The germ-cell cycle and method of reproduction in the daughter sporocysts of the bucephalid Cercaria bucephalopsis haimeana (Lacaze-Duthiers, 1854) and the gymnophallid Cercaria dichotoma Lebour, 1911 (non Müller) from the edible cockle, Cardium edule L., have been investigated by the examination of stained sections. A review is given of the principal interpretative theories which attempt to explain the nature of the reproduction and the homology of the germinal sacs of the Digenea. It is suggested that the life-cycle includes a cyclic alternation of homologous generations. Of these, the miracidium/mother-sporocyst and the daughter germinal-sac generations usually reproduce by apomictic parthenogenesis but polyembryony may also occur secondarily in some species.

We are very grateful to Dr Gwendolen Rees and Professor E. W. Knight-Jones for considerable help with the manuscript.

The structural relationship between cells of the parenchyma and those of the alimentary and excretory systems, and tegument are described.

Junctional complexes are formed in areas of close cell apposition. They occur laterally on invaginations of parenchymal processes which are embedded in the epithelial cells of the alimentary and excretory systems, and on flat plaques between parenchymal cells and the tegument.

The ultrastructure of the complexes has been resolved and a three-dimensional interpretation is put forward. It is suggested that the complex is a type of desmosome.

The findings reported here add support to the suggestion made in a previous paper that the parenchyma may function as a transport system in this fluke.

We wish to acknowledge grants from the Wellcome Trust and the S.R.C. for the purchase of an Akashi TRS 50 and an A.E.I. EM 6B electron-microscope respectively. This research and that reported in a previous paper were carried out while one of us (S.S.E.G.) was in receipt of a Training Studentship Grant from the Wellcome Trust.

We also wish to thank Mr W. Ferguson and Mr A. Lyness for photographic and technical assistance.

Additional ultrastructural features of the tegument of Fasciola hepatica, after glutaraldehyde fixation and high resolution, have been observed.

Two distinct types of tegumental cells are shown to be present, the type 1 cell having been previously described. Details of the fine structure of the nucleus, mitochondria, endoplasmic reticulum and Golgi complex of the type 2 cell are given. The type 2 cell is characterized by biconcave secretory bodies derived from the Golgi complex and the latter is intimately related to the granular endoplasmic reticulum, from which it is derived by blebbing. The type 2 cells are connected by protoplasmic tubes to the same surface syncytium as type 1 cells.

These new findings of the structure of the tegument of Fasciola are compared with previous reports on the ultrastructure of the tegument in other flukes.

I should like to express my sincere thanks to the following: the Wellcome Trust and S.R.C. for grants to purchase an Akashi TRS 50 and an A.E.I. E.M. 6B electron microscope respectively; the Royal Society for a personal grant to purchase a Reichert Ultramicrotome and a vacuum coating unit.

My thanks are also due to Mr W. Ferguson for photographic skill and to Mr A. Lyness for technical assistance and microscope maintenance, and my Research Associate, Mrs S. S. E. Dermott, for expert electron-microscopy.

Both histological and histochemical studies have been made of the food, mode of feeding, gut cell structure and digestive sequences in a selected number of digenetic trematodes.

The species investigated range from gut-dwelling trematodes feeding on the superficial epithelial tissues and associated mucoid secretions of the host to those forms living within the respiratory and circulatory system and feeding exclusively on blood.

The mode of feeding is suctorial, brought about by the muscular pharynx and normal attachment process of the oral sucker. In one case there is evidence that this purely mechanical process is supplemented with enzymic secretions produced by the trematode which have a histolytic effect upon host tissues.

On the basis of gut cell structure the species investigated are divided into those with cells and associated microvilli that vary in size and appearance so that the gastrodermal border is irregular in outline, and those with a gastrodermis comprising regular cuboidal or columnar cells bearing microvilli organized into a striated border. There is evidence to suggest that this difference in gut cell structure reflects differences in digestion. In all cases, gland cells are absent from the gastrodermis which is capable of both secretory and absorptive functions.

Digestion is predominantly an extracellular process, but the exact sequence varies according to the nature of the food ingested and different degrees of adaptation are shown by the trematodes to the blood-feeding habit. The tissue-feeding species show less modification.

I wish to thank Dr J. B. Jennings for helpful discussion and advice throughout the course of this work. The study was undertaken during the tenure of a Studentship from the Department of Scientific and Industrial Research.

