The effects of purified monoclonal immunoglobulins from control, or transmission-blocking anti-Pbs21 antibodies, upon the infection of Anopheles stephensi by ookinetes of Plasmodium berghei are compared. Anti-Phs21 antibody reduced mean intensity and prevalence of infection by 94·7 and 58·7% respectively if added to the infectious bloodfeed at a concentration of 100 μg/ml. Fab fragments were of similar efficacy. No transmission enhancement was detected with declining antibody concentrations. Addition to subsequent (second) feeds reduced mean oocyst intensity but not prevalence. The reduction in blockade declined from 41% at day 2, to 4% at day 8. Second bloodfeeds, containing control globulin taken 4 or 6 days (but not 2 days) after infection, increased sporozoite burden in the salivary glands. At all times anti-Pbs21 reduced sporozoite number in the thorax compared to time-matched controls, but again highest gland intensities were obtained when the second bloodfeed was given on day 4. We conclude that second bloodfeeds containing transmission-blocking antibody simultaneously serve two opposing roles, (1) inhibition of parasite development and (2) the supply of nutrients which permit more sporozoites to be produced by each oocyst.

The fine structure of the oncosphere of Hymenolepis nana has been investigated by transmission and scanning electron microscopy, together with light microscope observations of JB–4 embedded material. The outer surface of the oncosphere is covered by an epithelial layer, termed the embryonic epithelium. Cell types present within the oncosphere include the penetration gland cell, oncoblast, or hook-forming cells, nerve cells, muscle cells (both somatic and hook), and undifferentiated ‘stem’ cells. The penetration gland is a large, U-shaped structure, situated in the anterior region of the oncosphere, and filled with secretory granules of 2 distinct morphological types. Histochemically, the secretory material yields reactions characteristic of an acid mucopolysaccharide. A proteinaceous-substance and small amounts of glycogen are also present. Up to 4 pairs of ducts from the penetration gland have been observed. They pass through the basal lamina and the epithelial layer to open against the polar filament layer at the anterior end of the oncosphere. Nerve cells are described in a cestode oncosphere for the first time. The cells are paraldehyde-fuchsin-positive and show a high level of secretory activity, as evidenced by the large numbers of dense-cored vesicles produced by the Golgi apparatus in the perikarya; consequently, they are tentatively regarded as possible neurosecretory cells. The vesicles are transported down the axon to be stored in specialized swollen axon terminals, which form definite junctions with the muscle cells.

A detailed study of the structure of the tegument of D. phoxini has been carried out using probe and transmission electron microscope. The distribution of spines and sense organs on the forebody has been described, as well as the nature of the lappets in the everted and retracted state. The adhesive organ exhibits two different surfaces which also differ from the surface of the rest of the body. The use of the ‘Stereoscan’ has confirmed and extended the concept of surface specialization based on transmission studies.

In this paper the dynamics and control of nematode parasites of farmed ruminants are discussed via a qualitative analysis of a differential equation model. To achieve this a quantity, ‘the basic reproduction quotient’ (Q0), whose definition coincides with previous definitions of R0 for macroparasites, but extends to models with periodic time-varying transition rates between parasite stages or management interventions, is introduced. This quantity has the usual threshold property: if Q0 is less than one the parasite population cannot maintain itself in the host population, and in the long term becomes extinct; but if Q0 is greater than one the parasite can invade the host population. An alternative quantity, R(E), that is often easier to calculate is also introduced, and shown to have the same threshold property. The use of these two quantities in analysing models for the dynamics of nematodes in complex situations is then demonstrated, with reference to the dynamics of mixed parasite species in one host; the effects of breeding host animals for resistance to parasitism; and the development of parasite strains that are resistant to chemotherapy. Five examples are discussed using parameters for the dynamics of nematode infections in sheep, and some statements on control policies are derived.

A second species of the genus Endorchis is described—E. mandubé; also new species of the rare genera Monticellia, Amphoteromorphus and Nomimoscolex, viz. Monticellia piracatinga, M. rugosa, Amphoteromorphus parkarmoo, Nomimoscolex piracatinga and N. kaparari.

A new genus Manaosia is created to contain the anomalous species M. braço de moça. Proteocephalus lenha becomes Nomimoscolex lenha.

The species Spatulifer surubim, originally described by me as Peltidocotyle rugosa and later referred to the Amphilaphorchidinae, is now regarded as a species of the Monticelliinae.

The eight known species of the Monticelliinae are all referred to the single genus Monticellia, since, differences in vitellaria distribution not corresponding to differences of scolex form, the only alternative is to create an unnecessary number of genera. The genera Goezeella Fuhrmann and Spatulifer Woodland are thus regarded as superfluous.

