Methods for screening compounds against Eimeria tenella in cell culture and in the chicken are described. An analysis for incidence of anticoccidial activity and host cell toxicity has been made of 11550 compounds screened in vitro, and some correlation of activity in vitro and in vivo has been attempted. The results show that screening compounds in tissue (cell) culture is not a satisfactory or reliable alternative to screening in chickens, although in vitro studies undoubtedly have a part to play in the evaluation of an active drug.

The course of infection with a local strain of Schistosoma mansoni was investigated in Kenyan baboons (Papio anubis). A single exposure to 5000 cercariae per baboon (cpb) killed five 4–8 kg baboons; a sixth survived the full 30-week post-exposure period, with suppressed egg production after week 11. Lower doses were not lethal. A single exposure to 1000 cpb gave satisfactory adult worm recoveries, high and steady faecal egg excretion, and a high tissue egg recovery, predominantly from the large intestine. A similar but less consistent picture was obtained with a single exposure to 200 cpb. Baboons receiving 5 monthly exposures of 200 cpb (1000 cpb total) showed some evidence of the development of protective immunity. Three animals showed patterns resembling those for 200 and 1000 cpb. The other 3 had suppressed egg production (faecal and tissue) indicating, perhaps, some immunological process. They also had an abnormally high percentage of eggs in the liver.

On three occasions, antibody positive blood from wild red deer produced overt infections with Babesia when inoculated into splenecto-mized red deer. One of the deer also became infected with Eperythrozoon sp. Babesia divergens, B. capreoli and the Babesia of red deer are morphologically similar and the marginal position of the parasites in the host cell is characteristic. Babesia were not seen and no antibody was formed in five out of six splenectomized bovine calves which were injected with parasitaemic red deer blood. Two of these calves when challenged with B. divergens were fully susceptible. A transient infection with the deer Babesia may have occurred in the sixth calf since antibody was detected and the animal resisted challenge with B. divergens.In indirect fluorescent antibody tests there was little or no difference in the titre of sera from naturally or experimentally infected cattle and deer when reacted with B. divergens or the red deer Babesia antigens. Despite their similarities, specific status for B. divergens and the red deer Babesia is probably justified; at present there is insufficient evidence to justify separation of the red deer Babesia from B. capreoli.

A relatively simple, standardized hatching test was devised, tested and used to estimate the Schistosoma mansoni hatching rate for 88 St Lucian subjects selected by age, sex and intensity of infection.

The hatching rate was dependent on the intensity of infection and rose proportionately with it. The rate also decreased with increasing age of the subject. Sex alone had no direct effect but there was a suggestion of an interaction between sex and age.

These results suggest that several hatching tests are necessary, before and after treatment in schistosomicidal drug trials, to permit valid conclusions to be drawn. The hatching data are used in conjunction with survey results to calculate the contamination potential of different age groups in a population. School children (5–14 years old) are about twice as important as young adults (15–29 years old) who, nevertheless, contribute over a quarter of the total contamination potential. However, whereas school children are fairly accessible for mass chemotherapy control programmes, young adults often are not and, furthermore, involve problems associated with the treatment of women of child-bearing age.

Rhesus monkey erythrocytes when incubated in vitro under similar conditions to those used for the cultivation of Plasmodium knowlesi-infected erythrocytes in vitro, exhibit an increase both in their osmotic fragility and in the activity of their acetylthiocholinesterase. No effect was observed on the catabolism of glucose through the glycolytic pathway or through the primary dehydrogenases of the pentose phosphate pathway. The ATP content of normal monkey erythrocytes was also unchanged during incubation in vitro. These observations indicate that incubation of erythrocytes in vitro primarily causes membrane changes. Infection of normal erythrocytes by P. knowlesi was reduced markedly by preincubation in vitro at 37 °C for 24 and 48 h. These results suggest that the maintenance of integrity of the surface of the eythrocyte in vitro is a necessary prerequisite for an efficient culture system for the malaria parasite.

