In the Kingdom of Saudi Arabia (KSA), Old World cutaneous leishmaniasis (CL) is mainly caused by Leishmania major and Leishmania tropica parasites. Diagnosis of CL is predominately made by clinicians, who at times fail to detect the disease and are unable to identify parasite species. Here, we report the development of a chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) to measure the levels of anti-α-galactosyl antibodies in human sera. Using this assay, we have found that individuals infected with either Leishmania spp. had significantly elevated levels (up to 9-fold higher) of anti-α-Gal IgG compared to healthy control individuals. The assay sensitivity was 96% for L. major (95% CI; 94–98%) and 91% for L. tropica (95% CI; 86–98%) infections and therefore equivalent to restriction fragment length polymorphism-polymerase chain reaction analysis of parasite ITS1 gene. In addition, the assay had higher sensitivity than microscopy analysis, which only detected 68 and 45% of the L. major and L. tropica infections, respectively. Interestingly, up to 2 years following confirmed CL cure individuals had 28-fold higher levels of anti-α-Gal IgG compared to healthy volunteers. Monitoring levels of anti-α-Gal antibodies can be exploited as both a diagnostic tool and as a biomarker of cure of Old World CL in disease elimination settings.

Caffeine, which stimulates intracellular Ca2+ release channels known as ryanodine receptor (RyR) channels, induces contraction of individual muscle fibres dissociated from the trematode Schistosoma mansoni, and the turbellarians Dugesia tigrina and Procerodes littoralis. Caffeine is much more potent on S. mansoni fibres (EC50 0·7 mM) than those from D. tigrina or P. littoralis (3·2 mM and 4·6 mM, respectively). These caffeine-induced contractions are blocked by ryanodine, confirming the presence of functional RyR channels in these flatworm muscles. However, the contractions are not blocked by typical RyR channel blockers ruthenium red or neomycin, indicating that there may be important pharmacological differences between the RyR channels in this early-diverging phylum and those of later animals. These studies demonstrate that RyR channels are present in the muscle of these flatworms, and that the sarcoplasmic reticulum stores sufficient Ca2+ to support contraction.

Diseases and host dynamics are linked, but their associations may vary in strength, be time-lagged, and depend on environmental influences. Where a vector is involved in disease transmission, its dynamics are an additional influence, and we often lack a general understanding on how diseases, hosts and vectors interact. We report on the occurrence of six zoonotic arthropod-borne pathogens (Anaplasma, Bartonella, Borrelia, Coxiella, Francisella and Rickettsia) in common voles (Microtus arvalis) throughout a population fluctuation and how their prevalence varies according to host density, seasonality and vector prevalence. We detected Francisella tularensis and four species of Bartonella, but not Anaplasma, Borrelia, Coxiella or Rickettsia. Bartonella taylorii and B. grahamii prevalence increased and decreased with current host (vole and mice) density, respectively, and increased with flea prevalence. Bartonella doshiae prevalence decreased with mice density. These three Bartonella species were also more prevalent during winter. Bartonella rochalimae prevalence varied with current and previous vole density (delayed-density dependence), but not with season. Coinfection with F. tularensis and Bartonella occurred as expected from the respective prevalence of each disease in voles. Our results highlight that simultaneously considering pathogen, vector and host dynamics provide a better understanding of the epidemiological dynamics of zoonoses in farmland rodents.

