A novel tetrahydroquinolinyl ester, quinfamide, administered orally in multiple doses for 3 days had an ED50, of 0·25 mg/kg/day (total dose 0·75 mg/kg) for eradicating Entamoeba criceti in hamsters in several tests. It was significantly more active by direct comparison than 3 commercially available amoebicides and at least as active as 2 other esters of the parent compound, 1-(dichloroacetyl)-1,2,3,4-tetrahydro-6-quinonnol. After administration of a single dose, ED50, calculations for quinfamide averaged 0·9 mg/kg. Quinfamide was considerably more active than the other tetrahydroquinolinols, diloxanide furoate and teclozan, and it was approximately 1·5 times more active than etofamide; a statistical significance between the latter 2 drugs could be demonstrated in one of 4 tests. Administered prophylactically, quinfamide was shown to protect hamsters from re-infection with E. criceti. It also inhibited propagation of E. histolytica in vitro at a concentration of 20 /tg/ml. No adverse effects were noted in rodents after a single dose as high as 10 g/kg. Daily administration to monkeys of doses up to 500 mg/kg for as long as 37 days produced no pharmacological aberrations during or after medication; haematological studies and urine analyses were normal and no gross or microscopical tissue changes attributable to quinfamide were observed. No toxicity was revealed following acute (2 g/kg) and chronic (500 mg/kg/day × 31 days) administration of the drug to dogs and rats, respectively.

The surface topography of the tegument of adult Schistosoma mekongi was studied by scanning electron microscopy (SEM). In comparison to other species of human schistosomes the male tegument lacks tubercles and, except in the gynecophoral canal, also lacks spines; instead the surface is composed chiefly of trabeculae of highly perforated ridges which give it the ‘spongy’ appearance. In addition, there are 3 kinds of papillae interspersed on the surface among the ridges. The first is a doughnut-shaped papilla with a central crater which is most abundant on the ventral surface of the anterior part, on the floor of the gynecophoral canal and on the dorsal-lateral aspect of the tail. The second is a pleomorphic papilla with irregular shape and size, which is scattered throughout the dorso-lateral aspect of the middle part of the body. The third type of papilla has a uniform hemispherical shape, possesses a cilium projecting from its apex and probably corresponds to the ‘sensory papilla’ found in other species. The tegument of the female differs from that of the male by having numerous short spines over the whole surface; however, the pleomorphic papillae are much fewer in number and the ridges are much less developed than those of the male tegument; complex trabeculae are absent.

A description is given of the stages of sporogony of Hepatozoon sciuri, which have been found in three wild fleas (Orchopeas wickhami) out of a total of nearly sixty specimens collected from squirrels in England. Although an attempt to infect the fleas in the laboratory, by feeding them on infected squirrels, failed, the above findings indicate that these fleas may act as vectors of the squirrel parasite.

We wish to record our thanks to Professor P. C. C. Garnham for his kind interest and constructive suggestions regarding this work. We are specially indebted to Mrs M. Vizoso and to Dr J. F. D. Frazer for sending us fleas and squirrels from time to time. Our thanks are due to the authorities of the British Museum of Natural History for the identification of the flea.

The distribution of glycogen has been studied in P. minutus by means of histochemical methods. The strongest staining for glycogen was detected in the proboscis, the radial layer of the body wall and the non-contractile parts of the muscles.

Histochemical methods have also been used to study the localization of the activity of ten oxidoreductase enzymes. The enzymes are situated in the mitochondria of P. minutus, the distribution of which corresponds closely with that of glycogen.

I am grateful to Dr P. Tate for helpful discussions, and to Drs D. L. Lee and F. P. B. Wooding for criticizing the manuscript. Thanks are also due to Mr D. Barnard for technical assistance.

Changes in the ultrastructure of the daughter sporocyst and contained metacercariae of Microphallus pygmaeus during culture in artificial seawater and modified Medium 199 were examined and compared with variations in oxygen consumption. The very early commencement and rapid progression of autolysis in the sporocyst body wall as compared with little or no change in most metacercariae in both media, together with the elevated oxygen consumption in the nutrient medium, suggests that most of the oxygen is consumed by the contained metacercariae. The almost simultaneous onset of body wall degeneration in both media suggests that the nutrient medium is not suitable for the maintenance of healthy daughter sporocysts.

