Female accessory gland fluid promotes sperm survival in yellow dung flies

Female and male reproductive traits co-evolve through pre- and post-copulatory sexual selection and sexual conflict. Although males typically transfer many sperm during copulation, only a small proportion reach the fertilization site because females often actively or passively reduce sperm number in their reproductive tract. Males may transfer accessory substances to protect their ejaculates against female selective processes, which benefits males but can harm females. In turn, females may use accessory gland fluids to control paternity or sperm storage. Female yellow dung flies (Scathophaga stercoraria) have paired accessory glands that produce fluids involved in fertilization and egg laying. One proposed function for these fluids is spermicide. Alternatively, female accessory gland fluid may help keep sperm alive to avoid fertilization failure or encourage sperm competition. Using yellow dung flies, we investigated the interaction of female accessory gland fluid with sperm in vitro. Significantly more sperm remained alive when exposed to accessory gland fluid compared to buffer only (63% vs. 44%). We conclude that female accessory gland fluid in yellow dung flies can help nourish rather than kill male sperm, although selective nourishment of sperm is as consistent with cryptic female choice as is selective spermicide.

Energetic underpinnings of yellow dung fly mating success in the field

Foraging provides the basis for animal reproduction, but requires energy and time to be sustained, entailing a trade-off. Whereas females should maximize their time foraging for resources, males should minimize their foraging time by optimizing time budgets to maximize their access to mating partners. Mark-resight field studies are difficult and hence uncommon for small insects. Yellow dung flies (Scathophaga stercoraria L.) abound on pastures in cold-temperate regions across the northern hemisphere. Adult flies lick nectar from flowers for energy, but require small insect prey to produce eggs and sperm. Males wait for females around fresh cow dung, but at one point also need to replenish their energy and/or sperm reserves in the surrounding vegetation. Their foraging time budgets should depend on their body size, nutritional energy reserves, availability of sperm, competitor and female density. Marked male dung flies whose nutritional status was experimentally manipulated – water only (null control); water + sugar (energy replenishment); or water, sugar + Drosophila prey (energy and sperm replenishment) – were repeatedly observed on an experimental pasture for an entire day. Both nutrient types were expected to increase the mating success of especially large males. The total number of resighted males seen copulating was lowest for water-treated flies. Mating success was positively related to body size. The distance travelled between dung pats was greater for males fed sugar or prey and also increased with body size, while pat residence times decreased with size. No differences were found between the sugar- and prey-fed groups. Crucially however, there was no evidence in the field for a time budget or mating advantage of small males when nutrients were limited.

A fungal parasite selects against body size but not fluctuating asymmetry in Swiss subalpine yellow dung flies

Evidence for selective disadvantages of large body size remains scarce in general. Previous studies of the yellow dung fly Scathophaga stercoraria have demonstrated strong positive sexual and fecundity selection on male and female size. Nevertheless, the body size of flies from a Swiss study population has declined by ~10% 1993–2009. Given substantial heritability of body size, this negative evolutionary response of an evidently positively selected trait suggests important selective factors being missed. An episodic epidemic outbreak of the fungus Entomophthora scatophagae permitted assessment of natural selection exerted by this fatal parasite. Fungal infection varied over the season from ~50% in the cooler and more humid spring and autumn to almost 0% in summer. The probability of dying from fungal infection increased with adult fly body size. Females never laid any eggs after infection, so there was no fungus effect on female fecundity beyond its impact on mortality. Large males showed their typical mating advantage in the field, but this positive sexual selection was nullified by fungal infection. Mean fluctuating asymmetry of paired appendages (legs, wings) did not affect the viability, fecundity or mating success of yellow dung flies in the field. This study documents rare parasite-mediated disadvantages of large-sized flies in the field. Reduced ability to combat parasites such as Entomophthora may be an immunity cost of large body size in dung flies, although the hypothesized trade-off between fluctuating asymmetry, a presumed indicator of developmental instability and environmental stress, and immunocompetence was not found here.

