Haemonchus contortus worm populations isolated from naturally infected sheep at the Pastoral Research Laboratory, Armidale, N.S.W., were found to contain approximately 20% of worms resistant to a 50 mg/kg dose of thiabendazole. Following 3 generations of selection with 50 mg/kg thiabendazole the number of worms removed by the anthelmintic was too small to detect differences between treated and control groups. After more than 15 generations of selection, matings between males from the selected strain and non-resistant females produced resistant males and females in equal numbers. Thus, thiabendazole resistance does not appear to be sex-linked. A dose–response assay on the F2 adults indicated that worms from female resistant × male non-resistant crosses were more resistant than F2 adults of the reciprocal cross. An in vitro technique that identified thiabendazole-resistant eggs by their ability to hatch in a solution containing thiabendazole and 0·1% NaCl solution was also used to study the inheritance of resistance. F1 eggs had similar LC50's to the resistant parents. F2 and back-cross eggs from an original mating of thiabendazole-resistant females × non-resistant males had a higher LC50 than F2 and back-cross eggs from the reciprocal mating, indicating a degree of matroclinous inheritance of resistance. However, the resistant parents had tolerances to thiabendazole exceeding those of F2. F3 eggs had a resistance distribution that ranged from that of the resistant to the non-resistant parent. No significant deviation from linearity was observed in any of the dose–response lines. These results indicate that thiabendazole resistance in H. contortus worms is inherited as an autosomal and semi-dominant trait.

A membrane-bound monoamine oxidase (EC 1.4.3.4) was demonstrated in homogenates of Hymenolepis diminuta. The enzyme oxidized a variety of biologically active amines (in decreasing order: dopamine, adrenaline, noradrenaline, tryptamine, tyramine, octopamine), there was, however, no activity with 5-hydroxytryptamine or benzylamine. No diamine oxidase (EC 1.4.3.6) could be detected in H. diminuta (using histamine, cadaverine or putrescine as substrates). The monoamine oxidase from H. diminuta was not inhibited by azide, hydroxylamine or semicarbazide, but was inhibited by cupferron, α-α dipyridyl and iodoacetamide, and by the specific monoamine oxidase inhibitors pargyline, nialamide and iproniazid. Several anthelmintics were also found to be inhibitors of monoamine oxidase. The possible roles of monoamine oxidase in H. diminuta are discussed.

Two new species of Lepeophtheirus are described and illustrated based on a collection from the Orange County sewer outfall, California. They are: L. remiopsis sp.nov. from the branchial cavities and external body surfaces of Parophrys vetulus Girard, Hippoglossina stomata Eigenmann & Eigenmann, Pleuronichthys verticalis (Jordan & Gilbert), Chitonotus pugetensis (Steindachner), and Porichthys notatus (Girard); and L. rotundipes sp.nov. from the external body surfaces and occasionally the gill cavities of Scorpaena guttata (Girard) and Citharichthys stigmaeus Jordan & Gilbert.

A cell-sorting method is described for the analysis and separation of red blood cells in Plasmodium berghei-infected mouse blood based on their DNA content. This method involves a selective uptake of the bis-benzimidazole dye 33258 Hoechst, a DNA-binding dye, by red blood cells containing parasites. Infected blood is incubated at 37 °C with the dye then washed at 4 °C to remove unbound dye. Uninfected cells are then non-fluorescent at the characteristic wavelengths for 33258 Hoechst excitation and emission, whereas parasitized cells display fluorescence intensities in approximately direct proportion to the number of parasite nuclei (i.e. amount of parasite DNA) within the cell and can be sorted accordingly. Providing cells were incubated in a complex nutrient medium during dye uptake at 37°C, the sorted parasite cells produced lethal P. berghei infections when injected into BALB/c mice. The dyelabelling technique is simple and sufficient red blood cells at various stages of infection can be collected for biochemical or immunochemical studies by cell sorting.