In order to provide a better understanding of the interaction between the liver fluke (Fasciola hepatica) and the immune system of its mammalian host immunoreactive λ bacteriophage clones containing F. hepatica cDNA have been isolated. Plasmids from these clones were sequenced and found to encode a family of proteins containing certain common elementsThe nucleotide sequence data reported in this paper are available in the GenBank database with the accession number AF153056.. All the clones contained a coding repeating sequence (RRRXCA) which is conserved at the nucleic acid level followed by a non-repeating element coding for the C terminal used by the proteins which shows conservation of amino acids at certain positions. Antisera raised against a β-galactosidase fusion protein with one of these sequences as a terminal extension was used to localize the immunoreactive antigens. Binding was predominantly in the tegument of the juvenile fluke but was reduced in the adult tegument. The wall of the uterus showed strong reactivity in the adult. Rats immunized with the β- galactosidase fusion protein showed enhanced resistance to challenge infections. The role of these antigens in the host response to infection by F. hepatica is discussed.

Echinorhynchus gadi (Zoega) Müller has been found parasitic in various species of Pacific salmon. The amphipod Cyphocaris challengeri Stebbing serves as an intermediary host for this parasite in the region in which the investigations were carried out.

Echinorhynchus lageniformis sp.nov. and Corynosoma strumosum (Rudolphi) both parasitic in the Starry flounder (Platichthys stellatus (Pallas)) and the Two-lined flounder (Lepidopsetta bilineata (Ayres)) are recorded with data on their occurrence.

From lizards caught in the Codajaz region, Amazonas, Brazil, the following malaria parasites are described:

Plasmodium rhadinurum Thompson & Huff, 1944 from Iguana iguana L.

A species of Plasmodium from Ameiva ameiva L.

A species of Plasmodium from Cnemidophorus lemniscatus (Daudin).

I wish to express my thanks to the Instituto Nacional de Pesquisas da Amazonia (I.N.P.A.) at Manaus for their help in the collection of material; to Professor P. C. C. Garnham, C.M.G., F.R.S., for assistance in identification, and to Mr P. E. Nesbitt for advice on staining procedures. This work was carried out during the tenure of a Medical Research Council Scholarship for Training in Research Methods.

The aim of this study was to evaluate the effect of nerolidol free (N-F) and nerolidol-loaded in nanospheres (N-NS) on the hepatic antioxidant/oxidant status of mice experimentally infected by Trypanosoma evansi. In the liver it was measured: reactive oxygen species (ROS), thiobarbituric reactive acid substances (TBARS) and non-protein thiols (NPSH), catalase (CAT), superoxide dismutase (SOD) and glutathione-S-transferase (GST) and performed histopathological examination. In addition, seric levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured. Liver samples from mice infected by T. evansi showed increased (P < 0·05) ROS, TBARS, AST and ALT levels and SOD activity, and decreased NPSH levels and CAT activity (P < 0·05) compared with uninfected animals. N-NS treatment prevented (P < 0·05) ROS and TBARS increase, and increased NPSH levels, and ameliorate CAT and SOD activities on liver of infected mice. Moreover, N-NS treatment reduced (P < 0·05) AST and ALT levels, and prevented histopathological changes caused by the parasite. N-NS protected the liver from the oxidative stress caused by T. evansi, which might be due to its antioxidant properties. Nerolidol might be considered a promising therapeutic agent against oxidative stress, and nanotechnology is an encouraging approach to be explored.

