The species Phytomonas serpens is known to express some molecules displaying similarity to those described in trypanosomatids pathogenic to humans, such as peptidases from Trypanosoma cruzi (cruzipain) and Leishmania spp. (gp63). In this work, a population of P. serpens resistant to the calpain inhibitor MDL28170 at 70 µm (MDLR population) was selected by culturing promastigotes in increasing concentrations of the drug. The only relevant ultrastructural difference between wild-type (WT) and MDLR promastigotes was the presence of microvesicles within the flagellar pocket of the latter. MDLR population also showed an increased reactivity to anti-cruzipain antibody as well as a higher papain-like proteolytic activity, while the expression of calpain-like molecules cross-reactive to anti-Dm-calpain (from Drosophila melanogaster) antibody and calcium-dependent cysteine peptidase activity were decreased. Gp63-like molecules also presented a diminished expression in MDLR population, which is probably correlated to the reduction in the parasite adhesion to the salivary glands of the insect vector Oncopeltus fasciatus. A lower accumulation of Rhodamine 123 was detected in MDLR cells when compared with the WT population, a phenotype that was reversed when MDLR cells were treated with cyclosporin A and verapamil. Collectively, our results may help in the understanding of the roles of calpain inhibitors in trypanosomatids.

We report the discovery of a biological rhythm in the reproductive behaviour of the tick Bothriocroton hydrosauri that was absent in Amblyomma limbatum, a species that occurs on the same species of reptile host. Female B. hydrosauri mated in autumn or winter delayed oviposition until the following spring, while there was no diapause in conspecific females mated in spring or early summer. Initiation of ovipositional diapause in ticks is usually related to photoperiodic stimuli, but this was not the case for B. hydrosauri. The sinusoidal pattern in pre-oviposition times of B. hydrosauri females mated in different months in the laboratory suggests an internal seasonal time-keeping mechanism. We hypothesize that hormones imbibed by females during their bloodmeal may provide environmental cues associated with the induction of diapause. Irrespective of the mechanism underlying the rhythm, diapause by B. hydrosauri females mated during autumn or winter is of adaptive advantage because it synchronizes oviposition with favourable environmental conditions for egg hatching and increases the chance of larvae finding a host. The lack of a similar biological rhythm in A. limbatum may be a reflection of the different environmental conditions this species experiences throughout most of its range as compared with B. hydrosauri.

Theileria parva is an intracellular protozoan parasite transmitted by ticks that causes a fatal lymphoproliferative disease of cattle known as East Coast Fever. Vaccination against the disease currently relies on inoculation of the infective sporozoite stage of the parasite and simultaneous treatment with long-acting formulations of oxytetracycline. Sporozoites are maintained as frozen stabilates of triturated infected ticks and the method requires accurate titration of stabilates to determine appropriate dose rates. Titration has traditionally been undertaken in cattle and requires large numbers of animals because of individual variation in susceptibility to infection. An alternative tissue culture-based method is laborious and time consuming. We have developed a flow cytometric method for quantifying the infectivity of sporozoite stabilates in vitro based on the detection of intracellular parasite antigen. The method allows clear identification of parasitized cells with a high degree of sensitivity and specificity. Analysis of infected cells between 48 and 72 h post-infection clearly defines the potential transforming capability of different stabilates.

The trematode described in this paper was found by Dr Alexander Brown, British Central Africa, in the stomach of a Rhodesian hartebeest (Bubalis sp.). About 120 specimens were collected and placed at my disposal for investigation by Dr John Rennie, Lecturer on Parasitology in this University, to whom I am much indebted for the use of the interesting material. The specimens proved to belong to a new species to which the name Gastrothylax bubalis has been given. Although a fuller knowledge of recorded species rather than the recovery of new trematodes is to be desired, it is hoped that the description of this parasite may be interesting from the fact that the genus includes only ten other known species.

Whilst examining the contents of a number of insects from the neighbourhood of Pretoria (Transvaal), a flagellate was found occurring in the alimentary canal of a Hemipterous insect, Cletus varius Dall1. Unfortunately, no observations were made on the living parasite and the following notes on its morphology are only derived from the study of ordinary dried smears, fixed in alcohol and stained with Giemsa. In consequence of the method of preparation, the finer nuclear details have been destroyed, but nevertheless we have been able to make out the main points in the structure of this parasite, and as no further material is available we have decided to publish our results in their present form.

