Leishmaniasis is a vector-borne disease and an important public health issue. Glycosaminoglycan ligands in Leishmania parasites are potential targets for new strategies to control this disease. We report the subcellular distribution of heparin-binding proteins (HBPs) in Leishmania (Viannia) braziliensis and specific biochemical characteristics of L. (V.) braziliensis HBPs. Promastigotes were fractionated, and flagella and membrane samples were applied to HiTrap Heparin affinity chromatography columns. Heparin-bound fractions from flagella and membrane samples were designated HBP Ff and HBP Mf, respectively. Fraction HBP Ff presented a higher concentration of HBPs relative to HBP Mf, and SDS-PAGE analyses showed 2 major protein bands in both fractions (65 and 55 kDa). The 65 kDa band showed gelatinolytic activity and was sensitive to inhibition by 1,10-phenanthroline. The localization of HBPs on the promastigote surfaces was confirmed using surface plasmon resonance (SPR) biosensor analysis by binding the parasites to a heparin-coated sensor chip; that was inhibited in a dose-dependent manner by pre-incubating the parasites with variable concentrations of heparin, thus indicating distinct heparin-binding capacities for the two fractions. In conclusion, protein fractions isolated from either the flagella or membranes of L. (V.) braziliensis promastigotes have characteristics of metallo-proteinases and are able to bind to glycosaminoglycans.

An account is given of a composite virus disease of tobacco for which the name tobacco rosette has been suggested.

The two component viruses, named the mottle and vein-distorting viruses respectively, have been separated, and their symptomatology and methods of transmission described. The mottle virus is both sap and aphis-transmitted, but the vein-distorting virus is aphis-borne only.

The symptoms and histopathology of the complex disease in the tobacco plant are dealt with in some detail. There are three main types of symptoms: (1) intense rosetting, (2) splitting of the tissues, (3) formation of enations on the under-surface of the leaves.

The splitting of the tissues has been examined microscopically, and a number of photomicrographs are given illustrating the formation of the fissures. It is suggested that there is a concentration of virus in the cambium which prevents the formation of the normal xylem. Abnormal tissue and giant cells are formed in the cortex and pith. This appears to set up stresses which cause the splitting.

The insect vector of the complex disease is the aphis Myzus persicae Sulz. Another aphis, M. pseudosolani Theob., is also a vector but is less efficient than M. persicae.

The writer's thanks are due to Prof. F. T. Brooks, F.R.S., with whom he discussed the histopathology of the rosette disease, to Dr Roy Markham for taking the photographs illustrating Pl. I, figs. 2–5, and to Mr Charles Harpley of the Molteno Institute for his assistance in taking' the photomicrographs.

The tegument of the human parasite Schistosoma mansoni is critical for parasite survival within the mammalian host. The role of protein kinase C (PKC), a major effector molecule in the phosphoinositide pathway, in maintaining the structural organization of this syncytial layer was examined in adult worms. Phorbol 12-myristate, 13-acetate (PMA) and phorbol 12,13-dibutyrate (PDB), phorbol esters that activate PKC, induced formation of surface vesicles as determined by light and scanning electron microscopy. Similar results were seen with sn-2-dioctanoyl-glycerol, a synthetic analogue of diacylglycerol. No effect was seen in parasites incubated with 4-a-phorbol ester or a isomers of PMA or PDB, compounds that do not activate PKC. Vesicle formation was reversible in parasites treated with sn-2-dioctanoyl-glycerol but not with phorbol esters. The tegument of male worms was more sensitive to the effect of phorbol esters than females. Transmission electron microscopy revealed vacuolization of the tegument. These data suggest that signal transduction pathways may have a critical role in the maintenance of the structural integrity of the tegument of parasitic helminths.

It has been known for a long time that certain species of Mallophaga attach themselves to hippoboscid flies, the act taking place on the bird host. This has been referred to as Phoresy, ‘a type of interrelationship between insects in which one is carried on the body of another larger insect, but the former does not feed on the latter’, or, in other words, free passage without food.

