Sepsid Flies Research Articles

Nutritional effects on growth and development of sepsid flies

Nutritional effects on growth and development of sepsid flies

Geographic Variation in Genomic Signals of Admixture Between Two Closely Related European Sepsid Fly Species

The extent of interspecific gene flow and its consequences for the initiation, maintenance, and breakdown of species barriers in natural systems remain poorly understood. Interspecific gene flow by hybridization may weaken adaptive divergence, but can be overcome by selection against hybrids, which may ultimately promote reinforcement. An informative step towards understanding the role of gene flow during speciation is to describe patterns of past gene flow among extant species. We investigate signals of admixture between allopatric and sympatric populations of the two closely related European dung fly species Sepsis cynipsea and S. neocynipsea (Diptera: Sepsidae). Based on microsatellite genotypes, we first inferred a baseline demographic history using Approximate Bayesian Computation. We then used genomic data from pooled DNA of natural and laboratory populations to test for past interspecific gene flow based on allelic configurations discordant with the inferred population tree (ABBA–BABA test with D-statistic). Comparing the detected signals of gene flow with the contemporary geographic relationship among interspecific pairs of populations (sympatric vs. allopatric), we made two contrasting observations. At one site in the French Cevennes, we detected an excess of past interspecific gene flow, while at two sites in Switzerland we observed lower signals of past microsatellite genotypes gene flow among populations in sympatry compared to allopatric populations. These results suggest that the species boundaries between these two species depend on the past and/or present eco-geographic context in Europe, which indicates that there is no uniform link between contemporary geographic proximity and past interspecific gene flow in natural populations.

Size rather than complexity of sexual ornaments prolongs male metamorphosis and explains sexual size dimorphism in sepsid flies.

Male sexual ornaments often evolve rapidly and are thought to be costly, thus contributing to sexual size dimorphism. However, little is known about their developmental costs, and even less about costs associated with structural complexity. Here, we quantified the size and complexity of three morphologically elaborate sexually dimorphic male ornaments that starkly differ across sepsid fly species (Diptera: Sepsidae): (i) male forelegs range from being unmodified, like in most females, to being adorned with spines and large cuticular protrusions; (ii) the fourth abdominal sternites are either unmodified or are converted into complex de novo appendages; and (iii) male genital claspers range from small and simple to large and complex (e.g. bifurcated). We tracked the development of 18 sepsid species from egg to adult to determine larval feeding and pupal metamorphosis times of both sexes. We then statistically explored whether pupal and adult body size, ornament size and/or ornament complexity are correlated with sex-specific development times. Larval growth and foraging periods of male and female larvae did not differ, but the time spent in the pupal stage was ca 5% longer for sepsid males despite emerging 9% smaller than females on average. Surprisingly, we found no evidence that sexual trait complexity prolongs pupal development beyond some effects of trait size. Evolving more complex traits thus does not incur developmental costs at least in this system.

Developmental bias predicts 60 million years of wing shape evolution

The degree to which developmental biases affect trait evolution is subject to much debate. Here, we first quantify fluctuating asymmetry as a measure of developmental variability, i.e., the propensity of developmental systems to create some phenotypic variants more often than others, and show that it predicts phenotypic and standing genetic variation as well as deep macroevolutionary divergence in wing shape in sepsid flies. Comparing our data to the findings of a previous study demonstrates that developmental variability in the sepsid fly Sepsis punctum strongly aligns with mutational, standing genetic, and macroevolutionary variation in the Drosophilidae--a group that diverged from the sepsid lineage ca. 64 My ago. We also find that developmental bias in S. punctum wing shape aligns with the effects of allometry, but less so with putatively adaptive thermal plasticity and population differentiation along latitude. Our findings demonstrate that developmental bias in fly wings predicts evolvability and macroevolutionary trajectories on a much greater scale than previously appreciated but also suggest that causal explanations for such alignments may go beyond simple constraint hypotheses.

