Studies of population genetic structure of parasites can be used to infer which parasite genes are under selection. Here, the population structure of 4 genes associated with drug resistance of Plasmodium falciparum (the chloroquine resistance transporter, pfcrt, dihydrofolate reductase, dhfr, dihydropteroate synthase, dhps, and multi-drug resistance, pfmdr-1) were examined in parasite populations in 3 villages in eastern Sudan and in an urban area of Khartoum, the capital. In order to differentiate the effects of drug selection from neutral influences on population structure, parasites were also genotyped for 3 putatively neutral microsatellite loci (polyα, TA81 and pfg377), and for 2 antigenic loci that are either under balancing selection or neutral, merozoite surface protein 1 and 2, (MSP-1 and MSP-2). Cross-sectional surveys were carried out during the peak transmission (wet) season and in the ensuing dry season. No significant variation in frequencies of MSP-1 and MSP-2 alleles was seen among villages in the eastern region and between the villages and Khartoum, nor between the wet and dry season. However, the drug resistance genes, pfmdr-1, pfcrt and dhfr and to a lesser extent the microsatellite loci showed high FST values when comparing villages with Khartoum, indicating strong geographical differentiation at these loci. Moreover, variation in frequencies of the drug resistance genes, pfmdr-1, pfcrt and dhfr, was observed between the wet and dry season. These differences most probably reflect the variation in drug pressure between each region, and in drug usage between the wet and dry season in a given region.

Mutations in pfcrt K76T are associated with chloroquine resistance in Plasmodium falciparum. Previous studies of K76T mutations in Senegal reported the association of T76 with in vitro-resistant isolates, but this mutation was also prevalent in chloroquine-sensitive isolates. This suggests involvement of additional genetic loci in modulating chloroquine resistance. Additional pfcrt polymorphisms at codons A220S, Q271E, N326S and R371I have been found in chloroquine-resistant isolates. We wanted to test if sequential acquisition of mutations at these codons leads to in vitro chloroquine resistance. Stepwise accumulation of mutations was not detected, rather there was almost complete linkage between the pfcrt K76T mutation and polymorphisms in these codons. Therefore these additional polymorphisms do not enhance the correlation between pfcrt T76 and chloroquine resistance in Senegal. These data suggest that in vitro chloroquine resistance requires the genetic background of the pfcrt K76T mutation and additional mutations in genetic loci outside the pfcrt gene.

Genetic studies have demonstrated profound differences between the ‘porcine’ genotype of Cryptosporidium parvum, versus ‘human’ and ‘bovine’ genotypes. The study analysed infectivity and pathogenicity of the ‘porcine’ genotype (CPP-13 isolate) of C. parvum, and compared the results with published data on the ‘bovine’ genotype (CPB-0 isolate). This was investigated in calves and piglets from commercial herds. Piglets were mildly affected by the CPP-13 isolate, contrary to piglets infected with the CPB-0 isolate, which caused diarrhoea of a mean duration of 3·5 days. CPP-13 produced no or very mild clinical signs in piglets despite the excretion of high numbers of oocysts. Concomitant infection with rotavirus, however, caused a dramatic aggravation of the clinical signs, and 5 of 6 experimentally infected piglets died. CPP-13 appeared to be adapted to porcine hosts as illustrated by the lack of infectivity to 1 experimentally inoculated calf, and the absence of clinical signs, the long pre-patent period (15 days), and the excretion of very low numbers of oocysts following experimental infection of another calf. Thus, in accordance with other molecular studies, our results support the genetic evidence for the existence of a new species of Cryptosporidium adapted to pigs.

Cystic echinococcosis, caused by Echinococcus granulosus, is an emerging disease in many parts of the world and, in particular, in eastern Europe and the former Soviet Union. This paper examines the abundance of infection of E. granulosus in the definitive host in southern Kazakhstan. Observed data are fitted to a mathematical model in order to decide if the parasite population is partly regulated by definitive host immunity and to define parameters in the model. Such data would be useful to develop simulation models for the control of this disease. Maximum likelihood techniques were used to define the parameters and their confidence limits in the model and the negative binomial distribution was used to define the error variance in the observed data. The results indicated that there were 2 distinct populations of dogs in rural Kazakhstan which had significantly different exposures to E. granulosus. Farm dogs, which are closely associated with livestock husbandry, particularly sheep rearing, had a relatively high mean abundance of 631 parasites per dog and a prevalence rate of approximately 23%. The best fit to the model indicated that there was significant herd immunity in the dog at this infection pressure. In contrast, village dogs which were more likely to be kept as pets had a much lower mean abundance of parasites of only 27 parasites per dog and a lower prevalence of 5·8%. With this village population of dogs, the best fit indicated negligible herd immunity.

