinal tissues are viable, and some of these can be hatched to release miracidia that penetrate the intermediate snail host. Cercariae are sometimes liberated from infected snails, but in insufficient numbers to permit infection of naive guinea-pigs. The schistosome cycle cannot, therefore, be completed in this host. Collaterals are sometimes observed in the vasculature serving the rectum and kidneys of infected guinea-pigs and the fact that schistosome eggs are deposited in the lungs of these animals indicates that portal systemic anastomosis is a feature of the model. Pathology associated with cercarial invasion or egg deposition is not dissimilar to that described for other laboratory animals infected with S. mansoni, except that basophils participate in the inflammatory response observed in the skin and intestine.

Heterologous immunity, or protection by one invading organism against another across phylogenetic divides, has been recognised for decades. It was initially thought to operate largely through enhancement of phagocytosis, but this explanation became untenable when it was realised it worked extremely well against intraerythrocytic protozoa and killed them while they were free in the circulation. Clearly a soluble mediator was called for. This review summarises the logic that arose from this observation, which led to a wider appreciation of the roles of pro-inflammatory cytokines, and then nitric oxide, in the host's response against invaders, as well as the ability of these mediators to harm the host itself if they are generated too enthusiastically. This has led to a discernable pattern across heterologous immunity as a whole, and its lessons influence a range of areas, including vaccine development.

The insect fauna of tree wounds with sap-exudations or “slime flux,” as well as that of the water-filled hollows of trees, has already attracted the attention of entomologists and several interesting life histories of insects have been recorded from these media. In the present paper I propose to deal with the fauna of only one tree, a horse-chestnut standing on the Downing site between the School of Biochemistry and the Molteno Institute. For several years this tree has supplied me with very rich material for the study of Dipterous larvae and their parasites. Unfortunately, this study recently came to a natural end when this tree, with several others, was cut down to make place for a new building. This fact justifies the publication of the present records of observations collected during several years, although I do not consider them as complete. The most interesting feature of our horse-chestnut tree was the presence of two kinds of lesions each harbouring a different fauna: (A) a sap-exudating or “ slime flux ” wound, and (B) a waterfilled hollow or reservoir.

An immunochromatographic test (ICT), using recombinant truncated P50 (P50t), for the detection of antibodies to Babesia gibsoni was developed and evaluated. Whereas all sera from specific pathogen-free dogs were clearly negative, all sera from dogs experimentally infected with B. gibsoni were clearly positive in the ICT. In addition, the ICT detected no cross-reactivity with sera from dogs experimentally infected with closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi, or with Neospora caninum, and Leishmania infantum. Sequential sera from a dog experimentally infected with B. gibsoni were tested with the ICT; it was shown that the specific antibodies are detectable as early as 6 days post-infection (p.i.) and that strong antibody responses remained until the end of the experiment (144 days p.i.). To evaluate the clinical application of the ICT, a total of 54 serum samples collected from domestic dogs that had been identified as having signs of anaemia at veterinary hospitals in Japan, were tested with the ICT, the previously established enzyme-linked immunosorbent assay (ELISA) and with the indirect fluorescent antibody test (IFAT). Twenty-four of the tested samples (44·4%) were positive in both ICT and ELISA, and (51·8%) in IFAT. The concordance between ELISA and ICT was found to be 100%, and 85·7% with IFAT. Taken together, the results above suggest that the ICT using P50t is rapid, simple, accurate, and suitable for use at clinical sites for the diagnosis of B. gibsoni infection in dogs.

