We used the fluorescent dye monochlorobimane (MCB) which binds glutathione (GSH) to localize between 2 and 6 distinctly labelled nuclear and cytoplasmic GSH foci in recently excreted and aged, intact Cryptosporidium parvum oocysts and sporozoites. Buthionine sulfoximine (BSO), a potent and specific inhibitor of GSH, was used to determine whether GSH is synthesized in BSO-treated C. parvum oocysts, by labelling treated oocysts with MCB. Both visual and electronic quantifications were performed. At 5 mM BSO, a significant inhibition of MCB fluorescence, reflecting reduced MCB uptake, was observed in GSH-depleted oocysts (mean±S.D. 35±3·7) compared with controls (3·3±1·2, P=0). This clear reduction occurred only in viable oocysts. 1 mM BSO-treated oocysts exhibited weak or no MCB fluorescence, although they were viable (excluded propidium iodide, PI)), and intact and contained sporozoites by differential interference contrast microscopy (DIC). MCB was used in conjunction with PI to determine C. parvum oocyst viability. Oocysts labelled with MCB/PI or 4′6-diamidino-2-phenyl indole (DAPI)/PI produced comparable labelling patterns. Viable oocysts were labelled with MCB or DAPI whereas dead oocysts were labelled with PI only. The localization of GSH in viable, intact oocysts and excysted sporozoites and UV light-irradiated oocysts and sporozoites revealed no changes in MCB uptake at levels up to 40 mJ.cm−2 irradiation. Although GSH can be detected following MCB localization in both the nucleus and cytoplasm of sporozoites, and can be specifically depleted by BSO treatment, MCB is unlikely to be useful as a surrogate for detecting UV damage in UV-treated Cryptosporidium oocysts.

An attempt to differentiate by means of the electron microscope between particles of the top and bottom components of the turnip yellow mosaic virus is described. Since the particles of the top component are thought to be hollow spheres, it might be expected that they would collapse after drying and thus throw a smaller shadow than the particles of the bottom component which are thought to have a more solid centre. No difference could be perceived between the two types of particle, so that the apparently hollow particles can withstand the effect of drying without collapsing.

Sections were cut of leaves of Chinese cabbage (Brassica chinensis) and particles were observed inside the cells which could conceivably be virus particles.

Life histories of the Trombidiidae, or, for that matter, other families of mites have, with few exceptions, not been studied and very little information is available as a guide. This is not entirely the fault of the acarologists, as not one of them was employed in any country of the world entirely upon the study of mites. Most of them, if official, were entomologists who could spend only a fraction of their time on mites. Their time was spent therefore on systematics, an essential branch of acarology.

The Diptera larvae can be divided from an ethnological point of view into four categories: and the members of each, though of diverse systematic affinities, possess in common certain structural characters, which Keilin (1912) considers to be adaptations to the manner of life. The categories are as follows: (i) Saprophagous larvae: those which feed on decayed vegetable matter or the excrement of herbivorous animals; these have a buccal armature similar to that of Calliphora or Lucilia: the ventral surface of the basilar piece is feebly chitinized and has a number of longitudinal ridges projecting into the lumen of the pharynx. (ii) Phytophagous larvae, which feed on living plants, as leafminers, root-miners, etc.: these have the bucco-pharyngeal armature more concentrated, the basilar piece more chitinized, with ridges feebly developed or absent, the mandibles modified as several toothed scrapers. (iii) Parasitic larvae: their buccal armature resembles that of parasitic Tachinidae. (iv) Parasitic larvae—carnivorous larvae: It is now known from the researches of Portchinsky (1910), and later of Keilin himself, that a good number of larvae found in excrement are not really coprophagous, but live on other larvae or other small animals found with them in the excrement. These carnivorous larvae attack their prey, perforating its skin, and sucking its contents. For instance, the larvae of Graphomyia maculata, found in liquefied decomposing vegetable matter, feed on larvae of Eristalis and pupa of Tipulidae, those of Calliophris riparia, found among moss, feed on small oligochaete worms and larvae and pupae of Psychodidae. These carnivorous larvae have certain characters in common. Thus the mandibles are sharp, piercing hooks and the basilar piece of the armature is very long, narrow and strongly chitinized and entirely without the internal longitudinal ridges.” (Scott, 1915.)