“Ephedrocephalus” lobosus is removed from the Ephedrocephalinae (which retains its single species E. microcephalus) and, with the two species of “Amphilaphorchis” recently described by me, is placed in Fuhrmann's genus Rudolphiella, now re-established, in the subfamily Rudolphiellinae, thus replacing the Amphilaphorchidinae.

The complete list of known subfamilies, genera and species of the Proteocephalidae described from the Amazon, amended in accordance with the above conclusions, has been published in my last paper on the subject (Woodland, 1935).

Babesia divergens was cultivated in RPMI 1640 (25 mM HEPES) supplemented with 10% human serum (RPMI-10% HS) with a high percentage of parasitized erythrocytes (PPE) ([ges ]40%). Standardization of in vitro tests, purification of exoantigens, biochemical studies and the safety of the culture handler motivated the development of a serum-free defined medium. Removal of serum greatly reduced the PPE but, after a period of adaptation, the culture was continuous and the parasite was able to develop a 3% routine PPE. Addition of vitamins or reduced glutathione in basal medium (RPMI) did not improve the PPE. The supplementation of basal medium with lipidic carrier (Albumax I or bovine serum albumin–Cohn's fraction V) promoted the growth of B. divergens with high PPE (>30%) close to those obtained in RPMI–10% HS. Neither protein nor lipid fractions alone were able to restore the growth of B. divergens. Nevertheless, the whole lipid fraction from serum or Albumax I added to delipidated albumin partially restored the growth (7% PPE), indicating that the presentation of specific lipids by a carrier is crucial for the parasite. All the data indicate that Albumax I can replace human serum offering the advantages of safety, standardization for chemosensitivity tests, and exoantigen purification.

Ternidens deminutus (Strongylida) is a parasitic nematode infecting non-human and human primates in parts of Africa, Asia and the Pacific islands. The present study genetically characterized T. deminutus and defined genetic markers in nuclear ribosomal DNA (rDNA) as a basis for developing molecular-diagnostic tools. The sequences of the second internal transcribed spacer (ITS-2) of rDNA were determined for adult specimens of T. deminutus (Nematoda: Strongylida: Oesophagostominae) from the Olive baboon and the Mona monkey.Nucleotide sequence data used in this paper are available in the EMBL, GenBank and DDJB databases under the Accession nos. AJ888729, AJ888730, AF136576, Y10789, Y10790, Y11733, Y11735, Y11736, AJ001594, AJ001599, AJ006149 and AJ006150. The former 2 sequences represent original data reported in this paper. The length and G+C content of the ITS-2 sequences was 216 bp and ~43%, respectively. While there was no sequence variation among individual T. deminutus specimens from the baboon, 6 (2·8%) nucleotide differences were detected in the ITS-2 between the parasite from baboon and that of the Mona monkey, which is similar to the difference (3·2%) between 2 other species of Oesophagostominae (Oesophagostomum bifurcum and O. stephanostomum) from non-human primates, suggesting significant population variation or the existence of cryptic (i.e. hidden) species within T. deminutus. Pairwise comparisons of the ITS-2 sequences of the 2 operational taxonomic units of T. deminutus with previously published ITS-2 sequences for selected members of the subfamilies Oesophagostominae and Chabertiinae indicated that species from primates (including those representing the subgenera Conoweberia and Ihleia) are closely related, in accordance with previous morphological studies. The sequence differences (27–48·3%) in the ITS-2 between the 2 taxonomic units of T. deminutus and hookworms (superfamily Ancylostomatoidea) enabled their identification and delineation by polymerase chain reaction (PCR)-based mutation scanning. The genetic markers in the ITS-2 provide a foundation for improved, PCR-based diagnosis of T. deminutus infections and for investigating the life-cycle, transmission patterns and ecology of this parasite.

Ascaris suum possesses a large number of FMRFamide-related peptides (FaRPs) of which KNEFIRFamide (AF1), KHEYLRFamide (AF2) and KSAYMRFamide (AF8/PF3) have been shown to modulate the intrinsic, rhythmic activity of the vagina vera of A. suum in vitro. In the present study, the effects of the nematode FaRPs, SDPNFLRFamide (PF1), SADPNFLREamide (PF2) and KPNFIRFamide (PF4) (from Panagrellus redivivus) and AVPGVLRFamide (AF3) and GDVPGVLRFamide (AF4) (from A. suum) on the in vitro activity of the vagina vera were examined. The effects of each of the peptides were qualitatively and quantitatively distinct. All 3 FaRPs from P. redivivus were inhibitory, causing a cessation of contractions. PF2 was 3 times more potent than PF1, with a threshold of 1 nM. Although PF4 was the least potent (threshold, 10 nM), its effects at [ges ]10 nM were quantitatively the greatest. Both AF3 and AF4 (1 μM) induced complex, multiphasic responses consisting of an initial contraction and spastic paralysis followed by a return of contractile activity of increased amplitude. AF3 was 3 times more potent than AF4. The effects of these peptides had some similarities to those observed on A. suum somatic body wall muscle in vitro, with PF1, PF2 and PF4 being inhibitory and AF3 and AF4 being excitatory.

