Scathophaga Stercoraria Research Articles

Chromosome-level genome assembly of Scathophaga stercoraria provides new insights into the evolutionary adaptations of dung flies

The yellow dung fly Scathophaga stercoraria is a widely distributed species in high-altitude regions of the Northern Hemisphere. It plays important roles as a decomposer, predator, and pollinator in the ecosystem. As a staple model organism, S. stercoraria serves as a standard test species for assessing the toxicity of drug residues in livestock dung and has been the focus of numerous studies. The genetic mechanisms underlying the ecological adaptability of S. stercoraria remain poorly understood. To fill the gap, we first assembled a high-quality chromosome-level genome of S. stercoraria, resulting in a final assembly size of 549.64 Mb, with a contig N50 of 4.06 Mb, and 92.53 % of the sequence anchored to six chromosomes. Gene family analysis revealed an expansion of Toll (Toll1), GNBP3, Cyp303a1, Cyp4d14, Cyp6g1, OR67d, and yolk protein genes in the S. stercoraria genome. Transcriptome analysis indicated that most genes in the trypsin and carboxypeptidase gene families are predominantly expressed during the larval stage, whereas the α-Amylase gene family is mainly expressed during the adult stage. Additionally, PGRP-SC is highly expressed during the larval stage, OBPs are primarily expressed during the adult stage, and yolk protein genes exhibit female-biased expression. Our study not only provides a new resource for the dung flies genomic pool, but also identifies the expression patterns of key ecologically adaptative genes and gene families at the developmental stages, which provides new insights into the ecological adaptive evolution of dung flies.

Are yellow dung flies domesticated cow dung specialists?

The theory of niche differentiation implies some extent of specialization of species with regard to key resources, notably food. Coprophagous (dung-eating) insect larvae play a critical role in the decomposition of livestock dung in modern and traditional agricultural grasslands. The yellow dung fly (Scathophaga stercoraria L.; Diptera: Scathophagidae) is one of the largest, most common and abundant dung decomposers on pastures in cold-temperate regions across the entire northern hemisphere. As this fly is often associated with domesticated cattle or dairy cows, which are commonly kept for human nutrition worldwide (beef, milk, cheese, etc.), it is sometimes suspected to be a cow dung specialist. However, yellow dung flies are regularly active on and around other dung types, and must have reproduced on dung of wild vertebrates before the domestication of cattle. We therefore experimentally studied the performance of yellow dung fly larvae on dung of various large domestic vs. wild mammals (cow, horse, wild boar, red deer) in the laboratory in Switzerland. Larval performance in terms of juvenile survival, egg-to-adult development time, growth rate, and final adult body size, the major life history indicators of individual reproductive success, did not vary greatly among the various dung types tested. Thus, yellow dung flies can successfully reproduce on multiple types of mammal (vertebrate) dung, wild and domestic, and are therefore dung generalists rather than specialists. We conclude that yellow dung flies are common in European low- and highlands because they could plastically shift to dung of common herbivorous livestock after their domestication without losing the ability to reproduce on dung of common wild mammals.

Sperm competition in yellow dung flies: No consistent effect of sperm size.

The male competition for fertilization that results from female multiple mating promotes the evolution of increased sperm numbers and can impact sperm morphology, with theory predicting that longer sperm can at times be advantageous during sperm competition. If so, males with longer sperm should sire more offspring than competitors with shorter sperm. Few studies have directly tested this prediction, and findings are inconsistent. Here we assessed whether longer sperm provide a competitive advantage in the yellow dung fly (Scathophaga stercoraria; Diptera: Scathophagidae). Initially, we let brothers with different temperature-mediated mean sperm lengths compete - thus minimizing confounding effects of genetic background - and found no clear advantage of longer sperm. We then used flies from lines subjected to bidirectional selection on phenoloxidase activity that had shown correlated evolutionary responses in sperm and female spermathecal duct lengths. This experiment also yielded no main effect of sperm size on siring success. Instead, there was a trend for a shorter-sperm advantage, but only when competing in females with longer spermathecal ducts. Our data corroborated many previously reported findings (last-male precedence, effects of copula duration and body size), suggesting our failure to find sperm size effects is not inherently due to our experimental protocols. We conclude that longer sperm are not competitively superior in yellow dung flies under most circumstances, and that, consistent with previous work, in this species competitive fertilization success is primarily determined by the relative numbers of sperm competing.