The ultrastructural changes accompanying oocyte differentiation in the ovaries of the monogeneans, Diclidophora merlangi, Diplozoon paradoxum and Calicotyle kröyeri have been described. In each case, oogenesis in the ovary proceeds as far as meiotic prophase in the primary oocyte. A three-stage sequence of development based on oocyte morphology is proposed: (1) Oogonia and early, immature primary oocytes are typically undifferentiated, with chromatin-laden nuclei occupying most of the cell volume. The cytoplasm contains small clumps of mitochrondria and unattached ribosomal aggregates. There is evidence of mitosis and, in later stages, meiotic prophase is indicated by the appearance of nuclear synaptonemal complexes. (2) Maturing primary oocytes are characterized by increased nucleolar volume associated with the production of RNA for export to the cytoplasm. An organized GER and Golgi apparatus are established and involved in the synthesis and packaging of membrane-limited cortical granules. Annulate lamellae and nucleolus-like bodies appear in the cytoplasm and, with development, the cells increase in size and, peripherally, become interdigitated. (3) Mature primary oocytes represent a resting phase when cellular activity is minimal. Golgi disappear and the ER fragments or becomes reduced in dimensions. Mitochondria and free ribo-somes are numerous and cortical granules move to the cell periphery. The cells separate and, when mature, are released from the ovary. There are minor species differences in oocyte ultrastructure and development.

Fasciola hepatica miracidia labelled with radioselenium were used to study aspects of their host-finding capacity by determining radioactivity subsequently taken up by exposed ‘target’ snails (Lymnaea truncatula). Possible interfering effects exercised by a number of non-host snails and bivalves were examined in linear test channels. The infection rate (radioactivity) among ‘target’ snails was markedly lowered when non-host Lymnaea species (L. pereger, L. palustris, L. stagnalis) were interposed as ‘decoys’. The prosobranch Bithynia tentaculata and the bivalve Sphaeriwm corneum exhibited a slight decoy effect whereas pulmonate species like Anisus vortex, Gyraulus albus, Planorbis planorbis, Physa fontinalis did not interfere with miracidial host-finding.

Other experiments showed that miracidia are more strongly attracted towards L. truncatula than L. pereger. Miracidia are not able to penetrate intact egg clusters of L. truncatula.

The development of resistance by the Houghton strain (H) of E. tenella to robenidine has been studied, in the laboratory, by serially passaging the strain in chickens fed increasing concentrations of drug. Resistance to robenidine developed more readily in experiments using larger numbers of birds with higher numbers of oocysts in the inoculum. Both these factors increased the parasite population and increased the chance of selecting parasites resistant to the drug. E. tenella (H) was made resistant to 264 ppm robenidine and showed no cross-resistance to other anticoccidial agents. Resistance arose in a series of ‘steps’ as the concentration of drug was increased. E. tenella (H) was continuously passaged at concentrations ranging from 2 to 33 ppm of robenidine. After 16 passages, lines passaged at 2, 4 and 8 ppm were not resistant to 33 ppm robenidine, suggesting that the degree of resistance developed was dependent upon the drug selection pressure.

An attempt was made to determine the relative importance of the different life-cycle stages of Eimeria maxima in the induction of immunity and also those stages most affected by the immune response of the host. In one experiment the life-cycle was controlled by chemotherapy but in all other experiments partial life-cycles were induced by transfers of infected mucosa between hosts.

The results indicated that the second generation schizont stage is probably that most concerned in the induction of protective immunity and that sexual stages are most susceptible to immune inhibition. After initial inhibition in the immune host the earlier asexual stages were able to resume development when transferred to a susceptible host. The longer the period of exposure to the immune environment, the less able was the parasite to recover.

Microvilli are developed on the surface of Schistosoma mansoni schistosomula during penetration of the host skin; they form rapidly but are lost approximately 90 min after penetration. Identical microvilli are also formed on schistosomula which have penetrated a mouse skin preparation in vitro, and on schistosomula prepared by mechanical separation of the tail from the body of the cercaria. The microvilli, which are limited by the trilaminate tegumental membrane of the cercaria, eventually degenerate and are cast off from the surface of the tegument. There is little change in the surface area of the schistosomulum at this time, and the formation and loss of microvilli coincides with the replacement of the cercarial tegumental membrane by the new heptalaminate membrane. It is suggested that during the cercaria/schistosomulum transformation, some intramembraneous components of the original cercarial membrane may migrate into the new heptalaminate membrane and thus be retained, while other peripheral components such as the glycocalyx are almost certainly lost.

