The structure of the various stages of Eimeria tenella grown in the chorioallantoic membranes of the chick embryo has been shown to be similar to the stages in the caecum of the bird.The effects of the anticoccidial drug, Robenidine, on the fine structure of the various stages of E. tenella, grown in the chick embryo and in the caecum of the bird, have been described. The first noticeable change occurs in the endoplasmic reticulum, including the nuclear envelope, and in the Golgi complexes of all stages. The nucleus itself, and the mitochondria, do not appear to be affected by the drug until much later, and this effect is probably due to the degeneration of the cell.It is suggested that the drug may have a direct effect upon protein metabolism of the parasite, resulting in swelling of the granular endoplasmic reticulum and Golgi complexes, or that it may have a primary effect upon some other organelle or metabolic process, and that the swelling of the endoplasmic reticulum and Golgi complexes is a secondary or tertiary phenomenon.

Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro. Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.

Full development of Oswaldofilaria chlamydosauri (Breinl) was obtained in Culex fatigans Wied. and Culex annulirostris Skuse. Infections in four lizards were observed 4–6 months after inoculation of infective larvae from both species of mosquitoes.

1. Eleven new species of cercariae are described from China. Of this number ten have been obtained from species of bithynoid snails and one from Planorbis möllendorfi.

2. One of these species, Cercaria beatifica, a heterophyid larva, needs to be differentiated from the cercaria of Clonorchis sinensis, which develops experimentally in the same mollusc.

3. The forms described are much more common parasites of bithynoid snails than is the larva of Clonorchis.

4. Wherever it has been possible the larval characters, particularly those of the excretory system, have been utilised to show the relationship of the cercariae to adult groups.

During the eighteen months spent with the League of Nations International Commission on Human Trypanosomiasis, I had a unique opportunity for studying the transmissibility of the human trypanosomes by Glossina palpalis.

The rate of new emerging infectious diseases entering the human population has increased over the past century, with pathogens originating from animals or from products of animal origin accounting for the vast majority. Primary risk factors for the emergence and spread of emerging zoonoses include expansion and intensification of animal agriculture and long-distance live animal transport, live animal markets, bushmeat consumption and habitat destruction. Developing effective control strategies is contingent upon the ability to test causative hypotheses of disease transmission within a statistical framework. Broadly speaking, molecular phylogeography offers a framework in which specific hypotheses regarding pathogen gene flow and dispersal within an ecological context can be compared. A number of different methods has been developed for this application. Here, our intent is firstly to discuss the application of a wide variety of statistically based methods (including Bayesian reconstruction, network parsimony analysis and regression) to specific viruses (influenza, salmon anaemia virus, foot and mouth disease and Rift Valley Fever) that have been associated with animal farming/movements; and secondly to place them in the larger framework of the threat of potential zoonotic events as well as the economic and biosecurity implications of pathogen outbreaks among our animal food sources.

There are three species of true parasites in the genus Aphelenchus, viz.: A. fragariae R.B., 1891, A. olesistus R.B., 1893, and A. phyllophagus n.sp. A. ormerodis R.B., 1891, is a doubtful species, possibly a young form of A. fragariae (Marcinowski, 1908).

A. fragariae causes the disease strawberry bunch; A. olesistus and phyllophagus cause leaf disease in flowering plants and ferns. The diseases are widely distributed and of considerable economic importance.

From the study of the anatomy of the three species certain points of general importance emerge: (1) The salivary glands of Aphelenchus represent the oesophageal glands of other nematode genera, which have separated from the body of the oesophagus. This separation causes the reduction in size of the posterior portion of the oesophagus, and its lack of clear demarcation from the intestine. (2) The excretory organ of Aphelenchus, and of some other nematode genera, originates from the left lateral line. (3) The vaginae are ectodermal organs distinct in origin from the gonads proper.