A technique for the transplantation of Trichinella spiralis worms directly into the host intestine is described. Infections established by the direct transfer of adult worms were essentially normal both in terms of their survival and reproduction and in their stimulation of, and susceptibility to, host immune responses. Worms transplanted from NIH mouse donors at intervals after infection had an equal ability to survive in the recipient, even when taken from the donor shortly before or during the process of worm expulsion, showing that expulsion does not require worms to be irreversibly damaged. It was noted, however, that after 7 days in the donor the ability of the worm to reproduce in the recipient was temporarily impaired.

Ten variants of phosphoglucomutase were detected by starch-gel electrophoresis in extracts of the tick Ixodes ricinus. Agreement of phenotype frequencies with those predicted by the Hardy–Weinberg model indicated that the enzyme is coded by a single gene locus (Pgm) at which 10 alleles are segregating. Allelic proportions in 5 Irish tick samples indicated that both spatial and temporal genetic differentiation exist. It is suggested that this polymorphism may be of use as a marker for studying the relationships of I. ricinus populations in Europe, as well as in taxonomic studies. A hypothesis is advanced which offers a possible adaptive explanation of the very high Pgm heterozygosity in tick populations. It is suggested that if kinetic differences exist between the various enzyme variants, the polymorphism may act to maintain an array of individuals in each population with varying developmental rates and longevities. Such a genetic strategy could be one factor which determines that the host-infesting seasons are spread over several months, thus ensuring maximum feeding and reproductive rates in each population.

Vitamins A, E and K were incorporated into turkey starter rations at normal (1×), 5× and 50× the National Research Council recommended vitamin levels and evaluated for their effect on histomoniasis in turkeys. Infected as well as comparable uninfected poults were included in all trials. Each vitamin was also evaluated at normal and 50× the NRC level in combination with ipronidazole, a low-level antihistomonal agent. In addition to performance parameters, glutamate oxaloacetate transaminase (GOT) and lactate dehydrogenase (LDH) plasma enzyme levels were used to evaluate the vitamin effect. None of the vitamins tested alone were found to be effective in terms of reducing mortality and morbidity following a severe Histomonas meleagridis challenge, regardless of the vitamin dose. A significant improvement in performance was demonstrated with the vitamin E (50×)–ipronidazole combination and a numerical improvement with the vitamin A (50×)–ipronidazole combination over ipronidazole alone or the poults fed NRC recommended vitamin levels. The GOT and LDH plasma enzyme levels correlated well with the progressive pathological changes associated with the disease. At 11 days post-infection both enzymes were consistently and significantly elevated above those of the uninfected controls, irrespective of the vitamin levels provided. The presence of low level ipronidazole in most instances delayed the onset of clinical symptoms and was associated with a delay in enzyme elevation, indicating the excellent correlation and sensitivity of the enzyme method in assessing liver damage.

The ovary of Aspiculuris tetraptera has a prominent terminal cap cell. This is considered to be part of the ovarian epithelium. Oogonia detach from the short rachis and increase in size from 6 to 60 μm; accumulating hyaline granules, shell granules and glycogen. The hyaline granules persist in the egg cytoplasm after shell formation has been completed and are considered to be lipoprotein yolk. The shell granules contribute to the non-chitin fraction of the chitinous layer. A classification of the cytoplasmic inclusions of the nematode oocyte is proposed. Upon fertilization a vitelline membrane is formed which constitutes the vitelline layer of the egg-shell. The chitinous layer is secreted in the perivitelline space, between the vitelline layer and the egg oolemma. Upon completion of chitinous layer synthesis, the egg cytoplasm contracts away from its inner surface. The material of the lipid layer is secreted at the surface of the egg cytoplasm and adheres to the inner surface of the chitinous layer. During secretion of the chitinous and lipid layers by the egg cytoplasm, the uterine cells secrete the unit membrane-like external uterine layer and the crystalline internal uterine layer. A complex system of interconnecting spaces develops in the internal uterine layer. This system is open to the exterior via breaks in the external uterine layer. There is no direct involvement of the uterine cells in the formation of this structure.