Among the bird-parasites included in the family Hippoboscidae, are four small closely conneceted genera, in all of which the wings exhibit more or less extensive reduction and are apparently useless for the purpose of flight. In various parts of the world species belonging to one or other of three of these genera, namely Crataerina, v. Olf., Myophthiria, Rond., and Brachypteromyia, Will., infest Swifts (Micropus, Collocalia, etc.) and their nests; while the solitary representative of the fourth genus Stenepteryx, Leach, at present known is similarly parasitic on the House-Martin (Delichon urbica, L.), which, though in no wise related, is closely associated with the Common Swift in the popular mind.

We investigate the factors associated with the occurrence and abundance of external and blood parasites in African penguins (Spheniscus demersus), an endangered seabird that breeds exclusively on the coasts of Namibia and South Africa. External parasites were collected using the dust-ruffling method from 171 African Penguins admitted at a rehabilitation facility in the Western Cape, South Africa. Additionally, blood smears were obtained upon admission and weekly during rehabilitation and examined for blood parasites. Fleas Parapsyllus longicornis humboldti, ticks Ornithodoros capensis and lice Austrogoniodes demersus were recovered from 93, 63 and 40%, respectively, of the penguins upon admission to the centre. Rescue location and age group were identified as significant determinants of flea abundance, whereas month of admission was a significant determinant of tick abundance. Blood parasites were also common on admission, with Babesia being the most frequent (46% prevalence) whereas Borrelia was recorded sporadically (1.2%) and Plasmodium was recorded once. The prevalence and abundance of ticks on admission was positively associated with Babesia infection on admission. Our findings demonstrate the variability and contributing factor of parasite infections in an endangered species of penguin, and highlight the need for additional research on the parasite–host dynamics involving these potential disease vectors.

GLP-1 (abnormal germline proliferation) is a Notch-like receptor protein that plays an essential role in pharyngeal development. In this study, an orthologue of Caenorhabditis elegans glp-1 was identified in Meloidogyne incognita. A computational analysis revealed that the orthologue contained almost all the domains present in the C. elegans gene: specifically, the LIN-12/Notch repeat, the ankyrin repeat, a transmembrane domain and different ligand-binding motifs were present in orthologue, but the epidermal growth factor-like motif was not observed. An expression analysis showed differential expression of glp-1 throughout the life cycle of M. incognita, with relatively higher expression in the egg stage. To evaluate the silencing efficacy of Mi-glp-1, transgenic Arabidopsis plants carrying double-stranded RNA constructs of glp-1 were generated, and infection of these plants with M. incognita resulted in a 47–50% reduction in the numbers of galls, females and egg masses. Females obtained from the transgenic RNAi lines exhibited 40–60% reductions in the transcript levels of the targeted glp-1 gene compared with females isolated from the control plants. Second-generation juveniles (J2s), which were descendants of the infected females from the transgenic lines, showed aberrant phenotypes. These J2s exhibited a significant decrease in the overall distance from the stylet to the metacorpus region, and this effect was accompanied by disruption around the metacorporeal bulb of the pharynx. The present study suggests a role for this gene in organ (pharynx) development during embryogenesis in M. incognita and its potential use as a target in the management of nematode infestations in plants.

Efficacious antimalarial drugs are important for malaria control and elimination, and continuous monitoring of their efficacy is essential. The prevalence and distribution of Pfmdr1 were evaluated in African migrant workers in Henan Province. Among 632 isolates, 13 haplotypes were identified, NYSND (39.87%, 252/632), YYSND (2.85%, 18/632), NFSND (31.01%, 196/632), NYSNY (0.47%, 3/632), YFSND (13.77%, 87/632), NFSNY (0.32%, 2/632), YYSNY (2.06%, 13/632), YFSNY (0.16%, 1/632), N/Y YSND (1.90%, 12/632), N Y/F SND (6.17%, 39/632), N/Y Y/F SND (0.47%, 3/632), YYSN D/Y (0.16%, 1/632) and N/Y FSND (0.79%, 5/632). The highest frequency of NYSND was observed in individuals from North Africa (63.64%, 7/11), followed by South Africa (61.33%, 111/181), Central Africa (33.33%, 56/168), West Africa (28.94%, 68/235) and East Africa (27.03%, 10/37) (χ2 = 54.605, P < 0.05). The highest frequency of NFSND was observed in East Africa (48.65%, 18/37), followed by West Africa (39.14%, 92/235), Central Africa (26.79%, 45/168), South Africa (22.65%, 41/181) and North Africa (9.09%, 1/11) (χ2 = 22.368 P < 0.05). The mutant prevalence of codons 86 and 184 decreased. These data may provide complementary information on antimalarial resistance that may be utilized in the development of a treatment regimen for Henan Province.

