Distribution of Pomphorhynchus laevis s.l. (Acanthocephala) among fish species at a local scale: importance of fish biomass density.

Parasite distribution among hosts is a fundamental aspect of host-parasite interactions. Aggregated parasite distributions within and across host species are commonly reported and potentially influenced by many factors, whether host or parasite specific, or related to host-parasite encounter and compatibility. Yet, the respective role of each in observed parasite distributions are often unclear. Here, we documented the distribution of the acanthocephalan parasite Pomphorhynchus laevis sensu lato (s.l.) in two replicate fish host populations. Aggregated distributions were observed in both populations, within and across fish host species. We found a positive abundance-prevalence relationship across fish species, suggesting that resource availability (fish host biomass density) was the main driver of P. laevis s.l. distribution. This was supported by further positive associations between mean parasite load and fish biomass density. We found little evidence for intensity-dependent regulation within host (i.e. intra-host competition among co-infecting parasites). Furthermore, P. laevis s.l. infection had no detectable effect on fish condition indices, except on the body condition of female barbel (Barbus barbus). Therefore, P. laevis s.l. tended to accumulate with size/age within fish species, and with fish biomass density among fish species, with apparently negligible limitations due to intra-host intensity-dependent regulation of parasite, or to parasite-induced morbidity in fish. The relative availability of final hosts for trophic transmission thus appears to be the main driver of P. laevis s.l. distribution among fish.

Inter-population variation in the intensity of host manipulation by the fish acanthocephalan Pomphorhynchus tereticollis: are differences driven by predation risk?

Many trophically-transmitted parasites induce behavioural alteration in their intermediate hosts that tend to increase host vulnerability to predation. Inter-population variability in parasite-induced alterations is expected to arise from variable local opportunities for trophic transmission. Yet, this hypothesis has not been investigated so far. We addressed the issue in four populations of the fish parasite Pomphorhynchus tereticollis (Acanthocephala), using variable fish biomass density as a proxy for transmission opportunities. We found variation in the intensity of parasite-induced changes in phototaxis and refuge use among populations. Two of the populations with the lowest predator biomass exhibited the highest levels of behavioural manipulation and prevalence, as expected at low transmission opportunities. They also exhibited micro-habitat segregation between infected and uninfected gammarids in the field. In addition, infection had variable effects on two physiological defence systems, immunity and antioxidant capacity, and on total protein content. Overall, our study brings partial support to the prediction that host manipulation and prevalence should be higher at low predator biomass. Although stronger evidence should be sought by increasing population replicates, our study points to the importance of the ecological context, specifically transmission opportunities brought about by predation pressure, for the evolution of parasite manipulation in trophically-transmitted parasites.

Intraspecific conflict over host manipulation between different larval stages of an acanthocephalan parasite.

Competitive interactions between coinfecting parasites are expected to be strong when they affect transmission success. When transmission is enhanced by altering host behaviour, intraspecific conflict can lead to 'coinfection exclusion' by the first-in parasite or to a 'sabotage' of behavioural manipulation by the youngest noninfective parasite. We tested these hypotheses in the acanthocephalan parasite Pomphorhynchus laevis, reversing phototaxis in its intermediate host Gammarus pulex. No evidence was found for coinfection exclusion in gammarids sequentially exposed to infection. Behavioural manipulation was slightly weakened but not cancelled in gammarids infected with mixed larval stages. Therefore, coinfecting infective and noninfective larvae both suffered competition, potentially resulting in delayed transmission and increased risk of mortality, respectively. Consequently, noninfective larva is not just a 'passive passenger' in the manipulated host, which raises interesting questions about the selective pressures at play and the mechanisms underlying manipulation.

Infection with acanthocephalans increases the vulnerability of Gammarus pulex (Crustacea, Amphipoda) to non-host invertebrate predators.

