It has previously been shown that antibodies against the transmembrane proteins, band 3 and glycophorin A, inhibit entry of the merozoite into the red cell and, in the case of band 3, it was established that attachment of the parasite to the cell is not prevented. We have found that antibodies against the cytoplasmic domains of band 3 and of glycophorin A, when present in the interior of resealed ghosts of human red cells, also inhibit invasion by P. falciparum. It is inferred that attachment of the merozoite to the red cell causes structural effects that are transduced to the membrane cytoskeleton and the antibodies against transmembrane proteins interfere with the invasion sequence at this level.

Sixteen monoclonal antibodies (MABs) were selected for their reactivity with adult schistosome tegument. The distribution of target epitopes in different tissues of the adult and in various developmental stages was investigated by indirect immunofluorescence. The distinct patterns of reactivity of the MABs permitted their classification into 9 groups. The distribution of epitopes in larvae, particularly 3 h schistosomula, generally mirrored that in adults. A change in distribution of epitopes coincided with transformation from cercaria to schistosomulum providing a marker for this process. Two MABs reacted with surface membranes of intact cercariae, and 5 with the surfaces of intact 3 h schistosomula. The target epitopes of these 5 MABs were present in the tegument of adults, but not accessible externally. These observations suggest masking of antigens, not shedding, in the course of development. Indeed, no MAB reacted with intact lung or liver worms unless they were damaged. Three MABs reacted with membranes of the tegument and most other tissues, implying properties in common. Evidence for shared functions between epithelia was provided by the reactivity of 2 MABs with the tegument and protonephridia. Eight MABs bound both to the tegument and its cell bodies. Since the latter contains the machinery for protein synthesis, it is difficult to explain why 5 MABs bound to the tegument alone. Little cross-reactivity was found with the tissues of the tapeworm Hymenolepis, rather more with the closely related Schistosoma japonicum. It was concluded that tegument antigens are to a degree tissue specific but definitely not stage specific.

Promastigotes of Leishmania major were frequently detected in the urine droplets discharged by infected Phlebotomus papatasi and P. duboscqi females during feeding. Parasites were present in the urine of 37·5% P. papatasi and 16·1% P. duboscqi females, even in those with low intensity gut infections. Free-swimming forms (elongated nectomonads, short slender promastigotes and metacyclic forms) predominated in excreted droplets. Viability of excreted parasites was proved by cultivation on blood agar, and the presence of metacyclic forms in urine droplets was confirmed by specific fluorescence assay with 3F12 antibodies. While the release of promatigotes from the anus of the sandfly was frequent, these were rarely egested from the mouth-parts of sandfly females (1·3% for P. duboscqi and 0% for P. papatasi) fed on microcapillaries, even if the females were heavily infected. The possible role and significance of the discharge of parasites in sandfly urine are discussed.

A comparison has been made of the interactions between passively transferred and actively acquired immunity in regulating populations of Taenia hydatigena and T. ovis. When ewes were grazed prior to parturition under a high infection pressure, immunity was transferred to their offspring for up to 8 weeks. A qualititative difference between the species was the destruction of larval T. ovis prior to their establishment (‘pre-encystment immunity’) and that of T. hydatigena after they had become established (‘post-encystment immunity’) in the challenged lambs. The major difference in terms of population regulation between the two parasites was that infection occurred with T. hydatigena but not with T. ovis in those lambs reared from birth for 16 weeks under high infection pressure. Passive, like active immunity, is a density-dependent constraint. It plays an important role in the population regulation of T. ovis, but not of T. hydatigena. This is discussed in terms of transmission in the natural environment, an hypothesis on humoral protection and the need to elucidate pathways of protection when immunization schedules are being evaluated for controlling the taeniid zoonoses.

The insect parasites bred out of Psyllid hosts, in the course of studying their biology in Scotland, belong to four families of Hymenoptera and one of Diptera, the total number of species encountered being seven. Of these one and possibly two more of the hymenopterous species are hyperparasites. The hosts involved include four species, of which only one, Psyllia mali race peregrina, may be said to have been fairly heavily parasitised. As observed by Ferrière (1926), species of Psyllidae, in spite of their nymphs leading a comparatively sedentary life, are less susceptible to attack by parasites than are the other Homoptera. As a rule the hymenopterous parasites attack them in the nymphal stage, while the dipterous species parasites the adults as well.