The pathogenic trypanosomes Trypanosoma equiperdum, T. evansi as well as T. brucei are morphologically identical. In horses, these parasites are considered to cause respectively dourine, surra and nagana. Previous molecular attempts to differentiate these species were not successful for T. evansi and T. equiperdum; only T. b. brucei could be differentiated to a certain extent. In this study we analysed 10 T. equiperdum, 8 T. evansi and 4 T. b. brucei using Random Amplified Polymorphic DNA (RAPD) and multiplex-endonuclease fingerprinting, a modified AFLP technique. The results obtained confirm the homogeneity of the T. evansi group tested. The T. b. brucei clustered out in a heterogenous group. For T. equiperdum the situation is more complex: 8 out of 10 T. equiperdum clustered together with the T. evansi group, while 2 T. equiperdum strains were more related to T. b. brucei. Hence, 2 hypotheses can be formulated: (1) only 2 T. equiperdum strains are genuine T. equiperdum causing dourine; all other T. equiperdum strains actually are T. evansi causing surra or (2) T. equiperdum does not exist at all. In that case, the different clinical outcome of horse infections with T. evansi or T. b. brucei is primarily related to the host immune response.

Viable eggs produced weekly per infective stage was used as a measure of the reproductive success of Echinostoma friedi during the first 12 weeks of infection in hamsters. The weekly reproductive success was not constant during the experiment in relation to the egg output and the proportion of viable eggs produced. The egg release started during week 2 post-inoculation, attaining a maximum during week 3. A decline in egg output was observed from week 9. Viable eggs were only produced from week 3 post-inoculation and a maximum was attained at week 4 of the experiment. A decline in egg viability was observed from week 9. Considering together the egg output and the egg viability, the maximum weekly reproductive success was obtained during week 4 post-inoculation. The changes in the weekly reproductive success were not reflected in variations in worm numbers and body sizes during the course of the infection. The humoral immune response of golden hamsters during the infection with E. friedi was determined. Increases of IgG levels against somatic and excretory/secretory products of E. friedi were detected coinciding with the reduction in the reproductive success.

Echinostome larval stages in the snail have a great potential as effective competitors for the control of schistosomes and adult worms can cause painful intestinal diseases in humans. Ecology and transmission of the larval stages of trematodes are poorly understood, especially because their identification in field-collected samples by microscopy is difficult. We cloned, sequenced and analysed a 192 bp tandem repreated DNA sequence of Echinostoma caproni (EcSau3A), an often discussed antagonist of Schistosoma mansoni in Biomphalaria snails. PCR primers against this sequence can detect less than 10 fg of E. caproni DNA, 2 miracidia in snails 1 day p.i., 1 metacercaria in 50 mg snail tissue and 1 cercaria in 50 mg plankton with high specificity. Methods described in this study can support the discovery of fundamental ecological principles on distribution, host specificity and epidemiology of E. caproni larvae under field conditions.

Psoroptes ovis, the causative agent of sheep scab, is an important ectoparasitic mite infecting sheep, goats and cattle. Infection is characterized by an extensive dermatitis, scab formation and intense itching. Initial focal lesions spread outwards, coalesce and may extend over the whole body. The host response to infestation has all the characteristics of an immediate-type hypersensitivity reaction but the mite antigens and allergens which initiate this response are almost completely undefined. Here, 507 randomly selected cDNAs derived from a mixed population of P. ovis were sequenced and the resultant nucleotide sequences subjected to Cluster analysis and Blast searches. This analysis yielded 280 clusters of which 49 had >1 sequence with 24 showing significant Blast X homology to another protein in the databases. There were 231 sequences which appeared on one occasion and 109 of these showed significant Blast X homology to other sequences in the databases. This analysis identified homologues of 9 different types of allergens which have been characterized in other allergic conditions such as responses to house dust mites. It also identified a number of cysteine proteases which may contribute to lesion development as well as several free-radical scavenging enzymes which may protect the mite from host immune effector responses.

Most sylvatic species of Trichinella are known to have poor infectivity to rats, but in the present study oral administration of bile from other hosts appeared to modify this infectivity. A total of 75 rats were inoculated in groups of 25 with 3 species of Trichinella (T. spiralis, T. nativa, and T. nelsoni) and each group of rats was given per os daily doses of bile from pig, sheep, chicken and fox respectively (4×5 rats). As a control 1 group of 5 rats was given daily doses of water. Whereas, the addition of bile did not increase the establishment of T. spiralis, fox bile had a significant positive effect on the establishment of muscle larvae of T. nativa and T. nelsoni. Addition of bile to cultures of the same Trichinella species had an overall negative effect on the in vitro survival of larvae. The present observation that carnivore bile favours the establishment of sylvatic Trichinella may explain why carnivores are equally receptive to all Trichinella species.