To investigate larval development of Acanthocheilonema reconditum in the cat flea Ctenocephalides felis, fleas were fed through an artificial feeding system with dog blood containing different concentrations of microfilariae (i.e. low, group L = 250; medium, group M = 500; high, group H = 1500 microfilariae per mL) or no microfilariae (group C). Fleas were sampled at 12 different time-points throughout the study period (D1–D28) and A. reconditum was detected by dissection, PCR and histology. Of 2105 fleas fed with infected dog blood, 891 (38·7%) died during the study before being sampled whilst the remaining (n = 1214) were examined for A. reconditum. Upon dissection, first-stage larvae (L1) were identified after 2 days post infection (D2), second-stage (L2) at D13 and infective third-stage larvae (L3) at D15. Eighteen (30%) of 60 pools of fleas molecularly examined tested positive. Histologically, L2 were detected at D13 in the sub-cuticle region embedded in the back muscle of one female flea. This study provides original data on larval development of A. reconditum in C. felis and reports on the usefulness of the artificial feeding system.

A fish-infecting Microsporidia Potaspora morhaphis n. gen., n. sp. found adherent to the wall of the coelomic cavity of the freshwater fish, Potamorhaphis guianensis, from lower Amazon River is described, based on light microscope and ultrastructural characteristics. This microsporidian forms whitish xenomas distinguished by the numerous filiform and anastomosed microvilli. The xenoma was completely filled by several developmental stages. In all of these stages, the nuclei are monokaryotic and develop in direct contact with host cell cytoplasm. The merogonial plasmodium divides by binary fission and the disporoblastic pyriform spores of sporont origin measure 2·8±0·3×1·5±0·2 μm. In mature spores the polar filament was arranged into 9–10 coils in 2 layers. The polaroplast had 2 distinct regions around the manubrium and an electron-dense globule was observed. The small subunit, intergenic space and partial large subunit rRNA gene were sequenced and maximum parsimony analysis placed the microsporidian described here in the clade that includes the genera Kabatana, Microgemma, Spraguea and Tetramicra. The ultrastructural morphology of the xenoma, and the developmental stages including the spores of this microsporidian parasite, as well as the phylogenetic analysis, suggest the erection of a new genus and species.

Light and transmission electron microscopy revealed pericytes of brain capillaries of moles (Talpa europaea L., 1758) as parasitized intracellularly. These host cells were enlarged and of globular or ellipsoid shape, and incorporated a cell-within-cell sequence of primary, secondary and, rarely found, tertiary developmental stages of an eukaryotic organism. Other stages like spores were not discovered either in brain or in other organs. Due to the vertebrate host, and the parasitic cells showing the enveloped state this parasite can be classified as belonging to the Myxozoa rather than Paramyxea. Since spores, which would allow an exact identification of the parasite, could not be detected and mammals are very unusual hosts for Myxozoa, the parasite was designated a myxozoan-like organism.

In the Czech Republic, sera from 551 clinically healthy adult slaughtered pigs (females, 6–8 months old) were collected during the first half of June in 2010. Sera were tested for Toxoplasma gondii-specific IgG antibodies by an enzyme-linked immunosorbent assay; samples with more than 50% S/P were considered as positive. The same samples were also analysed for Neospora caninum antibodies using a commercial competitive-inhibition enzyme-linked immunosorbent assay; samples with more than 30% inhibition were considered as positive. Antibodies against T. gondii were found in 198 pigs (36%) in all districts with prevalences ranging from 18% to 75%. Antibodies against N. caninum were found in 16 pigs (3%); positive animals were found in 4 districts with prevalences ranging from 1% to 20%. Indication of mixed infections (concurrent presence of both N. caninum and T. gondii antibodies) was found in 8 (1·5%) pigs. The results of our study indicate that pigs in the Czech Republic have a relatively high seroprevalence for T. gondii, while they have only a low seroprevalence for N. caninum. Therefore, natural infection with T. gondii seems to be very common in Czech pigs. It is the first evidence of N. caninum antibodies in pigs in the Czech Republic. These results complete data about N. caninum infection in pigs in Europe.