The post-cercarial development of G. crumenifer (Creplin) has been experimentally elucidated.

Gyraulus convexiusculus (Hutton) serves as the intermediate host under natural conditions.

The adult stage of the fluke was raised experimentally in ruminants, the time taken by the parasite to attain maturity being 118 days in the kid and 114 in the buffalo calf.

Cercaria chungathi Peter & Srivastava, 1955, has been established to be the larva of Gastrothylax crumenifer. The cercaria, the redial parthenita and the metacercaria have been described.

The authors are grateful to the Director, Indian Veterinary Research Institute, for his keen interest in these studies.

Infective 3rd-stage larvae of three species of marine ascaridoid nematodes, Pseudoterranova (= Phocanema = Porrocaecum = Terranova) decipiens, Anisakis simplex and Phocascaris/Contracaecum sp. were cultured in vitro in 0·9% saline or Medium 199 at 37°C beneath an atmosphere of either air or 95% air:5% CO2 All three species responded to culture at 37°C by producing a new cuticle. The majority of P. decipiens completed ecdysis under all the culture conditions employed. Ecdysis in A. simplex was stimulated by a high concentration of CO2; this effect was particularly noticeable in saline cultures. Both A. simplex and Phocascaris/Contracaecum sp. developed more effectively in a nutrient medium than in saline.

The growth of an axenic strain of Entamoeba invadens in two media (Trypticase–liver broth and Trypticase–Panmede) is compared with that of a monobacterial strain in diphasic medium supplemented with starch. The axenic strain multiplied more slowly, and produced lower yields of amoebae than the monobacterial strain.

Amoebae multiplied more rapidly and produced higher yields in Trypticase–liver broth than in Trypticase–Panmede.

The effect of certain variations in the basic Trypticase–Panmede medium was investigated. Serum stimulated the growth of the axenic strain in Trypticase–Panmede, so that although the initial multiplication rate was not increased, the maximum yields of amoebae were increased to levels that varied according to the concentration of the serum.

The multiplication of the axenic amoebae was depressed by procedures which probably aerated the medium.

I thank Dr L. S. Diamond for axenic strain NIH:200 of E. histolytica, and Professor E. Meerovitch for axenic strain PZ of E. invadens. I am much indebted to Dr P. Whittlestone, of the Department of Animal Pathology, Cambridge, who examined cultures of strain N for the presence of mycoplasmata. The work was carried out while the author was a member of the scientific staff, Medical Research Council.

Plasmodium rhoptry neck protein 2 (RON2), which is released from the neck portion of the merozoite rhoptries and interacts with the microneme protein Apical Membrane Antigen 1 (AMA1), plays a crucial role in erythrocyte invasion. In this study, we sequenced the Plasmodium vivax RON2 gene from 19 P. vivax isolates collected in central China in order to establish whether this protein is under positive diversifying selection, which may occur as a result of protective host immune pressure†. In comparison with the P. vivax Sal-1 reference line, we found 10 amino acid substitutions dispersed throughout the open reading frame as well as indels caused by polymorphism in a repeat unit (21–23 repeats of (Q/E/K/N/H)(G/D)G(H/L/Y/P)G) in the second tandem repeat region located at amino acid positions 541–650. A McDonald-Kreitman test with RON2 sequences from the primate malaria parasite Plasmodium knowlesi, detected significant departure from neutrality in the PvRON2 3′ region (nucleotide positions 2668–6609). These results suggest that the PvRON2 gene has evolved under positive diversifying selection.

We analysed data on the abundance and distribution of 26 species of mesostigmate mites with different feeding habits collected from bodies of small mammalian hosts in 2 geographical regions (West Siberia and Argentina). We tested whether prevalence of a mite can be reliably predicted from a simple epidemiological model that takes into account mean abundance and its variance. We theorized that the difference between prevalence predicted from the model and observed prevalence would be smallest in obligatory haematophagous mites, intermediate in facultatively haematophagous mites and greatest in non-haematophagous mites. We also theorized that prevalence of mites from the region with sharp seasonality (Siberia) would be predicted accurately only if host number would be taken into account. We found that the success of a simple epidemiological model to predict prevalence in mites was similar to that reported earlier for other ectoparasitic arthropods. Surprisingly, the model predicted prevalence of obligatory exclusively haematophagous mites less successfully than that of mites with other feeding habits. No difference in the model performance between mites occurring in the 2 geographical regions were found independent of whether the model took the number of hosts into account.

