Bluetongue is a disease of major economic concern in Europe. Its causative agent, bluetongue virus (BTV), is transmitted by several Culicoides species (mainly Culicoides imicola and Culicoides obsoletus in Europe). The application of insecticides on animals may reduce transmission of BTV, however, no formulation is currently licensed specifically against Culicoides midges. The present study assesses the susceptibility of C. obsoletus to deltamethrin using an adapted World Health Organization (WHO) susceptibility test. Midges were exposed to different dosages of deltamethrin for 1 h, and mortality after 1 h and 24 h was recorded. Results indicated that deltamethrin is highly toxic to C. obsoletus since a dose of 1·33×10−4% was enough to kill 50% of the population (LD50) in 24 h. The deltamethrin concentration needed to kill 90% of the population (LD90) was 5·55×10−4%. The results obtained in the present work could help to create a system that can be used to assess insecticide resistance and susceptibility of Culicoides biting midges.

A new species of Microsporidia Microgemma carolinus n. sp. found in the marine teleost Trachinotus carolinus collected in Florianópolis, Brazil was described based on light, ultrastructural and phylogenetic studies. This parasite developed in the liver forming whitish xenomas that contained different developmental stages with monokaryotic nuclei. The periphery of the xenoma presented some vacuolization and possessed several small projections in the membrane. The mature spores, measuring 3·8 ± 0·4 μm in length and 2·4 ± 0·4 μm in width, were slightly pyriform to ellipsoidal and had rounded ends. The polaroplast was bipartite and the isofilar polar filament was coiled with 8 – 9 turns in a single or double row at the posterior end of the spore. The nucleus was voluminous and in a central position, measuring ∼0·9 μm in diameter. A large posterior vacuole appeared as a pale area, occupying about a third of the spore length. The SSU rRNA gene was sequenced and analysed using maximum parsimony, maximum likelihood and neighbour-joining methods. This study allowed us to conclude that this was a new species of the genus Microgemma, being the first description of this genus from among South America fauna.

The claim made for the first time by Barrett and Smith (1914a) some two years and a half ago, that the causative agent of Pyorrhoea Alveolaris is the Entamoeba gingivalis led the writers to enquire into the question; the results of which enquiry are set forth in the following paper.

The aim of this study was to evaluate the therapeutic effects of osteopontin neutralization treatment on schistosome-induced liver injury in BALB/C mice. We randomly divided 100 BALB/C mice into groups A, B, C, D and group E. Mice in all groups except group A were abdominally infected with schistosomal cercariae to induce a schistosomal hepatopathological model. Mice in group C, D and group E were respectively administered with praziquantel, praziquantel plus colchicine and praziquantel plus neutralizing osteopontin antibody. We extracted mouse liver tissues at 3 and 9 weeks after the ‘stool-eggs-positive’ day, observed liver histopathological changes by haematoxylin-eosin and Masson trichrome staining and detected the expression of osteopontin, alpha-smooth muscle actin (α-SMA) and transforming growth factor-beta (TGF-β1) by immunohistochemistry, RT-PCR and Western blot. We found that praziquantel plus neutralizing osteopontin antibody treatment significantly decreased the granuloma dimension, the percentage of collagen and the expression of osteopontin, α-SMA and TGF-β1 compared to praziquantel plus colchicine treatment in both the acute and chronic stage of schistosomal liver damage (P<0·05). So we believe that the combined regimen of osteopontin immunoneutralization and anti-helminthic treatment can reduce the granulomatous response and liver fibrosis during the schistosomal hepatopathologic course.

Interactions between infection and nutrition have been well recognized for several years now since they contribute directly to the health of individuals and communities. Malnourished individuals are specially prone to developing infections while infections themselves can lead to profound changes in the nutritional status of the individual. Health workers in developing countries in the tropics have long recognized the mutually aggravating interactions of malnutrition and infection. The importance of this synergistic relationship between infection and nu-tritional status has been studied extensively in the case of young children. The nutritional status of a young child is a critical determinant of both c morbidity and mortality resulting from a wide range of infections: bacterial, viral, or parasitic. Chandra (1983), in his review on the relationship of nutrition, immunity and infection has categorized the wide range of infectious agents (bacterial, viral, fungal and parasitic) into those that are definitely, variably or minimally influenced by the nutritional status of the child.

Various endocrine operations were performed on the adult female cockroach Blatta orientalis and the adult populations of the nematode Hammerschmidtiella diesingi were recorded 2 or 4 weeks later. Cautery of the median neurosecretory cells (m.n.c.) caused a significant reduction in total numbers of adult nematodes after 2 or 4 weeks; after 2 weeks, removal of the retrocerebral complex (corpora cardiaca plus corpora allata) caused a significant overall reduction in the adult nematode burden and this could not be prevented by reimplanting the corpora allata immediately after the retrocerebral complex had been removed.