Allozyme markers were used to study genetic variation in Lecithochirium fusiforme within a natural population of Conger conger. Six of 16 enzyme-coding loci studied were found to be polymorphic. These loci were surveyed in 12 infrapopulations of adult flukes. High levels of genetic variation were detected (P=0·375); Ho=0·048; He=0·085). However, the population did not conform to Hardy-Weinberg expectations, as it showed a significant deficit of heterozygotes. L. fusiforme also exhibited low differentiation between infrapopulations (FST=0·064). Despite significant linkage disequilibrium at Pgm-1 and Pgm-2 (P<0·05), mating system does not appear to be the principal reason for the deficit of heterozygotes detected, because some polymorphic loci were in Hardy-Weinberg equilibrium. Association between FIS and FST statistics suggests the existence of the Wahlund effect. However, all infrapopulations showed a strong deficit of heterozygotes for most polymorphic loci (FIS=0·409). Detection of significant genetic differentiation among temporal samples and the existence of paratenic hosts in the life-cycle suggests the Wahlund effect, caused by the mixture of genetically distinct temporal samples in the infrapopulations. Occasional temporal gene flow also might explain the high estimated genetic polymorphism.

The host specificity of a parasite is not merely a function of how many host species it can exploit, but also of how closely related these host species are to each other. Here, a new index of host specificity is proposed, one that takes into account the average taxonomic or phylogenetic distance between pairs of host species used by a parasite. The index is derived from measures of taxonomic distinctness used in biodiversity studies. It is easy to compute and interpret, ranging from a minimum value of 1 when all host species are members of the same genus, to a maximum of 5, when all host species belong to different classes. The variance of this measure can also be computed, and provides additional information on the taxonomic or phylogenetic structure of the host assemblage. Using data on helminth parasites of Canadian freshwater fishes, we show that the new index, unlike the mere number of known host species, is independent of study effort i.e. the number of published records of a parasite. Although the index and the number of known hosts are not entirely independent statistically, each captures a different aspect of host specificity. For instance, although acanthocephalans infect significantly more host species than trematodes, cestodes or nematodes, there is no difference in the average index value among these 4 helminth taxa, suggesting that the average taxonomic distances between the host species of a parasite do not vary among these higher taxa. We recommend the use of our new index in future comparative studies of host specificity, in particular when the focus is on the evolutionary history of parasites and of their past colonizations of host lineages.

The tapeworm Raillietina trapezoides was studied in a Tunisian population of the fat sand rat Psammomys obesus. Seasonal changes in the abundance of parasite and host were monitored in a longitudinal field survey lasting 20 months. In total 582 intestinal samples were collected during 10 trapping sessions between May 1995 and January 1997 and examined. The impact of abiotic and biotic factors on the prevalence and parasite burden were explored, using generalized linear models. R. trapezoides showed a seasonal pattern with most transmission occurring in summer and autumn. In August–September when the rodents were at low density and most adult (90–100%) were infected. In winter, infections with R. trapezoides were highly prevalent in the cohort of adults only. The worm burden by number was also high in late summer and autumn. Prevalence and parasite burden by number or by biomass were highly age dependent. Season and age act in synergy so that the rodents were highly infected in late summer and autumn. Unusually dry conditions in winter seemed to increase prevalence and to reduce the number of worms. There was no difference between males and females in prevalence or parasite burden (by number or by biomass). Prevalence was not correlated with the relative density of the hosts whereas parasite burden depended upon their densities. The more numerous the worms were, the smaller they were individually, reflecting a crowding effect. The possibility of an immune response regulating P. obesus is discussed, in regard to the very high natural prevalence and the variation in worm burden, and compared with other parasite–host systems.

The genetic structure of populations of the fish cestode, Bothriocephalus acheilognathi collected from Bailianhe Reservoir (BLH), Changshou (CSH) and Liangzi (LZH) Lakes was investigated by using 8 microsatellite loci. A total of 108 adult worms were genotyped at each of the 8 loci. For the 3 populations, the mean number of alleles per locus ranged from 2·38 to 5·5, and the mean expected heterozygosity ranged from 0·432 to 0·559. The average polymorphic information content (PIC) was from 0·384 to 0·492. The significant Fis values indicated non-random mating within LZH and BLH populations. On the other hand, when samples were further classified into subpopulations at the level of host fish species, no or little heterozygote deficiency was detected at most loci, showing that cross-fertilization, predominantly, but not exclusively, must have occurred within the subpopulations. Microsatellite markers also revealed an unexpected high level of genetic differentiation, as measured by Rst and Nm values or by δu2 genetic distance among subpopulations from different hosts. Factors influencing the population genetic structure and the parasite host specificity are discussed.