Infections in the chicken embryo have been used to study the development of drug resistance in an embryo adapted strain of E. tenella. Resistance was developed to decoquinate, clopidol and robenidine by serially passaging this strain, but evidence for the development of resistance to amprolium was inconclusive. Resistance to decoquinate developed more readily than to the other drugs. Attempts to increase resistance to clopidol, robenidine and amprolium by increasing the sporozoite inoculum and by the use of a mutagenic agent were unsuccesful. No cross-resistance was found between the 4 drugs.

Drug resistant lines of the Houghton strain (H) of E. tenella, made resistant to the 4 anticoccidial drugs by passage in chickens, were found to be resistant when evaluated using chicken embryo infections. Lines made resistant to decoquinate were not controlled by any concentration of this drug, suggesting that resistance, once developed, was absolute and not dependent on drug concentration. Lines made resistant to robenidine, clopidol and amprolium, however, were controlled by higher drug concentrations suggesting that in this case resistance was dependent on drug concentration.

Young, previously uninfected white mice became infected with Hymenolepis nana when exposed in small cages to parasite eggs which had been sprinkled in suspension onto the floor of their cage, or to faeces from mice with patent infections. The mean daily probabilities of infection for individual eggs under these two conditions were 2·9 × 10−6 and 5·4 × 10−6 respectively. If the mice were starved for 24 h prior to exposure to faeces, then the mean daily probability of infection increased to 2·6 × 10−4. These differences in transmission are interpreted as being due to differences in the extent of coprophagy; they are not due to any effect of starvation on the hatchability of eggs.

A primary infection with Schistosoma mansoni, of 5 or more weeks duration, stimulated a high level of resistance to a challenge infection in the WO outbred strain of golden hamster. In sharp contrast, the LGN strain showed no statistically significant immunity to reinfection in most experiments and where detected the level of resistance was only about half that observed in the WO strain. Immunity to reinfection was assayed with the lung recovery method which assesses resistance a few days after challenge and with the conventional perfusion assay which measures immunity 6 weeks after challenge. Wide differences in the immune response to S. mansoni in different strains of the same host species have not been reported previously.

Infections of Raillietina cesticillus in the small intestine of the fowl caused a mononuclear leucocyte infiltration, which occurred in the tissues around the worm scolices and was especially prevalent in challenge infections. There was no evidence of vascular congestion or of a neutro-philic response, and the cellular reaction seemed to consist of a local proliferation of lymphoid tissue. Mast cell and pyroninophylic cell numbers increased in the intestinal mucosa of birds with longstanding primary infections and with secondary infections, but very little eosin-lophil infiltration was detected and the globule leucocyte response was minimal. The infections had no effect on the numbers of thelio-lymphocytes and goblet cells. Challenge infections did not result in an increased level of serum antibodies, but more cells containing globulin seemed to be present in the intestinal mucosa of these birds. The interrelationships of the various cell responses to the cestode and the origin of the globule leucocyte are discussed in the light of previous work on helminth infections.

The anticoccidial action of 6 drugs was studied by treating cell cultures with drug for 24 h after inoculation of the cultures with sporozoites of Eimeria tenella. A monitoring technique was used to confirm that the drug was being removed. Only monensin was shown to be coccidiocidal. Amprolium, lasalocid, methyl benzoquate, nicarbazin and robenidine were coccidiostatic at their minimum effective doses. Misleading coccidiocidal effects were observed when higher concentrations of methyl benzoquate, nicarbazin or robenidene were used.

Proteocephalus ambloplitis was recovered from largemouth bass inhabiting different thermal regimes, ranging from normal to greater than 10 °C above normal, on the Energy Research and Development Administration's (ERDA) Savannah River Plant near Aiken, South Carolina. Possible alterations in the mechanisms initiating plerocercoid migration as proposed by Fischer & Freeman (1969) were noted. According to these authors, a rise in temperature from 4 to 7 °C and above stimulated migration of parenteric plerocercoids into the gut. The lowest water temperature recorded for the reservoir for the entire sampling period was 8·0 °C. Theoretically, this should have terminated the life-cycle of this tapeworm since migration temperature was never attained. At Savannah River, the appearance of adult tapeworms coincided with a decrease in water temperature, while the appearance of adult tapeworms in Canada was correlated with an increase in water temperature. It is postulated that the decline in water temperature in southern latitudes and the increase in water temperature in northern latitudes initiates the same response, that is, migration of plerocercoids from parenteric to enteric sites where maturation to the adult form ensues.

