An evaluation programme is presented for determining activity of unknown compounds on Ascaris suum larvae migrating into pulmonary tissues of animals. Data on gross lung lesions due to migrating ascarid larvae were obtained in a series of 44 screening tests consisting of 223 group observations from 1115 mice orally infected with A. suum. Lesion scores were graded in ascending order from 0 to 5 depending upon number and extent of pulmonary haemorrhages. In general, there was no significant difference in lesion scores between untreated mice and mice treated with unknown compounds in their ration. The programme is based on an initial 95% rejection of compounds with a 5% ability to retest compounds in groups of mice.

The grand mean score for 44 tests was 2·88. Ninety per cent confidence limits for means among tests established scores at 2·21 and 3·55, and among groups of mice at 1·76 and 4·00, When the test mean was less than 2·2, the ascarid ova used for infection were considered non-virulent. Compounds selected as active were in groups of mice having a mean lesion score of 1·76 or less. Banminth, Styquin and Tetramisole showed larvicidal activity. A reasonable compromise exists in this system wherein false leads are tolerated to avoid screening out effective materials. The host-parasite relationship is proposed as a valuable tool for anthelmintic evaluation.

The authors wish to thank Mr W. Henry and Mr M. Jackley of this laboratory.

1. Camallanus anabantis is recorded from Ceylon in Anabas testudineus, Ophicephalus punctatus, Puntius filamentosus and Rasbora daniconius. P. filamentosus, R. daniconius and O. punctatus are new hosts for this species.

2. Camallanus sweeti is recorded from Ceylon in Anabas testudineus, Ophicephalus punctatus, Ophicephalus striatus and Rasbora daniconius. One third stage larva was observed in each of O. punctatus and Clarias batrachus. O. punctatus, O. striatus and R. daniconius are new hosts for this species.

3. Procamallanus planoratus is recorded in Clarias batrachus, Ophicephalus punctatus and Ophicephalus striatus from Ceylon. O. punctatus and O. striatus are new hosts for this species.

4. Variations in the anatomy of the posterior ends of the three species are mentioned.

5. No Camallanidae were obtained from Macropodus cupanus, Puntius chola, P. dorsalis, P. vittatus, Panchax panchax, Nemacheilus botia botia, Etroplus macu-latus and Etroplus suratensis.

6. Reasons are given for suggesting direct infection of the host from the first intermediate host, although a ‘carrier host’ may exist.

Forty-one Trypanosoma cruzi stocks isolated from the Chilean vectors Triatoma infestans and Triatoma spinolai were characterized by pulse-field gel electrophoresis and Southern blotting with the cruzipain gene, and by schizodeme analysis of kinetoplast DNA with EcoRI and Msp I. Seven parasite groups were found by molecular karyotype which correlate with schizodeme and multilocus enzyme electrophoresis, supporting the concept of clonal propagation for Trypanosoma cruzi. A predominant T. cruzi stock was isolated from domiciliary T. infestans in several geographical areas of Chile. In contrast, other frequently found genotypes were circulating in the sylvatic and domestic transmission cycles of specific geographical areas. The greatest heterogeneity of T. cruzi stocks was found among sylvatic T. spinolai where at least 4 genotypes were obtained from a sample of 16 T. cruzi stocks.

A polymorphic set of 14 kDa excretory–secretory (E–S) antigen-encoding cDNAs, with similarity to a previously characterized 15 kDa E–S antigen of Haemonchus contortus, was cloned from Cooperia punctata. Five cDNAs encoding predicted proteins of 70–80% identity were sequenced. Nucleotide sequence data reported in this manuscript are available in the GenBankTM databases under the accession numbers AF352718, AF352719, AF352720, AF352721, AF352722, AF352723, AF352724, AF352724, AF352726 and AF352727. Genomic analyses of individuals proved the existence of three 14 kDa E–S antigen-encoding genes, excluding that the differences reflected polymorphisms between individuals in a population. Southern blots indicated the presence of additional members of this gene family. Thus, despite the fact that heterologously expressed C. punctata 14 kDa E–S products are shown to be recognized by immune sera, potential pitfalls in the development of a recombinant vaccine are presented by this genetic diversity. Vaccine design could be further rationalized by knowledge of the function, and possible redundancy in function, of the E–S products which is presently lacking. The limitations encountered in assigning a function to the 14/15 kDa family of E–S proteins that is thus far unique to the trichostrongyloid nematodes are discussed.