Anomalies appeared in the data when the sex of the parasite was examined and this may be related to a possible postembryonic mode of sex determination.

Both m.n.c. cautery and removal of the retrocerebral complex suppressed host terminal oocyte development and ootheca production, whilst reimplantation of corpora allata into ‘retrocerebralectomized’ cockroaches allowed reproductive processes to continue normally. Hence, the m.n.c. and corpora allata are essential for host egg development.

Histological studies showed a profound reduction in fuchsinophilic material in the corpora cardiaca of m.n.c. cauterized cockroaches.

Although the m.n.c. may affect the nematode directly or indirectly, a direct effect on the parasite by the corpora allata is unlikely.

Acanthocephalans have evolved a hooked proboscis and some taxa have trunk spines to attach to their definitive hosts. These structures are generated before being used, thus a key question is how investment in attachment could optimally be allocated through the ontogeny. The number and arrangement of hooks and spines are never modified in the definitive host, but it is unclear whether these structures grow during adult development. A comparison of the size of trunk spines between cystacanths and adults of Corynosoma cetaceum and C. australe indicated that spines grow in both species, but only in females, which also had significantly larger spines than males. This sexual dimorphism did not result from pure allometry because the body of females was smaller, and did not grow more than that of males. However, having a longer lifespan, females would need to withstand the extreme flow conditions prevailing in marine mammals for longer, inducing different investment and development schedules for spines. Patterns of spine growth also differed between species: fore-trunk spines grew in both species, but hind-trunk spines did only in C. cetaceum. In conclusion, investment strategies on attachment may differ, not only between congeneric species of acanthocephalan, but also between sexes of the same species.

Production of elevated haemolymph agglutination titres by Biomphalaria glabrata following exposure to Echinostoma paraensei miracidia was investigated, to characterize this parasite-induced response and to understand its functional relevance. Both the dose of infection (1, 10 or 100 miracidia per snail) or the number of separate exposures to infection (between one and three, over a 4 or 8 day interval) were varied, and assuming a threshold dosage (10 miracidia per snail or higher) was exceeded, titres of juvenile snails peaked at 8–16 times the values for unexposed control snails, regardless of the exposure regimen. Adult snails, which are relatively refractory to infection, have slightly higher resting titres than juveniles, but exhibit only a 2- to 4-fold increase in titre following exposure. Juveniles exposed to infection but lacking demonstrable infection had lower titres than snails with confirmed infections. Exposure to infection increased heterogeneity of plasma agglutinins and provoked production of unique specificities not found in unexposed snails. However, the overall pattern of agglutination responses for snails with successfully developed parasites did not differ from those in which parasite development was unsuccessful. Agglutinating activity was inhibitable by several different monosaccharides, although plasma from infected snails was relatively unaffected by N-acetyl-glucosamine or N-acetyl-galactosamine. Wounding of snails provoked no change in plasma agglutination activity. As the highest agglutination titres were produced in snails with successfully developing parasites and agglutinin composition did not differ between snails with successful or unsuccessful parasites, the functional relevance of the response remains enigmatic. The production of unique agglutinins following exposure deserves additional study.

The ultrastructure and development of Histomonas meleagridis in the reproductive tract of male Heterakis gallinarum has been described. In the testis the protozoon is a relatively large amoeboid organism which lies between the epithelial cells of the testis and the spermatogonia, spermatocytes and spermatids. It is similar in appearance to the invasive stage which is found in the final host and like the invasive stage, it also feeds upon the host cells by phagocytosis. In the vas deferens the protozoon lies between the host cells, where it is much smaller than the stages in the testis but still amoeboid, and inside the cells of the vas deferens.Intracellular stages are much smaller than the other stages, are rounded in appearance and cause little or no damage to the host cell. They are similar to the stages found in oocytes of the female nematode. The possible role of the male nematode in the life cycle of the protozoon is discussed.

I wish to thank Mrs B. Fisher for technical assistance and Mr B. Carter for assistance with the photography.

The trypanosome to be described in this paper was discovered in blood films taken from the snake Erythrolamprus aesculapii (Duméril and Bibron) of tropical America. For these films I am indebted to Dr Leiper. In addition to the trypanosome there was present in the blood a haemogregarine. Though haemogregarines are very common in snakes, especially in the Tropics, where nearly every snake examined is found to harbour these parasites, the reverse is the case with trypanosomes. Several observers have recorded the presence of trypanosomes in snakes but hitherto no one has given an accurate description of one of trypanosomes of the whole group of reptiles is very limited when campared with other groups of vertebrata. On this account it seems of interest to place on record the characters of this trypanosome as it appears in the blood films mentioned above.

