It was found that interference contrast microscopy gave good morphological differentiation of the living and intact salivary glands from adult ixodid ticks, Rhipicephalus appendiculatus and Amblyomrna variegatum, removed by dissection. In many cases the individual cell types of the acini could be recognized. When the salivary glands of infected adult ticks were examined Theileria parasites were detected. Theileria parva, T. taurotragi and T. mutans could be detected in the salivary glands after they reached the mature secondary sporoblast stage achieved even in unfed ticks. Parasites were detected only in type III acini. The morphology of tertiary sporoblasts and sporozoites of these Theileria spp. was clearly recognized in feeding ticks. Sporozoites of T. parva proved to be infective to cattle and to peripheral blood leucocytes of cattle in vitro after interference contrast examination. Interference contrast examination of living salivary glands was used for selection of salivary glands with high infections of T. parva sporozoites and the sporozoites could be separated in large numbers from these salivary glands. Using this technique it appeared possible to clone Theileria parasites from salivary glands with 1 infected acinus.

Groups of ten cattle were exposed to 0, 40 and 400 adult Rhipicephalus appendiculatus feeding once a week for 24 weeks. Although no differences in cumulative percentage weight gain were found at the end of the experiment, animals free of ticks performed better during the first 12 weeks. A marked recovery of the 400-tick group was recorded during weeks 13–24 and after ticks were removed. Haemoglobin and packed cell volume values were lowered in tick-infested animals but no clear trends were observed in red and white blood cells. No differences between the percentages of ticks feeding in the 2 tick-infested groups on day 2 were observed but higher numbers of females were feeding on the 40-tick group on day 5. More ticks were found on day 5 after tick application to the cattle during weeks 1–6 than in the rest of the experiment. Significant correlations were found between lymph node enlargement and ear damage when they were tested against tick load. R. appendiculatus nymphs feeding on the formerly tick-infested cattle were fewer, lighter and engorged in a shorter time than those on the control animals. The possible causes for the reduction in ticks feeding, the changes in cattle weights, blood composition and development of host resistance are discussed.

Host selection among humans by Anopheles punctulatus was studied in an area of Papua New Guinea endemic for malaria and filariasis. Blood films were made from the stomach contents of freshly engorged mosquitoes found resting on the walls of houses in which the parasite status of the occupants was known. Engorgement rates on humans were non-random but could not be consistently related to the parasite status of individuals in the houses for either malaria or filaria. In some households, anophelines preferentially fed on parasitaemic individuals while in other households aparasitaemic individuals were significantly more often selected. This finding is believed to reflect the fact that malaria and filarial infections in this endemic area are predominantly asymptomatic. There were no significant differences in axillary temperatures between malaria or microfilariae positive or negative individuals.

In this paper the Leishmania (Viannia) braziliensis complex is defined as containing all species of the actual subgenus Viannia. Organisms of the L. (V) braziliensis complex are the causative agents of localized human cutaneous and mucocutaneous leishmaniasis in South America, much of Central America and some areas of North America. In our search for better species and subspecies diagnostic probes we focused our research on repetitive DNA, since it provides a greater number of target sites for hybridization. In this work we report the isolation and sequencing of a 1·8 kb DNA region, LbJ38, which is probably tandemly repeated or dispersed at least 4 times along one chromosome and is naturally present in L. (V) braziliensis genomic DNA. This region contains microsatellites and simple repeat DNA sequences and was isolated by screening a genomic DNA cosmid library with complex- and species-specific probes. No homology was found with other Leishmania microsatellite or repetitive DNA. The utility of this repetitive sequence and primers derived from it in the identification of L. (V) braziliensis is demonstrated. As far as we are aware, this is the first report of sequence characterized repetitive microsatellite and GC rich simple repeat DNA from the nuclear genome of New World Leishmania.

Different stages of Dipetalonema viteae (males, females, microfilariae, and 3rd-stage larvae) have been iodinated in vitro under physiological conditions by chloroglycoluril, lactoperoxidase or chloramine T. The concentrations of the catalysts were correlated with the viability of the worms. Localization of the label with the different iodination methods had been visualized by electron microscopical autoradiography. Chloroglycoluril-mediated iodination is predominantly localized on the filarial cuticle. Lactoperoxidase-catalysed iodination is less specific and chloramine T catalyses iodination in a gradient decreasing from the cuticle to inner structures. It is necessary to visualize the labelling by electron microscopical autoradiography prior to biochemical and immunological experiments to avoid the extraction of structures iodinated by leakage of the catalyst into sub-cuticular regions.

