Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process of Ascaris suum larvae. This study describes moulting and development-specific protein expression profiles of A. suum lung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3–hydroxyacyl–CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.

The kinetoplastid parasite Trypanosoma brucei causes African trypanosomiasis in both humans and animals. Infections place a significant health and economic burden on developing nations in sub-Saharan Africa, but few effective anti-parasitic treatments are currently available. Hence, there is an urgent need to identify new leads for drug development. The T. brucei neutral sphingomyelinase (TbnSMase) was previously established as essential to parasite survival, consequently being identified as a potential drug target. This enzyme may catalyse the single route to sphingolipid catabolism outside the T. brucei lysosome. To obtain new insight into parasite sphingolipid catabolism, the substrate specificity of TbnSMase was investigated using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Recombinant TbnSMase was shown to degrade sphingomyelin, inositol-phosphoceramide and ethanolamine-phosphoceramide sphingolipid substrates, consistent with the sphingolipid complement of the parasites. TbnSMase also catabolized ceramide-1-phosphate, but was inactive towards sphingosine-1-phosphate. The broad-range specificity of this enzyme towards sphingolipid species is a unique feature of TbnSMase. Additionally, ESI-MS/MS analysis revealed previously uncharacterized activity towards lyso-phosphatidylcholine despite the enzyme's inability to degrade phosphatidylcholine. Collectively, these data underline the enzyme's importance in choline homoeostasis and the turnover of sphingolipids in T. brucei.

The magnetic resonance imaging (MRI) appearance of the brain and spinal cord in humans with neuroangiostrongyliasis (NA) due to Angiostrongylus cantonensis infection has been well reported. Equivalent studies in animals are lacking. This case series describes clinical and MRI findings in 11 dogs with presumptively or definitively diagnosed NA. MRI of the brain and/or spinal cord was performed using high-field (1.5 T) or low-field (0.25 T) scanners using various combinations of transverse, sagittal, dorsal and three-dimensional (3D) T1-weighted (T1W), transverse, sagittal and dorsal T2-weighted (T2W), T2W fluid-attenuated inversion recovery (FLAIR) and T2*-weighted (T2*W) gradient echo (GRE), dorsal T2W short tau inversion recovery (STIR) and post-gadolinium transverse, sagittal, dorsal and 3D T1W and transverse T2W FLAIR sequences. In 4/6 cases where the brain was imaged, changes consistent with diffuse meningoencephalitis were observed. Evidence of meningeal involvement was evident even when not clinically apparent. The spinal cord was imaged in 9 dogs, with evidence of meningitis and myelitis detected in regions consistent with the observed neuroanatomical localization. Pathognomonic changes of neural larva migrans, as described in some human patients with NA, were not detected. NA should be considered in the differential diagnosis of dogs with MRI evidence of focal or diffuse meningitis, myelitis and/or encephalitis, especially in areas where A. cantonensis is endemic. If not precluded by imaging findings suggestive of brain herniation, cerebrospinal fluid (CSF) collection for cytology, fluid analysis, real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) testing should be considered mandatory in such cases after the MRI studies.

Variability among samples analysed using the same ELISA protocol generates ambiguity in deciding which assay best quantifies the protein concentration. In this study, we propose a standardization method, called I-STOD (Improved STandardization method for Optical Density), for the transformation of OD values on different plates into relative concentrations of the antibody levels being assessed. We derived an equation relating OD values of different test samples to antibody levels according to the multi-stage reaction dynamics of the indirect-ELISA. Using serum samples from a Schistosomiasis japonica endemic area, we evaluated the fitness of the I-STOD model to experimental data of a standard reference serum in comparison with 5 other models. Calibration curves fitted by the I-STOD method judged to be superior, based on adjusted R2 (adjusted R2>0·99 on 22 out of 26 plates) values. The CV (coefficient of variation) value of the results between multi-well plates and the number of plates with OD values beyond the control range in Shewhart charts also demonstrate that the I-STOD method is a powerful tool which can greatly improve the comparability of results on different multi-well ELISA plates. We conclude that a standardization method is certainly necessary for antibody levels detected in order to properly illustrate clinical differences.

Freshwater gammarids infected with the acanthocephalan parasite Polymorphus minutus show behavioural alterations but also differ from uninfected individuals in their appearance because of the carotenoid-based colouration of the parasite visible through the cuticle. However, it's not clear whether this phenotypic alteration is an adaptation favouring parasite transmission to the definitive host. To test this hypothesis, we investigated the selective preference of mallard towards two prey types: uninfected gammarids on which we applied a dot of inconspicuous brown paint, and uninfected gammarids on which we applied a dot of bright orange paint to mimic the change in appearance due to P. minutus without changes in host behaviour. Mallards showed a significant preference for orange-painted gammarids regardless of how gammarids were distributed (isolated or aggregated). This suggests that parasite's colouration may play a role in enhanced transmission to definitive avian hosts. The role of P. minutus’ colouration in the conspicuousness of gammarids has however to be balanced by the extent to which mallards use visual cues to forage in the field. From the perspective of a multidimensional manipulation, this study suggests that the change in appearance may act synergistically with the changes in behaviour to promote transmission to waterbirds.