In the present paper, the phylogenetic relationships between genera, subfamilies and families of the Hemiuroidea are explored. Twelve new sequences of 28 rDNA and data taken from GenBank (NSBI) on 43 species affiliated to 34 genera were included in the analysis. Most of the hemiuroidean trematodes form two highly supported clades (A and B), which are sister groups to each other. Hemipera manteri joined with Gonocerca spp. with moderate statistical support. This clade is basal relative to the clades A and B. Сlade A is polytomic and contains representatives of the families Accacoeliidae, Syncoeliidae, Didymozoidae, Hirudinellidae and Sclerodistomidae, and derogenid subfamilies Derogeninae and Halipeginae. At the same time, the Syncoeliidae, Hirudinellidae and Accacoeliidae form a well-supported monophyletic group. The phylogenetic relationship between Derogeninae and Halipeginae is poorly resolved. Сlade B unites the isoparorchiid, bunocotylid, lecithasterid and hemiurid trematodes. Our data re-establishes the family Bunocotylidae, which consists of two subfamilies, Opisthadeninae and Bunocotylinae, and the Machidatrema chilostoma + Hysterolecithoides frontilatus group. The Bunocotylidae is the sister group to the Hemiuridae + Lecithasteridae group and the Isoparorchiidae is a basal relative to the representatives of these three hemiuroid families.

We investigated the molecular epidemiology of Cryptosporidium spp. in Estonia by testing fecal samples from 486 calves aged <2 months, raised on 53 cattle farms, for the presence of Cryptosporidium DNA. The parasites were identified and characterized by sequencing of the 18S rRNA gene and of the 60 kDa glycoprotein (gp60) gene. Moreover, using a questionnaire, we surveyed factors that could be relevant for animal-to-human and human-to-animal transmission of Cryptosporidium spp. on the farms. Cryptosporidium spp. were shed by 23% of the investigated calves and at least one shedding calf was found on 66% of the farms. Cryptosporidium parvum was the most common species shed, while C. bovis and C. ryanae were also detected. More than half of the calves aged 8–14 days shed C. parvum. Nine previously described C. parvum subtypes (IIaA14G1R1, IIaA16G1R1, IIaA17G1R1, IIaA18G1R1, IIaA19G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1 and IIaA16G2R1) and an apparently novel subtype IIlA21R2 were found. Calves from farms that reported spreading manure on fields during spring had 10 times higher odds to shed Cryptosporidium spp. in their feces than calves from farms that did not. Calves aged 8–14 days had higher odds to shed IIa18G1R1 as well as IIaA16G1R1 than younger calves.

The midgut epithelium of Glossina morsitans centralis, G. austeni, G. pallidipes, G. palpalis palpalis, G. p. gambiensis, G. fuscipes fuscipes, G. tachinoides and G. brevipalpis from ILRAD-bred colonies was examined, by electron microscopy, for the presence and distribution of Rickettsia-like organisms (RLOs). RLOs were present in the midgut epithelial cells of all non-teneral tsetse. In G. m. centralis, G. pallidipes and, to a much lesser extent, G. brevipalpis, RLOs were numerous and were present in all the specimens examined. RLOs were present in fewer numbers in the epithelial cells of tenerals of these three tsetse species. In contrast, RLOs occurred in very much lower numbers within the midgut cells of nonteneral G. austeni, G. p. palpalis, G. p. gambiensis, G. f. fuscipes and G. tachinoides; were not seen in every specimen, and were rarely observed in the midgut cells of teneral tsetse. The RLOs were typical rod-shaped bacteria with an inner and outer membrane, which occurred free within the host cell cytoplasm and appeared to cause no obvious pathology. The micro-organisms divided by binary fission and at least two distinct morphological forms plus a range of intermediate forms were seen in the midgut cells. A comparison of the presence and numbers of RLOs within the midgut cells and the midgut infection rates of both Trypanosoma congolense and T. b. brucei, both between Glossina species and also within the same stock of tsetse, clearly indicates that the ability of trypanosomes to establish and develop to mature infections is unlikely to be correlated solely with the presence of RLOs within the tsetse midgut.

Large numbers of electron-dense bodies, 700–1900 Å in diameter, usually seen each with its own limiting membrane, were found in axons of the ventral nerve of infective juveniles of H. contortus. Axons containing these granules merged, without limiting membranes, with a ‘light’, diffusely granular region, about 1·5–2 μm3. The ‘light’ area also merged without boundaries with the underlying cuticle which was different from the surrounding cuticle. This modified cuticle, of which the anterior border lies about 1·3 μm from the posterior edge of the excretory pore, extends slightly beyond the ‘light’ area. Otherwise it is a useful topographical marker for the extent of the ‘light’ area.