After penetrating through the body wall into the haemocoel of Bradysia paupera, the fertilized female of Tipius sciarae increased in size and slowly expelled the enlarging uterine cells through the vulva.

Within 7 days of penetration, the females were mature and began laying eggs into the haemocoel of the host. The eggs hatched in 3 days and, within 2 weeks, the host–s body was swarming with juvenile nematodes. The juveniles moulted three times in the body cavity of the host and 4th-stage forms emerged through ruptures in the intestine or body wall (in larval hosts) or were deposited on the surface of the soil (by adult female flies). They then moulted to adult forms while remaining ensheathed in their last juvenile cuticle, mated, and the fertilized infective females were ready to enter a new host.

Most parasitized fly larvae died before reaching the pupal stage but some emerged as adults, still carrying the nematodes within them. All parasitized adult flies were sterile. Infested larvae had smaller fat bodies and adult histoblasts than normal larvae and took twice as long to develop.

Preliminary tests suggested that this nematode may be useful in controlling sciarid gnats in glasshouses.

T. sciarae (Bovien) and T. gibbosus (Leuckart) were compared.

This work was done at Rothamsted Experimental Station, Harpenden, Herts, England, while the author held a postdoctoral grant from the National Institutes of Health, Bethesda, Maryland. I thank Mr F. G. W. Jones for a place in the Nematology Department, Dr Audrey Shepherd for supplying the New Blue R stain, Dr J. B. Goodey for advice, and Dr K. Lindhardt, Denmark, for the loaning of the late Dr Bovien–s slides of T. sciarae.

The aim of this study was to investigate the activities of important enzymes involved in the phosphoryl transfer network (adenylate kinase and creatine kinase (CK)), lactate dehydrogenase (LDH), respiratory chain complexes and biomarkers of cardiac function in rat experimentally infected by Trypanosoma evansi. Rat heart samples were evaluated at 5 and 15 days post-infection (PI). At 5 day PI, there was an increase in LDH and CK activities, and a decrease in respiratory chain complexes II, IV and succinate dehydrogenase activities. In addition, on day 15 PI, a decrease in the respiratory chain complex IV activity was observed. Biomarkers of cardiac function were higher in infected animals on days 5 and 15 PI. Considering the importance of the energy metabolism for heart function, it is possible that the changes in the enzymatic activities involved in the cardiac phosphotransfer network and the decrease in respiratory chain might be involved partially in the role of biomarkers of cardiac function of T. evansi-infected rats.

The identification of protective helminth antigens remains the most important challenge in the development of parasitic vaccines. To identify protective antigens of Ostertagia ostertagi, an important abomasal parasite of cattle, parasite-specific local antibodies from the abomasal mucus and from the draining lymph nodes were collected from calves immunized with multiple infections and from ‘primary infected’ animals. With these probes, Western blots of extracts and excretion/secretion (E/S) material from L3, L4 and adult life-stages as well as cDNA expression libraries were screened to identify antigens that were exclusively recognized by antibodies from ‘immunized’ calves. In the adult stage, a protein of 32 kDa was specifically detected on Western blot by mucus antibodies from ‘immunized’ animals. In the L3 and L4 larval stages, proteins situated in the regions of 28–29 kDa were recognized by mucus antibodies and a 59 kDa antigen was specifically recognized by lymph node antibodies from ‘immunized’ animals. Screening E/S material revealed no specific difference in recognition pattern between ‘immunized’ and ‘primary infected’ animals. Screening of the cDNA libraries revealed 26 relevant clones, coding for 15 proteins, among these several with potential protective capacity, immunodominant properties or functional and physiological importance e.g. metalloproteases, an aspartyl protease inhibitor and collagen.

1. A four- to eight-fold increase in resistance to pamaquin has been developed in a strain of Plasmodium gallinaceum in chicks.

2. Pamaquin resistance conferred no resistance to proguanil, sulphadiazine, mepacrine, chloroquine or sontochin, but it conferred some resistance to pentaquine and to quinine.

3. An appreciable loss in resistance to pamaquin was observed in the pamaquin-resistant strain after it had been maintained in the absence of the drug, in a patent state of infection, for a period of 6 months.

4. No synergism was observed between pamaquin and quinine when these drugs were tested, in combined doses, upon active infections of P. gallinaceum.