Phenotypic alterations induced by parasites in their intermediate hosts often result in enhanced trophic transmission to appropriate final hosts. However, such alterations may also increase the vulnerability of intermediate hosts to predation by non-host species. We studied the influence of both infection with 3 different acanthocephalan parasites (Pomphorhynchus laevis, P. tereticollis, and Polymorphus minutus) and the availability of refuges on the susceptibility of the amphipod Gammarus pulex to predation by 2 non-host predators in microcosms. Only infection with P. laevis increased the vulnerability of amphipods to predation by crayfish, Orconectes limosus. In contrast, in the absence of refuges, the selectivity of water scorpions, Nepa cinerea, for infected prey was significant and did not differ according to parasite species. When a refuge was available for infected prey, however, water scorpion selectivity for infected prey differed between parasite species. Both P. tereticollis- and P. laevis-infected gammarids were more vulnerable than uninfected ones, whereas the reverse was true of P. minutus-infected gammarids. These results suggest that the true consequences of phenotypic changes associated with parasitic infection in terms of increased trophic transmission of parasites deserve further assessment.

Increased susceptibility to predation and altered anti-predator behaviour in an acanthocephalan-infected amphipod.

According to the 'parasitic manipulation hypothesis', phenotypic changes induced by parasites in their intermediate hosts are effective means of increasing trophic transmission to final hosts. One obvious prediction, although seldom tested, is that increased vulnerability of infected prey to an appropriate predator should be achieved by the parasite altering the anti-predator behaviour of its intermediate host. In this study, we tested this prediction using the fish acanthocephalan Pomphorhynchus tereticollis and the freshwater amphipod Gammarus pulex. Firstly, we estimated the relative vulnerability of infected and uninfected gammarids to predation by the bullhead Cottus gobio in the field. Second, we investigated under experimental conditions how two common anti-predator behaviours of aquatic invertebrates, refuge use and short-distance reaction to predator chemical cues, were affected by infection status. We found that the prevalence of infection in the field was 10 times higher among gammarids collected from the stomach contents of bullheads compared with free-ranging individuals collected in the same river. In a microcosm uninfected gammarids, but not infected ones, increased the use of refuge in the presence of a bullhead. Finally, a behavioural experiment using an Y-maze olfactometer showed opposite reactions to predator odour. Whereas uninfected gammarids were significantly repulsed by the chemical cues originating from bullheads, infected ones were significantly attracted to the odour of the predator. Taken together, our results suggest that the alteration of anti-predator behaviour in infected G. pulex might enhance predation by bullheads in the field. Reversing anti-predator behaviour might thus be an efficient device by which parasites with complex life-cycles increase their trophic transmission to final hosts. Further studies should pay more attention to both the increased vulnerability of infected prey to an appropriate predator in the field and the influence of parasitic infection on the anti-predator behaviour of intermediate hosts.

Do distantly related parasites rely on the same proximate factors to alter the behaviour of their hosts?

Phylogenetically unrelated parasites often increase the chances of their transmission by inducing similar phenotypic changes in their hosts. However, it is not known whether these convergent strategies rely on the same biochemical precursors. In this paper, we explored such aspects by studying two gammarid species (Gammarus insensibilis and Gammarus pulex; Crustacea: Amphipoda: Gammaridae) serving as intermediate hosts in the life cycle of two distantly related parasites: the trematode, Microphallus papillorobustus and the acanthocephalan, Polymorphus minutus. Both these parasite species are known to manipulate the behaviour of their amphipod hosts, bringing them towards the water surface, where they are preferentially eaten by aquatic birds (definitive hosts). By studying and comparing the brains of infected G. insensibilis and G. pulex with proteomics tools, we have elucidated some of the proximate causes involved in the parasite-induced alterations of host behaviour for each system. Protein identifications suggest that altered physiological compartments in hosts can be similar (e.g. immunoneural connexions) or different (e.g. vision process), and hence specific to the host-parasite association considered. Moreover, proteins required to alter the same physiological compartment can be specific or conversely common in both systems, illustrating in the latter case a molecular convergence in the proximate mechanisms of manipulation.

Intergenomic interactions affect female reproduction: evidence from introgression and inbreeding depression in a haplodiploid mite.