Cercaria patialensis Soparkar, 1924, emerging from the snail Melanoides tuberculatus (Müller) has been encountered at Waltair. The cercariae begin to emerge from the snail at dusk and continue to do so during the period of darkness.

Stages resembling the adult but without the development of eggs have been encountered in nature on the fresh water fishes Panchax panchax (Ham. & Buch.) and Esomus danricus (Bleeker). These stages have also been raised experimentally by exposing P. panchax, E. danricus and fingerlings of Catla catla Ham. & Buch. to the cercariae.

The cercaria is referable to the genus Transversotrema Witenberg, 1944.

It is suggested that there may not be a metacercarial stage in the life-cycle of Transversotrema spp.

The effect of hexachlorophene, a fasciolicide, on some enzyme systems in the liver fluke was studied. The enzymes were adenosine triphosphatase, succinate oxidase and cholinesterase. Adenosine triphosphatase was activated, whereas succinate oxidase and cholinesterase were inhibited when homogenized flukes were incubated in some concentrations of hexachlorophene. Of the three enzyme systems studied in flukes killed by hexachlorophene, only the succinate system showed reduced activity, whereas the other two showed almost normal activities. The study indicates that biochemical changes are associated with the effect of hexachlorophene on the liver fluke.

Thanks are due to Dr C. Grant for critical examination of the language, to Miss Maarja Kippar for valuable help with the experiments and to Mrs Astrid Holmström for the typing of the manuscript.

This work was supported by grants A 1108/B 904 and A 1635/B 1299 from Jordbrukets Forskningsråd.

Large number of fishes of the family Cryprinodontidae collected from a stream at Hastings, Sierra Leone, were found with a diplostomulum encysting on the skin and in the musculature.

Infected pieces of fish were fed to day-old chicks and cattle-egrets, and adult stages of a worm, considered to be new species and named Diplostomum (Dolichorchis) leonensis, were obtained.

Morphological details of the diplostomulum and the adult worm are described.

The author wishes to express his gratitude to Dr G. Dubois for his interest and advice in connexion with the study, to Miss M. R. Coker who prepared the diagrams; and to Messrs F. Deen, T. Alhassan and M. Seisay for technical help.

Cercariae of Psilotrema oligoon and Notocotylus attenuatus encyst free to form multilayered, hemispherical cysts. The former consists of four layers in which the innermost (layer 4) is divided into two, chemically and structurally different, areas. Cysts of the latter have three layers in which the innermost (layer 3) is again divided into two chemically different parts. A third cyst, produced by the encystment of Cercaria tarda in the tissues of Limnophilus flavicornis, is spherical and contains three layers (of which the outermost was not studied).

All of the cysts studied contain a high proportion of protein, which in some layers occurs combined with carbohydrates. In both free-encysting species the two outer layers are produced by expansion of a non-cellular layer covering the integument and the innermost layer is produced by secretion of bâtonnets from a layer of cells lying beneath the body wall. Cysts of P. oligoon are divided into two separate parts, each having two layers, whereas those of N. attenuates consist of a single three-layered wall. The innermost layer in both consists of a protein with disulphide and sulphydryl linkages, and in the cyst of P. oligoon it has a circular ventral area containing carbohydrates. A similar carbohydrate-containing area occurs in the innermost layer of the cyst of N. attenuatus but as a ventro-lateral ring of material. The cyst of Cercaria tarda is also predominantly proteinaceous and stains intensely for RNA.

The present material of buffalo–dog origin could not be transmitted to the laboratory animals, such as white rats, white mice, rabbits, golden hamsters, guinea-pigs and chicks.

Secondary development took place only in mice.

The strobilar phase was infective to a cat but did not show sexual differentiation even at 30 days, and worms were further observed to be at a degenerative stage.

The buffalo was highly suited to the perpetuation of the parasite, forming larger cysts and massive production of scoleces; nearly 90% of the cysts were fertile.

The strobila had mostly two segments and not more than three.

The number and distribution of testes were more posterior to the genital pore and were also observed posterior to the vitellaria, occasionally.

The position of the genital pore in the gravid as well as mature segments was both anterior and posterior but with an average of 52.8%.

Uterine sacculations were observed to range from 28 to 54.

Echinococcus material of buffalo–dog origin does not appear to fall into a definite ‘subspecies’ pattern. It is suggested that it be merely recognized as a further ‘mutant’, the existence of which can be accounted for on theoretical grounds.