Material Properties and Morphology of Prestomal Teeth in Relation to the Feeding Habits of Diptera (Brachycera).

Simple SummaryThe prestomal teeth are structures on the mouthparts of some fly species that rasp surfaces to expose liquids for ingestion. Here, we investigated the material properties of prestomal teeth, including their hardness, elastic modulus, the extent of sclerotization, and elemental composition, and combined these results with morphology to determine their relationship to fly feeding habits. The results indicated that the prestomal teeth are heavily sclerotized and have hardness and elastic modulus values similar to the polymer polycarbonate. Although the presence of inorganic elements contributes to a harder cuticle in other insect species, the fly species studied here had only low concentrations of inorganic elements. We found evidence that the material properties and morphology of prestomal teeth relate to feeding habits, not phylogeny. In particular, flies that pierce tissues for liquids have larger prestomal teeth relative to their mouthpart sizes when compared to species that generally feed on exposed liquids. Flies are one of the most successful groups of organisms and their success likely relates to their ability to feed on a large array of nutritional liquids. Given their importance in ecology systems and relevance to medical and veterinary entomology, the functional morphology of fly mouthparts warrants additional studies.Prestomal teeth are cuticular projections on the mouthparts of some fly species that rasp surfaces when feeding. Although prestomal teeth morphology has been reported for several fly species, their material properties have not been investigated. Here we report the morphology, elemental composition, extent of sclerotization, hardness, and elastic modulus of prestomal teeth and relate these findings to feeding habits. Scanning electron microscopy revealed that species categorized as flower visitors have a large labellum with numerous pseudotracheae and lack prestomal teeth, generalist species have these same features but with prestomal teeth, and specialist species that feed on blood or other insects have a smaller labellum with few or no pseudotracheae and relatively large prestomal teeth. Confocal microscopy revealed that prestomal teeth are heavily sclerotized and the labellum contains resilin, an elastomeric protein. Hardness and elastic modulus were explored with nanoindentation and showed that the insectivorous Scathophaga stercoraria had the hardest prestomal teeth and the highest modulus. Energy dispersive x-ray spectroscopy revealed that prestomal teeth had low concentrations of inorganic elements, suggesting that hardness might be partially supplemented by inorganic elements. Our findings indicate that prestomal teeth morphology and material properties relate more to feeding habits than to phylogeny.

Growth rate mediates hidden developmental plasticity of female yellow dung fly reproductive morphology in response to environmental stressors.

Understanding how environmental variation influences even cryptic traits is important to clarify the roles of selection and developmental constraints in past evolutionary divergence and to predict future adaptation under environmental change. Female yellow dung flies (Scathophaga stercoraria) typically have three sperm storage compartments (3S), but occasionally four (4S). More spermathecae are thought to be a female adaptation facilitating sperm sorting after mating, but the phenotype is very rare in nature. We manipulated the flies' developmental environment by food restriction, pesticides, and hot temperatures to investigate the nature and extent of developmental plasticity of this trait, and whether spermatheca expression correlates with measures of performance and developmental stability, as would be expected if 4S expression is a developmental aberration. The spermathecal polymorphism of yellow dung fly females is heritable, but also highly developmentally plastic, varying strongly with rearing conditions. 4S expression is tightly linked to growth rate, and weakly positively correlated with fluctuating asymmetry of wings and legs, suggesting that the production of a fourth spermatheca could be a nonadaptive developmental aberration. However, spermathecal plasticity is opposite in the closely related and ecologically similar Scathophaga suilla, demonstrating that overexpression of spermathecae under developmental stress is not universal. At the same time, we found overall mortality costs as well as benefits of 4S pheno‐ and genotypes (also affecting male siblings), suggesting that a life history trade‐off may potentially moderate 4S expression. We conclude that the release of cryptic genetic variation in spermatheca number in the face of strong environmental variation may expose hidden traits (here reproductive morphology) to natural selection (here under climate warming or food augmentation). Once exposed, hidden traits can potentially undergo rapid genetic assimilation, even in cases when trait changes are first triggered by random errors that destabilize developmental processes.