The uptake of radioactive pyrimethamine by a sensitive and a resistant strain of Plasmodium falciparum, the metabolic fate of pyrimethamine inside these parasites and the kinetic properties of dihydrofolate reductase (DHFR) from both strains have been studied. Uptake of the drug was identical in both strains and no metabolite of pyrimethamine was found in either strain. DHFR from the resistant strain was 300 times less sensitive to inhibition by pyrimethamine than the enzyme from the sensitive strain, while the Michaelis constant for dihydrofolate remained unchanged and inhibition was competitive in both cases. Altered properties of plasmodial DHFR are apparently the only mechanism responsible for pyrimethamine resistance in the strain of Plasmodium falciparum studied.

Increased permeability of capillaries and venules, labelled with colloidal carbon, was observed in the superficial mucosa of the small intestine of sheep infected with Trichostrongylus colubriformis. The labelled vessels were restricted to infected portions of the gut, and the intensity of labelling appeared to be related to density of the worm population. Open junctions were seen between endothelial cells in permeable vessels. In moderately atrophic infected intestine, enterocytes were domed, had sparse, short, distorted microvilli and many polyribosomes. Tight junctions between enterocytes seemed to be intact and distended intercellular spaces were seen. In more severely atrophic mucosa, enterocytes had distended mitochondria and endoplasmic reticulum, bizarre shapes, and the continuity of the epithelial sheet was occasionally disrupted. There was oedema in the lamina propria and neutrophils were seen in migration to the lumen of the gut. The possible mechanisms inducing these changes and their significance in terms of plasma protein loss and intestinal function are discussed.

An African buffalo (Syncerus caffer), born in captivity and demonstrated to be Theileria-free, and 2 susceptible cattle were inoculated with a Theileria parva lawrencei sporozoite stabilate. The buffalo had a very mild disease reaction, while the 2 cattle died of acute theileriosis. It was possible to isolate T. p. lawrencei from the buffalo up to 888 days after infection by the application of non-infected Rhipicephalus appendiculatus nymphs and up to 657 days after infection by the establishment of lymphoblastoid cell lines infected with T. p. lawrencei schizonts from peripheral mononuclear blood cells. The infection rate and levels of Theileria in the resultant adult ticks varied from 11 to 70% with 0·3–11 acini infected/tick. Stabilates prepared from these tick batches caused fatal T. p. lawrencei infections in cattle.

We cloned and expressed Echinostoma caproni HSP70 in Escherichia coli. This molecule presents an open reading frame (ORF) of 655 amino acids, and a theoretical molecular weight of 71 kDa. E. caproni HSP70 protein showed a high homology to other helminth molecules, major differences being located in the C-terminal region of the molecule, with a hydrophobic portion. Studies of protein and messenger RNA (mRNA) expression revealed a distinct pattern, depending on the host (low- or high-compatible). Specific polyclonal antisera raised against the recombinant protein expressed in Escherichia coli demonstrated its selective presence in excretory/secretory products (ESP) of adult parasites obtained from high-compatible hosts. Immunological studies showed clearly the association of HSP70 with the parasite surface and other structures, including eggs.

The object of this paper is to develop the ideas put forward in previous publications (Duke, 1919, 1921).

The European earwig (Forficula auricularia L.) accidentally introduced some years ago into the United States, is now abundant in several of the Western and at least one of the Eastern States; it has become established also in New Zealand, where, according to Tillyard, it has become a major pest, eating into peaches, nectarines, and apricots and rendering them useless for sale.