The first mitochondrial genome of Sepsis monostigma (Diptera: Sepsidae)

The sepsid fly Sepsis monostigma belongs to the genus Sepsis of Sepsidae. We sequenced and annotated the mitogenome of S. monostigma which as the first representative of genus Sepsis with nearly complete mitochondrial data. This mitogenome is 14,887bp long, which contains of 22 transfer RNA genes, 13 protein coding genes (PCGs) and 2 ribosomal RNA genes and a part of the AT control region. ML phylogenetic outcome strongly supported the monophyly of Sepsidae, and the family Sepsidae is more close to the family Heleomyzidae. It also indicated that the genus Sepsis is the sister group to Nemopoda, and the genus Archisepsis is the sister group to Microsepsis.

Sexual size dimorphism is associated with reproductive life history trait differentiation in coexisting sepsid flies

Organismal life histories evolve as syndromes, resulting in correlated evolutionary differentiation of key traits that ultimately aid in discerning species. Reproductive success depends both on the absolute body size of an individual and its size relative to the opposite sex: sexual size dimorphism. In an attempt to further elucidate their coexistence and ecological diversification, we compared standard life history (first reproduction, clutch size, egg size) and associated reproductive trait differentiation of 15 widespread European sepsid fly species (Diptera: Sepsidae) under laboratory common garden conditions. Despite relatively uniform body sizes, sexual dimorphism ranged from female‐ to male‐biased, and development time varied twofold across species. We expected, and found, the abundant and relatively large species (Sepsis cynipsea, punctum, thoracica) with often male‐biased SSD to lay larger but fewer eggs and show fast‐developing, fast‐reproducing life histories with aggressive (coercive) mating behavior characterized by short mating latencies and male conflict. In contrast, the smaller and more dispersed species with female‐biased SSD (S. flavimana, orthocnemis, violacea) laid smaller but more eggs, showing a generally slower life history with long and delayed copulation and oviposition, high mating reluctance fostering extensive inter‐sexual conflict, and more elaborate male (pre‐)copulatory courtship. Two Saltella species were exceptional, being large, developing slowly, nevertheless copulating soon after adult emergence, profusely and briefly. The documented life history differentiation seems partly driven by sexual selection leading to male‐biased dimorphism, rather than undetermined ecological selection, but regardless appears insufficient to explain the coexistence and diversification of these sepsid species in European pastoral landscapes.

Usefulness and limitations of thermal performance curves in predicting ectotherm development under climatic variability.

Thermal performance curves (TPCs) have been estimated in multiple ectotherm species to understand their thermal plasticity and adaptation and to predict the effect of global warming. However, TPCs are typically assessed under constant temperature regimes, so their reliability for predicting thermal responses in the wild where temperature fluctuates diurnally and seasonally remains poorly documented. Here, we use distant latitudinal populations of five species of sepsid flies (Diptera: Sepsidae) from the temperate region (Europe, North Africa, North America) to compare estimates derived from constant TPCs with observed development rate under fluctuating temperatures in laboratory and field conditions. TPCs changed across gradients in that flies originating from higher latitudes showed accelerated development at higher temperatures, an adaptive response. TPCs were then used to predict development rates observed under fluctuating temperatures; these predictions were relatively accurate in the laboratory but not the field. Interestingly, the precision of TPC predictions depended not only on the resolution of temperature data, with daily and overall temperature summing performing better than hourly temperature summing, but also on the frequency of temperatures falling below the estimated critical minimum temperature. Hourly temperature resolution most strongly underestimated actual development rates, because flies apparently either did not stop growing when temperatures dropped below this threshold, or they sped up their growth when the temperature rose again, thus most severely reflecting this error. We conclude that when flies do not encounter cold temperatures, TPC predictions based on constant temperatures can accurately reflect performance under fluctuating temperatures if adequately adjusted for nonlinearities, but when encountering cold temperatures, this method is more error-prone. Our study emphasizes the importance of the resolution of temperature data and cold temperatures in shaping thermal reaction norms.

Does thermal plasticity align with local adaptation? An interspecific comparison of wing morphology in sepsid flies.

