Ecological and genetic interactions in Drosophila-parasitoids communities: a case study with D. melanogaster, D. simulans and their common Leptopilina parasitoids in south-eastern France.

Drosophila species are attacked by a number of parasitoid wasps, which constitute an important factor of population regulation. Since Drosophila melanogaster and Drosophila simulans share common parasitoid species, their ecology and evolution can hardly be understood without considering parasitoids. After a short review of data available on Drosophila-parasitoid interactions involving D. melanogaster and D. simulans as hosts, we report field and laboratory experiments investigating the ecological role of Leptopilina parasitoids in Drosophila communities of southern France. Seasonal survey of species abundance shows that strong interspecific interactions occur at both tropic levels. D. simulans progressively replaces D. melanogaster in southern areas suggesting competitive displacement. Parasitoids are responsible for very high Drosophila mortality (up to 90% in some fruits). Field data emphasize the importance of selective pressure that parasitoids exert on Drosophila communities. The two Leptopilina parasites (L. heterotoma and L boulardi) have different local abundances, which vary in time, and they also compete for hosts. We show that parasitoids can mediate the coexistence of D. melanogaster and D. simulans in the laboratory, and thus may contribute to their puzzling coexistence in the field. Conversely, hosts exert selective pressures on parasitoids, and development on either D. melanogaster or D. simulans strongly affects fitness of adult wasps in a temperature-dependent fashion. Local variation in host species abundance and diversity could thus account for the genetic differentiation we observed in one parasitoid species. Despite laboratory studies cannot fully explain complex field situations, it is clear that the ecology and evolution of Drosophila populations and communities, especially D. melanogaster and D. simulans, are strongly constrained by parasitoids, which should receive more attention.

Infection polymorphism and cytoplasmic incompatibility in Hymenoptera-Wolbachia associations.

Most cases of Wolbachia infection so far documented in haplodiploid Hymenoptera are associated with parthenogenesis induction. Only three examples of Wolbachia-mediated cytoplasmic incompatibility (CI) have been reported, resulting either in haploidisation of fertilised eggs, which develop into viable males, or in their death. To better document this variability, we studied two new Wolbachia-wasp associations involving Drosophila parasitoids. In Trichopria cf. drosophilae, individuals are infected by two different Wolbachia variants, populations are nearly totally infected, and Wolbachia induces incomplete CI resulting in death of the fertilised eggs. On the other hand, Pachycrepoideus dubius harbours only one bacterial variant, populations are polymorphic for infection, and Wolbachia has no detectable effect. These two cases show that the range of variation in Wolbachia's effects in Hymenoptera is as wide as in diploids, extending from complete CI to an undetectable effect. Cases so far studied show some parallel between the strength of incompatibility, the number of Wolbachia variants infecting each wasp, and the natural infection frequency. These empirical data support theoretical models predicting evolution of CI towards lower levels, resulting in the decline and ultimate loss of infection, and place multiple infections as being an important factor in the evolution of host-Wolbachia associations.

Insertion polymorphism of retrotransposable elements in populations of the insular, endemic species Drosophila madeirensis.

The insertion site numbers of the retrotransposable elements (TE) 412, gypsy and bilbo were determined in individuals of five distinct natural populations of the endemic species Drosophila madeirensis from the island of Madeira. The TE distributions were compared to those of the paleartic, widespread and phylogenetically closely related species, D. subobscura. In situ hybridization and Southern blots showed that in D. madeirensis the number of insertion sites ranged between 10 and 15, three and six, and 35 and 42 for elements 412, gypsy and bilbo, respectively. The corresponding values for D. subobscura were similar. Two of these elements, 412 and gypsy, had very few insertions in the heterochromatin, unlike bilbo, which displayed a high heterochromatic insertion number. The Southern band polymorphism was very high, leading to within-population variation of 97.2%, whatever the population and the TE concerned. Using the polymorphic TE insertion sites as markers to analyse population structure by AMOVA, adapted for RAPD (Randomly Amplified Polymorphic DNA) data, we found small but significant genetic differences between the populations on Madeira. This slight differentiation, coupled with similar copy numbers for each TE between populations, suggests that the D. madeirensis species consists of a single, only slightly subdivided population. These data also show that insular populations and endemic species of Drosophila can have as many copies of TEs as more widespread species.

Within-species diversity of Wolbachia-induced cytoplasmic incompatibility in haplodiploid insects.