Nuclear and cytoplasmic genomes can coevolve antagonistically or harmoniously to affect fitness. One commonly used test for nuclear-cytoplasmic coadaptation relies on the breakup of coadapted gene complexes by introgression, potentially resulting in an increased frequency of nuclear alleles in deleterious interaction with an alien cytoplasm. We investigated the phenotypic effect of such genes on female reproduction in outbred and inbred introgressed lines of the haplodiploid mite Tetranychus urticae. Introgression changed female lifetime fecundity and increased male production, in ways suggesting a control of fecundity by nuclear genes. Conversely introgression reduced the fertilization rate, possibly due to sperm-egg incompatibility or maternal effects. The intensity of inbreeding depression expressed as a reduction in fecundity was more severe in introgressed females than in nonintrogressed ones, giving evidence for recessive interacting alleles contributing to residual nucleo-cytoplasmic incompatibility. Overall, our data suggest recessive negative interactions between nuclear and cytoplasmic genes. This study is the first report of a contribution of nuclear polymorphism within a population to deleterious interactions with an alien cytoplasmic genome.

Genetic structure of a greenhouse population of the spider mite Tetranychus urticae: spatio-temporal analysis with microsatellite markers.

The genetic structure of a greenhouse population of the mite Tetranychus urticae was studied by the analysis of five microsatellite loci. Genetic variation was compared during a crop season between periods of population foundation and rapid population increase and was investigated in two consecutive years. The population displayed significant heterozygote deficiency at all the sampling periods. However, inbreeding tended to decrease with increasing density (FIS coefficient between 0.13 and 0.25). No significant genetic differentiation between samples was found either at a spatial scale within the greenhouse or at a temporal scale between two growing seasons (FST between 0.008 and 0.09). Estimations of the genetic relatedness between pairs of individuals indicated that the distances between pairs of sisters and unrelated mites in the greenhouse were not significantly different, suggesting that mites do not tend to form patches that reside close to the point of birth.

Meiotic and mitotic instability of two EMS-produced centric fragments in the haplodiploid wasp Nasonia vitripennis.

Terminal deletions that result in chromosomal fragments with centromeres (centric fragments) are relatively easy to generate and study in the haplodiploid insect Nasonia. We investigated the transmission stability of two chromosomal fragments generated by chemical mutagenesis. Visible mutations at the R locus (peach-233 and St-DR) and a linked body-colour mutant (purple) were used to track transmission of the centric fragments (which lack the purple locus and are wild-type at the R locus). Transmission rates in meiotic oogenesis were low (medians 0.15-0.18) and comparable to previous data on centric fragments in this species. The homologous chromosome genetic background strongly affected meiotic stability of one centric fragment (CF2) but not the other (CF1). Specifically, in peach/scarlet R locus heterozygous females, CF2 showed a normal segregation proportion with the chromosome bearing the scarlet allele (0.16), but near complete failure to segregate with peach (0.0002). Data show that this is due to loss of CF2 in eggs receiving peach, rather than to preferential segregation of CF2 with scarlet or mortality of CF2-bearing males. CF1 shows typical segregation ratios with both chromosomes. We hypothesize that deletions (or rearrangements) associated with the peach-233 mutant inhibit proper pairing and segregation of CF2. Consistent with the model, CF2 did segregate with chromosomes that had undergone recombination between peach and purple (a body-colour mutation 10 cM from peach), indicating that the domains inhibiting segregation are closely linked to peach. Mitotic instability also differed between the two fragments; reduced mitotic stability may relate to absence of telomeres on these centric fragments. Given the relative ease of generating and tracking terminal deletions in Nasonia, we propose this as a good system for studying mitotic and meiotic stability of centric fragments. Finally, results are discussed in relation to the evolution of B chromosomes from centric fragments.

Tracking paternal genes with DALP markers in a pseudoarrhenotokous reproductive system: biparental transmission but haplodiploid-like inheritance in the mite Neoseiulus californicus.