Although genetic and plastic responses are sometimes considered as unrelated processes, their phenotypic effects may often align because genetic adaptation is expected to mirror phenotypic plasticity if adaptive, but run counter to it when maladaptive. Because the magnitude and direction of this alignment has further consequences for both the tempo and mode of adaptation, they are relevant for predicting an organisms' reaction to environmental change. To better understand the interplay between phenotypic plasticity and genetic change in mediating adaptive phenotypic variation to climate variability, we here quantified genetic latitudinal variation and thermal plasticity in wing loading and wing shape in two closely related and widespread sepsid flies. Common garden rearing of 16 geographical populations reared across multiple temperatures revealed that wing loading decreases with latitude in both species. This pattern could be driven by selection for increased dispersal capacity in the cold. However, although allometry, sexual dimorphism, thermal plasticity and latitudinal differentiation in wing shape all show similar patterns in the two species, the relationship between the plastic and genetic responses differed between them. Although latitudinal differentiation (south to north) mirrored thermal plasticity (hot to cold) in Sepsis punctum, there was no relationship in Sepsis fulgens. While this suggests that thermal plasticity may have helped to mediate local adaptation in S.punctum, it also demonstrates that genetic wing shape differentiation and its relation to thermal plasticity may be complex and idiosyncratic, even among ecologically similar and closely related species. Hence, genetic responses can, but do not necessarily, align with phenotypic plasticity induced by changing environmental selection pressures.

The role of larval substrate specialization and female oviposition in mediating species diversity of closely-related sepsid flies (Diptera: Sepsidae)

Coprophagous insect communities play a critical role in the decomposition of vertebrate dung and provide ecosystem functions fundamental to modern agriculture. While the ecology of dung beetles is rather well understood, niche differentiation in coprophagous flies is poorly studied. Sepsid flies (Diptera: Sepsidae) are a vital part of the European community of coprophages, with 6-7 widespread species of Sepsis often found co-occurring in the same pasture. To advance our ecological understanding of the mechanisms that enable species to coexist, we investigated the oviposition preferences and larval performance of 7 common species of Sepsis in the dung of different large domestic and wild mammals. Substrate preferences and subsequent performance of larvae in laboratory experiments did not vary greatly. All species did very well on cow dung, the most common substrate in Central Europe, but also on dung of horse and wild boar. In contrast, flies did not prefer or grow well in dung of red and roe deer, two of the most common wild vertebrates. Thus there were only minor differences among the species tested along the specialist-generalist (dung) gradient, indicating that differences in the choice of oviposition sites by the adults of the different fly species and larval performance do not constitute a major axis of ecological differentiation. Nevertheless, there was a positive correlation between substrate choice and larval performance indicating the preference of gravid females for particular oviposition sites is adaptive. We conclude that sepsids are common in Europe because they are well adapted to the dung of herbivorous livestock rather than wild animals. Nevertheless, specialization on particular types of dung does not define the niche of Sepsis dung flies and hence plays a minor role in mediating their species diversity.

CRISPR/Cas9 deletions in a conserved exon of Distal-less generates gains and losses in a recently acquired morphological novelty in flies

SummaryDistal-less has been repeatedly co-opted for the development of many novel traits. Here, we document its curious role in the development of a novel abdominal appendage (“sternite brushes”) in sepsid flies. CRISPR/Cas9 deletions in the homeodomain result in losses of sternite brushes, demonstrating that Distal-less is necessary for their development. However, deletions in the upstream coding exon (Exon 2) produce losses or gains of brushes. A dissection of Exon 2 reveals that the likely mechanism for gains involves a deletion in an exon-splicing enhancer site that leads to exon skipping. Such contradictory phenotypes are also observed in butterflies, suggesting that mutations in the conserved upstream regions have the potential to generate phenotypic variability in insects that diverged 300 million years ago. Our results demonstrate the importance of Distal-less for the development of a novel abdominal appendage in insects and highlight how site-specific mutations in the same exon can produce contradictory phenotypes.