In vivo and in vitro parameters of immunity have been assessed in guinea-pigs sensitized with 500 normal or 500 radiation-attenuated cercariae of Schistosoma mansoni. High levels of resistance to a challenge infection developed in both the chronic and irradiated vaccine model, but immunity was expressed earlier (week 4) and reached higher levels (90%) in the latter case. Vaccinated guinea-pigs have thus been shown to achieve greater resistance than the more commonly used rodent hosts. In vitro cytotoxicity assays have demonstrated that antibodies capable of participating in complement-dependent (lethal antibody) or eosinophil-mediated schistosomular killing, develop in the serum of guinea-pigs immunized with either normal or irradiated cercariae. The time course of development of the eosinophil adherence promoting antibody approximated in both models, the development of immunity in vivo, but the lethal antibody response paralleled the immune status of the animal only in the irradiated vaccine model

The effect of platyhelminth FaRPs and selected classical neurotransmitters on the motility of intact Mesocestoides corti (syn. M. vogae) tetrathyridial larvae was studied in vitro using a micromotility meter. The effects of the test substances were temperature dependent and these were examined at 4, 23, 30 and 36 °C. At 36 °C all test substances had concentration-dependent excitatory effects, with thresholds for activity of: 100 nM (GNFFRFamide), 10 μM (YIRFamide), 30 μM (GYIRFamide), 100 nM (serotonin) and 100 μM (acetylcholine). At this temperature significant elevation of motility indices (MI) was recorded within 5 min of the addition of peptide or serotonin. The effect of acetylcholine was slower in onset and appeared 15–20 min post-addition. At 30 °C larval motility diminished more rapidly than that recorded at 36 °C, following the addition of 1 mM of each test substance. At 23 °C only serotonin (1 mM) significantly increased the MI, all other test substances having no apparent effect. Larval movement was completely arrested at 4 °C. The results demonstrate for the first time excitatory effects of platyhelminth neuropeptides and acetylcholine on muscle systems in cestode larvae. The fact that the only known cestode FaRP, GNFFRFamide, was more potent than any of the turbellarian FaRPs tested, suggests structural conservation of FaRPs and FaRP receptors within the cestodes.

Neospora caninum infection persists throughout the life of its intermediate host due to the conversion of tachyzoites to slowly dividing bradyzoites that encyst in the brain. This event results in persistent N. caninum infection in bovine herds and partially explains the poor efficacy of many chemotherapeutic agents and vaccine formulations. Thus, there is a need for greater understanding of the tachyzoite-to-bradyzoite conversion mechanisms. Here we studied for the first time the transcription kinetics of the N. caninum bradyzoite-specific gene NcSAG4 in brain samples from chronically infected mice by means of real-time RT-PCR. NcSAG4-messenger RNA (mRNA) levels increased significantly during the chronic phase but followed 2 different expression patterns depending on the isolate used for murine inoculation. NcSAG4-mRNA levels in brains from Nc-1-inoculated mice peaked during late chronic infection (on day 64 post-infection, p.i.), whereas those from Nc-Liv-inoculated mice peaked earlier during the chronic infection (on day 32 p.i.). This difference could be a reflection of the different abilities of these isolates to replicate and form cysts in parasitized brains. These results are consistent with our observations of anti-rNcSAG4 antibody production; low levels were present at seroconversion and slowly increased during the chronic phase. In contrast, NcSAG1 transcription levels, which mark the tachyzoite stage, were maintained without variation in both groups of mice. This suggests the presence of a significant amount of tachyzoites or intermediate zoites expressing NcSAG1 in the brain, even during the late chronic infection.

Although the literature on ticks is very extensive, we have failed to find any satisfactory description therein of the process of copulation. Most authors state that the male mouthparts are introduced into the female genital orifice, or vulva, and that this constitutes copulation, leaving it to the reader's imagination to determine how the seminal secretion gains access to the female generative organs. The mechanism of copulation, in other words, remains to be described, and we propose, in this paper, to give an account of what we have observed in Ornithodorus moubata. We shall describe the process in Ixodidae in a later paper, but may state here that, as far as our knowledge goes, it is essentially the same as in moubata, that is, impregnation takes place by means of spermatophores.

