Sporulated oocysts or oocyst hulls of Eimeria acervulina given via the crop, and oocyst juice or sporozoites inoculated directly into the in testinal lumen, were tested for their effects on the hypersensitivity of the intestines of immunized and susceptible birds. Only viable sporozoites consistently caused an increase in the permeability of the immunized intestine to macromolecules, shown by the leakage of intravenously injected dye. Repulsion of epithelium and disruption of the villi were seen more often in immunized than in susceptible chickens but there was no correlation with treatment or with dye leakage.

The hypersensitive response of immune intestines challenged with a heterologous organism was approximately half as intense as the response to homologous challenge.

The significance of these findings in relation to immunity to coccidiosis is discussed.

A method for the biochemical identification of protozoa belonging to the genus Eimeria is described for the first time. Starch gel electrophoresis of the enzymes lactate dehydrogenase, glucose phosphate isomerase, 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase from parasite extracts revealed both intra- and inter-species differences when 11 strains representative of 6 species of Eimeria were examined. Oocysts were the most accessible parasite stage for investigation but sporozoites and merozoites of an embryo-adapted strain of E. tenella were also examined for enzyme activity.

Populations of Schofield strain, random-bred mice were shown to have a bimodal variation in ability to bring about immune expulsion of the nematode Trichuris muris. This variation was genetically determined and independent of the size of infection experienced. The proportion of mice unable to achieve worm expulsion (non-responders) was relatively constant in various populations of the strain but was increased by selective breeding from mice of known status. Crosses made between non-responder and responder mice produced progeny that were almost all (92%) of responder phenotype, showing that the ability to achieve worm expulsion was inherited as a dominant characteristic. It is suggested that the genetic control involves a small number of genes; the possible immunological mechanisms by which control is mediated are briefly discussed.

A series of experiments is described in which two drug-resistant strains of Eimeria maxima were passaged together in untreated chicks. The resultant oocysts were then inoculated into chicks treated with both drugs. When strains resistant to methyl benzoquate and sulphaquinoxaline or clopidol and sulphaquinoxaline were used the resultant infections were not controlled by the double treatment, indicating the acquisition of resistance factors by one strain from the other. When strains resistant to clopidol and methyl benzoquate were used the phenomenon was not observed.

Soluble glycoproteins have been purified from a series of clones of Trypanosoma brucei 427. Each clone yielded a characteristic predominant glycoprotein which induced clone-specific immunity to trypanosome infection in mice. These glycoproteins were shown by specific labelling and enzyme digestion of cells to be the major components of the trypanosome surface coat. Each glycoprotein consisted of a single polypeptide chain having an apparent molecular weight of 65000 (as measured by SDS-polyacrylamide gel electrophoresis) and containing around 600 amino acid and 20 monosaccharide residues. Preliminary structural studies indicated large changes in amino acid sequence dispersed over a considerable length of the polypeptide chain. Proteolytic activity was demonstrated in semi-purified trypanosome extracts, providing one reason for the heterogeneity sometimes observed in surface glycoprotein antigen preparations.

Observations have been made on the hatching behaviour of the larva of the monogenean Entobdella soleae and on the effects of various enzymes on the opercular cement of the egg. These observations indicate that hatching is brought about by a proteolytic hatching fluid which is produced by two pairs of ventral head glands and spread over the inner surface of the opercular joint by rotation of the larva about its longitudinal axis. After hatching there is no trace of the ventral head glands. Similar glands are present before hatching in the oncomiracidium of E. hippoglossi.

The erythrocytic stages of Plasmodium falciparum from Aotus trivirgatus were grown in Mill Hill medium. Significant invasion of erythrocytes was obtained by a subculture technique. This invasion, by parasites grown in Aotus erythrocytes, was greater when human erythrocytes were added to the cultures than when parasites were allowed to invade Aotus erythrocytes. This difference appeared to be a function of the properties of the erythrocyte and was not serum dependent.