A Comparative Study of the Role of Sex-Specific Condition Dependence in the Evolution of Sexually Dimorphic Traits

Sexual selection can displace traits acting as ornaments or armaments from their viability optimum in one sex, ultimately giving rise to sexual dimorphism. The degree of dimorphism should not only mirror the strength of sexual selection but also the net viability costs of trait maintenance at equilibrium. As the ability of organisms to bear exaggerated traits will depend on their condition, more sexually dimorphic traits should also exhibit greater sex differences in condition dependence. While this has been demonstrated among traits within species, similar patterns are expected across the phylogeny. We investigated this prediction within and across 11 (sub)species of sepsid flies with varying mating systems. When estimating condition dependence for seven sexual and nonsexual traits that vary in their sexual dimorphism, we not only found a positive relationship between the sex difference in allometric slopes (our measure of condition dependence) and relative trait exaggeration within species but also across species for those traits expected to be under sexual selection. Species with more pronounced male aggression further had relatively larger and more condition-dependent male fore- and midlegs. Our comparative study suggests a common genetic/developmental basis of sexual dimorphism and sex-specific plasticity that evolves across the phylogeny-and that the evolution of size consistently alters scaling relationships and thus contributes to the allometric variation of sexual armaments or ornaments in animals.

Comparative effects of the parasiticide ivermectin on survival and reproduction of adult sepsid flies

Ivermectin is a veterinary pharmaceutical widely applied against parasites of livestock. Being effective against pests, it is also known to have lethal and sublethal effects on non-target organisms. While considerable research demonstrates the impact of ivermectin residues in livestock dung on the development and survival of dung feeding insect larvae, surprisingly little is known about its fitness effects on adults. We tested the impact of ivermectin on the survival of adult sepsid dung fly species (Diptera: Sepsidae) in the laboratory, using an ecologically relevant and realistic range of 69–1978 µg ivermectin/kg wet dung, and compared the sensitivities of larvae and adults in a phylogenetic framework. For one representative, relatively insensitive species, Sepsis punctum, we further investigated effects of ivermectin on female fecundity and male fertility. Moreover, we tested whether females can differentiate between ivermectin-spiked and non-contaminated dung in the wild. Adult sepsid flies exposed to ivermectin suffered increased mortality, whereby closely related species varied strongly in their sensitivity. Adult susceptibility to the drug correlated with larval susceptibility, showing a phylogenetic signal and demonstrating systemic variation in ivermectin sensitivity. Exposure of S. punctum females to even low concentrations of ivermectin lowered the number of eggs laid, while treatment of males reduced egg-to-adult offspring survival, presumably via impairment of sperm quality or quantity. The fitness impact was amplified when both parents were exposed. Lastly, sepsid flies did not discriminate against ivermectin-spiked dung in the field. Treatment of livestock with avermectins may thus have even more far-reaching sublethal ecological consequences than currently assumed via effects on adult dung-feeding insects.

CRISPR/Cas9 Deletions in a Conserved Exon of Distal-Less Generates Gains and Losses in a Recently Acquired Morphological Novelty in Flies

Distal-less has been repeatedly co-opted for the development of many novel traits. Here, we document its curious role in the development of a novel abdominal appendage (“sternite brushes”) in sepsid flies. CRISPR/Cas9 deletions in the homeodomain result in losses of sternite brushes, demonstrating that Distal-less is necessary for their development. However, deletions in the coding exon upstream of the homeodomain (Exon 2) produces both losses and gains of brushes. A dissection of Exon 2 reveals that the likely mechanism for gains involves a deletion in an exon-splicing enhancer site that leads to exon-skipping. Such contradictory phenotypes are also observed in butterflies which suggests that deletions in the conserved upstream regions have the potential to generate phenotypic variability in insects that diverged 300 million years ago. Our results demonstrate the importance of Distal-less for the development of a novel abdominal appendage in insects and highlight how site-specific mutations in the same exon can produce contradictory phenotypes.