Cyclophilins are a family of proteins found ubiquitously in eukaryotes, many of which bind to the immunosuppressive drug cyclosporin A (CsA). CsA has been found to have anti-parasitic effects against a variety of helminth and protozoan parasites and this activity could be mediated via cyclophilin. In this study we characterize a full length cyclophilin gene from Echinococcus granulosus, the associated natural gene and expression pattern, and investigate the functional properties of the recombinant E. granulosus cyclophilin protein. In addition, the effects of CsA were investigated on E. granulosus protoscoleces in in vitro culture. The full length E. granulosus cyclophilin cDNA encodes a protein of 20 kDa and is encoded by a single gene (EGCyP-1) comprising 2 exons separated by a 31 bp intron. The gene is expressed constitutively in all E. granulosus life-cycle stages examined. Recombinant E. granulosus cyclophilin (egCyP-1) exhibited functional enzyme activity as an isomerase. Treatment of in vitro cultures of E. granulosus protoscoleces with CsA was found to be lethal to the parasites. No protoscoleces survived treatment with 10 μg/ml of CsA over 7 culture days, as determined by observing motility and the uptake of toluidine blue dye. Untreated protoscoleces remained viable for the duration of experiments. The survival of protoscoleces was CsA dose dependent. A concentration of 10 μg/ml CsA was 100% lethal while doses of 8 μg/ml and 5 μg/ml resulted in 82% and 32% killing, respectively, after 7 days in culture. The anti-parasitic activity of CsA may have the potential to be developed as a new therapeutic agent for treatment of cystic hydatidosis in humans.

While working on the amphistomes in the Congo Collection of the American Museum of Natural History, New York, it has been necessary to review the literature on this group of trematodes. In an earlier paper (1917), the writer described certain amphistomes from reptiles and birds and submitted an arrangement to show the probable relationship of the reptilian and avian amphistomes to those of other vertebrates. The amphistomes of fishes had been studied by Daday (1907), those of frogs by Cohn (1904) and Johnston (1912), while the mammalian species had been the subject of extensive papers by Fischoeder (1903) and Stiles and Goldberger (1910). Many of the descriptions were fragmentary, knowledge of the forms was incomplete, and the classification of Stiles and Goldberger was received with severe criticism, especially by Braun (1911), Odhner (1911) and Looss (1912). In this paper Looss described the lymphatic systems of Schizamphistomum scleroporum and Paramphistomum gigantocotyle, and announced (p. 358) that for many years he had been engaged in preparing a classification of the amphistomes which would be based on the form of the lymph and excretory systems and the structure of the copulatory apparatus.

The fish fauna in the State of Acre represents 10·7% of all fish species recorded from Brazil, but, despite this, there are few fish parasite studies in this area. The recent expansion of fish farming in Acre prompted a need for helminthological studies of the most commonly consumed fish species in the area, Pimelodus blochii (Pimelodidae). The aim of this study was to analyse the helminth fauna of P. blochii from the Acre and Xapuri Rivers in Northwestern Brazil. Numerous nematodes were collected from the intestine and two species of the family Atractidae were identified: Rondonia rondoni Travassos, 1920 and Orientatractis moraveci n. sp. The new species is distinguished from its congeners mainly by having: 10 pairs of caudal papillae (3 pairs pre-cloacal, 2 pairs ad-cloacal and 5 pairs post-cloacal); unequal spicules of 161–198 and 69–100 µm long; and a gubernaculum 38–58 µm long with an antero-lateral process. Morphological and ultrastructural data on O. moraveci n. sp. and R. rondoni are presented, in addition to new genetic data based on partial 18S rDNA and 28S rDNA. The taxonomic status of Labeonema synodontisi (Vassiliadès, 1973) is discussed, suggesting that it should be returned to the genus Raillietnema.