Toxoplasma ‘Nicolle et Manceaux’ has been examined in the electron microscope in thin sections of infected tissue cultures. The intracellular forms, single or in rosette formations, show that the parasite has no flagella, cilia or other organelles for locomotion. It is covered by a double membrane which is especially well denned at the two poles. A ring-like formation, often slightly protruding, can be seen at one end of the parasite, and in connexion with this, long, homogeneous, dark-stained inclusions of still unknown nature. At the opposite pole a distinct opening has been found.

Mitochondria and strands of an endoplasmic reticulum are also present in the cytoplasm and a fine striation in the region of the periplast. No dividing forms have been observed. Near the nucleus, in the region of what may correspond to the centrosome and the Golgi apparatus, a few extremely fine granules and a spiral-like formation of fine striae or tubules have been observed.

This work has been partly supported by the National Research Council of Brazil. The cost of reproduction of figures was defrayed by the Institute de Biofísica da Universidade do Brasil.

Three species of furcocercariae are described. By means of direct animal experiment with secondary, intermediate and final hosts I have shown that one species develops into Tetracotyle typica Dies., the larva of Strigea tarda Steenstrup, and a connected detailed account of this life cycle is here given. Of the two other species, both of which are new, I have been able to trace the development of one into Tetracotyle n.sp. in Planorbis crista L., and P. albus Müll. Feeding experiments with this new Tetracotyle using ducklings as final hosts failed, however, to produce the corresponding adult.

Effect of the parasite on the host. The usual results of helminth infection of this nature are observable, namely, marked disruption of liver tissue and in many cases complete parasitic castration of the molluscs infected.

Double infection. In the course of my investigations on larval Trematodes I have met with one example of double infection with sporocysts, viz. Cercaria F.1 and F.2, within one and the same individual of Planorbis albus Müll. This condition was found only on one occasion, Cercaria F.1 being in predominance. Instances of double infection have been recorded by Faust (1971) and Sewell (1922), and from this country by Hesse (1923) and Brown (1926). The condition, however does not appear to be a common one.

Establishment, survival and distribution of Leptorhynchoides thecatus (Acanthocephala) were investigated in largemouth bass, Micropterus salmoides, fed 10, 25, or 40 cystacanths and examined at 1, 3 or 5 weeks post-infection. Worms established widely in the alimentary tracts of bass but by 5 weeks post-infection had localized in the pyloric caeca and intercaecal region. Other individuals moved to parenteral sites where they remained immature, though viable. In the 10- and 25-level exposures, establishment and survivorship in the alimentary tract were roughly proportional to the dose of cystacanths. After 1 week post-infection in the 40-level exposure class, numbers of worms in the alimentary tract decreased significantly and parenteral occurrence increased significantly. Total survival of L. thecatus appeared to be density-independent. Maturation of worms was retarded temporarily as intensity of infection increased, but by 5 weeks post-infection worms from all doses were at roughly the same stage of development within sex. The caeca and intercaecal area apparently did not differ in their suitability for maturation.

Analysis of natural populations of Trypanosoma brucei has shown that there is linkage disequilibrium between alleles at pairs of loci in isolates taken from the field. This disequilibrium can occur as a result of a low frequency of genetic exchange, the masking of frequent genetic exchange by the rapid expansion of a few genotypes or by the treatment of 2 (or more) genetically isolated populations as a single population. We have analysed stocks from 2 geographically separate locations using 3 minisatellite markers to determine the frequencies of the alleles in each area and the frequency and nature of the multilocus genotypes. The results show that many alleles and multilocus genotypes are unique to each geographical location, supporting the conclusion that these populations are genetically isolated with limited or no gene flow between them. This geographical substructuring needs to be taken into account in considering the origins of the linkage disequilibrium in a number of populations.

An epidemiological survey of intestinal parasitic infections was conducted in a sample of 203 children aged 3–5 years from a semi-urban and a rural community in Chiriqui, Panama, in 1983–4. On the basis of stool examinations, the prevalences of Ascaris lumbricoides, Trichuris trichiura, hookworm, Giardia intestinalis, Entamoeba spp. and Strongyloides stercoralis were found to be 27, 34, 14, 15, 5 and 4% respectively. The results from children from the two communities were compared. Polyparasitism occurred significantly more often in rural than semi-urban children. Following anthelmintic treatment with levamisole, the numbers of A. lumbricoides passed/child were recorded and the frequency distribution of the parasite was observed to be highly aggregated with a variance to mean ratio of 10·2. For A. lumbricoides, relationships between worm burden, worm biomass and egg production were investigated. In the data analysis, an attempt was made to explore the influence of numbers of male worms on egg production. The results are compared with those obtained during other recent studies on the epidemiology of A. lumbricoides infection in other countries.