An examination of 235 Plica umbra and 167 Uranoscodon superciliosa (Sauria: Iguanidae) from Belém, Pará, Brazil, has revealed a number of hitherto undescribed blood parasites. These include Fallisia audaciosa, F. simplex, Garnia multiformis, Plasmodium vacuolatum and Trypanosoma plicae spp.nov., all in P. umbra, and G. uranoscodoni sp.nov. in U. superciliosa. In addition, both species of lizards showed occasional Eperythrozoon and microfilaria infections. Among the haemosporines, F. audaciosa is particularly interesting in its restriction to the neutrophils; and G. multiformis for its early development in erythroblasts of the bone-marrow, and its variable morphology in subsequent stages of the infection in the peripheral blood. Mixed infections of the different haemosporines resulted in 18 combinations of parasites in P. umbra, indicating the importance of examining a large number of these animals when preparing descriptions of the individual organisms. T. plicae is a large, broad trypanosome, averaging ∼ 57·0 × 16·0 μm. Isolation of the parasite was made from four out of six P. umbra, after the culture of heart blood in NNN medium: the fact that none of the four positive lizards showed detectable trypanosomes in the peripheral blood suggests a high incidence of occult infections. The incidence among the remaining 229 P. umbra, as given by the direct examination of blood-films, was only 10·0%.

Skin penetration by infective Ancylostoma tubaeforme larvae has been investigated cinematographically and using in vitro techniques. The dermal tissue appears to cause little hindrance to larval migration but complete penetration through the skin from the dermal direction did not occur, although total penetration from the epidermal surface was frequently accomplished. No evidence could be found for enzymic secretions emanating from the worms under conditions that gave positive results from Necator americanus and Strongyloides fülleborni infective larvae. The results indicated that A. tubaeforme was able to penetrate without the use of enzymic secretions and an alternative, mechanical mechanism for penetration is advanced.

The humoral immune response of plaice to infestations of selected metacercariae was studied. Precipitating antibodies to both Cryptocotyle lingua and Rhipidocotyle johnstonei were demonstrated using agar geldiffusion techniques. Immunochemical investigations of these antibodies indicated they were macroglobulins, resembling immunaglobulin M(IgM) of mammals. The rate and magnitude of antibody production by the host was determined by the ambient temperature

Amblyospora sp. is a microsporidian parasite which is transovarially transmitted from an infected female Culex salinarius to her progeny. This transmission is dependent on synchronization of sporulation of the parasite and ovarian development of the host, both of which are initiated when the mosquito takes a blood meal. We induced sporulation in the absence of a blood meal and, with or without prior ovariectomy, by injection of 20-hydroxyecdysone, a hormone whose precursor, ecdysone, is produced by the ovaries of blood-fed mosquitoes. Sporulation in response to a blood meal was prevented by ovariectomy of the host prior to blood feeding or by decapitation immediately after feeding.

1.The morphology of Protopolystoma xenopi from Uganda is compared with specimens previously described by Williams. The chief differences are the slightly greater maximum size, the presence of a second pair of anchors on the opisthaptor, and the absence of a second large oviducal chamber in the uterus.

2. Encapsulation of the egg is described from living worms. A thin, plastic membrane forms around the ovum and yolk cells within the ootype before all the yolk cells have passed into the ootype. The remaining yolk cells pass from the ovo-vitelline duct into the sac formed by this membrane, which is attached to the distal walls of the ootype.

3. Eggs are produced at the rate of 4–15 eggs/worm/24 hr. They hatch in 24–27 days at 24–26° C. The opisthaptor of the larva bears sixteen hooklets and two pairs of anchors, which are in the same positions as in the adult.

4. One larva with one pair of suckers was obtained from the alimentary canal or kidney of a metamorphosing Xenopus tadpole. About sixty other tadpoles were examined for P. xenopi but none was found.

5. Twelve P. xenopi were recovered from the kidneys of a small frog that had been infected experimentally 10 days previously.

6. When naturally infected small frogs from Kajansi Fish Farm were examined, immature P. xenopi were found in the kidney of one frog, in the urinary ducts of two frogs, and in the bladder of one frog. The infection rate was much lower than that in mature Xenopus. Immature P. xenopi were also found in the kidneys of mature Xenopus mülleri.

7. P. xenopi matures more rapidly than its host. A frog weighing only 2·9 g. contained a nearly mature worm in the bladder, while one weighing 4·2 g. contained two mature worms, shedding eggs. Eggs are shed throughout the year. The life cycle of P. xenopi is therefore not regulated by that of its host.