The present paper deals with descriptions of some new species of parasitic mites, a fresh study of the forms of Eulaelaps stabularis (Koch), some observations on the occurrence of hypopial forms on other parasitic arthropods, and finally some observations on Radford's ‘Genera and species of parasitic mites’ (1943).

1. A brief outline of the life cycle of a species of Nosema parasitic in Saturniid moths is given. It is of the normal type.

2. The structure of the spore is described, and from electron microscopy evidence is presented which supports the theory that the sporoplasm is injected into the host's tissue.

3. The volume of the spore is affected by the ambient medium, by the pretreatments of fixation and of desiccation, by the site of development within the host and by the age of the host.

4. These results have been discussed with reference to the use of spore size as a diagnostic character in the classification of the Microsporidia. It is concluded that spore size is not a reliable character for taxonomic purposes.

The parasitaemia and associated blood picture following infections of Nuttallia rodhaini in rats and mice, and N. microti in rats are described. After inoculation of the blood stages of these parasites, it is evident that only further blood stages occur in the host animal. Infections with both piroplasms follow a similar course, there being a prepatent period followed by a patent infection which reaches a peak and then disappears. The prepatent period is an artifact due to the rarity of the parasites in the peripheral blood. The patent infection of N. rodhaini in rats and mice rapidly reaches a peak of up to 90% R.B.C. infected. That of N. microti develops more slowly and rarely reaches 20%. Rats show age resistance to both piroplasmoses but none is shown by mice aged up to 1 year against N. rodhaini infection. Increase of the parasite inoculum dose decreases the prepatent period and also the time from infection to the peak. In rats there is a corresponding increase of the peak but in mice this remains constant. N. microti infections in rats are enhanced by splenectomy, the peak being higher but the time from infection to the peak not altering. The parasitaemia produced by both piroplasms results in a temporary anaemia and reticulocytosis. The anaemia is not prolonged which does not suggest autoimmune reactions. Both piroplasms preferentially invade reticulocytes as compared with mature erythrocytes although this is apparently reversed at the onset of reticulocytosis.

An explanation of the mechanism of the parasitaemia of the piroplasms in rats and mice is suggested.

This paper is a report of work carried out in part for a thesis submitted for the degree of Ph.D. in the University of London. The work was aided by an S.R.C. research studentship. I would like to thank Dr F. E. G. Cox for his supervision and also for performing the splenectomies.

(1) Light-microscopic observations were made on B. pahangi larvae at various stages of development in stained serial sections of A. aegypti thoraces, and on 8-to 9-day 2nd-stage live, unstained larvae.

(2) Six-day larvae situated in A. aegypti flight muscle, and isolated 8- to 9-day 2nd-stage larvae, were examined with the electron microscope.

(3) Results showed that host-muscle mitochondria which appeared to be morphologically intact aggregated outside the larvae at the head and tail ends and were present from 6 days onwards in the oesophagus and in the midgut. Those in the oesophagus were unchanged; some of those in the midgut were intact, others showed varying degrees of disintegration suggestive of digestion. Host-muscle mitochondria are therefore likely to be a source of nutrients for B. pahangi larvae during the last third of their intramuscular developmental phase.

Dr W. W. Macdonald, of the Liverpool School of Tropical Medicine, kindly supplied the infected mosquitoes, and offered valuable advice in the course of this work. Miss M. A. Johnson assisted with the care of the mosquitoes.

We are particularly indebted to Mr David John for his patient work in preparing the filariae for electron-microscopic observation. Mrs J. Oliver assisted with some of the histological preparations.

We are pleased to acknowledge the financial assistance of the Royal Society towards the purchase of the ultramicrotomes used in the course of this work.