Hymenolepis citelli caused a marked mortality in Tribolium confusum during the first 14 days of infection. The mortality was higher when the beetles were starved for 6 days immediately prior to infection (73–93%) than when they were starved for 1 day (51%). Regardless of the starvation time, the majority of beetle deaths occurred between days 8 and 11 post-infection (p.i.) and the precise survival time for individuals tended to vary inversely with the number of parasites they contained. With few exceptions beetles that survived until day 15 p.i., when the experiments were terminated, contained 14 or fewer cysticercoids, whereas those that died on or before day 14 p.i. contained 20 or more parasites. Mortality was equally high in both sexes but the mean survival time was significantly shorter for female beetles (8·8 days) than for males (9·6 days). Also, parasite development rate varied with the sex of the host. When parasite populations recovered from beetles that died on the same day were compared, the degree of development attained by those from males was significantly higher than in the populations grown in female hosts. The line of H. citelli used in these experiments has been maintained in the laboratory for the past 5 years. The results obtained with the current generation of this line were compared with those obtained with it 2 years earlier (approximately 16 generations ago). Mortality was lower with the current generation (67%) than with the earlier generation (93%). However, with the latter generation, the mean parasite population size in beetles that survived to day 15 p.i. was significantly higher (14·1 cysticercoids/beetle) than in those infected with the recent generation (7·3–7·9) cysticercoids/beetle). These findings suggest that relatively high selection pressures have been applied by T. confusum to H. citelli, thereby reducing the number of parasite lines with high infectivity and consequently improving host survivorship. The host also responded to artificial, directed selection for reduced cysticercoid number (heritability estimates of 0·30 ± 0·12 and 0·26 ± 0·12) and increased survival (heritability estimates of 0·28 ± 0·12 and 0·15 ± 0·083). The realized heritability was 0·37 for cysticercoid number and 0·17 for increased survival.

Field and in vitro studies have shown that high salinities and temperatures promote the proliferation and dissemination of Perkinsus marinus in several environments. In Brazil, the parasite infects native oysters Crassostrea gasar and Crassostrea rhizophorae in the Northeast (NE), where the temperature is high throughout the year. Despite the high prevalence of Perkinsus spp. infection in oysters from the NE of Brazil, no mortality events were reported by oyster farmers to date. The present study evaluated the effects of salinity (5, 20 and 35 psu) and temperature (15, 25 and 35 °C) on in vitro proliferation of P. marinus isolated from a host (C. rhizophorae) in Brazil, for a period of up to 15 days and after the return to the control conditions (22 days; recovery). Different cellular parameters (changes of cell phase's composition, cell density, viability and production of reactive oxygen species) were analysed using flow cytometry. The results indicate that the P. marinus isolate was sensitive to the extreme salinities and temperatures analysed. Only the highest temperature caused lasting cell damage under prolonged exposure, impairing P. marinus recovery, which is likely to be associated with oxidative stress. These findings will contribute to the understanding of the dynamics of perkinsiosis in tropical regions.

Survival of Hymenolepis diminuta eggs (as measured by their infectivity to the intermediate host) was found to be age-dependent with an expected value of 11 days when the eggs were retained within the faecal pellet at 10 °C. The expected life-span of eggs under experimental conditions (i.e. extracted from faecal material and placed on filter paper at 30 °C) was estimated as 33 min. The mean parasite burden of populations of Tribolium exposed to known densities of H. diminuta eggs was found to rise to a plateau with increasing exposure time. The experimental results provided a method of estimating the instantaneous rate of parasite transmission: a value of 0·0004/egg/min/host/13 cm2 was obtained. The mean parasite burden of populations of T. confusum exposed to known densities of H. diminuta eggs was found to decrease exponentially with increasing host density. A second estimate of the instantaneous rate of parasite transmission of 0·004/egg/min/host/13 cm2 was obtained from the experimental results. No differences in susceptibility to infection between 2-week-old male and female beetles were found. A marked decrease, however, was apparent with increasing beetle age up to 14 weeks post-eclosion.