The responses of susceptible Ndama and Zebu cattle to needle challenge with Trypanosoma congolense were followed using parasitological, haematological and radio-isotopic methods and compared with those of corresponding uninfected animals. In both breeds, infection became patent at the same time but peak parasitaemias were significantly lower, were attained later and were of short duration in the Ndama. All infected animals became anaemic, the severity of which correlated with the level and duration of parasitaemia. However, even when parasites could no longer be detected in the blood, packed cell volumes showed little tendency to recover. The anaemia was due to increased intravascular red cell destruction and was more pronounced in the Zebu. Haemodilution was not a feature. Increased red cell synthesis occurred in infected animals of both breeds but particularly in the Zebu; this accounted for the capacity to maintain packed cell volume levels following the initial drop associated with parasitaemia. However, in most cases red cell synthesis was less than expected from the degree of anaemia, suggesting impairment of bone marrow function. Measurement of red cell iron utilization indicated that this was due to defective iron re-utilization from degraded red cells arising from reticulo-endothelial blockade. It is concluded that the anaemia in this disease and its underlying processes are broadly in line with the number of parasites in the blood and that the superior resistance of the Ndama cattle lies in their ability to control parasitaemia rather than their capacity to mount a more efficient erythropoietic response.

N, N′-bis (3,4 ditrifluoromethylphenyl) methylmalonamide (Sch18545) completely controlled a mild Eimeria necatrix infection at 50, 40 or 30 p.p.m. in the diet, and controlled E. tenella infections at 50 and 40 p.p.m. Slight oocyst passage was observed at each E. tenella treatment level with a marked increase at the 30 p.p.m. treatment level. Fifty p.p.m. were necessary to control E. acervulina infections; levels of 40 p.p.m. reduced E. acervulina oocyst production while 30 p.p.m. were ineffective. Evaluations of Sch 18545 using a mixed infection (Coccivac D®) further suggested that activity with this compound was weakest against E. acervulina. Weight gains decreased with increasing concentration of drug in the diet of treated, infected birds and thus the compound showed an insufficient safety margin to be of practical value. Sch 18545 administered at 35 p.p.m. in the diet was effective against amprolium, zoalene, aklomide or nicarbazin-resistant strains of E. tenella.

One therapeutic oral dose (400 mg/kg) of 153C51 administered to infested mice caused pathological changes in the dorsal region of the tegument of male Schistosoma mansoni during the period 3–24 h after treatment. These changes occurred prior to the ‘hepatic shift’. At the ultrastructural level they consisted of a gradual accumulation in the tegument epidermis of numerous membranous inclusions with the characteristics of residual lysosomes and changes in the localization of acid phosphatase, a lysosomal enzyme. It seemed likely that these changes were due to inhibition or exhaustion of enzyme in the epidermis, followed by re-synthesis of enzyme in the cell bodies and its export to the epidermis. The elimination of hydrolytic activity from lysosomes in the epidermis would explain the accumulation of residual lysosomes. Drug- treated parasites retained their disguise of host red blood cell ghost antigens as shown by indirect fluorescent antibody-labelling and, therefore, it seemed unlikely that immunological factors could be important in producing the tegument pathology.

The egg of Aspiculuris tetraptera is an ellipsoid measuring 93 × 40 µm. The shell consists of 5 layers: the external uterine layer, internal uterine layer, vitelline layer, chitinous layer and the lipid layer. This nomenclature is based upon the formation and histochemistry of the shell layers. The internal uterine layer contains a system of interconnecting spaces, partly filled by uterine secretion, which open to the exterior of the egg via breaks in the external uterine layer. The surface of the egg is covered by a system of interconnecting grooves. Freeze-etching reveals that the internal uterine layer is open to the exterior via pores, which open into the grooves. Rod-shaped particles are also revealed in the external uterine layer. The operculum of the egg consists of a modification of the uterine and chitinous layers of the shell.