In theory there should be a strong coupling between host and parasite population sizes. Here, we investigated population size and structure in 3 species of acanthocephalans, Corynosoma semerme, C. strumosum and C. magdaleni, in ringed seals (Phoca hispida) from the Bothnian Bay over a period of more than 20 years. During this period, seal numbers first decreased markedly and then increased steadily; at the same time, a paratenic fish host particularly important for C. strumosum has gradually disappeared from the bay due to decreasing salinity. We found no evidence that the mean abundance of any of the 3 acanthocephalan species changed significantly over time, nor was there any relationship between parasite abundance at any point in time and seal numbers in the corresponding year. Based on the proportion of sexually mature female worms per infrapopulation, and on relationships between the sex ratio of worms and infrapopulation size, both C. magdaleni and C. semerme appear to be doing well, independently of the population size of their seal definitive hosts. In contrast, perhaps because of the loss of its main paratenic host, C. strumosum appears more at risk in the Bothnian Bay. Our results show that in complex natural systems, there are not necessarily simple, direct links between definitive host population size or density, and parasite population dynamics.

The migration of subadult parasites to preferred sites within final hosts is well characterized. In contrast, the migration of larval stages of trematodes to specific sites within their second intermediate hosts is poorly understood. We used a serial necropsy approach to characterize the migration of Ornithodiplostomum ptychocheilus diplostomules from the point of cercarial penetration, to encystment within the outermost tissues of the brain of fathead minnows. Diplostomules utilized peripheral nerves to access the central nerve cord, or they used specific cranial nerves to directly access the brain. Within 3 h of exposure to cercariae, 46% of all diplostomules were observed within the medulla of the brain. Diplostomules subsequently utilized specific neural tracts to reach lateral regions of the outermost tissue layer of the optic lobes, the stratum marginale. Diplostomules remained in this layer during their 4-week growth phase, then shifted site to the adjacent meninges for encystment. Characterization of a habitat shift for developing versus encysted metacercariae helps explain the results of previous ecological studies that document transient changes in the effects of metacercariae on the surivival, behaviour, and anti-parasite defences of infected fish.

The development of the male reproductive system in Gyrodactylus gasterostei has been followed using parasites of known age maintained on isolated hosts. The penis develops shortly after the parasite has given birth for the first time (at an age of 24–30 h at 13°C) and the first active spermatozoa appear after 40–50 h. Spermatogenesis occurs more rapidly in G. gasterostei than in any other parasitic flatworm (including those from warm-blooded hosts) in which it has been measured, and the onset of male maturity is further hastened by a reduction in the number of pre-spermatogenic germ cell divisions. Spermatogonia have a diploid chromosome complement of 12, and spermatocytes undergo meiosis to produce haploid spermatozoa. No evidence of aneuploidy in spermatozoa was obtained. Although the development of haploid spermatozoa suggests that sexual reproduction can occur, production of embryos by isolated flukes which lack a mature male system indicates that other means of reproduction may also be employed.

Interactions between infections of Trichinella spiralis and Nippostrongylus brasiliensis were studied in the NIH strain of mouse which is known to react strongly to T. spiralis. The course of N. brasiliensis infection in this strain of mouse is described and expulsion is shown to be accelerated in immunized mice and inhibited in cortisone-treated mice. There was no evidence of inter-specific competition between the two species of worm in concurrent infections; the number and location of adults of both species were normal and T. spiralis was able to grow and reproduce normally. No evidence was found of direct immunological cross-reaction between N. brasiliensis and T. spiralis as assessed by the kinetics of adult worm numbers on heterologous challenge of immunized mice 90 days after the initiation of the last of 3 immunizing infections. Interaction was observed only when the timing of concurrent infections was such that one species was established in the intestine immediately before the beginning of expulsion of the second species. Interaction was manifested as a premature loss of worms and, in addition, as impairment of growth and fecundity of T. spiralis. These effects on T. spiralis were similar to those observed as a consequence of a specific immune response to T. spiralis. The rapidity of appearance of these effects and the lack of direct cross-immunity between the two species of worm suggest that the events involved in interaction were non-specific in action and possibly due to environmental changes in the gut caused by the immune response. These non-specific effects are therefore analogous, but not necessarily homologous, to the expulsion of these parasites in single species infections.