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.

Comprehensive thermal performance curves for yellow dung fly life history traits and the temperature-size-rule.

Ambient temperature strongly determines the behaviour, physiology, and life history of all organisms. The technical assessment of organismal thermal niches in form of now so-called thermal performance curves (TPC) thus has a long tradition in biological research. Nevertheless, several traits do not display the idealized, intuitive dome-shaped TPC, and in practice assessments often do not cover the entire realistic or natural temperature range of an organism. We here illustrate this by presenting comprehensive sex-specific TPCs for the major (juvenile) life history traits of yellow dung flies (Scathophaga stercoraria; Diptera: Scathophagidae). This concerns estimation of prominent biogeographic rules, such as the temperature-size-rule (TSR), the common phenomenon in ectothermic organisms that body size decreases as temperature increases. S. stercoraria shows an untypical asymptotic TPC of continuous body size increase with decreasing temperature without a peak (optimum), thus following the TSR throughout their entire thermal range (unlike several other insects presented here). Egg-to-adult mortality (our best fitness estimator) also shows no intermediate maximum. Both may relate to this fly entering pupal winter diapause below 12°C. While development time presents a negative exponential relationship with temperature, development rate and growth rate typify the classic TPC form for this fly. The hitherto largely unexplored close relative S. suilla with an even more arctic distribution showed very similar responses, demonstrating large overlap among two ecologically similar, coexisting dung fly species, thus implying limited utility of even complete TPCs for predicting species distribution and coexistence.

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.

Heritable responses to combined effects of heat stress and ivermectin in the yellow dung fly

In current times of global change, several sources of stress such as contaminants and high temperatures may act synergistically. The extent to which organisms persist in stressful conditions will depend on the fitness consequences of multiple simultaneously acting stressors and the genetic basis of compensatory genetic responses. Ivermectin is an antiparasitic drug used in livestock that is excreted in dung of treated cattle, causing severe negative consequences on non-target fauna. We evaluated the effect of a combination of heat stress and exposure to ivermectin in the yellow dung fly, Scathophaga stercoraria (Diptera: Scathophagidae). In a first experiment we investigated the effects of high rearing temperature on susceptibility to ivermectin, and in a second experiment we assayed flies from a latitudinal gradient to assess potential effects of local thermal adaptation on ivermectin sensitivity. The combination of heat and ivermectin synergistically reduced offspring survival, revealing severe effects of the two stressors when combined. However, latitudinal populations did not systematically vary in how ivermectin affected offspring survival, body size, development time, cold and heat tolerance. We also found very low narrow-sense heritability of ivermectin sensitivity, suggesting evolutionary constraints for responses to the combination of these stressors beyond immediate maternal or plastic effects. If the revealed patterns hold also for other invertebrates, the combination of increasing climate warming and ivermectin stress may thus have severe consequences for biodiversity. More generally, our study underlines the need for quantitative genetic analyses in understanding wildlife responses to interacting stressors that act synergistically and threat biodiversity.

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

Abstract 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.

Evolutionary insight from a humble fly: sperm competition and the yellow dungfly.

Studies of the yellow dungfly in the 1960s provided one of the first quantitative demonstrations of the costs and benefits associated with male and female reproductive behaviour. These studies advanced appreciation of sexual selection as a significant evolutionary mechanism and contributed to the 1970s paradigm shift toward individual selectionist thinking. Three behaviours in particular led to the realization that sexual selection can continue during and after mating: (i) female receptivity to remating, (ii) sperm displacement and (iii) post-copulatory mate guarding. These behaviours either generate, or are adaptations to sperm competition, cryptic female choice and sexual conflict. Here we review this body of work, and its contribution to the development of post-copulatory sexual selection theory. This article is part of the theme issue 'Fifty years of sperm competition'.

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.

Evolution of reduced minimum critical size as a response to selection for rapid pre-adult development in Drosophila melanogaster.