The life cycle of the parasitic Aphelenchi is divided into three definite stages: (A) That of residence in the definitive habitat, which for A. fragariae is in the stem and leaves of the strawberry plant, for A. olesistus and phyllophagus in the mesophyll spaces of the leaves of many plants. Nutrition and reproduction are most actively carried on in this situation, several or many generations succeeding each other, and here the pathological effects are manifest. (B) The resting stage in the soil. The worms reach the soil in fallen leaves, and the adults survive in a partially dried, dormant condition on the surface of the soil for prolonged periods. (C) The stage of immigration into the host plant. When revived by moisture the Aphelenchi may be attracted by a suitable plant, and wanders on to it; they may live as ectoparasites in the leaf axils, breeding in this situation, the larvae migrating to the definitive habitat; or they may enter the definitive habitat direct. They do not traverse the tissues of the stem during immigration. A. olesistus and phyllophagus enter the mesophyll spaces through the stomata.

It should be noted that this life cycle is that of the race, not of the individual. It comprises many generations, and in this respect resembles that of Tylenchus dipsaci and differs from that of T. tritici, which includes one generation only.

In the life of the individual Aphelenchus, embryonic development occupies not more than five days, embryonic and larval development not more than ten, and a complete generation not more than fourteen days.

High atmospheric humidity assists the migration of the worms, previous disease or injury of the host plant may do so. Some races of various species of plants are immune.

Treatment of the affected plants by chemical methods is of limited applicability. Treatment by immersion in water at 50–52° C. for five minutes is recommended by Marcinowski (1908). Prophylaxis should be based on burning of infected plants and leaves, and on the avoidance of infected surface soil in the filling of pots and boxes.

The present paper is concerned with the description of six species of gregarines which are all parasites of British earthworms; they are: Apolocystis herculea (Bosanquet, 1894), A. lavernensis sp.nov., A. perfida sp.nov., A. rotaria sp.nov., A. spinosa sp.nov., A. villosa (Hesse, 1909) Cognetti, 1923. The observations upon which these descriptions are based form part of a larger survey of the monocystid fauna of several British earthworm species from the Cardiff area. The methods used are fully described in a previous paper (Rees, 1962).

1. Large numbers of Plagiorchis (M.) megalorchis n.nom. were found in the intestine of four turkey poults of a total of thirteen which had died on a farm in Radnorshire. Presumably the presence of the parasites was the cause of death.

2. The anatomy of the adult worm is described.

3. The first intermediate host of the parasite is Lymnaea pereger, and the second intermediate host any of the following species of insect larvae: Chironomus riparius Meigen, Culicoides stigma Meigen, Culicoides nubeculosus Meigen and Anatopynia (Psectrotanypus) varius Fabr.

4. The larval stages of the worm are described.

5. The life cycle has been demonstrated experimentally by the feeding of insect larvae containing encysted cercariae to turkey poults.

6. The turkey is probably not the normal host of Plagiorchis (M.) megalorchis. It is most likely that it occurs naturally in some species of wild bird, but this is not yet known.

Interest in the long-term low temperature preservation of malaria parasites was initiated by Coggeshall's (1939) demonstration that the intra-erythrocytic stages could survive storage at −76°C for 70 days. Numerous studies have subsequently been published in which a variety of techniques have been used. In addition, several reviews have included sections on the cryopreservation of malaria, the most recent being by James (1980), Leef, Hollingdale, & Beaudoin (1981), Nguyen-Dinh, Chemangey-Masaba, Spencer, Campbell, Chin & Collins (1981) and a WHO memorandum (1981). Most of these reports and reviews have dealt with the problems of cryopreserving malaria parasites from a parasitological viewpoint. In an attempt to provide a cryobiologically orientated review of value both to cryobiologists and practicing parasitologists the published technical reports have been summarized in chronological order in Tables 1 and 2, and the important cryobiological parameters are discussed.

Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), one of the most threatening zoonoses in Eurasia. Human AE is widespread in the Erzurum region of Turkey, but the situation of the disease in intermediate and definitive hosts is unknown. A Eurasian lynx (Lynx lynx) was killed in a traffic accident in the north of Erzurum, and was taken to our laboratory. Sedimentation and counting technique (SCT), DNA isolation and polymerase chain reaction (PCR) analysis were performed. The SCT results showed that the lynx was infected with E. multilocularis with a medium (745 worms) worm burden. The DNA of adult worms obtained from the lynx was analyzed with a species-specific PCR, and the worms were confirmed to be E. multilocularis by 12S rRNA gene sequence analysis. This is the first report of E. multilocularis from Eurasian lynx in Turkey.

