The relationship between antigens associated with the surface of newly transformed schistosomula of Schistosoma mansoni and the tegumental surface membrane of adult S. mansoni worms has been further explored. Immunoprecipitation of detergent-solubilized 125I-tegumental surface membrane antigens of adult S. mansoni with antibodies from mice vaccinated with highly irradiated S. mansoni cercariae revealed major antigens of Mr 32, 20, 15 and 8K. The Mr 32 and 20K antigens have been previously demonstrated to be antigenically and electrophoretically identical to major antigens on the schistosomulum surface. The Mr 15 and 8K antigens, on the other hand, have not been identified by the immunoprecipitation of 125I-schistosomulum surface antigens, although a distinct schistosomulum surface antigen of Mr 15K is precipitated by antibodies from mice vaccinated with highly irradiated cercariae. Nevertheless, it was shown that antibodies to the Mr 15 and 8K antigens were specifically absorbed from vaccinated mouse serum by intact, live schistosomula, demonstrating that the Mr 15 and 8K antigens are exposed on or released from the schistosomulum surface. In contrast, absorption of the antiserum with eggs failed to remove antibody against any of the four tegumental membrane antigens examined. The Mr 15 and 8K antigens were shown to be recognized via polypeptide epitopes and not periodate-sensitive carbohydrate epitopes, further emphasizing the similarity of these to the well-characterized Mr 32 and 20K tegumental surface membrane antigens. A general relationship between schistosomulum surface, adult tegumental membrane and egg antigens was demonstrated by ELISA, using antibodies raised against the three antigenic fractions. It was shown that both the egg and adult tegumental membrane antigens cross-react with the schistosomulum surface, but that the egg and adult membrane antigens exhibit very little, if any, mutual cross-reactivity. This antigen divergence possibly enables the host to dissociate pathological, anti-egg responses from potentially protective anti-membrane responses during the course of natural infection. It also suggests that adult membrane antigens could be used in an anti-schistosome vaccine without the possible complication of inducing pathological responses.

Sixteen species of mites belonging to nine different families were found on Mus musculus and Apodemus hebridensis and in the nests of the former. Of these, eleven species may be regarded as parasitic in habit, two species have previously been recorded in association with rodents and moles, and three species may be described as accidental visitors.

I wish to thank Mr Elton and Prof. Munro of the Imperial College for their help, and I am also greatly indebted to Dr Caiman and Dr Finnegan for allowing me to make use of the facilities afforded by the British Museum (Natural History).

Plant cysteine proteinases (CPs) from papaya (Carica papaya) are capable of killing parasitic nematode worms in vitro and have been shown to possess anthelmintic effects in vivo. The acute damage reported in gastrointestinal parasites has not been found in free-living nematodes such as Caenorhabditis elegans nor among the free-living stages of parasitic nematodes. This apparent difference in susceptibility might be the result of active production of cysteine proteinase inhibitors (such as cystatins) by the free-living stages or species. To test this possibility, a supernatant extract of refined papaya latex (PLS) with known active enzyme content was used. The effect on wild-type (Bristol N2) and cystatin null mutant (cpi-1−/− and cpi-2−/−) C. elegans was concentration-, temperature- and time-dependent. Cysteine proteinases digested the worm cuticle leading to release of internal structures and consequent death. Both cystatin null mutant strains were highly susceptible to PLS attack irrespective of the temperature and concentration of exposure, whereas wild-type N2 worms were generally resistant but far more susceptible to attack at low temperatures. PLS was able to induce elevated cpi-1 and cpi-2 cystatin expression. We conclude that wild-type C. elegans deploy cystatins CPI-1 and CPI-2 to resist CP attack. The results suggest that the cpi-1 or cpi-2 null mutants (or a double mutant combination of the two) could provide a cheap and effective rapid throughput C. elegans-based assay for screening plant CP extracts for anthelmintic activity.