Adult body size in holometabolus insects is directly proportional to the time spent during the larval period. The larval duration can be divided into two parts: (i) pre-critical duration—time required to attain a critical size/critical weight that would result in successful completion of development and metamorphosis even under non-availability of nutrition beyond the time of attainment of critical size, and (ii) post-critical duration—the time duration from the attainment of critical size till pupation. It is of interest to decipher the relative contribution of the two larval growth phases (from the hatching of the egg to the attainment of critical size, and from the attainment of critical size to pupation) to the final adult size. Many studies using Drosophila melanogaster have shown that selecting populations for faster development results in the emergence of small adults. Some of these studies have indirectly reported the evolution of smaller critical size. Using two kinds of D. melanogaster populations, one of which is selected for faster/accelerated pre-adult development and the other their ancestral control, we demonstrate that the final adult size is determined by the time spent as larvae post the attainment of critical size despite having increased growth rate during the second larval instar. Our populations under selection for faster pre-adult development are exhibiting adaptive bailout due to intrinsic food limitation as against extrinsic food limitation in the yellow dung fly.

Fitness consequences of the combined effects of veterinary and agricultural pesticides on a non-target insect

Pesticides and veterinary products that are globally used in farming against pests and parasites are known to impact non-target beneficial organisms. While most studies have tested the lethal and sub-lethal effects of single chemicals, species are exposed to multiple contaminants that might interact and exacerbate the toxic responses of life-history fitness components. Here we experimentally tested an ecotoxicological scenario that is likely to be widespread in nature, with non-target dung communities being exposed both to cattle parasiticides during the larval stage and to agricultural insecticides during their adult life. We assessed the independent and combined consumptive effects of varying ivermectin and spinosad concentration on juvenile life-history and adult reproductive traits of the widespread yellow dung fly (Scathophaga stercoraria; Diptera: Scathophagidae). Larval exposure to ivermectin prolonged development time and reduced egg-to-adult survival, body size, and the magnitude of the male-biased sexual size dimorphism. The consumption by the predatory adult flies of spinosad-contaminated prey showed an additional, independent (from ivermectin) negative effect on female clutch size, and subsequent egg hatching success, but not on the body size and sexual size dimorphism of their surviving offspring. However, there were interactive synergistic effects of both contaminants on offspring emergence and body size. Our results document adverse effects of the combination of different chemicals on fitness components of a dung insect, highlighting transgenerational effects of adult exposure to contaminants for their offspring. These findings suggest that ecotoxicological tests should consider the combination of different contaminants for more accurate eco-assessments.

Sublethal effects of the parasiticide ivermectin on male and female reproductive and behavioural traits in the yellow dung fly.

The veterinary pharmaceutical ivermectin is commonly used against parasites of livestock. Excreted in dung it can have lethal and sublethal effects on non-target organisms developing in and living around cattle dung. Research in this realm typically investigates the impact of pharmaceuticals on dung-feeding insects by looking at juvenile development and survival, while fitness effects of adult exposure are largely neglected. We conducted laboratory experiments to assess combined effects of ivermectin on life history and reproductive traits of juvenile and adult yellow dung flies (Scathophaga stercoraria). Two treatments (12 and 24 μg ivermectin/kg wet dung) were used for the larvae reared in dung, and one much higher concentration (3000 μg ivermectin/kg sugar) for the adult flies (in addition to uncontaminated controls). Juvenile ivermectin exposure lead to smaller body size of male and female flies. Adult feeding on ivermectin-contaminated dung additionally resulted in adult male flies with smaller testes (and likely fewer sperm) that experienced reduced mating durations, resulting in lower probability of producing offspring. Exposure of adult flies to ivermectin lowered offspring production and survival for both sexes. Thus, treatment of livestock with pharmaceuticals such as ivermectin appears to have even more far-reaching sublethal ecological consequences than previously assumed by affecting not only flies at their larval stage but also adult mating behaviour and reproduction.