The effects of mitochondrial inhibitors on the in vitro development of Nippostrongylus brasiliensis have been studied in free-living and parasitic life-cycle stages. Mitochondrial inhibitors were chosen as being representative of established electron transport inhibitors and oxidative phosphorylation inhibitors and uncouplers of the classical mammalian respiratory chain. All mitochondrial inhibitors tested were highly effective in killing or retarding development of free-living stages of N. brasiliensis. Free-living stages were particularly susceptible to such inhibitors upon hatching of embryonated eggs to 1st-stage larvae. Concentrations of inhibitors effective against free-living stages were consistent with their level of inhibition against isolated mitochondria from embryonated eggs and 3rd-stage infective larvae. Results suggest an absolute requirement in the development of free-living stages for the mammalian-like respiratory chain and associated oxidative phosphorylation. Electron transport inhibitors were effective in retarding at least the initial development of 4th-stage larvae to adults, but only antimycin A and azide produced a lasting effect leading to worm death. Oxidative phosphorylation inhibitors and uncouplers were ineffective against developing parasitic stages of N. brasiliensis. Experiments on whole-worm respiration indicated that most electron transport inhibitors were able to penetrate the adult worm, but oxidative phosphorylation inhibitors were without effect on whole-worm respiration. Results suggest that the mammalian-like electron transport chain is a necessary requirement to adult N. brasiliensis, but oxidative phosphorylation in the adult worm may not be required for development and survival in vitro although it could be necessary to support the parasite in vivo.

In dibacterial cultures of E. invadens the size of the cysts was significantly influenced by different species of concomitant bacteria, and by the composition of the media. These factors appear to influence cyst size as a result of their effect on the growth rate of the amoebae; a higher multiplication rate was associated with a decrease in the size of the cysts.

I wish to thank Dr A. Bishop for criticizing the manuscript, and Mr W. McConnachie, jun., for assistance with the statistical methods. The work has been financed by the Medical Research Council.

Protease expression among TCI and TCII field isolates was analysed. Gelatin-containing gels revealed hydrolysis bands with molecular masses ranging from 45 to 66 kDa. The general protease expression profile showed that TCII isolates presented higher heterogeneity compared to TCI. By utilizing protease inhibitors, we showed that all active proteases at acid pH are cysteine-proteases and all proteases active at alkaline pH are metalloproteases. However, the expression of cruzipain, the T. cruzi major cysteine-protease, did not reproduce a heterogeneous TCII cysteine zymogram profile. Dendogram analyses based on presence/absence matrices of proteases and cruzipain bands showed a TCI separation from the TCII group with 50–60% similarity. We suggest that the observed cysteine protease diversification contributes to differential host infection between TCI and II genotypes.

The primary larva of the porocephalid pentastomid Subtriquetra subtriquetra, which infects the nasopharynx of Caiman sclerops, is uniquely free-living. Eggs hatch within the nasal passages and the emergent larvae can remain infective to the fish intermediate host for 4–5 days. During this period larvae exhibit very stereotyped behaviour, alternating between a characteristic stationary ‘fishing’ posture and very brief periods of movement. Larval activity increases significantly at 3 days, probably effecting dispersal to a more favourable site for host invasion. Host location is passive and invading larvae penetrate to the swim-bladder where subsequent larval development, which involves 7 moults, occurs. All 8 larval stages are freely mobile in the swim-bladder and possess hooks. Sensilla, disposed in characteristic patterns, are present throughout development, increasing in number to the infective stage; the majority of sensilla are located on sensory papillae.

While working on the life cycle of Cercaria mehrai Faruqui, 1930, the author obtained the adults from the intestine of experimentally infected white rats. The trematodes thus obtained were reported as belonging to the genus Paryphostomum Dietz, 1909. In view of the facts that the collar spines are 43 in number, the vitellaria are confluent in the post-testicular region and the cirrus-sac extends far behind the acetabulum, P. mehrai is transferred to the genus Artyfechinostomum Lane, 1915, which is, in author's opinion, a valid genus (Jain, 1959).