Lactoperoxidase-catalysed radio-iodination was used to compare the surface proteins on red cells from Plasmodium yoelii-infected and normal BALB/c mice. The profile of radio-iodinated proteins separated by SDS-polyacrylamide gel electrophoresis was different for infected blood of similar parasitaemia from mice inoculated with different doses of the parasite. Inoculation with the lower dose resulted in the appearance of a major radio-iodinated protein of apparent molecular weight (Mr) 76000 which was labelled to a similar extent on uninfected red cells from infected blood and purified multinucleate infected cells. Several minor radio-iodinated bands, with identical mobilities to the minor bands on normal BALB/c erythrocytes, were also present on red cells from this infected blood. In contrast, the higher inoculation dose produced changes in the minor labelled bands, and the band with Mr of 76000 was absent. In this case, the minor radio-iodinated proteins of the normal BALB/c erythrocyte (with Mr of 65000, 57000, 48000, 38000 and 32000) were replaced by a series of bands with Mr of 60000, 50000, 43000 and 28000 on both uninfected and infected red cells. These differences with inoculation dose may be related to the different duration of these infections, the development of anaemia and the extent of pathological changes at the erythrocyte surface. P. yoelii infection caused a marked loss in periodate-dependent labelling of sialoglycoproteins on most, if not all, red cells in infected blood. There was also a large decrease in galactose oxidase-dependent glycoprotein labelling with or without neuraminidase treatment. These changes in the carbohydrate groups on red cell membrane glycoproteins may be linked to the excessive loss of both uninfected and infected red cells during some malaria infections.

Visceral leishmaniasis (VL) is a major public health problem in the Indian subcontinent where the Leishmania donovani transmission cycle is described as anthroponotic. However, the role of animals (in particular domestic animals) in the persistence and expansion of VL is still a matter of debate. We combined Direct Agglutination Test (DAT) results in humans and domestic animals with Geographic Information System technology (i.e. extraction maps and scan statistic) to evaluate the exposure to L. donovani on these 2 populations in a recent VL focus in Nepal. A Poisson regression model was used to assess the risk of infection in humans associated with, among other factors, the proportion of DAT-positive animals in the proximities of the household. The serological results showed that both humans and domestic animals were exposed to L. donovani. DAT-positive animals and humans were spatially clustered. The presence of serologically positive goats (IRR=9·71), past VL cases (IRR=2·62) and the proximity to a forest island dividing the study area (IRR=3·67) increased the risk of being DAT-positive in humans. Even if they are not a reservoir, domestic animals, and specially goats, may play a role in the distribution of L. donovani, in particular in this new VL focus.

In most parts of the world helminth infestations are responsible for such a heavy wastage of sheep that their control has undoubtedly become one of the most important charges that devolve upon the flock-owner. It might be thought that in tropical regions suited to the raising of sheep the problem of helminth control would cease to be urgent; and that climatic conditions, the general aridity of land chosen in such countries for sheep-rearing, and its consequently smaller carrying capacity, would contribute to form an environment distinctly unfavourable both to the survival of the free-living stages of parasitic worms and to their chances of reaching a suitable intermediary or definitive host. Such areas are not usually well watered, and it certainly does result from the paucity of permanent streams and standing water that few species of trematodes are represented. Those species that do occur, e.g. Fasciola hepatica, F. gigantica, are restricted to particular localities and are easily subjected to control.

The importance of various species of ticks in relation to the propagation of protozoal diseases, is so generally recognised that it has appeared to us eminently desirable to make a detailed study of one of the common species. Such a study has seemed to us specially necessary for the reason that our knowledge of these parasites is very imperfect, in spite of the fact that they possess an economic interest of the first order. Some of the diseases which ticks transmit, notably those due to the haematozoal parasites belonging to the genus Piroplasma, are among the most devastating affections of domesticated animals in many parts of the world, the useful animals which suffer from piroplas-mosis being cattle, sheep, goats, horses, and dogs. The disease known as “Heart-water,” occurring in South Africa and affecting sheep, goats and cattle, is likewise tick-transmitted. A disease of the domesticated fowl, analogous to relapsing fever in man, likewise of economic importance and occurring in different parts of the world, has also been demonstrated to be transmitted from animal to animal through the agency of ticks. The fowl disease is due to a Spirochaeta which is conveyed by ticks; the same holds for human “tick fever” and a spirochaete infection in cattle occurring in parts of Africa. Recent investigation appears to have clearly established the fact that a tick conveys spotted or Rocky Mountain Fever to man. Moreover it has been claimed that a Nematode worm, the Filaria perstans, parasitic in man, undergoes its development in a tick which is capable of conveying the parasite from one human host to another. There can be no doubt but that ticks will be found, upon further investigation, to be associated in the transmission of an increasing number of diseases in animals.