Musculature of the Male Abdominal Segments and Terminalia of Mydaea urbana (Meigen, 1826) and Graphomya maculata (Scopoli, 1763) (Diptera, Muscidae: Mydaeinae)

The structure of the abdominal and pregenital segments and genitalia was studied in males of Mydaea urbana (Meigen, 1826) and Graphomya maculata (Scopoli, 1763) (Muscidae, Mydaeinae). Reduction tendencies were revealed in the pregenital segments and musculature in Muscinae as compared with Mydaeinae, and also in the genitalia of Muscidae as compared with Scathophagidae (Scathophaga stercoraria). The genus Graphomya was found to be similar to Mydaea but different from the tribe Muscini in the characters of the sclerites and muscles of the male genitalia and pregenital segments; this confirms the position of Graphomya in the subfamily Mydaeinae.

Replicated latitudinal clines in reproductive traits of European and North American yellow dung flies

Geographic variation in phenotypic traits is commonly correlated with spatial variation in the environment, e.g. seasonality and mean temperature, providing evidence that natural selection generates such patterns. In particular, both body size and egg size of ectothermic animals are commonly larger in northern climates, and temperature induces plastic responses in both traits. Size‐independent egg quality can also vary with latitude, though this is rarely investigated. For the widespread yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae) we investigated whether there are latitudinal clines in reproductive traits (clutch size, egg size and egg composition), whether these clines are due to variation in body and/or egg size, and whether such clines replicate across independent experiments performed on different continents (North America and Europe). Egg size generally increased with latitude (especially in Europe), an effect largely explained by body size of the mother, while clutch size did not; overall reproductive effort thus increased with latitude. Both the absolute and relative (correcting for egg size) amount of egg protein increased with latitude, egg glycogen decreased with latitude, while latitudinal trends for egg lipids and total egg energy content were complex and non‐linear. Altitude sometimes showed relationships analogous to those of latitude (egg proteins and glycogen) but occasionally opposite (egg size), possibly because latitude and altitude are negatively related among populations of this cold‐adapted species. There was no evidence of a tradeoff between egg size and number across latitudinal populations; if anything, the relationship was positive. All traits, including body and egg size, varied with rearing temperature (12°C, 18°C, 24°C), generally following the temperature–size rule. Clines based on common garden rearing, thus reflecting genetic differentiation, were qualitatively but not always quantitatively consistent between continents, and were similar across rearing temperatures, suggesting they evolved due to natural selection, although the concrete selective mechanisms involved require further study.

Musculature of the Male Abdominal Segments and Terminalia in Musca autumnalis De Geer, 1776 and Pyrellia rapax (Harris, 1780) (Diptera, Muscidae: Muscini)

The structure of the abdominal and pregenital segments and genitalia of males was studied in Pyrellia rapax (Harris, 1780) and Musca autumnalis De Geer, 1776 (Muscidae, Muscini). The musculature of these species is compared with that in Musca domestica Linnaeus, 1758 (Muscidae) and Scathophaga stercoraria (Linnaeus, 1758) (Scathophagidae), previously investigated by O.G. Ovtshinnikova. Morphological analysis of sclerites and muscles of abdominal and pregenital segments and genitalia of males in Pyrellia rapax, Musca autumnalis, and M. domestica demonstrates processes of reduction in Musca as compared with P. rapax and S. stercoraria.

Geographic clines in wing morphology relate to colonization history in New World but not Old World populations of yellow dung flies.

Geographic clines offer insights about putative targets and agents of natural selection as well as tempo and mode of adaptation. However, demographic processes can lead to clines that are indistinguishable from adaptive divergence. Using the widespread yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae), we examine quantitative genetic differentiation (QST ) of wing shape across North America, Europe, and Japan, and compare this differentiation with that of ten microsatellites (FST ). Morphometric analyses of 28 populations reared at three temperatures revealed significant thermal plasticity, sexual dimorphism, and geographic differentiation in wing shape. In North America morphological differentiation followed the decline in microsatellite variability along the presumed route of recent colonization from the southeast to the northwest. Across Europe, where S. stercoraria presumably existed for much longer time and where no molecular pattern of isolation by distance was evident, clinal variation was less pronounced despite significant morphological differentiation (QST >FST ). Shape vector comparisons further indicate that thermal plasticity (hot-to-cold) does not mirror patterns of latitudinal divergence (south-to-north), as might have been expected under a scenario with temperature as the major agent of selection. Our findings illustrate the importance of detailed phylogeographic information when interpreting geographic clines of dispersal traits in an adaptive evolutionary framework.