Dung Fly Research Articles

Genetic and environmental sources of covariance among internal reproductive traits in the yellow dung fly

Substantial inter- and intraspecific variation is found in reproductive traits, but the evolutionary implications of this variation remain unclear. One hypothesis is that natural selection favours female reproductive morphology that allows females to control mating and fertilization and that diverse male reproductive traits arise as counter adaptations to subvert this control. Such co-evolution predicts the establishment of genetic correlations between male and female reproductive traits that closely interact during mating. Therefore, we measured phenotypic and genetic correlations between male and female reproductive tract characteristics in the yellow dung fly, Scathophaga stercoraria (Diptera: Scathophagidae), using a nested half-sib breeding experiment. We found significant heritabilities for the size of most reproductive tract traits investigated in both females (spermathecae and their ducts, accessory glands and their ducts) and males (testis size but not sperm length). Within the sexes, phenotypic and genetic correlations were mostly nil or positive, suggesting functional integration of or condition-dependent investment in internal reproductive traits. Negative intrasexual genetic correlations, potentially suggestive of resource allocation trade-offs, were not evident. Intersexual genetic correlations were mostly positive, reflecting expected allometries between male and female morphologies. Most interestingly, testis size correlated positively with female accessory gland size and duct length, potentially indicative of a co-evolutionary arms race. We discuss these and alternative explanations for these patterns of genetic covariance.

First records of flower biology and pollination in African Annonaceae: Isolona, Piptostigma, Uvariodendron, Monodora and Uvariopsis

Seven species from five genera of Annonaceae were studied with regard to their flower biology and pollination in the Southwest Province of Cameroon, West Africa. They have protogynous hermaphroditic flowers, with exception of Uvariopsis species, which are monoecious. Fused petals of Isolona campanulata remain apically spreading and open during anthesis but form a deep basal urceolate tube around the reproductive organs. At anthesis the yellow pendent flowers emit a fruit-like scent and attracted small beetles, the likely pollinators. Piptostigma sp. flowers also emit a fruit-like scent but provide a closed pollination chamber formed by the three inner petals. Small staphylinid beetles attracted during the female stage of anthesis are released from the flowers at the end of the male stage 2–3 days later. Both species have diurnal anthesis, attracting and releasing the flower visitors during daytime. In contrast, Uvariodendron connivens and U. calophyllum have nocturnal anthesis with floral thermogenesis, produce spicy, aromatic and fruity scents and attract large Scarabaeidae beetles, the pollinators, along with many curculionid beetles, which were principally predators of the thick petals. The very large flowers of Monodora tenuifolia have yellowish petals which are spotted with dark red markings. Together with the sweetish, slightly disagreeable scent the flowers attract flies, principally dung flies. The two investigated Uvariopsis species are monoecious with pistillate and staminate flowers being functional at the same time. The violet red flowers of U. bakeriana visually seem to mimic the fruiting body of certain stinkhorn fungi (Phallaceae) although without producing their strong unpleasant carcass stench. Flower-visiting dung flies were rare. Conversely, U. congolana has a strong fungus-like scent, its flowers are presented at litter height and dung flies living in the litter were the flower visitors, albeit sporadic. The 4–5 days lasting anthesis of both Uvariopsis species appears to be an evolutionary consequence of their diffuse pollinator spectra. The studied African Annonaceae therefore have either cantharophilous or myiophilous/sapromyiophilous flowers with, in part, respectively, remarkably long anthesis, thermogenesis, and widely open, large flowers – all attributes unknown or rare in the hitherto better studied Neotropical Annonaceae.

<p><strong>Double tracked dispersal strategy in Splachnaceae</strong><br /><strong>Doppelte Ausbreitungsstrategie bei Splachnaceae</strong></p>

Some species of Splachnaceae smell like berries when the spores are already ripe but the capsules still closed. It is supposed that this is a way to attract animals to eat the capsules, so that the spores reach suitable substrates directly with the faeces. This first strategy is little known, as the smell can be observed for a short time only. The second strategy, attraction of dung flies by dung smell of the open capsules, which transport the spores from one dunghill to another, is well-known.