The effects of the size of Cynoscion guatucupa on the size and demographic parameters of their parasitic copepod Lernanthropus cynoscicola were evaluated. Prevalence of copepods increased with host size up to fish of intermediate length, then it decreased, probably because changes in size of gill filaments affect their attachment capability, enhancing the possibility of being detached by respiratory currents. Body length of copepods was significantly correlated with host length, indicating that only parasites of an ‘adequate’ size can be securely attached to a fish of a given size. The absence of relationship between the coefficient of variability in copepod length and both host length and number of conspecifics, together with the host-size dependence of both male and juvenile female sizes, prevent to interpret this relationship as a phenomenon of developmental plasticity. Therefore, the observed peak of prevalence could reflect the distribution of size frequencies in the population of copepods, with more individuals near the average length. Concluding, the ‘optimum’ host size for L. cynoscicola could merely be the adequate size for most individuals in the population, depending, therefore, on a populational attribute of parasites. However, its location along the host size range could be determined by a balance between fecundity and number of available hosts, which increases and decreases, respectively, with both host and parasite size.

For the material herein dealt with, I am indebted principally to Mr G. A. H. Bedford of the Division of Veterinary Services of South Africa. Other material collected by the writer from bats in the United States National Museum some years ago and a specimen received from Dr C. R. Kellogg of Foochow, China, are also recorded.

Doses from 1 to 10 sporulated oocysts of a cloned strain of Eimeria hungaryensis were used to infect laboratory-reared, coccidia-free wood mice (Apodemus sylvaticus). Total oocyst production, reproductive indices (output/dose) and daily patterns of oocyst production were compared. At doses of 10 oocysts or less, observed yields exceeded expected yields calculated using details of endogenous stages. At doses above 10 oocysts, the converse occurred. From infections initiated with 10 oocysts, total outputs were not significantly different in male and female wood mice, host age had no apparent effect and repeated infections, at 10-day intervals, of both sexes, resulted in progressive decreases in output at each successive infection, suggesting development of resistance.

The clinical manifestation and course of Leishmania infections depend on factors such as species, virulence and host-immunity. Although trypanosomatids are considered to have clonal propagation, genetic hybridization has produced successful natural hybrid lineages. Hybrids displaying strong selective advantages may have an impact on pathogenesis and the eco-epidemiology of leishmaniasis. Thus, characterization of phenotypic properties of Leishmania hybrids could bring significant insight into the biology, infectivity, pathogenicity and transmission dynamics of these atypical strains. The present study focuses on phenotypic features and survival capacity of Leishmania infantum/Leishmania major hybrid isolates as compared with representative putative parental species, L. infantum and L. major. In vitro assays (growth kinetics, susceptibility to different conditions) and in vivo infection (parasite detection and histopathological alterations) showed that hybrids present higher growth capacity and decreased susceptibility to reactive oxygen species. Furthermore, evaluation of infected spleen tissue suggests that hybrids induce a stronger immune reaction than their putative parents, leading to the development of white pulp hyperplasia in B-lymphocyte compartments. Overall, these hybrids have shown high plasticity in terms of their general behaviour within the different phenotypic parameters, suggesting that they might have acquired genetic features conferring different mechanisms to evade host cells.

1. On the assumption that the limit of significant difference is 1%, it is necessary, when concentration methods are not used, to repeat the faecal examination 3 times for the egg of Ascaris lumbricoides, 7 times for hook worm and 11 times for Trichuris trichiura in order to obtain the diagnostic results obtained by the acid-ether concentration technique using 3 g. of faeces.

2. The coefficient of the identification by a single smear examination is 83·3% for eggs of Ascaris lumbricoides, 46·5% for hook-worm eggs and 33·6% for eggs of Trichuris trichiura.