Climatic factors shape plastic trade‐offs in the polyphenic black scavenger fly Sepsis thoracica (Diptera: Sepsidae)

AbstractAimTrade‐offs allow individuals to optimize their fitness by tailoring the investment into different traits to variable environmental conditions, such as along geographical gradients. Trade‐offs thus can help in adjusting to changing thermal and insolation profiles, especially in small ectotherms, whose body temperature typically follows environmental temperatures closely. Two traits usually involved in latitudinal adaptation are body size and melanism. Since both traits are costly, individuals need to optimize investment into each trait. Here, we studied how environmental factors influence this trade‐off in the short and long‐term.LocationEurope.MethodsWe raised flies from 15 latitudinal populations at three constant temperatures in a laboratory common garden to differentiate plastic and evolutionary responses to temperature. We further analysed how the different insolation components of the populations’ habitats influenced the evolution of the trade‐off.ResultsMale Sepsis thoracica (Diptera: Sepsidae) feature a sigmoid relationship between melanism and body size, defining two strikingly different male morphs: obsidian (small and black) and amber (large and orange). This relationship was altered by the developmental temperature, documenting its plasticity. The relationship further evolved across populations in response to the environmental characteristics of their habitat, notably temperature, insolation and UV radiation, suggesting that plasticity also has an underlying genetic basis. Nevertheless, melanism, but not body size, merely slightly increased with latitude.Main conclusionsAs the plastic and evolutionary responses of the relationship to temperature differed, plasticity does not necessarily follow the direction of evolution of this trade‐off, but rather adds to it. Our study evinces the role of several environmental factors in shaping the evolution of a plastic melanism—body size relationship defining a rare male polymorphism in temperate sepsid flies.

The evolution of male‐biased sexual size dimorphism is associated with increased body size plasticity in males

Abstract Sexual size dimorphism (SSD) can vary drastically across environments, demonstrating pronounced sex‐specific plasticity. In insects, females are usually the larger and more plastic sex. However, the shortage of taxa with male‐biased SSD hampers the assessment of whether the greater plasticity in females is driven by selection on size or represents an effect of the female reproductive role. Here, we specifically address the role of sex‐specific plasticity of body size in the evolution of SSD reversals to disentangle sex and size effects. We first investigate sex‐specific body size plasticity in Sepsis punctum and Sepsis neocynipsea as two independent cases of intraspecific SSD reversals in sepsid flies. In both species, directional variation in SSD between populations is driven by stronger sexual selection on male size. Using controlled laboratory breeding, we find evidence for sex‐specific plasticity and increased condition dependence of male size in populations with male‐biased SSD, but not of female size in populations with female‐biased SSD. To extend the comparative scope, we next estimate sex‐specific body size plasticity in eight additional fly species that differ in the direction of SSD under laboratory conditions. In all species with male‐biased SSD we find males to be the more plastic sex, while this was only rarely the case in species with female‐biased SSD, thus suggesting a more general trend in Diptera. To examine the generality of this pattern in holometabolous insects, we combine our data with data from the literature in a meta‐analysis. Again, male body size tends to be more plastic than female size when males are the larger sex, though female size is now also generally more plastic when females are larger. Our findings indicate that primarily selection on size, rather than the reproductive role per se, drives the evolution of sex‐specific body size plasticity. However, sepsid flies, and possibly Diptera in general, show a clear sexual asymmetry with greater male than female plasticity related to SSD, likely driven by strong sexual selection on males. Although further research controlling for phylogenetic and ecological confounding effects is needed, our findings are congruent with theory in suggesting that condition dependence plays a pivotal role in the evolution of sexual size dimorphism. A plain language summary is available for this article.

Sex ticklers and dirty flies: The development and evolution of a novel abdominal appendage in male sepsid flies

Sex ticklers and dirty flies: The development and evolution of a novel abdominal appendage in male sepsid flies

Selection on bristle length has the ability to drive the evolution of male abdominal appendages in the sepsid fly Themira biloba.