The authors are thankful to the Director, Indian Veterinary Research Institute, and to Dr H. D. Srivastava, Head of Division of Parasitology, I.V.R.I., for providing the necessary laboratory facilities.

The effect of an amino acid imbalance in the diet on the growth of Hymenolepis diminuta in rats was reinvestigated. The claims of earlier workers that growth of the tapeworm is retarded was not substantiated.

The size frequency distribution of 575, 13-day-old H. diminuta recovered from 9- and 10-worm infections was determined, to reveal the shape of the distribution and total variance. The total variance in worm weights was analysed to determine the amount attributable to variance ‘between rats’ (comparison of the mean weight of the worms recovered from rats) and variance between worms within individual rats. ‘Between rats’ variance was found to account for > 80% of the total variance.

Assuming variance equal to that in the experiments described, the degree to which experiments can determine a difference in effect between two diets on the growth of tapeworms is analysed, and the degree to which this is altered by increasing the number of rats in dietary groups and/or decreasing the ‘between rats’ variance is estimated.

The suitability of various procedures for host dietary experiments on tapeworms is discussed; these include the duration of the experiment, feeding regime, age of worm and the relative advantages of weighing worms singly or in groups.

We gratefully acknowledge the technical help we have received from Miss Patricia Grant and Mr Jack Keys. Our thanks are also due to Mr Robert Elton of the University of Glasgow Statistics Department who has advised us on statistical procedures and calculated Table 6.

Two new species of Ardeicola Clay, 1925 (Mallophaga, Ischnocera), A. keniensis and A. africanus, ectoparasites of African ibises, Lampribis olivacea akleyorum and L. rara respectively, are described. These species are closely related to each other, and the characters which distinguish them have been enumerated.

We are most grateful to Dr Theresa Clay, Department of Entomology, British Museum (Natural History), for the loan of material and for reading the typescript.

Cercaria helvetica XXXIII Dubois and the cercaria of Sphaerostoma bramae (Müll.) are redescribed and their metacercariae, which occur in various invertebrates, are described. Additional observations are made on C. parvus Khan and C. tarda Khan and their metacercariae, occurring mainly in arthropods, are described. C. wentloogensis sp.nov. has four pairs of penetration glands, a slender, thickened stylet 0·03 mm long and the flame-cell formula

2[(2 + 2 + 2) + (2 + 2 + 2)].

C. rumniensis sp.nov. has four pairs of penetration glands lying dorsally to the acetabulum, a slender thickened stylet 0·03 mm long and at least 20 flame cells arranged in pairs. C. octoglandulata sp.nov. is a virgulate species with four pairs of penetration glands, a partially thickened stylet 0·029–0·032 mm long and the flame-cell formula 2[(3 + 3 + 3) + (3 + 3 + 3)]. Each of these new species develops in sporocysts in the digestive gland of the prosobranch mollusc Bithynia tentaculata. A cercaria occurring in the digestive gland of B. tentaculata and assigned to the Microphallidae on the structure of its metacercaria is considered to encyst in the digestive gland without emerging. It bears no stylet or penetration glands and the cysts occur in the snail in exceedingly large numbers.

All of these cercariae were found in freshwater molluscs collected at the Wentloog level near Cardiff, South Wales.

A tenfold enhancement of resistance to primaquine was obtained by maintaining a strain of Plasmodium gallinaceum in a state of acute infection by serial passage of infected blood through young chicks treated with gradually increasing doses of the drug.

No loss in resistance to primaquine was observed when the resistant strain was transmitted through mosquitoes, though there was some loss in resistance to the maximum tolerated dose, but not to lower doses, when the strain was maintained in a state of acute infection through untreated chicks for 41 weeks.

The primaquine-resistant strain was cross-resistant to lower effective doses of pamaquin, and slightly less sensitive to quinine than the parent strain but the loss in sensitivity to chloroquine was only marginal. Sensitivity to proguanil, dihydrotriazine and pyrimethamine was normal.

Attempts were made to produce a chloroquine-resistant strain of P. gallinaceum using different doses of the drug but no change in sensitivity was observed though the experiments were continued for more than a year. An attempt to produce a chloroquine-resistant strain by treatment of a proguanil-resistant strain with chloroquine also failed.

The problem of resistance to quinoline compounds in different species of Plasmodium is discussed.