Wolbachia-induced cytoplasmic incompatibility (CI) can have two consequences in haplodiploid insects: fertilized eggs either die (female mortality, FM) or they develop into haploid males (male development, MD). Origin of this diversity remains poorly understood, but current hypotheses invoke variation in damage suffered by paternal chromosomes in incompatible eggs, thus intermediate CI types should be expected. Here, we show the existence of such a particular CI type. In the parasitoid wasp Leptopilina heterotoma, we compared CI effects in crosses involving lines derived from a single inbred line with various Wolbachia infection statuses (natural tri-infection, mono-infection, or no infection). Tri-infected males induce a FM CI type when crossed with either uninfected or mono-infected females. Crossing mono-infected males with uninfected females results in almost complete CI with both reduced offspring production, indicating partial mortality of fertilized eggs, and increased number of sons, showing haploid male development of others. Mono-infected males thus induce an intermediate Cl type when mated with uninfected females. The first evidence of this expected particular CI type demonstrates that no discontinuity separates MD and FM CI types, which appear to be end points of a phenotypic continuum. Second, different CI types can occur within a given species and even within offspring of a single pair. Third, phenotypic expression of the particular CI type induced by a given Wolbachia variant depends on other bacterial variants that co-infect the same tissues. These results support the idea that haplodiploids should be helpful in clarifying evolutionary pathways of insect-Wolbachia associations.

Modification, by the insecticide chlorpyrifos, of the behavioral response to kairomones of a parasitoid wasp, Leptopilina boulardi.

Parasitoids are key species involved in the regulation of natural populations. Host parasitization is realized via some important steps in which kairomone perception is essential. Due to the wide use of insecticides and their contribution to environmental pollution, the determination of their sublethal effects on behaviors involved in the reproduction of parasitoids becomes a necessity. In this work, we analyzed the effects of a lethal dose 20% of chlorpyrifos, an organophosphorus insecticide, on the behavior of L. boulardi toward the kairomone of its host. The insecticide significantly modified the kinetics of the residence time of parasitoids on a patch marked by kairomones. Females exposed to the insecticide were less efficient in finding the kairomone patch than control females, and they tended to stay on that patch, whereas control females rapidly left it. These effects are discussed considering the mode of action of the insecticide. Because search time allocation of host is an important component for parasitism efficiency, this modification of behavior could interfere with the reproduction capacity of parasitoids. Therefore, in a context of environmental pollution, the repercussions of such sublethal effects on the population biology of insects need to be considered.

Partial compensation of the sublethal effect of deltamethrin on the sex pheromonal communication of Trichogramma brassicae.

Pyrethroid insecticides are widely used and lead to a sizable environmental pollution that could interfere with the population biology of insects. Trichogramma is a beneficial insect used in biological control and which natural populations contribute to the control of Lepidopterus pests. In this work, we determined the effect of a sublethal dose of deltamethrin on the sex pheromonal communication of Trichogramma. The dose used (LD 0.1) induces no detectable mortality (the theoretical mortality is only one insect over 1000) and can be a good representation of contamination by this insecticide from environmental pollution. The insecticide was shown to have opposite effects on the sex pheromonal communication of Trichogramma, depending on which sex was exposed (Delpuech, J.M., Legallet, B., Terrier, O., Fouillet, P., 1999. Chemosphere 38, 729-739). We show that, when both sexes are simultaneously exposed to the insecticide, this effect is only partially neutralized. The mean response of treated males responding to the sex pheromone from treated females is not significantly different from that of controls, but the kinetics of their response is not the same. When both sexes are treated, the response of males to the sex pheromone is lower at the beginning but their response does not decrease during time contrary to controls and becomes finally higher than that of controls. Therefore, the sublethal effect of deltamethrin in the field can be either advantageous or disadvantageous depending on the difficulty in finding females and their scarcity.

Physiological cost induced by the maternally-transmitted endosymbiont Wolbachia in the Drosophila parasitoid Leptopilina heterotoma.