The complexity of some sexual reproductive systems in arthropods still leaves both their genetic and epigenetic determinism and their evolutionary significance poorly understood. Pseudoarrhenotoky is characterized by obligate fertilization and differential inactivation and/or elimination of paternal chromosomes in embryos that develop into males. Here, we investigate how the paternal genome is transmitted in a pseudoarrhenotokous mite, Neoseiulus californicus, using codominant genetic markers detected by DALP (direct amplification of length polymorphism). Transmission patterns of parental alleles through one and two generations are reported at four or five loci corresponding to four linkage groups. Our data provide strong evidence for selective elimination of the paternal genome among male tissues. Sperm contained maternal genes exclusively, whereas some male somatic tissues retained most if not all paternal chromosomes. No recombination between parental genomes prior to paternal genome elimination from the embryonic germ line was observed. These data allow a reinterpretation of previous phenotypic and cytogenetic observations in these mites, from which we suggest some relevant mechanistic and evolutionary implications. In addition, this is the first published study using polymorphic codominant loci detected by the recently developed DALP method.

Genetic differentiation in Tetranychus urticae (Acari: Tetranychidae): polymorphism, host races or sibling species?

Based on allozyme electrophoresis at the Pgm locus and nuclear ribosomal DNA (ITS2) sequences, we studied the genetic variation of the two-spotted spider mite Tetranychus urticae Koch collected on rose bay, Nerium oleander L. (Apocynaceae), from several localities around the Mediterranean basin. In addition, we compared these results with those of Navajas et al. (1998) and Tsagkarakou (1997) who collected from several other host plants from the Mediterranean. In the western part of this area (Spain, France, Tunisia), we found the individuals collected from rose bay to be clearly genetically differentiated from other samples. No evidence of such host-associated differentiation was detected in the eastern Mediterranean (Italy and Greece). The genetic differentiation of mites collected on rose bay was investigated further by studying the reproductive incompatibilities between populations in Greece and in France and a laboratory strain reared on bean, Phaseolus vulgaris, in France. Reciprocal crosses performed between these strains revealed variable levels of incompatibility, spanning from partial to complete reproductive isolation. In all cases incompatibility was asymmetric. We designed a test based on double-mating to establish the fertilization status of females in fully incompatible crosses. These crosses showed that the females had been inseminated, which suggests that the barrier to reproduction is not of a prezygotic behavioral nature. The data raises the question of the relative role of ecological factors (host plant) and geographical distance, in the ongoing differentiation process potentially leading to speciation.

Single and double infections with Wolbachia in the parasitic wasp Nasonia vitripennis: effects on compatibility.

Wolbachia are cytoplasmically inherited bacteria responsible for reproductive incompatibility in a wide range of insects. There has been little exploration, however, of within species Wolbachia polymorphisms and their effects on compatibility. Here we show that some strains of the parasitic wasp Nasonia vitripennis are infected with two distinct bacterial strains (A and B) whereas others are singly infected (A or B). Double and single infections are confirmed by both PCR amplification and Southern analysis of genomic DNA. Furthermore, it is shown that prolonged larval diapause (the overwintering stage of the wasp) of a double-infected strain can lead to stochastic loss of one or both bacterial strains. After diapause of a double-infected line, sublines were produced with AB, A only, B only or no Wolbachia. A and B sublines are bidirectionally incompatible, whereas males from AB lines are unidirectionally incompatible with females of A and B sublines. Results therefore show rapid development of bidirectional incompatibility within a species due to segregation of associated symbiotic bacteria.

Biparental inheritance of RAPD markers in males of the pseudo-arrhenotokous mite Typhlodromus pyri.

In pseudo-arrhenotokous mites, haploid males develop from fertilized eggs that undergo paternal genome loss (PGL) during early embryogenesis. We present evidence that some of the paternal genome may be retained in males of the predatory mite Typhlodromus pyri Scheuten (Acari: Phytoseiidae). Two reproductively compatible populations were differentiated by two random amplified polymorphic DNA markers and the inheritance pattern in the offspring was analysed. Maternal transmission rates are variable and independent of the sex of the offspring and of the marker. These data suggest a nuclear origin and independent segregation of the markers. One marker (330 base pairs (bp)) was paternally transmitted to male as well as female offspring, the other (990 bp) was paternally transmitted to all females and some of the male offspring. We propose that the paternal set of inactivated chromosomes may be partially retained in some tissues of the haploid males or, alternatively, that a B chromosome does not follow the process of PGL in male embryos, thereby segregating with the maternal set. The possible mechanisms controlling the condensation and the segregation of the chromosome(s) retained are discussed on the basis of current hypotheses on chromosome inactivation in insects.