We describe a laboratory model using Eimeria falciformis and E. pragensis to investigate some of the interactions in double-species infections of Eimeria. Mice were given trickle infections by oral inoculation of 100 sporulated oocysts of one species at 3 or 4 day intervals throughout the experiments and, once immunity had developed, as indicated by cessation of oocyst production, the animals were challenged with a single inoculation of the other species. A trickle infection of E. falciformis gave a significant enhancement of oocyst output from E. pragensis infection as compared with animals that had not received E. falciformis. Histological examination of the mice infected with E. pragensis showed significantly more asexual parasites and a significantly higher female/male ratio in the mice that had received trickle E. falciformis infections than in those that had not. There was no evidence to suggest that extra asexual generations were occurring in these mice. In the converse experimental protocol, trickle E. pragensis infections had no significant effect on a single E. falciformis infection. We discuss the possible mechanisms for the interactions and also how these interactions may influence multiple species infections in animals in their natural habitats.

The salivary glands (Fig. 1) consist of two pairs situated between the anteriorly protruding coeca of the midgut. The one pair are bean- or kidney-shaped (b) the other horseshoe-shaped (h) and extend further backward close to the gut. The four ducts of these glands run forward along the oesophagus, unite, and form a common duct emptying between the dorsal and ventral chitinous stylets of the mouth-parts.

Nosomma monstrosum was originally described from a male specimen under the name of Hyalomma monstrosum by Nuttall & Warburton (1908), and the female was described some ten years later by Warburton (1918). Three other males were also seen by him at this time, and he was somewhat hesitant to assign these ticks to the genus Hyalomma. The genus Nosomma was proposed by Schulze (1919) to accommodate this monotypic species, but this appears to have been done without seeing the species and his reasons for establishing the genus appear to be insufficient. Chodziesner (1924) and Sharif (1928) concur on the validity of the genus and the latter defines the genus as follows: ‘eyes flat, scutum, legs and dorsal surface of the capitulum ornate, festoons present, palps short and broad, basis rectangular dorsally in both sexes, coxae I bifid, other coxae with two short spurs on each, ventral shields in the male represented by adanal, accessory and trilobed subanal shields; spiracles short, comma-shaped in the male and subtriangular in the female’. Delpy (1936) agreed in the separation of Cosmiomma and Nosomma from Hyalomma, and in respect of Nosomma stated that ‘Caractérisé par des palpes dont l'article III est plus large que long et par des ecussons subanaux tres volumineux’. Generic status was also accepted by Toumanoff (1944) and Santos Dias (1954).

Fischer rats infected with Fasciola hepatica showed significant resistance to Schistosoma mansoni challenge, and vice versa, whereas immunization with 20 Krad-irradiated S. mansoni cercariae failed to protect against F. hepatica challenge, but did protect against homologous challenge. When groups of rats received intraperitoneal implants of newly excysted juvenile flukes, 20- to 22-day-old juveniles, or 8- to 10-week-old flukes, none was significantly protected against S. mansoni challenge, whereas juvenile implants did protect against homologous F. hepatica challenge. In passive transfer experiments in rats, serum from F. hepatica-infected rats or rabbits protected recipients against homologous, but not heterologous challenge, and serum from rats vaccinated with 20 Krad-irradiated S. mansoni cercariae protected recipients against homologous, but not heterologous challenge. These experiments provide evidence that the mechanisms involved in homologous and heterologous resistance are different, the latter lacking immunological specificity. Microsphere injections in F. hepatica-infected rats demonstrated ‘shunting’ from the portal system to the systemic circulation. If migrating schistosomula are also ‘shunted’ in Fasciola-infected rats, this, rather than immunologically specific effector mechanisms, might explain their failure to establish in the portal system.

Cholesterol is a major component of the erythrocyte membrane. In the present study, we investigated the effects of cholesterol reduction in host bovine erythrocytes (RBC) on the growth of Babesia bovis, a major bovine haemoprotozoon. An in vitro growth assay with bovine RBC that had been prepared by pre-treatment with a cholesterol depletion agent (methyl-β-cyclodextrin, MCD) showed that the culture with 5 mm MCD-treated RBC inhibited the growth of B. bovis significantly as compared with that with the control RBC. In further experiments, the treatment with 5 mm MCD was proved to suppress both activities of the parasite, erythrocyte invasion and replication within the infected RBC. In contrast, a slight reduction in the membrane cholesterol by 1 mm MCD treatment promoted both their growth and erythrocyte invasion activity. These results indicate that erythrocyte invasion and replication by B. bovis are affected by the amount of cholesterol in the host erythrocyte membrane.