Dientamoeba fragilis is an intestinal protozoan in humans that is commonly associated with diarrhoea and other gastrointestinal complaints. Studies conducted to investigate the biology of this parasite are limited by methods for in vitro cultivation. The objective of this study was to improve a biphasic culture medium, based on the Loeffler's slope, by further supplementation in order to increase the yield of trophozoites in culture. The current in vitro culture of D. fragilis is a xenic culture with a mix of bacteria. Three different liquid overlays were evaluated including Earle's balanced salt solution (EBSS), PBS and Dulbecco's modified PBS (DPBS), for their ability to support the in vitro growth of D. fragilis trophozoites. Out of these 3 overlays EBSS gave the highest increase in the trophozoite numbers. The effect of supplementation was analysed by supplementing EBSS with ascorbic acid, ferric ammonium citrate, L-cysteine, cholesterol and alpha-lipoic acid and quantification of in vitro growth by cell counts. A new liquid overlay is here described based upon EBSS supplemented with cholesterol and ferric ammonium citrate that, in conjunction with the Loeffler's slope, supports the growth of D. fragilis trophozoites in vitro. This modified overlay supported a 2-fold increase in the numbers of trophozoite in culture from all 4 D. fragilis isolates tested, when compared to a PBS overlay. These advances enable the harvest of a larger number of trophozoites needed for further studies on this parasite.

The status of bluegill (Lepomis macrochirus) and green sunfish (Lepomis cyanellus) as homologous hosts for the acanthocephalan Neoechinorhynchus cylindratus was experimentally determined. It was found that the adult parasite did not establish in bluegills, but that these fish could serve as paratenic host. In contrast, complete growth and development to the adult stage occurred in the green sunfish. When green sunfish were intubated with 10 cystacanths/fish, the parasite exhibited a clear preference for the anterior half of the intestine; when 50 cystacanths/fish were intubated, the parasites showed a preference for the posterior half of the intestine. With repeated exposure of cystacanths, the parasites were distributed throughout the intestine. The extent of histopathology induced by N. cylindratus was related to the numbers of parasites present. In light infections (10 cystacanths), the parasite penetrated deeply into the intestinal wall and connective tissue developed around the proboscis. In infections with 50 cystacanths, the proboscis penetration was shallow and little if any connective tissue accumulated. There was also an indication that in crowded areas, the parasites appeared to change their sites of attachment frequently. In both heavy and repeated infections, the parasites evoked significant goblet cell hyperplasia and substantial quantities of mucus covered the intestinal wall. It is suggested that the sticky covering and the presumed presence of antibodies in the mucus combined to create a protective barrier thereby reducing the numbers of parasites that could attach and become established.

The parasitic nematodes described in this paper were collected by Dr E. C. Faust during his stay in China, and sent to Prof. Hoeppli for identification and study. The total collection amounts to about 500 vials, the majority of which were collected in Peiping (Peking), mainly from birds.

The plerocercoid larvae of Grillotia heptanchi (Vaulle-geard) used in the present study were obtained from the right supra-orbital lateral line canal of a specimen of Merluccius merluccius (L.). Two larvae were located in the canal in the region of the frontal bone and two in the region of the nasal. All were alive and were found to expand and contract slowly on removal. Unfortunately, the exact locality from which the fish came is not known as it was obtained from a fishmonger at Cardiff, but it is fairly certain that it was caught off the west coast of the British Isles.

Most of the new species herein described were collected by me from skins in the Transvaal Museum, Pretoria, for the loan of which I am deeply indebted to Mr Austin Roberts.

Variability of immune responses is an essential aspect of ecological immunology, yet how much of this variability is due to differences among parasite genotypes remains unknown. Here, variation in immune response of the crab, Macrophthalmus hirtipes, is examined as a function of experimental exposure to 10 clonal cercarial lineages of the trematode Maritrema novaezealandensis. Our goals were (1) to assess the variability of the host immune reaction elicited by 10 parasite clones, (2) to test if the heterozygosity–fitness correlation, whereby organisms with higher heterozygosities achieve a higher fitness than those with lower heterozygosities, applies to heterozygous parasites eliciting weak immune responses, and (3) to see how concomitant infections by other macroparasites influence the crab's immune response to cercariae. Parasite clones were distinguished and heterozygosities calculated using 20 microsatellite markers. We found that exposure to cercariae resulted in increased haemocyte counts, and that although interclonal differences in immune response elicited were detected, parasite heterozygosity did not correlate with host immune response. Additionally, the presence of other pre-existing parasites in hosts did not influence their immune response following experimental exposure to cercariae. Overall, the existence of variability in immune response elicited by different parasite clones is promising for future ecological immunology studies using this system.