1. History of the classification of the family Metastrongylidae is briefly reviewed.

2. The probable origin of the family is discussed. It is believed that the metastrongylids arose, in common with the family Strongylidae, from a primitive group of eustomatous, cloacinin-like, gastro-intestinal strongylines that were symbiotic with an ancestral mammalian group, possibly the pantotheres. One group of these symbiotes established itself in the mammalian respiratory tract and originated the metastrongylid line.

3. The family Trichostrongylidae is regarded as of common origin with the family Ancylostomatidae from eustomatous ancestors. Thus meiostomy in the metastrongylids and trichostrongylids is convergent.

4. The Metastrongylidae consist of three main lines, each characteristic of a placentate order: the Filaroidinae in the Carnivora, the Pseudaliinae in the Cetacea, and the Protostrongylinae in the Artiodactyla. These lines probably have evolved from a common ancestor, symbiotic with a host at the base of the carnivore-cetacean-ariodactyl radiation. Certain forms have become secondarily adapted to symbiosis with members of other placentate orders.

5. Morphological and biological evidence is given for each of the foregoing phylogenetic conclusions.

6. Metastrongylus Molin, 1861, is regarded as a relict genus. With it, in a subfamily Metastrongy-linae, is placed the genus Heterostrongylus Travassos, 1925.

7. Heterostrongylus is represented by a single known species, H. heterostrongylus Travassos, 1925, from an opossum (order Marsupialia). Morphologically it is primitive with cloacinin-like cephalic features. In view of the phylogenetic status of its host it may well represent an ancestral type that has evolved along with the Marsupialia from a common ancestry with the other metastrongylids, probably in the panthotheres. As yet unknown lungworms of marsupials may shed light on this interesting point.

8. Diagnoses are given for the family Metastrongylidae and its subfamilies; the twenty-two component genera are listed under the latter. New synonymies are made: Neometastrongylus Kreis, 1944, a synonym of Neostrongylus Gebauer, 1932; Neometastrongylus buechii Kreis, 1944, of Neostrongylus linearis (Marotel, 1913) Gebauer, 1932; and Otophocaenurus Skriabin, 1942, of Pharurus Leuckart, 1848. Otophocaenurus oserskoi Skriabin, 1942, becomes Pharurus oserskaiaae (Skriabin, 1942, n.emend.) n.comb.; Metathelazia massino becomes M. massinoi (Davtian in Skriabin, 1933, n.emend.) Dougherty, 1943.

The host–parasite interaction between the rainbow trout Oncorhynchus mykiss and the fish louse Argulus japonicus was investigated by administering low levels of dietary cortisol before infecting the fish with low numbers of the parasite. After 24 h, the dietary cortisol treatment elevated blood cortisol and glucose levels and stimulated the synthesis of secretory granules in the upper layer of skin cells. Infection with 6 lice per fish caused skin infiltration by lymphocytes, also in areas without parasites. The lymphocyte numbers in the blood at 48 h post-parasite infection were reduced. Other changes, typical for exposure to many stressors and mediated by cortisol, were also found in the epidermis of parasitized fish, although neither plasma cortisol nor glucose levels were noticeably affected. Glucocorticoid receptors were localized immunohistochemically and found in the upper epidermal layer of pavement and filament cells, and in the leucocytes migrating in these layers. Cortisol-fed fish had reduced numbers of parasites and the changes in the host skin are likely involved in this reduction. Thus a mild cortisol stress response might be adaptive in rejecting these parasites. Further, the data suggest that this effect of cortisol is mediated by the glucocorticoid receptor in the skin epidermis, as these are located directly at the site of parasite attachment and feeding in the upper skin cells that produce more secretory granules in response to cortisol feeding.