In a paper published last year, I described the female, nymph and larva of Ixodes caledonicus. Having regard to the views expressed in my recent paper on the adaptation of ticks to the habits of their hosts, I wrote to Mr William Evans, in Edinburgh, asking him, if possible, to kindly search the dove-cot at Dunipace, Stirlingshire, for the missing male which he would probably find in the habitat of the host, the domestic pigeon. On 25 May, 1911, Mr Evans discovered two ticks, a male and female in copula, in the dove-cot, and he sent the specimens to me for identification. The female accords with my published description of caledonicus. The male is here described for the first time, and it is interesting to find that it agrees in structure and habits with what was to be expected according to the theory advanced by me in the second paper I have referred to.

In order to learn more about the nature of the chemical bonds stabilizing the surface membranes of the cercariae and schistosomula of Schistosoma mansoni, these organisms were treated with a variety of reagents. The cercarial surface was dissolved by 8 M urea alone (pH 9·0) but was protected from such dissolution if a certain critical concentration of cation or anion was included in the 8 M urea solution. Divalent cations were more effective than monovalent cations in this protective effect. The 8 M urea was also unable to dissolve the cercarial surface at any pH below 5·8. The cercarial surface was very rapidly dissolved by sodium hydroxide solutions at pH 11·4 and above. The divalent cation calcium, if included in the sodium hydroxide solution, slowed down the rate of surface solution by sodium hydroxide. The schistosomular surface was stable to 8 M urea at pH 9·0 but was readily dissolved in 8 M urea at pH 10·7 and 11·7. The schistosomular surface was stable in aqueous sodium hydroxide solutions at pH 12·0. Anionic detergents were very effective in dissolving the cercarial surface at concentrations above 0·001%. Cationic and non-ionic detergents were ineffective at 0·01% or below. A saturated solution of digitonin if diluted 1:100 caused destruction of the cercarial surface. At higher concentrations it caused expansion and stabilization of this surface.

It is suggested that in both the cercarial and the schistosomular surface, bonds between carboxyl (—COO′) and amino (—NH3+) groups are very important in maintaining structure. The origin of the —NH3+ group is possibly from —NH2 lysyl group in cercariae, and a guanido group in schistosomula. The suggestion that cations are incorporated into the cercarial surface and stabilize it during and after penetration has been tested by treating schistosomula with EDTA. The EDTA had no visible effect on the surface, nor did it labilize the surface to other reagents.

I should like to thank very warmly Dr S. A. Ibrahim, in whose Department this work was carried out. My grateful thanks are given to the Medical Research Council (U.K.) for a grant. I should also like to thank Mr J. R. Lauder, Mr Ahmed Gandour and Mr Saleh for their help during the work. I am very grateful to Dr J. A. Clegg and Dr S. R. Smithers for helpful discussions.

This gregarine was found inhabiting the alimentary canal, and especially the midgut, of both larvae and imagines of Anobium paniceum. All the larvae examined, and most of the adult beetles, were found to be very heavily infected, and in some of the larvae almost every epithelial cell harboured one or more of the cephalont stages of the parasite (Fig. 1). The lumen of the midgut of these larvae was densely packed with the freely moving sporonts, and a number of newly formed cysts were visible in the hindgut. In the imago only the cephalont stages were observed, and no sporonts or cysts have been found.

RJLs represent a recently described family of the Ras-related GTP-binding proteins. The Trypanosoma cruzi orthologue, TcRjl, was isolated and its locus was characterized in a region of almost 5 kb. Its 660 bp orf, predicting a protein of 24·13 kDa, is present as a single copy gene in T. cruzi I lineage, and from 1–2 copies in T. cruzi II lineage. TcRjl shares 73% aa sequence similarity with its closest identified orthologue, T. brucei TbRjl. RT–PCR experiments revealed that TcRjl is transcribed in mRNA in the 3 main life forms of the parasite, while Northern hybridization demonstrated that TcRjl is transcribed in T. cruzi epimastigotes as at least 2 transcripts, one of around 950 nt and the other of 1500 nt. Splice-leader addition was mapped to a single site at −69 bp upstream of TcRjl orf indicating that the two mRNA types may derive in differences at the 3′ of TcRjl mRNA. TcRjl locus presents considerable synteny with Rjl loci from Trypanosoma brucei and Leishmania major as available from their respective genome projects.