Endosymbiotic bacteria of the genus Wolbachia infect a number of invertebrate species in which they induce various alterations in host reproduction, mainly cytoplasmic incompatibility (CI). In contrast to most other maternally transmitted parasites, manipulation of host reproduction makes the spread of Wolbachia possible even if they induce a physiological cost on their hosts. Current studies have shown that fitness consequences of Wolbachia infection could range from positive (mutualist) to negative (parasitic) but, in most cases, Wolbachia do not have strong deleterious effects on host fitness and the status of association remains unclear. Here, we show that in the Drosophila parasitoid wasp Leptopilina heterotoma, Wolbachia infection has a negative impact on several host fitness traits of both sexes. Fecundity, adult survival and locomotor performance are significantly reduced, whereas circadian rhythm, development time and offspring sex-ratio are not affected. Although the cost of bacterial infection can be overcome by effects on host reproduction i.e. cytoplasmic incompatibility, it could influence the spread of the bacterium at the early stages of the invasion process. Clearly, results underline the wide spectrum of phenotypic effects of Wolbachia infection and, to our knowledge, Wolbachia infection of L. heterotoma appears to be one of the most virulent that has ever been observed in insects.

Evidence for female mortality in Wolbachia-mediated cytoplasmic incompatibility in haplodiploid insects: epidemiologic and evolutionary consequences.

Until now, only two Wolbachia-mediated cytoplasmic incompatibility (CI) types have been described in haplodiploid species, the first in Nasonia (Insect) and the second in Tetranychus (Acari). They both induce a male-biased sex ratio in the incompatible cross. In Nasonia, CI does not reduce fertility since incompatible eggs develop as haploid males, whereas in Tetranychus CI leads to a partial mortality of incompatible eggs, thus reducing the fertility of females. Here, we study Wolbachia infection in a Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae). A survey of Wolbachia infection shows that all natural populations tested are totally infected. Crosses between infected males and cured females show complete incompatibility: almost no females are produced. Moreover, incompatible eggs die early during their development, unlike Nasonia. This early death allows the parasitized Drosophila larva to achieve its development and to emerge. Thus, uninfected females crossed with infected males have reduced offspring production consisting only of males. Evidence of this CI type in insects demonstrates that the difference in CI types of Nasonia and Tetranychus is not due to specific factors of insects or acari. Using theoretical models, we compare the invasion processes of different strategies of Wolbachia: CI in diploid species, the two CI types in haplodiploid species, and parthenogenesis (the classical effect in haplodiploid species). Models show that CI in haplodiploid species is less efficient than in diploid ones. However, the Leptopilina type is advantageous compared to the Nasonia type. Parthenogenesis may be more or less advantageous, depending on the infection cost and on the proportion of fertilized eggs. Finally, we can propose different processes of Wolbachia strategy evolution in haplodiploid species from Nasonia CI type to Leptopilina CI type or parthenogenesis.

Adaptive significance of a circadian clock: temporal segregation of activities reduces intrinsic competitive inferiority in Drosophila parasitoids.

Most organisms show self-sustained circadian oscillations or biological clocks which control their daily fluctuations in behavioural and physiological activities. While extensive progress has been made in understanding the molecular mechanisms of biological clocks, there have been few clear demonstrations of the fitness value of endogenous rhythms. This study investigated the adaptive significance of circadian rhythms in a Drosophila parasitoid community. The activity rhythms of three sympatric Drosophila parasitoids are out of phase, the competitively inferior parasitoid species being active earlier than the superior competitor. This temporal segregation appears at least partially determined by endogenous periods of the clock which also vary between species and which correlate the time of activity. This earlier activity of the inferior competitor significantly reduces its intrinsic competitive disadvantage when multiparasitism occurs, thus suggesting that natural selection acting on the phase of the rhythm could substantially deviate the endogenous period from the optimal ca. 24 h period. This study demonstrates that temporal segregation of competing species could be endogenously controlled, which undoubtedly favours their coexistence in nature and also shows how natural selection can act on biological clocks to shape daily activity patterns.

Phylogenetic evidence for horizontal transmission of Wolbachia in host-parasitoid associations.

Endosymbiotic Wolbachia infect a number of arthropod species in which they can affect the reproductive system. While maternally transmitted, unlike mitochondria their molecular phylogeny does not parallel that of their hosts. This strongly suggests horizontal transmission among species, the mechanisms of which remain unknown. Such transfers require intimate between-species relationships, and thus host-parasite associations are outstandingly appropriate for study. Here, we demonstrate that hymenopteran parasitoids of frugivorous Drosophila species are especially susceptible to Wolbachia infection. Of the five common European species, four proved to be infected; furthermore, multiple infections are common, with one species being doubly infected and two triply infected (first report). Phylogenetic statuses of the Wolbachia infecting the different species of the community have been studied using the gene wsp, a highly variable gene recently described. This study reveals exciting similarities between the Wolbachia variants found in parasitoids and their hosts. These arguments strongly support the hypothesis of frequent natural Wolbachia transfers into other species and open a new field for genetic exchanges among species, especially in host-parasitoid associations.