The ultrastructure of the seminal vesicle, ejaculatory duct, cirrus sac and cirrus is described. The epithelium of the seminal vesicle consists of a single layer of squamous to cuboidal cells. The apical ends of the cells have thin polymorphic lamellae and long narrow pits, both of which enclose normal spermatozoa. The cells have a moderate amount of GER and Golgi complexes which produce a lucid secretory body. The ejaculatory duct epithelium is composed of cuboidal to columnar cells between or through which project the terminal parts of the ducts of the unicellular prostate glands. The apical surfaces of the epithelia are extended into triangular or filiform projections having thin sinuous lamellae. The cytoplasm contains GER cisternae and Golgi complexes which synthesize a dense ovoid secretion. The cirrus sac and cirrus are covered by a thin modified tegument. The cirrus has many spines and the normal ratio of T1 to T2 type of secretory bodies, whereas the cirrus sac has few spines and the T2 type of secretory body predominates over the T1 type. The significance and possible functions of the structures observed in the three tissues are discussed.

Protopolystoma xenopodis (Price, 1943) Bychowsky, 1957 is a parasite of the African clawed toads, species of Xenopus. Adult worms in the host's urinary bladder produce up to 15 eggs/24 h; these are voided with the urine and hatch after 20–24 days (at 26 °C). The swimming oncomiracidia enter the cloaca of the toad and travel via the urinary ducts to the kidneys. Larval development in the kidneys lasts 8–12 weeks (at 22 °C) and the juvenile parasites then migrate to the urinary bladder where maturity is reached after a further 3–4 weeks, i.e. 3–4 months after invasion. Egg production is continuous and parasite longevity has been found to exceed1½ years. Rarely, the adult worms may undertake a return migration from bladder to kidneys.

The morphology of the oncomiracidium is described and compared with available accounts of other polystomatid genera. Consideration of data for the genus Polystoma suggests that the arrangement of the epidermal ciliary cells is relatively constant at the inter-specific level. Comparison of Protopolystoma with Polystoma, Diplorchis and Oculotrema reveals that whilst the ciliary cell patterns are clearly related there are important differences in both cell number and disposition at the inter-generic level. Differences are also evident in the distribution of epidermal sensillae in Protopolystoma and Polystoma. Protopolystoma is distinguished from all other polystomatids infecting anuran hosts by the presence of two pairs of haptoral hamuli.

Oncospheral agglutination given by sera immunized with Hymenolepisnana eggs is described as a new way of assessing H. nana infection. All sera of mice which possessed acquired protective immunity against reinfection by H. nana eggs had the potency to induce oncospheral agglutination in vitro. Only oncospheres, which had been hatched, agglutinated, no agglutination occurred in sera from uninfected mice. Oncospheral agglutination was carried out by mixing 0·1 ml of serial two-fold dilutions of serum and 0·1 ml of Hanks' balanced salt solution containing about 600 hatched oncospheres. Titre of agglutinins was indicated as a reciprocal of the final dilution capable of giving agglutination clusters made of three or more oncospheres. Agglutinins developed within 14 days after a primary infection with 500 shell-free eggs. There was no rapid increase of agglutinins within 4 days following a secondary infection. The titre increase coincided with the increase in dosages of eggs. Agglutinins were thought to be immunoglobulins, because the potency of the serum to agglutinate oncospheres was extinguished after absorption of globulins with rabbit anti-mouse globulin serum.

Agglutinins were produced in rabbits by intravenous injections of shell-free eggs. The titres of the rabbit sera were much higher than those of mouse sera.

A culture chamber fitted with a polycarbonate sieve has been used to isolate Plasmodium knowlesi merozoites as they are released from schizonts. A 3 μm pore-size sieve allows passage of normal erythrocytes and red cells containing rings and trophozoites and can be used to concentrate schizonts from a mixed cell population. A 2 μm pore-size sieve retains normal and parasitized cells and provides uncontaminated merozoites in high yield (5 × 1010 merozoites per ml schizonts). Merozoite viability diminishes rapidly during 30 min after isolation. These preparations should prove valuable for studies of the biochemical, physiological and antigenic properties of this transient extracellular phase of the malaria parasite.

The structure and invasive behaviour of extracellular erythrocytic merozoites prepared by a cell sieving method have been studied with the electron microscope.

Free merozoites contain organelles similar to those described in late schizonts of Plasmodium knowlesi. Their surface is lined by a coat of short filaments. On mixing with fresh red cells, merozoites at first adhere, then cause the red cell surface to invaginate rapidly, often with the formation of narrow membranous channels in the red cell interior. As the merozoite enters the invagination it forms an attachment by its cell coat to the rim of the pit, and finally leaves this coat behind as it is enclosed in a red cell vacuole. Dense, rounded intracellular bodies then move to the merozoite periphery, and apparently rupture to cause further localized invagination of the red cell vacuole. The merozoite finally loses its rhoptries, the pellicle is reduced to a single membrane and the parasite becomes a trophozoite. Invasion is complete by 1 min after adhesion, and the trophozoite is formed by 10 min.

