Christophe Bressac

Heat stress affects male reproduction in a parasitoid wasp.

In insects, reproductive success and survival are affected by temperature. Reproduction is more sensitive than other physiological traits. While the effects of heat stress on females are well known, the effects on males are less clear. Hymenopteran parasitoids are valuable for studying the consequences of heat stress on male reproduction. In these species, through arrhenotoquous parthenogenesis, the sex ratio of the offspring is directly dependent on the sperm stock acquired by females during copulation. In the lab, heat temperature treatments (32-44°C) were applied for 3 days in the pupal stage of Anisopteromalus calandrae males, and development was completed at 30°C. Three different effects were observed depending on the temperature: mortality above 42°C, sterility of emerging males at 40°C, and sub-fertility at 38°C. This sub-fertility is characterized by a dramatic decrease in male sperm supplies, of up to 7% compared to control males. In the course of ageing, the sperm stock of sub-fertile males increases but never reaches the level of control males. Survival was significantly higher in control (30°C) males than those treated at 38°C. Male mating ability was similar whatever the treatment (control and 38°C), but females mated with 38°C-treated males stored 100 times less sperm on average than those mated with control males. The offspring sex ratio of females mated with 38°C-treated males was strongly male biased. The physiological mechanisms are as yet unknown. The relationship between temperature, sperm stock and sex ratio should be taken into account in the management of parasitoids for integrated pest management.

Qualitative aspects of sperm stock in males and females from Eupelmus orientalis and Dinarmus basalis (Hymenoptera: Chalcidoidea) as revealed by dual fluorescence

Abstract The quality of a sperm population can be characterized physiologically and its fecundity predicted by its viable : non‐viable sperm ratio. To improve the knowledge of reproductive strategies in two ectoparasitoid hymenopteran species, Eupelmus orientalis Crawford (Hymenoptera: Eupelmidae) and Dinarmus basalis Rondani (Hymenoptera: Pteromalidae), the assessment of sperm viability using the dual fluorescence staining procedure SYBR‐14 : propidium iodide was developed. The aim of the study was to provide a comparative test in vitro applicable to both sexes to study the evolution of sperm quality at various stages of the reproductive processes. The reliability of propidium iodide to detect non‐viable sperm (stained in red) was confirmed in both species on the basis of two stress tests (ethanol and Triton X‐100) but our study also revealed that propidium iodide concentrations must be adequately adjusted for each single species. This experiment also demonstrated the physiological heterogeneity of sperm populations in E. orientalis and D. basalis males and females. In both species, 40% of the sperm in the seminal vesicles was found to be non‐viable. By contrast with E. orientalis, the populations of non‐viable sperm estimated from the seminal vesicles of D. basalis were found to be strongly different from those observed in the spermatheca. From the present results, the population of viable sperm detected in the spermatheca of females from both species proved a reliable predictor of fertilization achieved in ovipositing females.

Microgynous queens in the Paleartic ant, Manica rubida: Dispersal morphs or social parasites?

Abstract In many ant species, queen size is dimorphic, with small microgynes and large macrogynes, which differ, for example, in size, insemination rate, ovary development, and dispersal tactics. These polymorphic queens often correspond with alternative reproductive strategies. The Palearctic ant, Manica rubida (Latreille) (Hymenoptera: Formicidae), lives mostly in mountainous regions in either monogynous colonies, containing one macrogynous queen or polygynous colonies, containing a few large macrogynous queens. In 1998, a colony of M. rubida was discovered containing macrogynes and many small alate microgynes that did not engage in a nuptial flight but, instead, stayed in the home nest the following winter. These microgynes were studied more closely by investigating their size, behavior, and spermatheca in relation to M. rubida macrogynes and workers. Mitochondrial DNA of macrogynes, microgynes and workers from four nests was sequenced to detect possible genetic differences between them. The microgynes were significantly smaller than the macrogynes, and the head width of the gynes was completely bimodal. The microgynes behaved like workers of the macrogynes in every experiment tested. Furthermore, the microgynes had a normal spermatheca and could be fecundated, but rarely (only one in several years). Finally, all the individuals were genetically identical, except three workers that differed by only one codon position. Because these microgynes have features of both queens and workers, their functional significance in the colony is not yet clear.