Infective 3rd-stage larvae of Dictyocaulus viviparus were filmed whilst climbing sporangiophores of the coprophilous fungus, Pilobolus and invading sporangia in numbers up to about 50, shortly before the sporangia were explosively discharged. Like the sporangiophores of Pilobolus the nematodes reacted positively towards light. Robinson (1962) demonstrated the spectacular way in which D. viviparus can be dispersed by Pilobolus, although he saw the nematodes accumulating only on top of the sporangia. Hazards including desiccation would offset the advantage of dispersal unless protection is somehow provided. Free D. viviparus larvae are susceptible to desiccation and discharged sporangia appear to dry quickly, so what protection, if any, operates? Several other nematodes that are susceptible to desiccation survive because structures surrounding them dry quickly (for example, an ensheathing cuticle, egg-shell, gelatinous ‘egg-sac’ matrix) and in doing so form barriers which limit the rate of water loss from the nematode within. Such dried sporangia of Pilobolus could aid survival of the ensheathed D. viviparus larvae inside them by retarding desiccation of the nematodes, in much the same way that within cysts of Globodera and Heterodera spp. the cyst wall, egg matrix and outer eggs protect the inner eggs by reducing the rate, if not the extent, of water loss. The sporangium wall may also protect the nematodes from harmful ultraviolet irradiation and from being washed by rain into less advantageous situations.

1. A method, based on that devised for E. histolytica by Dobell (1947), was worked out for testing the action of drugs upon E. invadens.

2. The sensitivity of di- and monobacterial strains of E. invadens to emetine was determined, and of a dibacterial strain to the quinaldines A 352, A 356 and A 404. E. invadens was killed by emetine at concentrations of 1/50,000–1/100,000, A 352 or A 356 at 1/100,000, and A 404 at 1/10,000. The strains of E. invadens used in the present tests are thus less sensitive to emetine than is E. histolytica, but the sensitivity of the two species to the quinaldines is similar.

3. A strain of E. invadens which was maintained for 9 months in the presence of increasing concentrations of emetine, did not become resistant to the drug.

I am indebted to Dr A. Bishop for much advice and criticism; to Dr J. D. Fulton for a culture of Bacterium coli A'; and to Dr J. Walker for supplies of the compounds A 352, A 356 and A404. The work has been financed by the Medical Research Council.

Metacestodes of Hymenolepis diminuta affect several aspects of female reproductive physiology in Tenebrio molitor and such effects are mediated via the endocrine system. The effects on male reproduction are less well known and were studied with respect to the Bean-Shaped Accessory Glands (BAGs). The size and wet and dry weight of BAGs from infected and uninfected beetles were compared and rose to a plateau from 0–6 days post-emergence in uninfected beetles but in infected individuals continued to increase in both size and weight. These effects were density independent. Glands from both infected and uninfected beetles were assayed for trehalase activity measured by its ability to convert the sugar trehalose to glucose. The activity of this enzyme, per mg wet weight, was not affected by the parasite. However, total activity per gland increased in infected males. Total protein content and electrophoretic profiles of BAGs from infected and uninfected individuals showed no change in profile but showed an increase in all protein subunits per gland over a broad molecular weight range.

Six stocks of Trypanosoma congolense were cloned from 17 stocks isolated from Eastern Zambia and used to initiate insect-form in vitro cultures producing metacyclic trypanosomes. Serological assays were then developed using these in vitro-derived metacyclics as a reference collection of antigens. Monoclonal antibodies recognized 8 metacyclic variable antigen types (M-VATs) of one stock, T. congolense TREU 1885, representing 70–80% of that stock's M-VAT repertoire, and in an indirect fluorescent antibody test (IFAT) there were no cross-reactions between them and the metacyclic trypanosomes of the other 5 stocks. Cross-protection assays between the 6 stocks in mice showed that the stocks cultured in vitro were serologically distinct. In order to facilitate serological typing for serodeme characterization, an IFAT was developed using formalin-fixed metacyclic trypanosomes to identify VAT specific immune responses using 21 day post-infection antisera. The cultured stocks reacted only with their homologous antisera thus confirming the results obtained in the cross-protection assays. No cross-reactions were observed with the 6 cloned stocks and antisera against the 11 stocks of T. congolense isolated in the same area at the same time suggesting that these stocks were different from the reference collection of cultured metacyclics. Hence, at least 7 serodemes of T. congolense have been identified from the 17 stocks isolated.