We address the effect of spatial scale and temporal variation on model generality when forming predictive models for fish assignment using a new data mining approach, Random Forests (RF), to variable biological markers (parasite community data). Models were implemented for a fish host-parasite system sampled along the Mediterranean and Atlantic coasts of Spain and were validated using independent datasets. We considered 2 basic classification problems in evaluating the importance of variations in parasite infracommunities for assignment of individual fish to their populations of origin: multiclass (2–5 population models, using 2 seasonal replicates from each of the populations) and 2-class task (using 4 seasonal replicates from 1 Atlantic and 1 Mediterranean population each). The main results are that (i) RF are well suited for multiclass population assignment using parasite communities in non-migratory fish; (ii) RF provide an efficient means for model cross-validation on the baseline data and this allows sample size limitations in parasite tag studies to be tackled effectively; (iii) the performance of RF is dependent on the complexity and spatial extent/configuration of the problem; and (iv) the development of predictive models is strongly influenced by seasonal change and this stresses the importance of both temporal replication and model validation in parasite tagging studies.

Parasites competing over limited host resources are faced with a tradeoff between reproductive success and host overexploitation jeopardizing survival. Surprisingly little is known about the outcome of such competitive scenarios, and we therefore aimed at elucidating interactions between the trematodes Himasthla elongata and Renicola roscovita coinfecting the periwinkle first intermediate host. The results show that the success of Himasthla colonies (rediae) in terms of cercarial emission is unaffected by Renicola competition (sporocysts), whereas deteriating host condition decreases fitness. Furthermore, double infection has no bearing on Himasthla’s colony size but elevated the proportion of non-reproductive rediae that play a decisive role in colony defence. Opposite, the development of the Renicola colony (size/maturity), and in turn fitness, is markedly reduced in presence of Himasthla, whereas the nutritional state of the host appears less important. Hence, the intramolluscan competition between Himasthla and Renicola is asymmetrical, Himasthla being the superior competitor. Himasthla not only adjusts its virulence according to the hosts immediate nutritional state, it also nullifies the negative impact of a heterospecific competitor on own fitness. The latter is argued to follow in part from direct predation on the competitor, for which purpose more defensive non-reproductive rediae are strategically produced.

Entamoeba histolytica is the protozoan parasite causative of human amoebiasis, disease responsible for 40 000–100 000 deaths annually. The cysteine proteinase-adhesin complex of this parasite (EhCPADH) is a heterodimeric protein formed by a cysteine protease (EhCP112) and an adhesin (EhADH) that plays an important role in the cytopathic mechanism of this parasite. The coding genes for EhCP112 and EhADH are adjacent in the E. histolytica genome, suggesting that their expression may be co-regulated, but this hypothesis has not yet been confirmed. Here, we performed the knockdown of EhCP112 and EhADH using gene-specific short-hairpin RNAs (shRNA), and the effect of these knockdowns on the expression of both complex components as well as on the in vitro and in vivo virulence was analysed. Results showed that the knockdown of one of the EhCPADH components produced a simultaneous downregulation of the other protein. Accordingly, a concomitant reduction in the overall expression of the complex was observed. The downregulation of each component also produced a significant decrease in the in vitro and in vivo virulence of trophozoites. These results demonstrated that the expression of EhCP112 and EhADH is co-regulated and confirmed that the EhCPADH complex plays an important role in E. histolytica virulence.

Malaria is endemic over a wide area of the earth's surface. Although popularly associated with tropical or subtropical countries, it occurs as far north as the south of Sweden and Lake Ladoga in Russia and as far south as Bechuanaland, Swaziland and Natal in South Africa and the Argentine in South America. Though most frequent in low-lying districts it may be found at altitudes as great as 6000–9000 ft.

We analysed the intragenomic variation in codon usage in Echinococcus spp. by correspondence analysis. This approach detected a trend among genes which was correlated with expression levels. Among the (presumed) highly expressed sequences we found an increased usage of a subset of codons, almost all of them G- or C- ending. Since an increase in these bases at the synonymous sites is against the mutational bias (these genomes are slightly A+T- rich), we conclude that codon usage in Echinococcus is the result of an equilibrium between compositional pressure and selection, the latter acting at the level of translation, mainly on highly expressed genes. This is the first report where translational selection for codon usage is detected among Platyhelminthes.