Many exaggerated and novel traits are strongly influenced by sexual selection. Although sexual selection is a powerful evolutionary force, underlying genetic interactions can constrain evolutionary outcomes. The relative strength of selection vs. constraint has been a matter of debate for the evolution of male abdominal appendages in sepsid flies. These abdominal appendages are involved in courtship and mating, but their function has not been directly tested. We performed mate choice experiments to determine whether sexual selection acts on abdominal appendages in the sepsid Themira biloba. We tested whether appendage bristle length influenced successful insemination by surgically trimming the bristles. Females paired with males that had shortened bristles laid only unfertilized eggs, indicating that long bristles are necessary for successful insemination. We also tested whether the evolution of bristle length was constrained by phenotypic correlations with other traits. Analyses of phenotypic covariation indicated that bristle length was highly correlated with other abdominal appendage traits, but was not correlated with abdominal sternite size. Thus, abdominal appendages are not exaggerated traits like many sexual ornaments, but vary independently from body size. At the same time, strong correlations between bristle length and appendage length suggest that selection on bristle length is likely to result in a correlated increase in appendage length. Bristle length is under sexual selection in T.biloba and has the potential to evolve independently from abdomen size.

Across the Baltic: a new record for an enigmatic black scavenger fly, Zuskamira inexpectata (Pont, 1987) (Sepsidae) in Finland.

Specimens of the enigmatic, monotypic European genus Zuskamira Pont, 1987 (Sepsidae) were initially collected only from the lower central Swedish provinces of Darlana, Uppland and Västmanland. However, the same species was subsequently found much more south in Lower-Saxony and Schleswig-Holstein although Germany is overall well sampled for sepsid flies. Here we report a further (longitudinal) range expansion based on new localities in Southern Finland. New localities for Finland and Sweden are here added and we discuss briefly the habitat requirements of the species.

The first mitochondrial genome of the sepsid fly Nemopoda mamaevi Ozerov, 1997 (Diptera: Sciomyzoidea: Sepsidae), with mitochondrial genome phylogeny of cyclorrhapha.

Sepsid flies (Diptera: Sepsidae) are important model insects for sexual selection research. In order to develop mitochondrial (mt) genome data for this significant group, we sequenced the first complete mt genome of the sepsid fly Nemopoda mamaevi Ozerov, 1997. The circular 15,878 bp mt genome is typical of Diptera, containing all 37 genes usually present in bilaterian animals. We discovered inaccurate annotations of fly mt genomes previously deposited on GenBank and thus re-annotated all published mt genomes of Cyclorrhapha. These re-annotations were based on comparative analysis of homologous genes, and provide a statistical analysis of start and stop codon positions. We further detected two 18 bp of conserved intergenic sequences from tRNAGlu-tRNAPhe and ND1-tRNASer(UCN) across Cyclorrhapha, which are the mtTERM binding site motifs. Additionally, we compared automated annotation software MITOS with hand annotation method. Phylogenetic trees based on the mt genome data from Cyclorrhapha were inferred by Maximum-likelihood and Bayesian methods, strongly supported a close relationship between Sepsidae and the Tephritoidea.

Distribution, diversity gradients and Rapoport's elevational rule in the black scavenger flies of the Swiss Alps (Diptera: Sepsidae)

Abstract Elevational gradients influence the distribution and abundance of species drastically and can lead to variation in community composition. Although coprophagous flies are of ecological and economic importance, their biodiversity and distribution are largely neglected. We studied the impact of steep elevational gradients and geography on the distribution of sepsid flies in the Swiss Alps. Sepsidae are a family of acalyptrate flies strongly associated with decaying organic matter and vertebrate dung, and characterised by a high extent of sympatry in their breeding substrates. Historical, haphazardly sampled specimens from 116 locations covering an elevational range from 200 to 2000 m were available in ethanol collections of various Swiss museums. Nineteen species encompassing all native genera (Meroplius, Nemopoda, Saltella, Sepsis and Themira) were recorded. Local species richness increased linearly with elevation, while area‐corrected regional species richness of elevational belts increased asymptotically. Species occurring at higher altitudes had lower wing loadings and greater elevational ranges than lowland species, supporting Rapoport's elevational rule. Despite compositional similarities, the sepsid communities of the northern lowlands differed significantly from the alpine sepsid fauna. The southern lowlands were particularly differentiated in community composition due to a number of presumably thermophilic species that predominantly occur south of the Alps. Relative abundances of several species were thus strongly affected by elevation and climatic variables. We illustrate the impact of elevational gradients and geography on a community of closely related, often sympatric species, and discuss potential mechanisms of niche partitioning via temporal succession, thermal adaptation and differential resource use.