Modifications of the sex pheromonal communication of trichogramma brassicae by a sublethal dose of deltamethrin

Pyrethroid insecticides, by their intensive use and their persistence that can exceed 3 months, largely contribute to the environmental pollution. In this work, we determined the effects of a very low dose of deltamethrin on the sex pheromonal communication of Trichogramma brassicae. The dose used was a dose that would theoretically kill one insect over 1000 (an LD 0.1). We found that this dose slightly but very significantly increased the arrestment behavior of treated males responding to the female pheromone. On the other hand when females were exposed to the same dose of insecticide, the response of males to their pheromone was very significantly decreased. In Trichogramma, like in other insects, sex pheromonal communication probably involves nervous transmissions both for the reception and the emission of the pheromone. Then, the sublethal effects of deltamethrin, observed in this work, are certainly due to multiple actions of this insecticide on nervous transmissions. Trichogramma is a beneficial insect that contributes to the control of pest populations of moths. Actions of this insecticide at a dose that can correspond to environmental pollution could be a real threat to the equilibrium of these populations.

A chromosomal analysis of the phenotypic plasticity of some life history traits in relation to developmental temperature.

The capacity to delay egg deposition in D. melanogaster females in the absence of a sexual partner is genetically determined and opposite types can be artificially selected. In natural populations, the relative frequency of these genotypes varies geographically and seasonally, with temperature as a selective factor. However, the retention duration of these genotypes can be modified by developmental temperature change. To study the genetic control of this response, chromosome substitution between opposite types of line was carried out in order to produce every possible homozygous chromosomal combination of the three major chromosomes (X,2,3). Eggs of these eight constructed lines were developed at two different temperatures (25 degrees C and 14 degrees C). Low temperature development directly affected the number of ovarioles but also modified the subsequent expression of adult characteristics such as retention duration and fecundity. The comparison of the eight lines revealed that, although the 3 chromosomes were involved in the genetic determinism of each trait, only one or two of them were sensitive to temperature change, and these differed according to the trait. For retention duration and fecundity, the effect of chromosome 3 from the long retention strain was particularly affected by low temperature, showing antagonism between the selective effect detected in natural populations and the effect on phenotypic plasticity studied here.

Male orientation to trail sex pheromones in parasitoid wasps: does the spatial distribution of virgin females matter?

We studied male locomotory response to trails and patches of sex pheromone (left respectively by free-ranging females and females constrained to stay on a small area) in the two parasitoids Aphelinus asychis (Hymenoptera: Aphelinidae) and Trichogramma brassicae (Hymenoptera: Trichogrammatidae). Under the hypothesis that the spatial distribution of virgin females differs between these species (scattered among host plants in A. asychis, gregarious at emergence sites in T. brassicae), we predicted that male locomotory response to their sex pheromones should also differ: A. asychis males should follow pheromone trails on plants in order to encounter the females along these trails, whereas T. brassicae males should stay on pheromone patches, at emergence sites, and mate the females on these patches. Using an improved video-tracking system, we found that males of both species respond to conspecific sex pheromone trails and patches, but that the response does not differ much between species. Males released on marked substrates walked in a more convoluted pattern (i.e. higher path fractal dimension and higher number of crossings within tracks) than males released on unmarked substrates. On pheromone patches, males turned persistently in the same direction when leaving the patch, which explains a higher number of visits on marked patches than on unmarked patches, and possibly, higher track convolution on pheromone trails. Contrary to our hypothesis, male A. asychis did not follow female trails more accurately than male T. brassicae, and male T. brassicae did not stay longer on pheromone patches than male A. asychis. We argue that these discrepancies between our predictions and the observed responses originates from discrepancies between the assumed spatial distribution of virgin females and their actual distribution in the wild.

Genetic variation in locomotor activity rhythm among populations of Leptopilina heterotoma (Hymenoptera: Eucoilidae), a larval parasitoid of Drosophila species.