The parasitic nematode Anguillicola crassus was recently introduced into populations of the European eel, Anguilla anguilla. We investigated, under experimental conditions, the regulation of A. crassus infrapopulations. We tested the effects of (1) the resource-limited habitat of the parasite and (2) the coexistence of several developmental stages in its niche (the swim-bladder) on the composition of the infrapopulations. The results revealed that the respective effects of these factors differed substantially during the course of the infection. Third-stage larvae (L3s) establishment would not be constrained by the size of the swim-bladder. Their moult to fourth-stage larvae (L4s) would be accelerated as the number of L3s increased. The moulting time of L4s to adults would be reduced by males and would be constrained by the size of the swim-bladder. However, the moult of L4s to adults and their further development would be synchronized with those of the opposite sex. At the time of mating, the number of males and the body weight of adults would depend on the size of the swim-bladder. Soon after the laying of eggs, the developmental constraint on the late L3s would decrease. When adults die, constraints would cease and late larval stages would moult to become adults.

AF3 (AVPGVLRFamide) and AF4 (GDVPGVLRFamide) are endogenous RFamide-like peptides isolated from the parasitic nematode Ascaris suum. Here the actions of these peptides on the somatic musculature of Ascaris have been investigated and compared to the action of acetylcholine (ACh), the excitatory transmitter at the neuromuscular junction. ACh, AF3 and AF4 contracted muscle with EC50s of 13±1 μM, 24±6 nM and 37±2 nM, respectively (n=6). The muscle cells were depolarized by ACh (3 μM; 5·2±0·4 mV, n=42), AF3 (1 μM; 2·6±0·3 mV, n=19) and AF4 (1 μM; 3·3±0·4 mV, n=19). EC50s were 681±329 nM (AF3) and 901±229 nM (AF4), but an estimate could not be made for ACh due to muscle contraction at concentrations greater than 10 μM. The depolarization to 3 μM ACh was abolished by the nicotinic receptor antagonist mecamylamine (10 μM; n=5) but the responses to the peptides were not (111±7% and 108±17% with respect to control; n=5). The depolarization elicited by ACh was reduced to a greater extent by a 50% reduction in extracellular Na+ concentration than the response to AF3 and AF4 (P<0·02). Cobalt was more effective at blocking the AF3 and AF4 depolarizations than those to ACh. These observations suggest that AF3 and AF4 contract Ascaris muscle without an action at the Ascaris nicotinic receptor. Furthermore, the ionic mechanism through which AF3 and AF4 depolarize Ascaris muscle is different from that for ACh. ACh, AF3 and AF4 were also found to contract Ascaridia galli somatic muscle with EC50s of 13±3 μM, 721±236 nM and 371±177 nM, respectively (n=7). The muscle cells were depolarized by ACh (EC50=14±5 μM, n=5), AF3 (EC50=5±3 μM, n=4) and AF4 (EC50=10±5 μM, n=4). Therefore the response to these peptides is not unique to Ascaris and they may subserve a functional role in the motor nervous system of parasitic nematodes.

The present paper includes two sections of a proposed series on the lungworms of the Odontoceti or toothed whales. Only members of the family Delphinidae among the Odontoceti have so far been reported as hosts to these lungworms.

The first section deals with the systematic relationships of the lungworms of the Odontoceti. They are held to constitute a family Pseudaliidae in the superfamily Metastrongyloidea. The nature of the female reproductive system and specifically of the ovejectoral apparatus in the suborder Strongylina is discussed and the concept expressed that the organization of the latter structure is of phylogenetic and taxonomic importance in this group. On the basis of variations in the ovejectoral apparatus three families of strongyline lungworms (except for Syngamus v. Siebold, 1836) are recognized—Dictyocaulidae Skriabin, 1941, Metastrongylidae Leiper, 1909, and Pseudaliidae Railliet, 1916.

The second section deals with the genus Halocercus Baylis & Daubney, 1925. Previous literature is reviewed. A key and table of measurements for the known species are presented. H. invaginatus (Quekett, 1841) Dougherty, 1943, is redescribed, and H. kirbyi n.sp. named and described.