Bucephalus haimeanus is shown to be an estuarine species. The cercaria from Cardium edule is briefly redescribed to include details of the nephridial system. The metacercaria is described for the first time, having been obtained experimentally in Pomatoschistus microps and Pleuronectes platessa. In Pomatoschistus microps it survives at least 10 months, development within this host being completed within 2 months. The main site of infection is the liver. Migration is completed within 1 h and in some instances within 10 min. During this period the metacercaria actively feeds on host tissue, the gut being greatly distended with food on encystment. A hyaline cyst wall is maintained throughout the remaining life of the metacercaria. ‘O’ group plaice are highly susceptible to invasion by B. haimeanus during early summer and 100% infections were recorded in catches from Ynys-las, Dovey Estuary, in June. Experiments showed that only 2% of the metacercariae survived more than two weeks and that ‘l’ group plaice were non-susceptible to infection. Plaice are therefore considered accidental hosts. The metacercaria was linked with the adult stage from Morone labrax on the basis of comparative morphology and the ecology of the hosts, the bass entering the estuarine habitat during the summer months. The adult is considered synonymous with Bucephalus minimus Stossich, 1887.

Sporozoites of Eimeria necatrix inoculated into the allantoic cavities of embryonating eggs completed their life-cycle in the chorio-allantoic membranes and produced fully viable oocysts. Subsequently, as a result of repeated passage in this host, an egg-adapted line was developed and was found to be markedly less pathogenic for chickens than the nonpassaged parent strain. Antigens capable of inducing protective immunity against challenge with low doses of the parent strain were retained, however, indicating a possible use for this egg-adapted (attenuated) line of E. necatrix in the immunization of comercially reared chickens.

The effect of chemical abbreviation of the primary infection dose (PID) of 160 infective larvae of Metastrongylus apri on the immune status of the guinea-pig host was studied. The criteria used for assessing the status of immunity consisted of clinical manifestations following administration of a challenge infection dose (CID) of 800 infective larvae of M. apri, the rate of worm recovery 15 days post-CID and the rate of mortality following administration of CID.

Among the guinea-pigs of the main experimental group, where 15-day-old PID was abbreviated by two parenteral doses of levamisole, a strong immunity to CID given 35 days post-PID was built-up. Against this, all the guinea-pigs of a control group, which did not receive PID, died between 16 and 22 days post-CID.

The increase in serum gamma-globulin level of the guinea-pigs, where the PID was abbreviated chemically, suggested that the rise of this globulin fraction in the serum could be in some way related to the resistant state of guinea-pigs.

The Piraeeba, or Pirá-aïba (Piraïba) as Goeldi spells it, is a large Siluroid (my largest specimen measured 135 cm. in length) which is locally plentiful in the Amazon and has been described by Goeldi (1902). Goeldi named this species Piratinga pirá-aïba and it is the modern Brachyplatystoma filamentosum (Lichtenstein). I am indebted top Mr J. R. Norman of the British Museum (Natural History) for the modern name.

The formation and development of monogenean communities on the gills of roach fry was followed in 1992 from early June to October (size range 9 to 47 mm). Roach fry (n=291) were sampled weekly from the small, humic River Rutajoki in central Finland. A further 209 roach fry were reared in a fish farm supplied by water from the river. Four Dactylogyrus species were found: D. nanus, D. crucifer, D. micracanthus and D. suecicus. Other species found on the gills were Gyrodactylus sp. and Paradiplozoon homoion. The first Dactylogyrus juvenile occurred on a 12 mm long fish fry in late June and the first adult (D. nanus) 1 week later in Tank 1. D. nanus was also the most common parasite in the river. Young fry had high numbers of Gyrodactylus sp. on the gills compared with adult roach in previous studies. Signs of site preference were found; D. nanus and D. micracanthus preferred the 2nd gill-arch and Dactylogyrus juveniles the outmost one. Abundances of monogenean infracommunities increased until the middle of August when, on average, 1·9 species and 3·7 worms per fry were noted and decreased then as the water temperatures fell. A lack of competition between species is suggested; for example, D. crucifer appeared on the gills at that time when abundance of D. nanus was at its highest. The roach fry hatched and reared in the fish farm revealed that larvae of the most common dactylogyrid species were able to disseminate and colonize over 120 m from the river to the farm. Influence of change colonization events was shown by the appearance and development of Gyrodactylus sp. population on roach fry in only 1 of the 2 tanks at the farm.