Comparisons were made between the protein concentrations and protein profiles of uninfected Gammarus pulex and those infected with cystacanths of Pomphorhynchus laevis. Comparisons of the extractable haemolymph volumes from uninfected and infected G. pulex, when related to host tissue weight, showed that there was no difference in the host tissue/haemolymph volume ratio between infected and uninfected individuals. SDS-PAGE of haemolymph samples resolved 36 bands of protein, the major components being band 6(19 kDa) and 21(33 kDa). Densitometric analysis of haemolymph samples showed that infected gammarids had total protein concentrations elevated by 84%. The (88 kDa) subunit was identified as the monomer of the respiratory protein haemocyanin. This band was significantly elevated in infected individuals (one-way ANOVA P < 0.001); increasing from 30 to 45% of the total haemolymph proteins.

The tick Boophilus microplus is a 1-host tick that causes important losses to bovine herds, and protective antigens are being investigated in order to develop vaccines that avoid the use of acaricides. Paramyosins are multi-functional invertebrate muscle proteins, whose roles may include host immunomodulation, and seem to be a prominent candidate in a schistosomiasis vaccine. We report here the cloning, expression and characterization of a B. microplus paramyosin (BmPRM). Sequence analysis of the full length coding sequence cDNA shows high identity to other arthropod paramyosin sequences, and the predicted molecular weight, pI and secondary structure are consistent with a typical paramyosin. Western-blot expression analysis indicates the presence of BmPRM in all tissues and developmental stages tested, but not in saliva. The recombinant protein (rBmPRM) was shown to bind both IgG and collagen. Possible implications of these activities with host evasion mechanisms are discussed.

Taenia cestodes were obtained from 5 different definitive host species in Kenya and 175 different samples were examined by classical morphological methods and by isoenzyme analysis using isoelectric focusing in agarose. Gels were stained for 17 different enzymes and 3 of these were used in the construction of isoenzyme profiles. The samples fell into 25 zymodemes, and no zymodeme contained more than 1 species of Taenia, indicating that isoenzyme analysis can reliably be used for the identification of species of this genus.

Data from a long-term study of the intestinal helminth parasite community of eels, Anguilla anguilla, stocked into the shallow eutrophic Neusiedler See, Austria, were collected over an 8 year period (1994–2001). In total, 720 eels from 2 sampling sites were examined. The parasite community showed characteristics similar to those in the natural eel populations in rivers of the UK and mainland Europe: it was species poor, with only 5 species (Acanthocephalus lucii, Acanthocephalus anguillae, Raphidascaris acus, Proteocephalus macrocephalus, Bothriocephalus claviceps) comprising the component community and a maximum infracommunity richness of 4 species. Over the period, the intestinal parasite community of the sampling site in Illmitz, which was originally dominated by A. lucii, changed. As levels of A. anguillae increased to a point at which it dominated the community, diversity increased whilst dominance of a single species decreased. By contrast the community in the southern sampling site remained rather constant with a continuous high infection level of A. anguillae and low abundance of A. lucii. Both acanthocephalan species exhibited higher infection levels in larger eels and in different seasons of the year and the infection parameters were significantly different between the years of study. The significant differences in the infection levels of the 2 acanthocephalan species at the 2 sampling sites were surprising as both acanthocephalan species use the same intermediate host, Asellus aquaticus, and the sampling sites were in close proximity and were similar in terms of water quality, host size and invertebrate abundance. Differences in the fish communities of the 2 sampling sites and eel movements rather than interspecific competition are discussed as possible explanations for the differences in the parasite communities of the 2 sampling sites.