Jameson (1926) has described from the caeca of cattle a ciliate belonging to the family Isotrichidae to which he has given the name Buxtonella sulcata. The most prominent character of this ciliate is a dorsal ridge running in a wide sweeping curve from one end of the body to the other with a groove running down the middle. Other characters of importance are a peculiar indentation near the mouth and the not uncommon occurrence of the macronucleus in two separate rounded portions. Roundish oval cysts of this ciliate, 80 to 100 microns in length by 60 to 80 microns in width, were also found by him.

The animal under consideration, or one very like it, has been observed by most collectors of parasitic Copepoda since the publication by Grant appeared in 1827. All authors subsequent to Grant have placed it in the genus Lernaeopoda. Whilst there is a singular unanimity that the area of fixation is the eye, there is a considerable divergence with regard to the host: thus, while Grant originally found it upon the Greenland shark, Scott (1912) records the principal host as Lamna cornubica (which he also calls the Greenland shark), and quotes Norman as having obtained a specimen from Mustelus vulgaris at Polperro. Wilson (1915), an accurate observer, has no record of the host for most of the specimens he examined but stated that one came from a shark. It does not however follow that all these animals are of the same species. The following description is based on two specimens taken from the Arctic shark Squalus borealis = Somniosus microcephalus (Syn. Laemargus and Scymnus) off Greenland on 11. viii. 1923, by W. S. Bristowe.

The underlying mechanism of predisposition to Ascaris infection is not yet understood but host genetics are thought to play a fundamental role. We investigated the association between the Intelectin-2 gene and resistance in F2 mice derived from mouse strains known to be susceptible and resistant to infection. Ascaris larvae were isolated from murine lungs and the number of copies of the Intelectin-2 gene was determined in F2 mice. Intelectin-2 gene copy number was not significantly linked to larval burden. In a pilot experiment, the response to infection in parental mice of both sexes was observed in order to address the suitablity of female F2 mice. No overall significant sex effect was detected. However, a divergence in resistance/susceptibility status was observed between male and, female hybrid offspring. The responsiveness to Ascaris in mice is likely to be controlled by multiple genes and, despite a unique absence from the susceptible C57BL/6j strain, the Intelectin-2 gene does not play a significant role in resistance. The observed intra-strain variation in larval burden requires further investigation but we hypothesize that it stems from social/dominance hierarchies created by the presence of female mice and possibly subsequent hormonal perturbations that modify the intensity of the immune response.

Parasites are detrimental to host fitness and therefore should strongly select for host defence mechanisms. Yet, hosts vary considerably in their observed parasite loads. One notable source of inter-individual variation in parasitism is host sex. Such variation could be caused by the immunomodulatory effects of gonadal steroids. Here we assess the influence of gonadal steroids on the ability of guppies (Poecilia reticulata) to defend themselves against a common and deleterious parasite (Gyrodactylus turnbulli). Adult male guppies underwent 31 days of artificial demasculinization with the androgen receptor-antagonist flutamide, or feminization with a combination of flutamide and the synthetic oestrogen 17 β-estradiol, and their parasite loads were compared over time to untreated males and females. Both demasculinized and feminized male guppies had lower G. turnbulli loads than the untreated males and females, but this effect appeared to be mainly the result of demasculinization, with feminization having no additional measurable effect. Furthermore, demasculinized males, feminized males and untreated females all suffered lower Gyrodactylus-induced mortality than untreated males. Together, these results suggest that androgens reduce the ability of guppies to control parasite loads, and modulate resistance to and survival from infection. We discuss the relevance of these findings for understanding constraints on the evolution of resistance in guppies and other vertebrates.

Two interesting and unusual Spiruroid parasites have been reported from the intestines of marine fish. These are referred to the new genera and species, namely, Bulbocephalus inglisi and Cestocephalus serratus. The former is characterized by a globular head and the muscular oesophagus has two bulbs in its most anterior part. The glandular portion of this oesophagus is 20–25 times as long as the muscular part. The diganosis of Cestocephalus serratus is mainly based on the presence of an oblong head which is covered anteriorly by lobe-like structures in the manner of bothridia in some Cestodes. A new subfamily (family Spiruridae) has been proposed to accommodate Bulbocephalus gen.nov. The family or subfamily diagnosis of Cestocephalus gen.nov. has been restricted at present due to the paucity of material.

The writer wishes to express her gratitude to Professor J. J. C. Buckley for his interest throughout this work.

Haematoloechus (Haematoloechus) exoterorchis sp.nov. and H. (H.) micrurus sp.nov. have been found in the lungs of Rana occipitalis (Gunther) in South Ghana.

62·5% of the frogs were infested with one or other or with both species.

The two species occupy different positions in the lungs of the host. H. (H.) exoterorchis is cylindrical and is wedged, anterior end foremost, into the alveoli. H. (H.) micrurus is flattened and remains free in the central lumen of the lung.

In the former case the wall of the alveolus, containing the worm, becomes stretched to a thin membrane.

The morphology of both species is described with reference to their different habitats.