There is a gap in our understanding of the relative and interactive effects of different parasite species on the same host population. Here we examine the effects of the acanthocephalan Acanthocephalus galaxii, an unidentified cyclophyllidean cestode, and the trematodes Coitocaecum parvum and Microphallus sp. on several fitness components of the amphipod Paracalliope fluviatilis, using a combination of infection surveys and both survival and behavioural trials. In addition to significant relationships between specific parasites and measures of amphipod survival, maturity, mating success and behaviour, interactions between parasite species with respect to amphipod photophilia were also significant. While infection by either A. galaxii or C. parvum was associated with increased photophilia, such increases were negated by co-infection with Microphallus sp. We hypothesize that this is due to the more subtle manipulative effect of A. galaxii and C. parvum being impaired by Microphallus sp. We conclude that the low frequency at which such double infections occur in our sampled population means that such interactions are unlikely to be important beyond the scale of the host individual. Whether or not this is generally true, implying that parasitological models and theory based on single parasite species studies do generally hold, requires cross-species meta-analytical studies.

Additional biomarkers are essential for control of Trypanosoma evansi (T. evansi) infection in dromedary camels. Two groups of 30 camels each, one naturally infected with T. evansi and other normal healthy were executed. The basis for the infection was the positive findings of clinical examination, blood smears and latex agglutination test. Blood samples of both groups and its harvested serum were used for the estimation of present serobiochemical parameters. The present findings revealed significant decrease (P ⩽ 0·05) in triacylglycerol, cholesterol, high density lipoprotein cholesterol with significant increase (P ⩽ 0·05) in low density lipoprotein cholesterol, beta hydroxyl butyric acids, non-esterified fatty acids, haptoglobin, serum amyloid A, ceruloplasmin, fibrinogen, interleukins, tumour necrosis factor-α, interferon gamma, malondialdehyde, superoxide dismutase, reduced glutathione and catalase of infected camels compared with the control. The present study suggests lipid profile, acute phase proteins, proinflammatory cytokines and oxidative stress parameters as biomarkers for T. evansi infection in camels.

The drive to control neglected tropical diseases (NTDs) has had many successes but to reach defined targets new approaches are required. Over the last decade, NTD control programmes have benefitted from increased resources, and from effective partnerships and long-term pharmaceutical donations. Although the NTD agenda is broader than those diseases of parasitic aetiology there has been a massive up-scaling of the delivery of medicines to some billion people annually. Recipients are often the poorest, with the aspiration that NTD programmes are key to universal health coverage as reflected within the 2030 United Nations sustainable development goals (SDGs). To reach elimination targets, the community will need to adapt global events and changing policy environments to ensure programmes are responsive and can sustain progress towards NTD targets. Innovative thinking embedded within regional and national health systems is needed. Policy makers, managers and frontline health workers are the mediators between challenge and change at global and local levels. This paper attempts to address the challenges to end the chronic pandemic of NTDs and achieve the SDG targets. It concludes with a conceptual framework that illustrates the interactions between these key challenges and opportunities and emphasizes the health system as a critical mediator.

Trypanosoma cruzi is the causative agent of Chagas’ disease, a parasitic disease that remains a serious health concern with unsatisfactory treatment. Drugs that are currently used to treat Chagas’ disease are partially effective in the acute phase but ineffective in the chronic phase of the disease. The aim of the present study was to evaluate the antitrypanosomal activity and morphological, ultrastructural and biochemical alterations induced by a new molecule, 4-nitrobenzaldehyde thiosemicarbazone (BZTS), derived from S-(-)-limonene against epimastigote, trypomastigote and intracellular amastigote forms of T. cruzi. BZTS inhibited the growth of epimastigotes (IC50 = 9·2 μm), intracellular amastigotes (IC50 = 3·23 μm) and inhibited the viability of trypomastigotes (EC50 = 1·43 μm). BZTS had a CC50 of 37·45 μm in LLCMK2 cells. BZTS induced rounding and distortion of the cell body and severely damaged parasite mitochondria, reflected by extensive swelling and disorganization in the inner mitochondrial membrane and the presence of concentric membrane structures inside the organelle. Cytoplasmic vacuolization, endoplasmic reticulum that surrounded organelles, the loss of mitochondrial membrane potential, and increased mitochondrial O2•ˉ production were also observed. Our results suggest that BZTS alters the ultrastructure and physiology of mitochondria, which could be closely related to parasite death.