In complex organisms, apoptosis is a constitutive cell death process that is involved in physiological regulation of cell numbers and that can also be induced in the course of inflammatory and immune responses. Neutrophils are among the first cells recruited during inflammation. Neutrophils constitutively die by apoptosis at inflamed sites, and are ingested by macrophages. Recent studies investigated how phagocytic clearance of senescent neutrophils influences the survival of intracellular protozoan parasites that have been phagocytosed by, or have invaded phagocytes. The results indicate that neutrophil clearance plays an unexpected role in regulation of intramacrophagic protozoan parasite infection.

Amoebae of the genus Endolimax have been found naturally inhabiting the large intestine of 3 species of Macacus (M. sinicus [= radiatus], M. rhesus [= mulatto], M. nemestrinus).

In all these hosts the amoebae are morphologically and culturally identical, at all stages of development, and are indistinguishable by any structural or cultural character from Endolimax nana of Man—with which they have been carefully compared. The several forms have been studied in “pure” culture (i.e., free from all other protozoa, but accompanied by bacteria suitable for their growth).

The Endolimax of M. sinicus, isolated in “pure” culture, has been experimentally transmitted to an uninfected man by the natural route (cysts per os): and in this human subject it has produced an infection indistinguishable from a natural one with E. nana. The induced infection has now persisted unchanged for close on 8 years.

From the experimentally infected man the amoeba has been artificially transmitted to an uninfected M. rhesus, in which it gave rise to an infection—likewise indistinguishable from a natural simian infection—which persisted for the rest of the monkey's life (2½ years).

Some further information is given about the distribution, cultivation, emetine-resistance, and other peculiarities of the forms of Endolimax found in Macaques and Man; and the findings of other workers with similar organisms are briefly considered. From all the evidence available it is concluded that E. nana naturally lives not only in Man but also in at least 3 different species of Macacus, and probably in monkeys of this genus generally. As yet there is no evidence that more than one species of Endolimax inhabits any of these hosts.

Infections with E. nana, whether in Man or Macaques, have usually been found to endure indefinitely; but a case (M. rhesus) has been observed in which the infection appears to have died out spontaneously after persisting for a known period of over 6 years.

Attempts to infect kittens with E. nana have all been unsuccessful hitherto, but there is evidence (from the work of others) that this amoeba may inhabit some other domestic mammals. Its host-distribution in nature needs further investigation.

The calcium-dependent protein kinases (CDPKs) are unique enzymes found only in plants, green algae, ciliates and apicomplexan parasites. In this study, a novel CDPK gene of Eimeria tenella, designed EtCDPK3, was cloned using rapid amplification of cDNA ends (RACE) based on the expressed sequence tag (EST). The entire cDNA of EtCDPK3 contained 1637 nucleotides encoding 433 amino acids and the deduced EtCDPK3 protein had canonical characteristic domains identified in other CDPKs, including a well-conserved amino-terminal kinase domain and a carboxy-terminal calmodulin-like structure with 4 EF-hand motifs for calcium binding. The expression profiles of the EtCDPK3 gene in different development stages were investigated by real-time quantitative PCR. Messenger RNA levels from the EtCDPK3 gene were higher in sporozoites than in other stages (unsporulated oocysts, sporulated oocysts and merozoites). Western blot analysis showed that rabbit antiserum against recombinant EtCDPK3 could recognize a native 49 kDa protein band of parasite. Indirect immunofluorescent antibody labelling revealed dispersed localization of EtCDPK3 during the first schizogony and intense specific staining. EtCDPK3 was located at the apical end of the sporozoites after early infection of DF-1 cells and the protein was highly expressed. Inhibition of EtCDPK3 function using specific antibodies reduced the ability of E. tenella to invade host cells. These results suggested that EtCDPK3 may be involved in invasion and survival of the parasite intracellular stages of E. tenella. Because this kinase family is absent from hosts, it represents a valid target that could be exploited for chemotherapy against Eimeria spp.