In a previous paper (Johnson & Mellanby, 1942) an account was given of the numbers ot adult female Sarcoptes and their distribution in 886 cases of scabies. These cases were all adult males and it was desired to complete the study by a similar analysis of the parasitic infection in women and children. The following is an account of the numbers and distribution of mites in 119 women and eighteen children. Owing to the small number of children these figures are only suggestive. As described in the previous paper the mites are first located with the help of a watchmaker's lens and the parasite is then extracted with a mounted needle. Only adult females are extracted. The number of parasites per case is called ‘the parasite rate’.

Plasmodium knowlesi (W1) shows a high level of innate resistance to mefloquine as shown by recrudescence of blood-induced infection in rhesus monkeys, following oral mefloquine treatment with up to 80–160 mg/kg total dose over 3–4 days. Ketoconazole, at a dose of 75 mg/kg/day×10 days given orally to P. knowlesi-infected rhesus monkeys, exerted a suppressive antimalarial effect in 1 of the 2 monkeys as evidenced by a delayed recrudescence (on day 30) and curative action in case of the other monkey, whereas a lower dose of ketoconazole (25 mg/kg/day×10 days) failed to suppress infection effectively. When ketoconazole at a low dose (25 mg/kg/day×10 days) treatment was combined with mefloquine (20 mg/kg/day×4), 1 of the 2 infected monkeys was cured, while the combination of a higher dose of ketoconazole (75 mg/kg/day×10 days) with mefloquine (20 mg/kg/day×4), had a complete curative effect on P. knowlesi infection in all the treated monkeys (7/7). Besides exerting direct antimalarial action against a mefloquine-resistant P. knowlesi, ketoconazole also acts as an inhibitor of hepatic microsomal CYP4503A, which regulates and slows down mefloquine biotransformation to carboxymefloquine, resulting in a possible elevation of the effective plasma drug level, which seems to be responsible for elimination of the resistant P. knowlesi. The study shows a resistance reversal effect of a ketoconazole – mefloquine combination that could be exploited for the control of mefloquine-resistant malaria infection. The study shows that ketoconazole can reverse mefloquine resistance of P. knowlesi. These findings can also be applied to reverse mefloquine resistance of P. falciparum in areas where mefloquine resistance is already established. The precise role of parasite CYP450 in mefloquine resistance and its possible role, if any, in expelling the intracellular mefloquine from the resistant plasmodia still remains to be elucidated.

Genetically determined variation in host capacity to express resistance to a given parasite plays a major role in determining the outcome of infection. It can be assumed that the same is true of variation in parasites, but very much less is known of its influence on the host–parasite relationship. Phenotypic and genotypic variation within species of intestinal worms is now well documented, detailed studies having been made of parasites such as Ascaris in humans and trichostrongyles in domestic animals. However, the extent to which this variation affects the course of infection or the host immune response in these hosts is limited. Of the nematodes used as experimental models in laboratory rodents, detailed data on phenotypic or genotypic variation are limited to Strongyloides and Trichinella. Parasite variation is known to be subject to host-mediated selection, the emergence of anthelmintic resistance being a good example. Repeated passage has been used to select lines of parasite that survive in abnormal hosts or which show adaptation to host immunity. Experimental studies with Trichinella genotypes in mice have demonstrated the extent to which parasite variation influences the nature and degree of the host's immune and inflammatory responses, the complex interplay between immunogenicity and pathogenicity influencing both partners in the relationship. Recent studies with isolates of Trichuris muris have shown how parasite variation influences the capacity of mice to express the T helper cell responses necessary for resistance. Molecular differences between T. muris isolates have been shown in their excreted/secreted products as well as at the level of their DNA. Knowledge of the functional consequences of parasite variation will add to our understanding of host-parasite evolution as well as providing a rational basis for predicting the outcome of controls strategies that rest on the improvement of host resistance through vaccination or selective breeding.