Cysticercosis caused by infection with the larval stage of Taenia solium is an important cause of neurological disease worldwide and immunodiagnosis is important for the control and elimination of cysticercosis. In the present study, we established a simple and reliable preparation of immunodiagnostic low-molecular-weight antigens (LMWAgs) from T. solium cyst fluids by a cation-exchange chromatography (CEC). Banding patterns of LMWAgs on SDS-PAGE were different between isolates from Ecuador and China. All cysticercosis patient sera and some echinococcosis patient sera recognized both LMWAgs by enzyme-linked immunosorbent assay (ELISA), but sera from healthy persons were not positive. There was no statistical difference in immunodiagnostic performance of LMWAgs prepared from different geographical isolates. These results indicated that these novel immunodiagnostic antigen preparations could contribute the control and prevention of cysticercosis in endemic areas, especially developing countries.

The Jamaican iguanid lizard Anolis lineatopus is the host of 4 species of helminth parasite: an acanthocephalan (Centrorhynchus spinosus), a digenean (Mesocoelium danforthi), and 2 nematodes (Cyrtosumum scelopori and Thelandros (?cubensis). Prevalance of C. spinosus was unrelated to host age but was considerably higher in males (43%) than in females (10%). The intensity increased with host age: (range: 1–23, median: 3). Prevalence of C. scelopori increased with host age in both sexes: juveniles (<3 months) were uninfected while infection occurred in almost all lizards older than 9 months. Intensity was higher in males (range: 1–403, median: 158) than in females (range: 1–297, median: 86). The infection patterns of C. spinosus and C. scelopori differed significantly from each other and were not consistent with a simplistic direct relationship between time of exposure and infection prevalence. Acanthocephalan and digenean infections occurred at low prevalence and intensity although prevalence in older, larger lizards tended to be higher than in younger, smaller ones.

Toxocara canis infective stage larvae continually produce excretory–secretory (TES) glycoproteins in long-term in vitro culture. The kinetics of synthesis and secretion were studied by metabolic labelling with radioactive [35S]methionine, [14C]serine and [14C]threonine. Maximal incorporation rates required overnight pre-incubation of parasites in medium depleted of the appropriate amino acid. Larvae rapidly incorporated isotope into their somatic tissues, but there was a minimum delay of 10 h before secretion of labelled antigens. Labelling with [14C]serine and [14C]threonine demonstrated a relative abundance of these amino acids in the major surface/secreted glycoproteins of this nematode (TES-32 and 120). Pulse-chase experiments suggested that TES-120 may be derived from a 58 kDa precursor, reflecting extensive post-translational glycosylation. Inhibition of N-glycosylation with tunicamycin and digestion with N-glycanase provided evidence of N-glycosylation in the lower molecular weight ES components (TES-32, 55 and 70). These agents had no effect on the higher molecular weight components (TES-120 and 400) implying that for these molecules glycosylation is predominantly O-linked. The largest ES component (TES-400) was unusual, in incorporating serine and threonine but not methionine, and by exhibiting increased apparent molecular weight following pronase digestion; it is suggested that this molecule is a proteoglycan.

Cryptosporidium is a protozoan parasite of humans and animals and has a worldwide distribution. The parasite has a unique epidemiology in Middle Eastern countries where the IId subtype family of Cryptosporidium parvum dominates. However, there has been no information on Cryptosporidium species in Yemen. Thus, this study was conducted in Yemen to examine the distribution of Cryptosporidium species and subtype families. Fecal samples were collected from 335 patients who attended hospitals in Sana'a city. Cryptosporidium species were determined by PCR and sequence analysis of the 18 s rRNA gene. Cryptosporidium parvum and C. hominis subtypes were identified based on sequence analysis of the 60 kDa glycoprotein (gp60) gene. Out of 335 samples, 33 (9·9%) were positive for Cryptosporidium. Of them, 97% were identified as C. parvum whilst 1 case (3%) was caused by C. hominis. All 7 C. parvum isolates subtyped belonged to the IIaA15G2R1 subtype. The common occurrence of the zoonotic IIa subtype family of C. parvum highlights the potential occurrence of zoonotic transmission of cryptosporidiosis in Yemen. However, this postulation needs confirmation with future molecular epidemiological studies of cryptosporidiosis in both humans and animals in Yemen.