How biases in sperm storage relate to sperm use during oviposition in female yellow dung flies

Precise mechanisms underlying sperm storage and utilization are largely unknown, and data directly linking stored sperm to paternity remain scarce. We used competitive microsatellite PCR to study the effects of female morphology, copula duration and oviposition on the proportion of stored sperm provided by the second of two copulating males (S2) in Scathophaga stercoraria (Diptera: Scathophagidae), the classic model for sperm competition studies. We genotyped all offspring from potentially mixed-paternity clutches to establish the relationship between a second male’s stored sperm (S2) and paternity success (P2). We found consistent skew in sperm storage across the three female spermathecae, with relatively more second-male sperm stored in the singlet spermatheca than in the doublet spermathecae. S2 generally decreased with increasing spermathecal size, consistent with either heightened first-male storage in larger spermathecae, or less efficient sperm displacement in them. Additionally, copula duration and several two-way interactions influenced S2, highlighting the complexity of postcopulatory processes and sperm storage. Importantly, S2 and P2 were strongly correlated. Manipulation of the timing of oviposition strongly influenced observed sperm-storage patterns, with higher S2 when females laid no eggs before being sacrificed than when they oviposited between copulations, an observation consistent with adaptive plasticity in insemination. Our results identified multiple factors influencing sperm storage, nevertheless suggesting that the proportion of stored sperm is strongly linked to paternity (i.e., a fair raffle). Even more detailed data in this vein are needed to evaluate the general importance of sperm competition relative to cryptic female choice in postcopulatory sexual selection.

The position of the Azeliinae in the Muscidae (Diptera) based on musculature of the male terminalia

The male genital and pregenital skeleton and musculature were studied in males of the following species of the Muscidae subfamily Azeliinae: Drymeia firthiana (Huckett, 1965), Drymeia longiseta Sorokina & Pont, 2015, Drymeia segnis (Holmgren, 1883), Thricops nigritellus (Zetterstedt, 1838), Thricops hirtulus (Zetterstedt, 1838), Hydrotaea dentipes (Fabricius, 1805), Muscina stabulans (Fallén, 1817), and Muscina levida (Harris, 1780). Descriptions and figures of the genital sclerites and muscles of D. firthiana and M. stabulans are given. A comparison was made between the genital segments and muscles of previously studied species of Mydaeinae and Muscinae and those of the Azeliinae. Based on the structure of the skeleton and muscles of syntergosternite VII + VIII and the phallapodeme muscles, significant differences were found between the subfamily Azeliinae and the subfamilies Mydaeinae and Muscinae. The basal position of the Azeliinae within the family Muscidae was confirmed. A comparison of the genital segments and muscles of the Muscidae with those of the Scathophagidae (Scathophaga stercoraria (Linnaeus, 1758)) and Anthomyiidae (Delia platura (Meigen, 1826)) was made. Tendencies in reduction of the pregenital segments and musculature, as well as of the phallapodeme muscles in the evolution of the Muscoidea have been revealed. The complete set of phallapodeme muscles in the Scathophagidae and Anthomyiidae corresponds to the basal state, and therefore the structure of the genital sclerites and muscles in the Muscidae shows a certain degree of reduction. The progressive changes in the Muscidae from the Azeliinae through the Mydaeinae to the Muscinae were traced.

Selection on morphological traits and fluctuating asymmetry by a fungal parasite in the yellow dung fly

A Preprint reviewed and recommended by Peer Community Evolutionary Biology: http://dx.doi.org/10.24072/pci.evolbiol.100027Evidence for selective disadvantages of large body size remains scarce in general. Previous phenomenological studies of the yellow dung fly Scathophaga stercoraria have demonstrated strong positive sexual and fecundity selection on male and female size. Nevertheless, the body size of flies from a Swiss study population has declined by almost 10% from 1993 to 2009. Given substantial heritability of body size, this negative evolutionary response of an evidently positively selected trait suggests important selective factors being missed (e.g. size-selective predation or parasitism). A periodic epidemic outbreak of the fungus Entomophthora scatophagae allowedassessment of selection exerted by this parasite fatal to adult flies. Fungal infection varied over the season from ca. 50% in the cooler and more humid spring and autumn to almost 0% in summer. The probability of dying from fungal infection increased with adult body size. All infected females died before laying eggs, so there was no fungus impact on female fecundity beyond its impact on mortality. Large males showed the typical mating advantage in the field, but this pattern of positive sexual selection was nullified by fungal infection. Mean fluctuating asymmetry of paired appendages (legs, wings) did not affect the viability, fecundity or mating success of yellow dung flies in the field. This study demonstrates rare parasite-mediated disadvantages of large adult body size in the field. Reduced ability to combat parasites such as Entomophthora may be an immunity cost of large size in dung flies, although the hypothesized trade-off between fluctuating asymmetry, a presumed indicator of developmental instability and environmental stress, and immunocompetence was not found here.