An official control programme against Echinococcus granulosus and Taenia hydatigena has been in operation in New Zealand for more than 28 years and against Taenia ovis for more than 18 years. This unique effort to control three metazoan parasites at the same time has led to a change from endemic to extinction status for E. granulosus but only a change from hyperendemic to endemic status for T. hydatigena and T. ovis. This has presented problems in determining the most cost-effective future control strategies. To facilitate this, a benefit/cost analysis of 20 options for the combined control of E. granulosus, T. hydatigena and T. ovis in New Zealand was undertaken. This showed that for E. granulosus a future change from the current non-targeted to a targeted approach is strongly indicated. For T. ovis 6 options were cost-effective using a discount rate of 10%. These were (1) a targeted control package using a vaccine in the non-targeted attack phase; (2) a targeted control package using a larvicide in the attack phase; (3) the transfer of all losses due to and responsibility for the control of T. ovis to the producer who administers a larvicide to sheep to be killed for dog food; (4) the transfer of all losses due to and responsibility for the control of T. ovis to the producer who administers praziquantel every 6 weeks to dogs; (5) and (6) two options involving the discontinuation of control. Control of T. hydatigena was assumed to be an incidental outcome of the policies for the other two parasites.

Most epidemiological models assume that disease is the inevitable outcome of infection (see Bailey, 1957). Yet as Dubos (1965) has said; ‘Throughout nature, infection without disease is the rule rather than the exception’. There are, in fact, many diseases whose distribution cannot be explained solely by a consideration of the probabilities of host parasite encounters. In these cases, a diseased state is only one possible outcome of an interaction between parasite phenotypes, host phenotypes and the external environment. Haemonchosis is an example of such a disease and has been studied extensively in quantitative terms (see Whitlock & Georgi, 1968).

A point mutation in codon 540 of the dihydropteroate synthetase (dhps) gene affecting sulfadoxine resistance has previously been found in parasites from patients with Plasmodium falciparum infection. Here, we investigated 4 methods of identifying this mutation in clinical specimens and established a reliable quantitative assay to estimate the percentage of resistant type in mixed infections. A diagnostic PCR assay based on allele-specific amplification was developed, which clearly typed the clinical specimens examined. The mutation in codon 540 introduces an additional FokI cleavage site which provided a second method to differentiate mutant from wild type, where the former gives rise to 2 characteristic fragments of 538 and 326 bp that are absent from the latter. To calibrate quantitatively the ratio of alleles in mixed samples, we constructed artificial mixes containing 2 plasmid DNAs, one carrying the mutation and the other a wild-type insert. When 32P-labelling was employed, the allele-specific PCR assay could detect the level of resistant type in a mixture down to 0·1–1%, while for the restriction enzyme/PCR analysis, the figure was approximately 10%. Furthermore, neither fluorescent dye-labelled terminator nor dye-labelled primer cycle sequencing was able to detect the mutant allele if it was present at less than 20–30%. We conclude that the allele-specific PCR assay is the most sensitive method of detecting the codon 540 mutation in P. falciparum dhps, and the method of choice for estimating the composition of mixed samples.

Golden hamsters infected with Spirometra erinacei plerocercoids develop a hypertriglyceridaemia characterized by an increase in very low-density lipoprotein. Acyl-CoA synthetase activity, the rate of fatty acid synthesis in the liver and triglyceride production studies showed that the activation and synthesis of fatty acid and production of triglyceride in plerocercoidinfected hamsters were not significantly different from the controls. Lipase activity in post-heparin plasma was found to be suppressed in plerocercoid-infected golden hamsters. The presence of plerocercoids also resulted in a significant reduction in serum thyroxine, but the hypertriglyceridaemia associated with plerocercoid infection was not reversed by injecting the infected hamsters with 2 μg/ day doses of L-thyroxine. The levels of serum immunoreactive insulin in plerocercoid-infected hamsters were not significantly different from the controls. We conclude that the hypertriglyceridaemia, associated with plerocercoid infection in hamsters, results predominantly from a suppression of triglyceride degradation, and that the suppression of lipase activity is probably not the result of hypothyroidism or the lack of insulin, but the result of secretion of growth hormone-like substances.