The species of Oesophagostomum here described has been collected quite frequently from sheep and goats in certain parts of the colony, always accompanied by the commoner oesophagostome O. columbianum.

Understanding factors that influence the spatial and temporal distributions of blood parasites is important to help predict how host species and their parasites may respond to global change. Factors that may influence parasite distributions are land cover and host dispersal patterns, which may result in exposure of a host to novel parasites, or escape from parasites of their origin. We screened golden-winged warblers from across the United States and Canada for blood parasites, and investigated whether land-use patterns or host dispersal affected the prevalence and composition of haemosporidian assemblages. Parasite prevalence varied strongly with study area, and areas with high agricultural cover had a significantly higher prevalence of Leucocytozoon and Parahaemoproteus parasites. Lineages of Parahaemoproteus and Leucocytozoon were genetically differentiated among study areas, and prevalence and composition of parasite assemblages indicated an increase in parasite prevalence and accumulation of unique parasite lineages from the southeast to the northwest. This matches the historical range expansion and natal dispersal patterns of golden-winged warblers, and suggests that golden-winged warblers may have been sensitive to novel parasites as they dispersed. The high prevalence and diversity of parasite lineages in the north-west extent of their breeding range (Manitoba) indicates that this population may face unique pressures.

The in vivo effects of the anthelmintics taenifugin, VÚFB 14170 and VÚFB 15269 on the tegument of Hymenolepis fraterna have been examined by SEM, TEM and cytochemistry. The drugs were given to H. fraterna-infected mice on the 14th day post-infection in a single oral dose of 150, 200 and 200 mg/kg, respectively. By 72 h post-treatment, the drug-induced pathomorphological changes in the tegument indicated that all three drugs had a significant effect. Changes were most pronounced on the brush border and in the intercellular material. On the apical surface, there was blebbing as well as accumulation of membrane fragments over the microthrix tips and erosion of the brush border. The intercellular material was changed in structure, showing increased electron density in some areas and oedema of the intercellular spaces in other areas. There were also fractures of the tegument of variable depth, sometimes reaching to the parenchyma. These results suggest altered tegumental integrity and, occasionally, complete disruption of the selective permeability barrier created by the normal tegument. This suggestion is further supported by the penetration of ruthenium red into some tegumental areas and its distribution into the intercellular spaces, down to the parenchyma. The intrategumental lysosomes also appeared to be significantly activated. There was evidence of autophagy in both distal cytoplasm and tegumental cells. Mature and gravid proglottides were more susceptible to drug damage than those in the anterior strobila and neck.

The intracellular development of Eimeria tenella sporozoites in in vitro cultured Madin–Darby Bovine Kidney (MDBK) cells was inhibited when parasite-infected MDBK cells were incubated with peripheral blood lymphocytes (PBL) from infected chickens. The inhibition mediated by PBL was quantified by [3H]uracil uptake and increased during the course of a series of oral infections of chickens with E. tenella. This was mirrored by the development of immunity in these birds, as assessed by counting the oocyst output following each re-infection. Similar levels of inhibition were observed using PBL from 3 inbred lines of chickens which differ in their relative susceptibility to infection with E. tenella, indicating that the genetic background of the host does not influence the production of this inhibitory activity. The inhibition could be transferred to freshly infected MDBK cells using supernatants prepared from parasite-infected monolayers incubated for 48 h with PBL from immune chickens. However, there was no inhibition using either supernatants from infected MDBK cells incubated with PBL from uninfected chickens, or supernatants from uninfected MDBK cells incubated with PBL from immune chickens. Experiments using Transwell plates showed that direct contact of PBL from immune birds with infected MDBK monolayers was not required to produce supernatants with inhibitory activity. Thus production of soluble inhibitory factor(s) by PBL from immune chickens can be specifically induced by soluble antigens present in the culture media of parasite-infected MDBK cells. These factors inhibit the intracellular development of sporozoites in in vitro culture.