Hymenolepis microstoma from secondary infections in mice were found to grow initially more slowly than H. microstoma from primary infections. Eventually a similar rate of growth was attained by both kinds of worm. After transplantation, young worms (≤4 days old) grew more slowly in previously infected than in naive mice. In contrast, 10-day-old worms grew equally well in naive or resistant mice. The administration of cortisone to mice during a secondary infection of H. microstoma inhibited the stunting of growth in young worms which were able to grow as well as those in a primary infection. The effects of the cortisone persisted for less than 4 days. Worms in a secondary infection given 4days after cortisone treatment were stunted in growth. These experiments suggest that the susceptibility of the worms to the resistance of the mouse applies to the first 4 days within the host. The loss or reduction of this susceptibility might be associated with the worms' entry into the bile duct which occurs 3–4 days after infection.

Oesophagostomum dentatum stages were investigated for glutathione S-transferase (GST) expression at the protein and mRNA levels. GST activity was detected in all stages (infectious and parasitic stages including third- and fourth-stage larvae of different ages as well as males and females) and could be dose-dependently inhibited with sulfobromophthalein (SBP). Addition of SBP to in vitro larval cultures reversibly inhibited development from third- to fourth-stage larvae. Two glutathione-affinity purified proteins (23 and 25 kDa) were detected in lysates of exsheathed third-stage larvae by SDS-PAGE. PCR-primers were designed based on peptide sequences and conserved GST sequences of other nematodes for complete cDNA sequences (621 and 624 nt) of 2 isoforms, Od-GST1 and Od-GST2, with 72% nucleotide similarity and 75% for the deduced proteins. Genomic sequences consisted of 7 exons and 6 introns spanning 1296 bp for Od-GST1 and 1579 and 1606 bp for Od-GST2. Quantitative real-time-PCR revealed considerably elevated levels of Od-GST1 in the early parasitic stages and slightly reduced levels of Od-GST2 in male worms. Both Od-GSTs were most similar to GST of Ancylostoma caninum (nucleotides: 73 and 70%; amino acids: 80 and 73%). The first three exons (75 amino acids) corresponded to a synthetic prostaglandin D2 synthase (53% similarity). O. dentatum GSTs might be involved in intrinsic metabolic pathways which could play a role both in nematode physiology and in host-parasite interactions.

Trypanosoma evansi is the aetiological agent of trypanosomosis in domestic animals. In this pathology, an inflammatory response can be observed and, as a consequence, the increase of extracellular adenine nucleotides such as ATP. These nucleotide concentrations are regulated by ectoenzymes such as NTPDase (EC 3.6.1.5, CD39), which catalyses the hydrolysis of ATP and ADP into AMP. In this study, the activity of NTPDase in lymphocytes of rats experimentally infected with T. evansi was evaluated. The animals were inoculated with the parasite and monitored by blood smear on a daily basis. The animals were then were divided into 4 groups according to the degree of parasitaemia and period of infection. The blood collections for enzyme analysis and lymphocyte count were performed on the 3rd (beginning of infection), 5th (acute infection) and 15th (chronic infection) days post-infection (p.i.). The control group was composed of non-infected animals. In the infected group a decrease in ATP hydrolysis (36%) was observed on the 3rd day p.i. and a decrease in ADP hydrolysis (62%) was observed on the 5th day p.i. when compared to the control. On the 15th day p.i., an increase in ATP (94%) and ADP (50%) hydrolysis was observed in the infected group. Considering these data it is suggested that NTPDase activity is altered on the surface of lymphocytes of rats infected with T. evansi at different time-points of infection.

Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that mediate the fast actions of the neurotransmitter, acetylcholine. Invertebrate nAChRs are of interest as they are targets of widely-selling insecticides and drugs that control nematode parasites. Here, we report the cloning of ShAR2β, a candidate nAChR subunit from the blood fluke, Schistosoma haematobium, which is the third trematode nAChR subunit to be characterized. While ShAR2β possesses key structural features common to all nAChRs, its amino acid sequence shares considerably low identity with those of insect, nematode and vertebrate nAChR subunits. In particular, the second transmembrane domain of ShAR2β, which lines the ion channel, bears unusual amino acid residues which will likely give rise to a receptor with distinct functional properties. Phylogenetic analysis shows that ShAR2β is a divergent nAChR subunit that may define a clade of trematode-specific subunits. We discuss our findings in the context of potentially exploiting this receptor as a target for controlling schistosome parasites.