A direct life-cycle in Reighardia sternae, a cephalobaenid pentastomid of gulls was investigated: the work was prompted by a report of eggs and larvae recovered from the stomach and intestine of a naturally infected gull.

Infective pentastomid eggs were obtained by surgically transplanting maturing female Reighardia, taken from freshly shot wild gulls, into captive recipients. Faecal material from birds thus artificially infected was collected daily and examined for eggs. Eggs were force fed to 33 hand-reared (from eggs or nestlings) juvenile gulls which were selected at random and sacrificed at intervals thereafter and examined for pentastomids.

One hour after infection, primary larvae appear in the body cavity where they moult immediately. They grow steadily and by 27–35 days are sexually differentiated, and by 66 days have copulated. Fertilized females take a further 116 days to produce eggs by which time they are 7·6 cm long.

The complex migrations undertaken by developing larvae in the gull, and the problems of the mechanism of direct transmission, are discussed.

Worm free lambs 2–4 months old were given a single infection of 50 000 Haemonchus contortus larvae. The changes in packed red cell volume were compared with those of uninfected sheep and the infected sheep were divided into two groups according to the rate of change of packed cell volume. In group 1, packed cell volume fell sharply from the 10th day of infection and the sheep eventually died. In group 2, the fall was less marked and the packed cell volume returned to normal after about 40 days. The rates of increase of parasite body length and dry weight were compared for the two groups. Parasites in group 1 sheep grew more slowly than their group 2 contemporaries. Accurate planning of metabolic studies on H. contortus was made possible by using this information to predict the course of an infection and the size of the worms. Parasite development was also measured in sheep for which the packed cell volume was not recorded. Computer analysis showed that adult worm length increase did not follow a simple growth pattern starting from the last ecdysis, but consisted of a rapid elongation followed by a simple negative exponential increase. Dry weight increase also followed a negative exponential during the second phase. Measurements of dry weight as a percentage of wet weight indicated that the rapid elongation was possibly linked with cell enlargement by the uptake of water.

When measured 56 days postinfection the length, wet weight and dry-weight of Hymenolepis diminuta were all found to decrease with increasing number of cysticercoids given up to 20. The mean position of the worms in 10, 12 and 20 worm infections is significantly posterior to that of 1, 2 and 5 worm infections and the worms are attached over a wider area of the intestine.

Egg production by the worms was followed up to day 56 postinfection; the number of eggs produced per worm and even per rat decreased with increasing population density. Thus the best way to get most eggs and to maintain the parasite in the laboratory is to have rats infected with only one tapeworm.

Rats given 1–20 cysticercoids showed a mean recovery of 100–65%, while rats given 40–200 cysticercoids showed a mean recovery ranging from 13 to 2%. In addition to ‘normal’ worms, defined as worms > 10 mm, small, most probably destrobilated, worms were found. In the 50 and 100 cysticercoid infections, worm recoveries were, respectively, 8% ‘normal’, 16% small, and 2% ‘normal’, 5% small. From the significantly lower recovery from heavy infections it is concluded that a deleterious factor is operating during the 8 weeks after the infection.

Opaque or darkened areas (DA) of variable size and position occur on Hymenolepis diminuta in mice and rats. In mice DA normally first appear in the neck region of the worm but subsequently they appear elsewhere and increase in number until destrobilation or worm expulsion. The posterior of destrobilated worms is often darkened. In the more immunogenic infections with six cysticercoids there are more DA per worm than in infections with one cysticercoid. DA are areas of the tegument with a homogeneous increase in electron density; abnormal mitochondria; reduced granular endoplasmic reticulum, Golgi complexes and discoidal secretory bodies; and accumulation of lipid droplets. DA disappear from worms maintained for up to 4 h in Hanks' balanced salt solution and can be induced by mechanical damage to the worms.

As the numbers of DA increase with the duration and intensity of infection and have similarities with types of cell injury, they are probably sites of worm pathology induced by host immunity.