Cytochemical staining with diaminobenzidine (DAB) has been used to locate cytochrome oxidase activity in mitochondria from major tissues of three intestinal parasitic nematodes, Nippostrongylus brasiliensis, Haemonchus contortus and Ascaridia galli. Deposition of DAB reaction product was uniform and relatively intense in mitochondria from all tissues studied in N. brasiliensis. However, mitochondria from different tissues of H. contortus and A. galli varied considerably in their cytochemical staining, decreasing in the order hypodermis < muscle < gut < reproductive tissue. Mitochondria in gut and reproductive tissue of A. galli possessed little or no cytochrome oxidase activity. These results are compatible with respect to oxygen availability and the extent and localization of the mammalian-type aerobic mitochondrial electron transport pathway in intestinal parasitic nematodes.

The larval output of Angiostrongylus cantonensis at different initial densities in rats was examined throughout the reproductive life-span of the worms using a modified dilution technique. With an initial density of 2 worms the mean duration of larval output was 414·2 days. The output was at a level above 50000 larvae/female/day from 15 to 45 weeks after infection. The mean total output was 18·9×106 larvae/female. The larval output/female decreased with an increase in the worm burden and also with the passage of time after infection. The longevity of host rats also decreased with increasing worm density and many of the heavily infected rats died early during infection. These results suggest that worm density has an effect on the fecundity of this parasite, and that the effects may be due to fibrous changes in host lung tissue but not changes in the physiology of the worms. It is also suggested that density-dependent effects on fecundity play a role in the population dynamics of both the parasite and host as regulatory mechanisms in the field.

Reciprocal crosses (Schistosoma intercalatum male Zaire×S. intercalatum female Cameroon: S. intercalatum female Zaire×S. intercalatum male Cameroon) were produced in 10 mice by exposing each mouse to 60 male cercariae of one isolate and 60 female cercariae of the other isolate, and vice versa. Hybrid generations originating from the two crosses were established. The infectivity of the F1, F2, F3 and F4 hybrid generations were evaluated after exposing snails individually to 5 miracidia. A comparative histological study of snails infected with F2 and F4 hybrid sporocysts from both crosses was made to assess abnormalities in the intramolluscan development of the hybrids. The worm recovery rate and fecundity were measured by comparing the fitness of the mid-parents with that of the hybrids. S. intercalatum Cameroon was compatible with Bulinus forskalii and incompatible with B. globosus whereas S. intercalatum Zaire was compatible with B. globosus and incompatible with B. forskalii. In the case of S. intercalatum male Cameroon×S. intercalatum female Zaire, hybrid miracidia develops in both B. forskalii and B. globosus in F1, F2 and F3 generations. The infection rate was much lower for B. globosus and F2 and F3 generations produced few cercariae (less than 30 cercariae in overall per snail) and F4 miracidia were only infective to B. forskalii again producing few cercariae. At 40 days post-infection the sporocyst contains masses of acidophilic granules originating from the breakup of pycnotic nuclei. Similarly the F1, F2 and F3 miracidia of the reciprocal cross (S. intercalatum male Zaire×S. intercalatum female Cameroon) exhibited a dual infectivity for B. forskalii and B. globosus, but cercarial productivity was low (less than 30 cercariae overall per snail for F2 and F3 generations). Histological studies demonstrated sporocyst degeneration in snails infected with F4 generation. In the definitive host, the F1 generation (both crosses) exhibited hybrid vigour in that the worm return was greater than that of the mid-parent, F2 and F3 generations. No significant difference in fecundity was demonstrated between the parental strains and the F1 and F2 generations, yet egg production of the F3 generation was significantly lower. It is apparent that there is a post-zygotic barrier in the crosses of S. intercalatum Zaire and S. intercalatum Cameroon from the F2 generations onwards, and sterility of the F4 hybrid sporocyst supports the concept of the existence of 2 distinct taxa.