Leishmaniasis develops after parasites establish themselves as amastigotes inside mammalian cells and start replicating. As relatively few parasites survive the innate immune defence, intracellular amastigotes spreading towards uninfected cells is instrumental to disease progression. Nevertheless the mechanism of Leishmania dissemination remains unclear, mostly due to the lack of a reliable model of infection spreading. Here, an in vitro model representing the dissemination of Leishmania amastigotes between human macrophages has been developed. Differentiated THP-1 macrophages were infected with GFP expressing Leishmania aethiopica and Leishmania mexicana. The percentage of infected cells was enriched via camptothecin treatment to achieve 64·1 ± 3% (L. aethiopica) and 92 ± 1·2% (L. mexicana) at 72 h, compared to 35 ± 4·2% (L. aethiopica) and 36·2 ± 2·4% (L. mexicana) in untreated population. Infected cells were co-cultured with a newly differentiated population of THP-1 macrophages. Spreading was detected after 12 h of co-culture. Live cell imaging showed inter-cellular extrusion of L. aethiopica and L. mexicana to recipient cells took place independently of host cell lysis. Establishment of secondary infection from Leishmania infected cells provided an insight into the cellular phenomena of parasite movement between human macrophages. Moreover, it supports further investigation into the molecular mechanisms of parasites spreading, which forms the basis of disease development.

Trichodinids are parasites generally found on gills and skin of a broad number of aquatic animals. Only a small number of endozoic species has been reported from the urinary tract, intestine and urogenital system in some fish, amphibians and molluscs. This is the first report on the presence of endozoic trichodinids in the spleen, kidney and liver of shorthorn sculpin (Myoxocephalus scorpius). In the present study, trichodinids displayed some of the typical morphological characteristics of endozoic trichodinids with narrow blades and straight rays of adhesive disc denticles. The parasites were observed at a relatively high prevalence (23.9%). There was a positive correlation between intensity of endozoic trichodinids in the internal organs and ectozoic trichodinids on the gills (R = 0.5, n = 46, P < 0.001) whereas there was no correlation between intensity of endozoic trichodinids and the host's body length, body weight or liver weight. Infection levels of endozoic trichodinids were not dependent upon sculpin sex and there was no effect of sampling locations on prevalence and intensity of endozoic trichodinids.

Seasonal life cycle of body cavity dwelling (BCD) Philometra ovata (Nematoda: Philometridae) has been reported in southern and central European countries, but its swim bladder dwelling (SBD) stage and northern populations have remained unstudied. In this study, we investigated the seasonal life cycle and infection ecology of P. ovata in both swim bladder and body cavity in the European minnow (Phoxinus phoxinus) in Finland. The larval P. ovata infected the swim bladder of minnows mainly in August. Female SBD P. ovata emigrated to body cavity mostly in September, grew to their full size by the end of the next June, and evacuated from minnows in July. In addition, female SBD P. ovata retarding their development and staying in swim bladder were found commonly in minnows, thus the mean monthly prevalence (6·7 ± 3·9%) and mean intensity (1·4 ± 0·8) of BCD P. ovata was lower than that of SBD P. ovata (37·8 ± 15·1% and 2·0 ± 1·5, respectively). Finally, despite the large size of BCD individuals, infection of P. ovata did not impair body condition and relative gonad size of minnows, but increased the mortality and caused physical damages in their hosts during the evacuation period.

Distomum incommodum (Leidy 1856) from mouth of Alligator mississippiensis is found to belong to the family Clinostomidae. A new genus Homoscaphis Canavan 1933 is erected for it, a diagnosis of which is given, with a redescription and illustration of the species.

Extraordinary as any of the Mallophaga are when compared with conventional insect forms, there are some species of the order which are still more peculiar than the majority of their fellows. Among these unusually strange forms is that here to be dealt with, a species that seems to combine within itself the characteristics of two otherwise trenchantly separated groups, that possesses striking specific peculiarities and that in addition is decidedly aberrant in its choice of host. It has but once been recorded. The availability of further specimens makes possible additions to our knowledge of its structure and a reconsideration of its relationships.