The use of tritiated leucine as a marker for protein synthesis and of tritiated glucosamine as a marker for polysaccharide/glycoprotein synthesis, is described. Adult worms were pulse-labelled by incubation in medium containing the substrate. Labelled worms were then incubated in chase medium, without labelled substrate, for varying lengths of time before fixation. The distribution of label which had been incorporated into macromolecules in the worm tissues, was examined by light and electron microscope autoradiography. It was estimated that the tegument and tegument cell bodies were the source of 67–80%, and the gut epithelium of 20–33%, of exportable leucine-containing protein. Conversely, the gut epithelium was the source of 72%, and the tegument cells 28%, of exportable glucosamine-containing polysaccharide. The specific activity of labelled protein reached a peak in the tegument cytoplasm after 1.5 h of chase incubation. Half of the labelled protein was secreted into the worm's environment by 3 h of chase incubation. The half-life of secretory protein in gut cells appears to be around 2 h. Labelled protein disappears from the gut lumen relatively rapidly but labelled polysaceharide remains in the lumen at high specific activity for at least 24 h. The major carbohydrate labelled may be the glycocalyx on the luminal surface of the gut epithelial cells. The results suggest that the bulk of worm secretions have a rapid turnover with a half-life of a few hours. Against this background of rapid mass secretion a slower process of membrane turnover would be difficult to detect and quantitatively small.

Eimeria dispersa was isolated from young, intensively housed turkeys in Britain. The parasite has morphological features in common with those of the original description. At least two generations of schizonts occur before gametogony and one or two more schizont generations are concurrent with gametogony. Four generations of schizonts are needed to account for the observed reproduction index of 2·08 × 106/oocyst inoculated. The first oocysts were discharged by 105 h and these sporulated. within 30 h at 30 °C. A single dose of between 1 × 102 and 1 × 106 oocysts depressed body weight gain of 3-week-old poults and 2 poults died 6 and 12 days after receiving 1 × 104 or 1 × 106 oocysts. A single dose of oocysts protected against a challenge inoculation given 14 days later. Poultsimmunized against E. dispersa were resistant to challenge with a strain of E. dispera obtained from the United States. Electrophoretic examination of 2 enzymes from both strains of E. dispersa showed that they had similar mobilities. Attempts to infect chickens, guinea-fowl and Japanese quail with E. dispersa were unsuccessful. The strain of E. dispersa isolated here is best referred to as E. dispersa (Briston).

Propagation of Nosema eurytremae, a microsporidian pathogen of trematode larvae, was investigated by inoculation of spores into the haemocoele of insects and by growth in tissue cultures. Locusts and the larvae of three lepidopteran species were good hosts but cockroaches were not. Low replication was obtained in one lepidopteran species after per os infection. Antibiotics controlled bacterial growth in suspensions of microsporidian spores but fungi were unaffected by all antibiotics tested, except at concentrations detrimental to the microsporidia. All stages of the microsporidium developed in cell lines of Xenopus laevis and Aedes pseudoscutellaris when spores were induced to hatch in contact with cell monolayers: the Aedes culture was contaminated by yeasts. Repeated washing of the Xenopus cells with fresh medium, after the sporoplasms of N. eurytremae had penetrated the cells, removed yeast contaminants and sterile cultures were obtained. Replication during 4 passages over 53 days was only 100 to 200-fold compared with the original inoculum but spores harvested from the cultures were infective to a fresh culture and to Pieris brassicae by inoculation into the haemocoele.