Species of the phylum Myxozoa are common parasites of fish and can cause severe losses in cultured species. Although a number of myxozoan life-cycles have now been elucidated, little is known about the biology of these organisms in the fish host. Monoclonal antibody B4 raised to the myxozoan Tetracapsuloides bryosalmonae has been previously noted to react with a number of species infecting fish kidney. We present the results of a survey of 55 myxosporean species that determined that this antibody detects an antigen on the spore surface of 33 of these species in the genera Myxobolus, Sphaerospora and Thelohanellus. However, there appears to be no clear relationship between those spores that contain the MAb B4 reactive antigen and the host or organ in which they are detected. The antigen appears to be synthesized in the plasmodial cytoplasm and is intimately associated with the surface of the spore capsules and, where present, the mucus envelope. The nature of this envelope is further discussed in relation to its formation and distinctive properties.

Dictyocotyle coeliaca is recorded for the first time from Raja fullonica and R. batis. It is suggested that normally D. coeliaca may be restricted to waters deeper than about 50 fathoms and that this should be considered in attempts to incubate its eggs in the laboratory. No Calicotyle kroyeri were present in female R. radiata which contained eggs in their uteri and a brief account is given of the apparent differential distribution of the species in male and female rays. Up to 64 specimens of C. kroyeri per fish are recorded from the cloacae of R. radiata (previous records refer to an average of about four specimens per host). In a brief discussion of the controversial papers on these two monogeneans reasons are given for retaining the two in separate genera.

I am most grateful to the following for providing excellent facilities for the work. The Director and staff, in particular Dr B. B. Rae and Dr Z. Kabata, Marine Laboratory, Aberdeen; the Director and Staff, Plymouth Marine Laboratory; Professor James Brough, University College, Cardiff; the Director, Commonwealth Bureau of Helminthology; and Mr H. Atkinson of Messrs J. Marr and Son, Fleetwood. Additional specimens were provided by Mr J. Slinn, Marine Biological Station, Port Erin, and by Mr J. W. Smith of this Bureau.

Trophically transmitted parasites may use multiple intermediate hosts, some of which may be ‘key-hosts’, i.e. contributing significantly more to the completion of the parasite life cycle, while others may be ‘sink hosts’ with a poor contribution to parasite transmission. Gammarus fossarum and Gammarus roeseli are sympatric crustaceans used as intermediate hosts by the acanthocephalan Pomphorhynchus laevis. Gammarus roeseli suffers higher field prevalence and is less sensitive to parasite behavioural manipulation and to predation by definitive hosts. However, no data are available on between-host differences in susceptibility to P. laevis infection, making it difficult to untangle the relative contributions of these hosts to parasite transmission. Based on results from estimates of prevalence in gammarids exposed or protected from predation and laboratory infections, G. fossarum specimens were found to be more susceptible to P. laevis infection. As it is more susceptible to both parasite infection and manipulation, G. fossarum is therefore a key host for P. laevis transmission.

During my work on the mites I have often wished that a book were available in which all the known and valid genera and species of parasitic mites could be found. The literature on the Acarina contains numerous genera and species which are synonyms of older identifications.

Endogenous transplacental transmission (EnTT) of Neospora caninum is the most common route of infection in cattle and occurs as a consequence of a reactivation of N. caninum infection that may lead to abortion or to the birth of congenitally infected calves. The reactivation of N. caninum infection was studied during the gestation of chronically infected dams and, for the first time, in their congenitally infected pups. BALB/c mice were infected with Nc-Spain 7 (Group 1) or Nc-Spain 3H (Group 2), high virulence isolates and low-to-moderate virulence isolates, respectively. The mice were mated after 90 days post-infection, and the morbidity, mortality, vertical transmission and humoral immune responses were recorded for 2 consecutive generations. In the first generation, higher morbidity and mortality rates were observed in G1 before mating than in G2 (P < 0·0001). In the second generation, low vertical transmission rates were observed in both inoculated groups (7·7% and 17·1% in G1 and G2, respectively) and were significantly diminished in the third generation (8·7% in G2 versus 0% in G1). Low rates of reactivation of N. caninum infection were induced in chronically infected mice and decreased in subsequent generations regardless of the isolate employed in the inoculations. Thus, further studies are needed to improve this reactivation mouse model.

The serial passage of 3 strains of Eimeria from the chicken in embryonating turkey embryos is reported here. All 3 strains increased their reproduction in turkey embryos; reproduction being equal to or better than the same lines maintained in chicken embryos. Pathogenicity and immunogenicity trials conducted in chickens using these 3 strains of Eimeria showed that all 3 turkey embryo-adapted lines were significantly less pathogenic than the parent lines. Only 1 of the lines (E. tenella (PS) turkey embryo-adapted) maintained its immunogenic potential