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.

Plasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200000 and 180000 were detected only after extraction with deoxycholate or SDS.

Culture characteristics of two species of piscine trypanosomes were compared. Trypanosoma catostomi from Catostomus commersoni was isolated in hypo-osmotic blood agar media but not in hyper-osmotic media. However, T. phaleri from Amia calva was isolated in both media. Only T. catostomi was stimulated to divide by diluting infected blood with water. The maximum number of T. phaleri and percentage of trypomastigotes were not affected by NaCl concentration. The maximum number of T. catostomi was inversely related to osmotic pressure and the percentage of trypomastigotes was directly related to osmotic pressure. This was confirmed by using equi-osmotic concentrations of NaCl, NaBr, Na acetate, KCl, choline-Cl or sucrose. Supplementation with urea had no effect on the number or morphology of trypanosomes. The survival of T. catostomi following cryopreservation was enhanced by prior incubation in hyper-osmotic media. Differential culture characteristics of these species may reflect adaptation to different host–vector systems.

The antigenic differences and similarities between culture forms of Trypanosoma (Schizotrypanum) hedricki, Trypanosoma (Schizotrypanum) myoti, Trypanosoma (Schizotrypanum) vespertilionis and Trypanosoma (Schizotrypanum) dionisii, all isolated from bats, were compared using the double-diffusion technique. Antiserum to each trypanosome species was produced in rabbits by inoculating them with sonicated culture forms. Two isolates of T. hedricki were antigenically identical to each other, as were 2 isolates of T. myoti. There were several common antigens between the species. However, at least 1 antigen of each species was distinct from those of the other species.

In September, 1908, an opportunity was afforded me of examining some of the larger food-fishes brought in to Aberdeen fish market. For this I have to thank the courtesy of Professor J. Arthur Thomson, who kindly provided the necessary laboratory accommodation, and was of great assistance to me in various other directions. I have also to thank the Government Grant Committee of the Royal Society for defraying the expenses of procuring specimens.

A comparison was made of the immunogenicity of normal worms, derived from a primary infection, and immune-adapted worms, derived from a trickle infection, after surgical transfer into previously uninfected rats. Exposure to 300 normal worms for 2 days was sufficient to induce resistance to challenge infection 3 weeks later in four of eight rats. The immune-adapted worms were, however, significantly less immunogenic, only one rat in ten developing resistance to challenge after 21 days exposure to 300 worms.These results offer further evidence that the protective antigens produced by normal worms are in some way altered in adapted worms, so that adapted worms have a reduced immunogenicity.

The inhibitory effect of niridazole on hydrogen production by metronidazole-resistant (CDC-85) and susceptible (Cl-NIH) Trichomonas vaginalis strains was investigated. The results show that niridazole is more effective than metronidazole in inhibiting hydrogen production by the resistant isolate. In CDC-85 aerobic inhibition requires a 4-fold increase in metronidazole concentration compared with that required anaerobically, but the corresponding factor for niridazole is only 1·5-fold. Reduction of the drug by a hydrogenosome-enriched preparation gave rise to a multiline electron spin resonance detectable signal, which is due to a nitrogen-centred radical.

The distribution of helminth parasites within their host population is usually overdispersed and can be described by the negative binomial distribution. The causes of this overdispersion are poorly understood, but heterogeneity in the distribution of infective stages within the environment has been implicated as a possible factor. Here we describe the distribution of infective stages of the rat intestinal nematode parasite Strongyloides ratti among the faecal pellets of its host. The distribution of infective stages between faecal pellets is overdispersed and well described by the negative binomial distribution. This overdispersion increases during the course of infection and occurs over a range of infection intensities. Overdispersion of nematode infective stages among faecal pellets may result in increased spatial heterogeneity of the infective stages in the environment and thus may contribute to the generation of overdispersion of adult parasitic stages. In addition, these findings raise important issues regarding the accurate quantification of helminth egg counts from faecal samples.