A combined approach to heat stress effect on male fertility in Nasonia vitripennis: from the physiological consequences on spermatogenesis to the reproductive adjustment of females mated with stressed males.

In recent years, several studies have shown a decline in reproductive success in males in both humans and wildlife. Research on male fertility has largely focused on vertebrates, although invertebrates constitute the vast majority of terrestrial biodiversity. The reduction of their reproductive capacities due to environmental stresses can have strong negative ecological impacts, and also dramatic consequences on world food production if it affects the reproductive success of biological control agents, such as parasitic wasps used to control crop pests. Here Nasonia vitripennis, a parasitic wasp of various fly species, was studied to test the effects of 24h-heat stress applied during the first pupal stage on male fertility. Results showed that only primary spermatocytes were present at the first pupal stage in all cysts of the testes. Heat stress caused a delay in spermatogenesis during development and a significant decrease in sperm stock at emergence. Females mated with these heat-stressed males showed a reduce sperm count stored in their spermatheca. Females did not appear to distinguish heat-stressed from control males and did not remate more frequently to compensate for the lack of sperm transferred. As a result, females mated with heat-stressed males produced a suboptimal lifetime offspring sex ratio compared to those mated with control males. This could further impact the population dynamics of this species. N. vitripennis appears to be an interesting biological model to study the mechanisms of subfertility and its consequence on female reproductive strategies and provides new research perspectives in both invertebrates and vertebrates.

Delayed sexual maturity in males of Vespa velutina

Vespa velutina var nigrithorax (Lepelletier, 1835) is an invasive predator of bees accidentally introduced in France in 2004, and it is having a serious impact on apiculture and ecosystems. Studying the reproduction of an invasive species is key to assess its population dynamic. This study explores the sexual maturation of V. velutina males and the evolution of their fertility. The main studied parameters were physiologic (spermiogenesis, spermatogenesis) and anatomic (testes size and structure, head width). Two populations of males were described based on their emergence period: early males in early summer or classic males in autumn. Each testis has an average of 108 testicular follicles. Spermatogenesis is synchronous, with only 1 sperm production wave, and completed, on average, at 10.3 d after emergence with the degeneration of the testes. The sperm counts in seminal vesicles of mature males are 3 × 106 in October/November and 0.8 × 106 in June. In comparison, females store 0.1 × 106 sperm in their spermathecae. The early males emerged from colonies made by fertilized queens. The reproductive potential of these early males seemed limited, and their function in the colony is discussed. The sperm stock evolution in autumn males suggests the occurrence of a reproductive pattern of male competition for the access to females and a single copulation per male. The synchronicity of male and foundress emergences and sexual maturation is of primary importance for the mating success and the future colony development.

The spatio-temporal partitioning of sperm by males of the prospermatogenic parasitoid Nasonia vitripennis is in line with its gregarious lifestyle

Male fitness depends on the number of lifetime progeny of their mates and could be constrained by the chance of finding a mate, lifespan and temporal patterns of sperm production and allocation. Here, we used the parasitic wasp Nasonia vitripennis with a two-week lifespan and a gregarious lifestyle, to analyze how the reproductive system is organized to allocate spermatozoa over consecutive matings. Results show that spermatogenesis is synchronized and completed one day before emergence so that males emerge with a full sperm complement. We also found a regulation of spermatozoa transfer between testis and seminal vesicles that allows males to partition small ejaculates over multiple matings. Overall, this study shows that for N. vitripennis, male fertilization potential is determined (1) at the pupal stage, when spermatogenesis takes place to generate a complete life-long stock, (2) on emergence, when transport of spermatozoa from testes to seminal vesicles is initiated and (3) in adulthood, during which spermatozoa are partitioned over successive copulations. Such life history-traits are consistent with the gregarious lifestyle of N. vitripennis.