The locomotor activity rhythm of Leptopilina heterotoma, a parasitoid insect of Drosophila larvae, was investigated under laboratory conditions. Under LD 12:12, the locomotor activity of females shows a clear rhythm which persists under continuous darkness (circadian rhythm). However, comparative study of five populations indicates that both the rate of activity and the profile of the rhythm vary according to the origin of females. The Mediterranean populations (Tunisia and Antibes) show two peaks of activity, at the beginning and at the end of the photophase, whereas more northern populations (Lyon and the Netherlands) are mostly active during the afternoon. Females originating from the area of Lyon have a very low level of activity. Reciprocal crosses (F1 hybrids and backcrosses) between the French and the Tunisian strains demonstrated the genetic basis of these variations and the biparental inheritance of the trait. This genetic variability is interpreted as a consequence of selective pressures and suggests a local adaptation of natural populations in host foraging behavior. The selective factors which could act on the daily organization of parasitoid behaviors are discussed.

[Terminal constipation: priority for anorectal manometry]. / Constipation terminale: priorité à la manométrie ano-rectale.

Terminal constipation is a common ailment which may be readily suspected by appropriate questioning of the patient. Investigation is by function tests such as anorectal manometry and measurement of colonic transit time. The goal of this investigation was to explore the relative merits of these two methods in terminal constipation. Measurement of colonic transit time did not make it possible to distinguish terminal from other forms of constipation, whereas anorectal manometry allowed this distinction and, in cases of terminal constipation, provided information on the physiopathological mechanisms, thereby permitting appropriate anorectal reeducation by biofeedback. We conclude that, once an organic cause has been excluded by coloscopy, anorectal manometry is the investigation of choice in terminal constipation.

[Constipation and incontinence: significance of colonic transit time, anorectal manometry and defecography]. / Constipation et incontinence: intérêt du temps de transit colique, de la manométrie ano-rectale et de la défécographie.

Constipation and fecal incontinence are frequent motives of gastroenterological consultation. An etiological diagnosis can often be suspected from the history and can be confirmed by functional testing. We here report our experience with the measurement of colonic transit time (TTC), anorectal manometry (MAR) and defecography (D). Whilst TTC was unhelpful, MAR revealed abdomino-pelvic asynchrony (anismus) in 60 constipated patients and 7 (47%) of 15 incontinent patients. Perineal descent was suspected in 25 constipated patients and confirmed by defecography, which also revealed associated static pelvic disorders. Our experience confirms the role of functional exploration in the investigation of constipation and fecal incontinence and permits a more precise therapeutic approach.

Chromosomal analysis of initial retention capacity in virgin Drosophila melanogaster females.

In natural temperate populations, virgin Drosophila melanogaster females present highly variable periods of preoviposition duration (from 2 to 25 days). Strains were selected for long and short initial retention capacity. Chromosome substitution between two of these lines produced, by appropriate mating procedures, every possible homozygous chromosomal combinations of the X, II and III chromosomes. Analyses of these lines demonstrate that both X and III chromosomes are involved in this egg-laying control, and have complementary effects.

[Genetic variability of the circadian activity pattern of Drosophila in a natural population]. / Variabilité génétique du profil circadien d'activité de la Drosophile dans une population naturelle.

With an actographic recorder (IR emitter), the genetic variability in the circadian rhythm of locomoter activity has been analysed within a natural population of D. melanogaster. The activity pattern shows differences between sexes and environmental conditions (photoperiod LD or continuous darkness). In free running conditions, the period and the amplitude of the activity peaks differ between isofemale lines.

Biliary lithiasis: combined endoscopic and surgical treatment.

A combined endoscopic and surgical procedure was performed in 69 patients suffering from gallbladder lithiasis with suspected associated lithiasis of the common bile duct (CBD). The procedure was spread over two days: Day 1 - Endoscopic retrograde cholangiopancreatography (ERCP) revealed choledocholithiasis in 50 cases and papillary sclerosis in 4 cases. In 15 patients, the common bile duct was found to be normal. Fifty-four endoscopic sphincterotomies were performed. Among the 50 cases of choledocholithiasis, endoscopic removal of the stones was judged complete in 44 cases and incomplete in the other 6. Day 2 - Cholecystectomy was carried out in all the patients. Peroperative cholangiography confirmed the results of the pre-operative ERCP with respect to the CBD in all the patients except one, and a choledochotomy was necessary in 6 patients. Results were as follows: The average length of hospitalization was 12.4 days. No residual choledocholithiasis was noted. The morbidity of the procedure was 7.2%, and the mortality 1.4% unrelated to the technique. The diagnostic and therapeutic significance of the procedure is discussed.