A brief discussion of the genus is included.

I am most grateful to the University of Ghana and to the staff of the Zoology Department at Legon for the facilities which they placed at my disposal and without which the work could not have been done.

Intact Angiostrongylus cantonensis is able to hydrolyse glucose-phosphate esters, mononucleotides and p-nitrophenyl phosphate as well as β-glycerophosphate in vitro. Reciprocal inhibition studies suggest that the hydrolysis of such substrates is due to a non-specific phosphomonoesterase. Molybdate ions, which exert no effect on either the uptake of glucose or the production of lactate, inhibit the hydrolysis of glucose-1- phosphate in the external medium and simultaneously lower the production of lactate by the intact worms in vitro.

Adelphocystis aeikineta, a new species of monocystid gregarine, is found in the coelom of the earthworm Keffia variabilis in which the trophozoites occur in large numbers in permanently associated pairs. They are large, measuring 750–1500, μm in length, and exhibit continual peristaltic motion. The ectosarc of the trophozoite is markedly striated and there is no evidence of any anterior differentiation. The gametocytes are rounded and the presumed bipolar sporocysts measure 12–14·5 μm in length. Some associated pairs occur in groups enclosed in a membrane and there is evidence that gametocytes may form under these conditions. A. aeikineta is placed in the subfamily Zygocystinae and separated from Zygocystis on its elongate shape. The site of infection of this gregarine, and others recently described, suggests that the morphological basis on which the family Monocystidae is divided into subfamilies and genera is inadequate and a sounder classification should take into account these sites of infection.

The material on which this paper is based was collected while I was in receipt of a Wellcome Trust Travelling Grant, and I wish to thank the Trust for this support. It is also a pleasure to thank Mr R. W. Sims of the British Museum (Natural History) who identified the earthworms, Dr Keith Vickerman who helped me to collect them and Mr R. B. Freeman who advised me on the nomenclature.

Lactating and nulliparous outbred Swiss (CF-1 strain) mice were infected at 12–16 weeks of age with Nippostrongylus brasiliensis or Trichinella spiralis. Lysophospholipase B levels in the intestinal tissue and faecal pellets were greatly depressed in infected lactating mice in contrast to infected nulliparous mice. Correlated with these depressions in lysophospholipase levels were markedly reduced numbers of bonemarrow eosinophils in infected lactating mice. Although the peak levels of lysophospholipase in the intestinal tissues occurred in both nulliparous and lactating mice by days 9 and 14 after infection with N. brasiliensis and T. spiralis, respectively, lactating mice had significantly lower lysophospholipase peak levels than nulliparous mice. The peak of luminal levels of the enzyme coincided with peak levels of the enzyme in the intestinal tissues and with theexpulsion of the majority of the worms from the small intestines of nulliparous mice. However, lactation delayed the temporal relation between the peak of lysophospholipase levels in the intestinal lumen and worm expulsion. These results suggest that lactation depressed the levels of the enzyme and interfered with its release into the intestinal lumen.

Anti-thymocyte serum (ATS) was administered to CD-1 mice infected with 100 Trichinella spiralis larvae and its effects on intestinal lysophospholipase (EC 3.1.1.5) activity, peripheral blood lymphocytes and bone marrow, peripheral blood and intestinal eosinophilia were assayed in the same experimental animal. The ATS caused a significant suppression of both the tissue lysophospholipase response and eosinophilia, in all three compartments, when compared to the values found in infected mice that were either treated with normal rabbit serum (NRS) or untreated. The suppressed eosinophil response and reduced lysophospholipase activity demonstrated a close temporal relation throughout the experiment. These findings support the hypothesis that helminth parasite-induced eosinophilia is the cause of increased lysophospholipase activity present in parasitized tissue and that the responses are thymus cell dependent.

Irradiation doses of 800 rad. or less had no detectable effect on the subsequent development of unfed nymphal Rhipicephalus appendiculatus ticks. Doses of 1600 rad. or above impaired feeding and moulting and delayed the subsequent development of unfed nymphs. At 6400 rad. no ticks completed their moult. Unfed nymphal R. appendiculatus irradiated at 0, 400, 800 and 1600 rad. were applied to 3 cattle infected with T. parva (Kiambu 4). The Theileria infections in the salivary glands of the resultant adult ticks were assessed and it was found that irradiation doses of 800 rad. or less had little effect on the Theileria infections, but an–irradiation dose of 1600 rad. greatly reduced the level of infection. The numbers of salivary gland acini in ticks of this group were reduced but the percentage of infected acini was similar.

Erythrocytes of Tropidurus torquatus (Sauria, Iguanidae) infected with Plasmodium (Sauramoeba) tropiduri show acid phosphatase activity in Golgi vesicles, multivesicular bodies, secondary lysosomes and autophagic vacuoles. The functional relations between these structures and exocytotic activity is discussed.

The authors express their gratitude to: Professor A. G. E. Pearse for his helpful criticism of the work and during the preparation of the revised manuscript; to Professor P. C. C. Garnham for his advice and for his provision of facilities. We are also indebted to Mr Ian McLure for his help in translating the original Spanish into English.