Life-history attributes of the capsalid monogenean Benedenia lutjani, a parasite of Lutjanus carponotatus from the Great Barrier Reef, Queensland, Australia, were investigated from experimental infections. Oncomiracidia of B. lutjani invaded and attached at any site on the fish, but more commonly invaded body surfaces. Immature specimens then migrated to the pelvic fins. Development of the reproductive organs of B. lutjani corresponded with migratory movements on the host. Parasite aggregation on the pelvic fins coincided with the development of functional male reproductive organs and some protandrous worms that possessed a vagina appeared to be inseminated. Migration to, and aggregation on, the branchiostegal membranes (membranous folds posterior to the opercula) coincided with the onset of sexual maturity and commencement of egg production by parasites. The rate of parasite development and the timing of migratory events on the host were influenced by water temperature. All specimens of B. lutjani reached sexual maturity between 12 and 14 days p.i. at 24 °C and between 8 and 10 days p.i. at 27 °C. Anterior hamuli grew continually during a 16-day experiment at 27 °C and 25-day experiment at 23 °C and their length appeared to provide a suitable index to estimate parasite age. The possible adaptive significance of the migratory behaviour, site-specificity and its link with changes in parasite development are discussed.

The phylogenetic relationship of 5 genera, i.e. Diplozoon Nordmann, 1832, ParadiplozoonAchmerov, 1974, InustiatusKhotenovsky, 1978, SindiplozoonKhotenovsky, 1981, and EudiplozoonKhotenovsky, 1985 in the subfamily Diplozoinae Palombi, 1949 (Monogenea, Polyopisthocotylea) was inferred from rDNA ITS-2 region using neighbour-joining (NJ), maximum likelihood (ML) and Bayesian methods. The phylogenetic trees produced by using NJ, ML and Bayesian methods exhibit essentially the same topology. Surprisingly, freshwater species of Paradiplozoon from Europe clustered together with species of Diplozoon, but separated from Chinese Paradiplozoon species. The results of molecular phylogeny and lower level of divergence (4·1–15·7%) in ITS-2 rDNA among Paradiplozoon from Europe and Diplozoon and, on the other hand, high level of divergence (45·3–53·7%) among Paradiplozoon species from Europe and China might indicate the non-monophyletic origin of the genus Paradiplozoon. Also, the generic status of European Paradiplozoon needs to be revised. The species of Paradiplozoon in China is a basal group in Diplozoinae as revealed by NJ and Bayesian methods, and Sindiplozoon appears to be closely related to European Paradiplozoon and Diplozoon with their relationship to Eudiplozoon and Inustiatus being unresolved.

A deterministic, dynamic simulation model is developed to account for the interactions between gastrointestinal parasitism and host nutrition, and predict their consequences on performance and level of parasitism of sheep. Larval intake and established adult worms are assumed to result in nutrient loss for the host. In order to reduce this loss the host will mount an immune response, which will affect the establishment rate of incoming larvae, mortality rate of adult worms, and fecundity of female worms, as well as nutrient loss caused by larval intake per se. Host anorexia is modelled as a function of worm mass. Parasitism is also assumed to affect the allocation of ingested nutrients to the host's bodily functions, with maintenance getting absolute priority, and protein allocated to immunity and production proportionally to their requirements. Inputs to the model include the expected growth attributes of the animal, feed quality, various parasitological parameters and daily larval intake. Outputs include feed intake, growth rate and body composition, as well as worm burden and faecal egg counts. The model allows exploration of the consequences of gastrointestinal parasitism on sheep of different growth characteristics, kept under environments that vary in the provision of nutrients and exposure to parasites.