It is considered that the ‘light’ region has a special role in the physiology of the cuticle.

I wish to thank Mrs M. Ross for expert technical assistance and Mr R. I. Sommerville for valuable advice. This work was supported by a U.S. Public Health Service Grant, AI 04093–05. The use of the Siemens electron microscope was made possible by a grant from the Australian Research Grants Committee. Financial help from Parke, Davis and Co. (Australia) is also gratefully acknowledged.

After hamsters have been infected with Dipetalonema viteae for about 4 months, microfilariae disappear from the circulation but the adult worms remain alive and active. Such hamsters are said to harbour a ‘latent’ infection. Additional adult worms implanted into such hamsters produce no microfilariae, but adult worms recovered from hamsters with latent infections resume microfilariae production when surgically transplanted into new hosts. In vitro, worms from hamsters with latent infections renewed microfilariae production in the presence of serum from uninfected but not infected hamsters. Serum from infected animals had a marked suppressive effect on the microfilariae production of adult worms implanted into passively immunized hamsters. This suggests that microfilarial release is regulated by a serum factor. Microfilarial production in in vitro and in vivo experiments was more strongly inhibited by serum taken from hamsters with latent infections and hamsters infected for only 30 days (pre-patent) than by serum taken 90 days after infection (whilst microfilariae were in the circulation of the serum donors). A cytotoxic effect of sera from infected hamsters on the microfilariae could not be demonstrated.

In animals immunized by extracts of adult worms or microfilariae, microfilaraemia was considerably reduced. In contrast, animals immunized repeatedly with the extract from small numbers of microfilariae during the pre-patent phase of an infection had an enhanced microfilaraemia. Adult worm numbers were not affected by immunization except in animals given extract from a single female worm which harboured fewer worms than controls.

1. The geographical distribution of cattle lice in Britain is recorded in detail. Bovicola bovis is the commonest and most widely distributed species in Britain.

2. The incubation period for the eggs was found to be: Haematopinus eurysternus, 9–19 days (av. 12); Bovicola bovis, 7–10 days (av. 8); Linognathus vitula, 10–13 days; Solenopotes capillatus, 10–13 days. With eggs of H. eurysternus it was found that the higher the minimum air temperature the shorter was the incubation period.

3. In H. eurysternus the average length of the instars was: 1st, 4 days; 2nd, 4 days; 3rd, 4 days; pre-oviposition period, 3–4 days. The average time for the complete life cycle, egg to egg, was 28 days.

4. The maximum longevity of H. eurysternus on the host was: males, 10 days; females, 16 days. No males or females of H. eurysternus survived a starvation period of 72 hr. at 20° C. and R.H. 70 or 0–10° C. and R.H. 70–85; but some nymphs survived this period at 20° C. and R.H. 70, but none survived 96 hr. starvation.

5. The maximum number of eggs recorded for one female was 24; and eggs were laid at the rate of 1–4 a day.

6. The threshold of development of the eggs of H. eurysternus appears to be about 27·5° C.

The thermal preferences of two strains of the snail Biomphalaria glabrata, one resistant to, and one susceptible to, the parasite Schistosoma mansoni were determined in an aquatic thermal gradient. Snails were tested without exposure to the parasite, and 2 h and 5 weeks after exposure to trematode miracidia. The mean temperature selected by susceptible strain snails 2 h post-exposure tended to be lower than that of unexposed controls, although this was not statistically significant. In this strain, at 5 weeks post-exposure, the preferred temperature dropped by 1.9±0.5°C. The resistant strain displayed a significant drop of 1.8±0.6°C 2 h post-exposure. These results are consistent with the hypothesis that a drop in mean temperatures selected by snails is due to altered levels of endogenous cytokines such as IL-1 or TNF in association with parasite activation of the snail internal defense system.

Experimental studies were conducted on the proportion of Steinernema feltiae Filipjev (Site 76 strain) infective stages that infected Galleria mellonella L larvae, in sand tube assays. A full statistical analysis of the sand assay tubes was also undertaken. The analysis showed that the sand tubes were not a significant source of variation. Consequently the observed behaviour of the infective stages was considered to be due to a biological effect. The study shows that the proportion of infective stages that became parasitic did not increase with increases in the density of applied infective stages, nor did the proportion increase with an increase in the number of hosts. It is argued that these findings are consistent with the presence of a proportion of individual infective stages that are capable of causing infection.