Miltefosine was investigated for its activity against Neospora caninum tachyzoites in vitro, and was shown to inhibit the proliferation of N. caninum tachyzoites cultured in human foreskin fibroblasts (HFF) with an IC50 of 5·2 μM. Treatment of infected cells with 25 μM miltefosine for a period of 10 h had only a parasitostatic effect, while after 20 h of treatment parasiticidal effects were observed. This was confirmed by transmission electron microscopy of N. caninum-infected and miltefosine-treated HFF. Administration of miltefosine to N. caninum-infected Balb/c female mice at 40 mg/kg/day for 14 days resulted in 6 out of 10 mice exhibiting weight loss, ruffled coat and apathy between days 7 and 13 post-infection. In the group that received placebo, only 2 out of 8 mice succumbed to infection, but the cerebral burden was significantly higher compared to the miltefosine treatment group. In a second experiment, the time-span of treatment was reduced to 5 days, and mice were maintained without further treatment for 4 weeks. Only 2 out of 9 mice in the miltefosine treatment group exhibited signs of disease, while 8 out of 10 mice succumbed to infection in the placebo group. These results showed that miltefosine hampered the dissemination of parasites into the CNS during experimental N. caninum infection in mice.

A Theileria parva lawrencei isolate in the form of a sporozoite stabilate, derived by feeding clean Rhipicephalus appendiculatus nymphal ticks on an African buffalo (Syncerus caffer) captured in the Laikipia District, Kenya, was inoculated into groups of cattle at dilutions between 100 and 10-3. Groups of 3 cattle infected with 1 ml inocula at 100, 10-1 and 10-2 dilutions were treated with 2·5 mg/kg body weight of buparvaquone on day 0 and similar groups were left untreated to act as controls. An additional group, given 100 dilution of the stabilate, was treated with buparvaquone on day 8 post-inoculation. It was found that all control cattle inoculated with the stabilate at dilutions between 100 and 10-2 became infected, but only 2 out of 3 cattle developed patent infections at 10-3 dilution. All 3 control cattle receiving 100 dilution died of theileriosis, 2 at 10-1 and 10-2 dilutions, and 1 at 10-3 dilution died. Buparvaquone treatment on day 0 at 100 dilution resulted in the survival of 2 of 3 cattle and all the cattle at 10-1 and 10-2 dilutions. All the surviving cattle eventually developed a significant serological response against T. parva in the indirect fluorescent antibody test, except 1 in the 10-3 dilution group, and were immune to homologous challenge when tested 3 months later with a lethal inoculum of stabilate, except 2 cattle in the 10-3 dilution group. As a result of a theileriosis problem at about day 60 after inoculation in 2 cattle given 10-2 dilution of stabilate and buparvaquone treatment on day 0, an additional 5 cattle were given 10-2 dilution of stabilate and developed a good immunity after buparvaquone treatment. None was shown to develop the carrier state. Treatment with buparvaquone on day 8 after infection with 100 dilution of stabilate was not successful since 2 died. The stabilate used was shown to produce reproducible infection in cattle at different dilutions.

1 Populations of Babesia rodhaini were isolated which were ‘putative’ variants of a reference population.

2. They were shown to have some antigenic dissimilarity to the reference population by comparing the response of rats immunized with irradiated parasites of the reference population, to challenge with either the reference population or with the variant populations. To the homologous challenge the immunized rats were strongly resistant. After a heterologous challenge, however, for the first 3 or 4 days the immunized rats showed no resistance but thereafter rapidly controlled the infection.

3. Five ‘variation’ populations were tested and all were shown to have some antigenic dissimilarity to the reference population. Two variant populations were compared and were not found to be identical with each other. There appeared, however, to be less dissimilarity between them and any one variant and the reference population.

Mrs J. Page and her successor Miss D. Leader, gave technical assistance. I thank my colleagues for helpful criticism of the manuscript.