CBA mice deprived of their T-cells by means of thymectomy and administration of rabbit anti-mouse thymocyte serum before a primary infection of Schistosoma mansoni, failed to resist re-infection to the same extent as immunologically intact controls. However, little difference was made to the degree of resistance to re-infection acquired by thymectomized S. mansoni-infected mice when the anti-thymocyte serum was administered after the primary infection and just before the challenge. Hydrocortisone acetate, when administered to primarily infected mice just before reinfection appeared to reduce the degree of acquired resistance, but the drug also had a schistosomicidal effect on the challenge-derived organisms. An attempt to specifically suppress the acquisition of resistance against S. mansoni by administering serum from homologously infected, resistant donors before and during the course of a primary infection failed, and partial depletion of complement activity with cobra venom factor that was administered just before challenge also failed to affect resistance. Serum obtained from mice with chronic S. mansoni infections and injected intradermally at the site of subsequent administration of a homologous percutaneous challenge infection failed to markedly inhibit maturation of the challenge in the recipient. Recipients of 3 ml of serum from heavily infected donors were also not significantly more resistant to challenge than untreated controls. However, isolated skin from heavily infected mice allowed fewer cercariae to penetrate and emerge transformed as schistosomula than did isolated skin from uninfected controls.

The frontal and sub-parietal glands of the pentastomid Reighardia sternae elaborate lamellate secretion which is poured on to the cuticle. The entire surface of the cuticle, including the mouth, hook pits and reproductive apertures, is coated with secretion. Electron microscope studies indicate that the glands are continuously active, which implies a turnover of surface membranes. The postulated function of these membranes is to protect certain vital areas of the host–parasite interface, notably the pores of ion-transporting cells, from the host immune response. The available evidence suggests that pentastomids do evoke a strong immune response but since most are long-lived they must circumvent it. We believe the surface membrane system to be instrumental in this. Studies on another pentastomid, Porocephalus crotali in rats have shown that an immune response stimulated by a primary infection will kill subsequent infections and that the surface membranes are strongly immunogenic. Obvious parallels between this situation and that of schistosome infections in mammals are discussed. An alternative explanation of concomitant immunity is proposed.

Very variable levels of immunity to a second infection with Schistosoma mansoni were recorded in 7 strains of mice, 12–15 weeks following a small primary infection. When 2 or more strains of mice were assayed at the same time, less variation occurred within the experiment than between different experiments. This evidence suggested variation between pools of cercariae as the main cause of variability in immunity. In direct experiments in one strain of mouse, 2 different pools of cercariae stimulated widely different levels of immunity to the same challenge. Conversely, challenge infections drawn from different pools showed different susceptibility to immunity stimulated by the same primary infection. Individual clones of cercariae, from snails infected with single miracidia, showed a high level of susceptibility to immunity stimulated by a small bisexual infection, or were not susceptible at all. Antigenic polymorphism is the most likely explanation for the differences observed between clones of cercariae. However, indirect immunofluorescence showed the presence of at least 1 common antigen on the surface of schistosomula derived from different clones of cercariae and clone-specific antigens have not been detected.

The effect of the intestinal changes brought about by the expulsion of Trichinella spiralis in rats was studied in relation to the growth and survival of a concurrent infection with Hymenolepis diminuta, a cestode not normally rejected by the rat in low-level infections. Growth of H. diminuta was stunted in rats given T. spiralis just before, or after, infection with H. diminuta, the stunting being more pronounced when the cestode was given closer to the period of inflammation. There was no loss of the cestode from dual-infected rats and no evidence for destrobilation was found. Lower T. spiralis burdens had a correspondingly weaker effect on growth of H. diminuta, and stunting was abolished by administration of the anti-inflammatory drug cortisone acetate. It is concluded that the stunting of H. diminuta is probably due to the non-specific inflammatory component of the rat's response to T. spiralis infection.

Common marmosets were shown to be susceptible to Plasmodium knowlesi malaria. The morphology of the parasite was indistinguishable from that observed in the natural host (Macaca fascicularis) and the common laboratory model (Macaca mulatta). A differential susceptibility to P. knowlesi was observed in the 8 marmosets studied. Multiplication rates of parasites were variable over 24 h periods. Five animals died of a fulminating infection within 12–17 days after challenge. Three animals recovered spontaneously from infection and were subsequently resistant to challenge with homologous and heterologous variants and strains of P. knowlesi. This resistance was maintained for intervals up to 100 days between challenge infections.