Hamsters (WO strain) with a primary infection of Schistosoma mansoni or S. haematobium rapidly developed immunity to homologous challenge judged by the lung recovery assay. Immunity was detected at 4–5 weeks and reached a plateau 6 weeks after infection.

Using this information, hamsters with an 8-week primary infection with S. mansoni or S. haematobium were tested for resistance to homologous reinfection and resistance to a challenge with the other species of schistosome. Primary infection with S. mansoni or S. haematobium conferred a high level of immunity to reinfection with either species of schistosome judged by the perfusion assay, involving recovery of adult worms 6–10 weeks following challenge. Estimation of the level of immunity with the lung recovery assay, 5 days after challenge, indicated that immunity due to a primary infection with S.mansoni acted at or before migration of the challenge through the lungs but immunity stimulated by a primary S. haematobium infection was only partially effective at the lung stage and substantial destruction of challenging organisms occurred at a later stage of development.

Antibodies in immune sera of hamsters with a primary S. mansoni or S. haematobium infection were shown to bind to common antigens on the surface of young schistosomula of either species by u.v. microscopy using as detecting agent a fluorescein-labelled rabbit antiserum directed against hamster globulins.

Features of the anticoccidial activity of nicarbazin, amprolium, zoalene, sulphadimidine, diaveridine, Darvisul, spiramycin, chloramphenicol and Oxytetracycline have been re-investigated both in vivo and in cell culture using Eimeria tenella. Of the drugs studied, only spiramycin was appreciably coccidiocidal, although nicarbazin and amprolium showed possibly slower coccidiocidal activity. In order to show activity against a particular stage in the life-cycle, higher concentrations of drug than those usually recommended for field usage had in most cases to be used. Under these conditions, parasites were usually inhibited as multinucleate 1st generation schizonts. With delayed medication, effects against 2nd generation parasites were in most cases found, and in many cases, although the parasites never matured to give viable merozoites, the large degenerating forms produced were able to cause extensive tissue destruction and haemorrhage. Methodology in this type of study is discussed in relation to more active and more recent anticoccidials, and some further experiments with robenidine reported.

By means of conventional light microscopical and counting procedures, estimations have been made of the average number and size of ovarian balls from unfertilized and fertilized female Moniliformis dubius. The observations were made on worms recovered from experimental primary infections lasting 1, 2, 3, 4, 5, 8, 11, 14, 17 and 20 weeks in male rats. The number of ovarian balls rises to about 6300 per worm, with the maximum being attained more rapidly in unfertilized than in fertilized females. On average more ovarian balls were present in unfertilized than in fertilized females, but a marked decline in number occurred in both groups of worms towards the end of the experimental period. Similarly, on the basis of the area within the elliptical profiles of fixed ovarian balls, it appeared that those from unfertilized females were generally larger on average than those from fertilized worms.

The equilibrium, of vulvar phenotypes in Haemonchus contortus cayugensis was changed by infecting lambs with either a selected strain of smooth or of linguiform worms. Subsequently the lambs grazed the same pasture and observations were made on changes in worm populations in the lambs, and on pasture it appeared that the seasonal change in the relative frequencies of A and B linguiform worms was related to density-dependent factors in the host. In the spring, when lambs first become infected, the worm population is small and the majority of linguiform worms are A-type. As the season progresses the density of the parasite population increases. Once the population becomes crowded selection begins to favour the B-type females. The effects of crowding on linguiform A females can be seen as a positive correlation between the percentage of linguiform A and their weight. Population density also affected the survival of A-type worms since there was a negative correlation of percentage A-type on total worm numbers. There was a similar negative correlation between percentage A-type and total worm numbers in both the continuously infected and tracer lambs.

Population density did not affect the proportion of smooth worms. However, smooth worms increased in the tracers from, approximately 10% at the beginning of the experiment to 55% at its conclusion 18 weeks later. The latter percentage is almost within range of the local subspecies, which has 60% or more smooth worms. It appears therefore that the proportion of smooth to linguiform worms is a stable equilibrium maintained by natural selection.