Comparison of reproductive traits of foundresses in a native and an invasive hornet in Europe

The population dynamics of annual social hymenoptera such as vespids depend largely on the fertility of the foundresses, which, in turn, is a key factor in the context of biological invasions. The native European hornet Vespa crabro (Vc) and the invasive Asian hornet Vespa velutina (Vv) have generally similar ecological traits, e.g. nesting and feeding habits, although they differ in colony size, which is higher in Vv. Furthermore, in contrast to Vc, Vv is more specialized in its predatory habits, intensively hunting honey bees at the hive. Comparing the morphological and reproductive traits of two closely related species occupying the same ecological niche, one of which is a native species and the other an alien, can help us to gain an understanding of the invasion process. To this end, we here compare reproductive (ovarian size and maturation, fat level, spermatheca size and sperm stock, fecundity) and morphological traits (head size, weight) of the foundresses of these two hornet species. We observed that ovarian maturation began approximately one month earlier in Vv than in Vc, and that the fat level in the former was lower. We found twice the number of sperm in the mated foundresses of Vv than in those of Vc (more than 100 × 103 and less than 50 × 103 sperm, respectively), in a 16% smaller spermatheca in Vc. Furthermore, the sperm of Vv was found to be 65% shorter than that of Vc. The precocity and higher potential fecundity of Vv queens may have favoured this species over Vc in terms of predatory behaviour, and thereby contributed to its invasiveness.

Sperm limitation affects sex allocation in a parasitoid wasp Nasonia vitripennis : Sex allocation in sperm-limited wasps

Insect reproduction is influenced by various external factors including temperature, a well‐studied constraint. We investigated to what extent different levels of sperm limitation of males exposed to different heat stresses (34 and 36 °C) affect females’ offspring production and sex allocation in Nasonia vitripennis. In this haplodiploid parasitoid wasp attacking different species of pest flies, we investigated the effect of the quantity of sperm females received and stored in their spermatheca on their sperm use decisions, hence sex allocation, over successive ovipositions. In particular, we compared the sex allocation of females presenting three levels of sperm limitation (i.e., mated with control, 34 °C heat‐stressed or 36 °C heat‐stressed males) on each host they parasitized. To disentangle the potential reduction of sperm quality after a heat stress exposure from that of sperm quantity, we also explored the clutch size and sex ratio produced by females that were partially sperm limited after copulating with multiply mated males. Independently of their sperm numbers, all types of females produced a similar total number of offspring, but the more limited ones had fewer daughters. Sperm limitation further affected the distribution of daughters’ production across time. In addition to constraints acting on female physiology, male fertility should therefore be considered in studies measuring reproductive outputs of insects submitted to heat stresses.

Centrioles without microtubules - a new morphological type of centriole

ABSTRACT The centrosome is the organizing center of microtubules in the cell, the basis for the origin of cilia and flagella and a site for the concentration of a regulatory proteins multitude. The centrosome comprises two centrioles surrounded by pericentriolar material. Centrioles in the cells of different organisms can contain nine triplets, doublets or singlets of microtubules. Here, we show that in somatic cells of male wasp larvae Anisopteromalus calandrae, centrioles do not contain microtubules and are composed of nine electron-dense prongs, which together form a cogwheel structure. These microtubule-free centrioles can be the platform for procentriole formation and form microtubule-free cilia-like structures. In nymph and imago cells centrioles have a microtubule triplet structure. Our study describes how centriole structure differs in a development-stage-dependent and a cell-type-dependent manner. The discovery of a centriole without microtubules casts a new light on the centriole formation process and the evolution of this organelle. Summary: Centrioles without microtubules in larvae cells of parasitoid wasps duplicate and form primary cilia without microtubules, in nymph and imago cells centrioles acquire a ‘normal’ structure with nine microtubules triplets.