Tubulin is the predominant phosphoprotein in Trypanosoma cruzi epimastigotes and is phosphorylated by a protein kinase CK2. Interestingly, the presence or absence of divalent cations affected the solubilization of a pool of the parasite tubulin and the CK2 responsible for its phosphorylation. This fraction of tubulin and its kinase co-eluted using phosphocellulose, DEAE-Sepharose and Sephacryl S-300 chromatographies. Anti-α tubulin antibodies co-immunoprecipitated both tubulin and the CK2 responsible for its phosphorylation, and anti-CK2 α-subunit antibodies immunoprecipitated radioactively labelled α and β tubulin from phosphorylated epimastigote homogenates. Additionally, native polyacrylamide gel electrophoresis of the purified and radioactively labelled fraction containing tubulin and its kinase demonstrated the phosphorylation of a unique band that reacted with both anti-CK2 α-subunit and anti-tubulin antibodies. Together, these results establish a strong interaction between a pool of the heterodimeric α/β tubulin and a CK2 in this parasite. Hydrodynamic measurements indicated that the T. cruzi tubulin-CK2 complex is globular with an estimated size of 145·4–147·5 kDa.

1. The distribution of filarial larvae in the dorsoventral and dorsal-longitudinal muscle fibre groups was studied in serial sections of mosquitoes of a susceptible strain of Aedes aegypti infected with Brugia pahangi or subperiodic B. malayi and of wild-caught Mansonia uniformis mosquitoes containing the latter parasite.

2. The experiments showed that more larvae were situated in the dorsal-longitudinal muscle than in the dorsoventral and that the difference in numbers was highly significant.

A comparison has been made of the attenuating effect on N. brasiliensis larvae of X-rays (HVL, 8 mm Al) and γ-rays from 60Co over the range 10–80 kr. In the dose range where attenuation was manifested as a decrease in total worm burden the γ-rays had an RBE (relative biological effect) of 0-7 relative to the X-rays as unity.

Because of the enhancement of infectivity occurring at low radiation doses, attenuation in this region is better assessed in terms of alteration to the sex-ratio of the parasite rather than effect on total population.

This work was supported by grants from the Agricultural Research Council and the Horse Race Betting Levy Board, and was carried out during the tenure by one of the authors (T. Kassai) of a British Council exchange scholarship.

Thanks are due to Dr T. Martin Young and Mr J. S. Orr of the Radiotherapy Department, Western Infirmary, Glasgow, for access to radiation sources and helpful advice.

1. The morphology and life history of a new haplosporidian parasite, Nephri-diophaga xenoboli sp.nov. from the gut of a millipede, Xenobolus carnifex are described.

2. The earliest stage hatched from the spore is a free uninucleate amoebula which penetrates the gut epithelium and grows into a multinucleate plasmodium.

3. The uninucleate gametes are formed by a process of endogeneous budding in the centre of the plasmodium and the sporoblast is formed as a result of fusion of two such gametes.

4. A characteristic cyst membrane enclosing about forty to fifty sporoblasts is formed immediately after fertilization is completed which is accompanied by a disintegration of the peripheral layer of the cytoplasm of the plasmodium.

5. Each sporoblast develops into a bivalved simple spherical spore, 3·0μ in diameter.

The presence of Ellobiopsis chattoni is recorded on Undinula vulgaris var. major for the first time.

The presence of this parasite in shallow tropical water, within a temperature range of 25·92–26·28° C. (salinity range 35·43–36·38‰) is recorded for the first time.

It is noted that the parasite was completely species specific to its host.

Table l analyses the incidence of the parasite in a population of Undinula vulgaris.

It was found that the parasite affected egg production in the ovaries. The suggestion is that it produces a starvation effect on the copepod, this effect being reversible.