Males of Nematodirus battus have a body wall composed of a longitudinally ridged cuticle made up of 8 layers, a hypodermis and a layer of longitudinal muscle cells which are divided into 4 sectors by the hypodermal cords. There is a characteristic cephalic inflation of the cuticle in both sexes. The structure of the body wall in the anterior half of the female is similar to that of the male. In the posterior half of the female the cuticle is not ridged and has 7 layers; the longitudinal muscles are reduced in number and the lateral hypodermal cords increasingly extend dorsally and ventrally along the length of the posterior part of the nematode and come to occupy most of the lateral sectors of the body. The possible significance of these modifications in the structure of the female nematode are discussed.

Synthesized lipophilic tyrosyl ester derivatives with increasing lipophilicity were effective against Leishmania (L.) major and Leishmania infantum species in vitro. These findings prompted us to test in vivo leishmanicidal properties of these molecules and their potential effect on the modulation of immune responses. The experimental BALB/c model of cutaneous leishmaniasis was used in this study. Mice were infected with L. major parasites and treated with three in vitro active tyrosyl esters derivatives.

Among these tested tyrosylcaprate (TyC) compounds, only TyC10 exhibited an in vivo anti-leishmanial activity, when injected sub-cutaneously (s.c.). TyC10 treatment of L. major-infected BALB/c mice resulted in a decrease of lesion development and parasite load. TyC10 s.c. treatment of non-infected mice induced an imbalance in interferon γ/interleukin 4 (IFN-γ/IL-4) ratio cytokines towards a Th1 response. Our results indicate that TyC10 s.c. treatment improves lesions’ healing and parasite clearance and may act on the cytokine balance towards a Th1 protective response by decreasing IL-4 and increasing IFN-γ transcripts. TyC10 is worthy of further investigation to uncover its mechanism of action that could lead to consider this molecule as a potential drug candidate.

The human malaria parasite, Plasmodium falciparum, is currently being actively studied by molecular biologists. It is hoped that the use of recombinant DNA techniques in this area will give new insights into the biology of the organism and, at the same time, provide new approaches to diagnosis and vaccine development.

Our own studies employ the blood stages of the parasite and cover three main areas: enzymes of importance in parasite metabolism; antigens of potential use in a subunit vaccine; and repetitive DNA as a probe able to distinguish genetically different isolates of P. falciparum and as a species-specific diagnostic tool in human and mosquito infections.

Two new species of Lepeophtheirus are described and illustrated based on a collection from the Orange County sewer outfall, California. They are: L. remiopsis sp.nov. from the branchial cavities and external body surfaces of Parophrys vetulus Girard, Hippoglossina stomata Eigenmann & Eigenmann, Pleuronichthys verticalis (Jordan & Gilbert), Chitonotus pugetensis (Steindachner), and Porichthys notatus (Girard); and L. rotundipes sp.nov. from the external body surfaces and occasionally the gill cavities of Scorpaena guttata (Girard) and Citharichthys stigmaeus Jordan & Gilbert.

Specimens of Stylops advarians were sampled by collecting foraging bees of Andrena milwaukeensis along the South Saskatchewan River within Saskatoon, Saskatchewan. As the foraging season progressed from early May till late June over three consecutive years (2016–2018), most stylopized bees possessed endoparasitic adult (neotenic) females of S. advarians protruding from the bee gaster's dorsum. In contrast, very few adult bees stylopized by male puparia, and no free-living males, were encountered. Over the sampling period, prevalence remained around 22% each year; mean intensity was 1.2 (range of 1–3 female parasites per bee); and parasite abundance was 0.27. Also newly reported for Stylops is the occurrence of one bee bearing four Stylops (two neotenic females and two males with puparia), plus another bee with a male puparium extruded from its gaster's sternites. Around 2 May each year, a high proportion of the earliest captured female bees were stylopized. However, non-stylopized female bees typically were not encountered until about 10 days later, suggesting the parasites manipulate female bee hosts to emerge earlier, in close synchrony to male bee emergence. First-instar larvae of S. advarians appeared from 22–25 May, indicating that adults of S. advarians matured and mated at similar times each season.