Assessment of risk of monepantel faecal residues to dung fauna

To evaluate the safety of monepantel and its major metabolite for dung fauna. Monepantel is a new oral anthelmintic drug for use in sheep at a dose of 2.5 mg active ingredient/kg body weight. Hazard (toxicity) is related to the expected exposure. The methodology was based on the Organisation for Economic Cooperation and Development (OECD) draft guideline for testing of chemicals, so dung from cattle, not sheep, and eggs of the dung fly, Scathophaga stercoraria and larvae of the dung beetle Aphodius constans were used. Monepantel or its sulfone metabolite were mixed into bovine faeces in which either dung fly eggs or dung beetle larvae were placed and their development observed. The primary endpoint was survival. Real exposure data (faecal concentrations) for comparison with the generated laboratory data were taken from a kinetics study in sheep dung. The no-observed effect concentration (NOEC) for monepantel was >1000 mg/kg substrate for dung flies and 250 mg/kg for dung beetles. The sulfone metabolite was slightly more toxic, with a NOEC of 500 mg/kg for dung flies and 125 mg/kg for dung beetles. A comparison of the results to the maximum concentrations of 15 mg monepantel and 4.5 mg sulfone per kg dung observed under an exaggerated dosing regime in sheep indicates that monepantel poses no risk to insect dung fauna when used as recommended. The study is considered valid because representatives of both genera were able to develop in bovine or ovine dung.

A cost of being large: genetically large yellow dung flies lose out in intra-specific food competition

Most life history traits are positively influenced by body size, while disadvantages of large size are poorly documented. To investigate presumed intrinsic costs of large body size in yellow dung flies (Scathophaga stercoraria; Diptera: Scathophagidae), we allowed larvae from replicate lines artificially selected for small and large body size for 21 generations to compete directly with each other at 20°C (benign) and 25°C (stressful) and low and high food (dung) availability. Greater mortality of large line flies was evident at low food independent of temperature, suggesting a cost of fast growth and/or long development for genetically large flies during larval scramble competition under food limitation. Our results are congruent with a previous study assessing mortality when competing within body size lines, so no additional mechanisms affecting scramble or contest behavior of larvae need be invoked to explain the results obtained beyond the costs of longer development and faster growth. Thus, artificial selection producing larger yellow dung flies than occur in nature revealed some, albeit weak mortality costs of large body size that otherwise might have remained cryptic. We conclude, however, that these costs are insufficient to explain the evolutionary limits of large body size in this species given persistently strong fecundity and sexual selection favoring large size in both sexes.

Effects of maternal and offspring environmental conditions on growth, development and diapause in latitudinal yellow dung fly populations

Extensive phenotypic plasticity can allow populations to persist in changing environ- ments. Maternal effects represent one important but often neglected source of phenotypic plasticity. Mothers and offspring of 2 high- (northern Norway and central Sweden) and 2 low- (northern and southern Spain) latitude yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae) popula- tions were exposed to cold (12°C) and warm (18°C) temperatures and to short (8 h light:16 h dark) and long (16 h light:8 h dark) photoperiods in a fully blocked, split-brood common garden design of 8 treatment combinations. We also considered the role of energy content and size of the eggs in pro- ducing cross-generational maternal effects on offspring diapause, development time and growth rate. The incidence of diapause strongly declined towards the south, and the northernmost popula- tion grew and developed faster in response to perceived seasonal time constraints. There was strong population-specific phenotypic plasticity of all traits in response to offspring temperature and, more weakly, to offspring photoperiod, indicating a genetic basis of plasticity as well as genetic differenti- ation among populations. There were additional subtle cross-generational maternal effects exerted primarily by the lipid content of the eggs, largely independent of maternal treatment and population. Phenotypic plasticity of life-history traits in the yellow dung fly is predominantly influenced by the growing conditions during larval development, but populations can also respond to changing envi- ronments via trans-generational maternal effects.

The Karyotype of the Yellow Dung Fly,Scathophaga stercoraria, a Model Organism in Studies of Sexual Selection

Knowledge of karyotypical characteristics of a species is essential for understanding how sexually selected and sexually antagonistic traits evolve. The yellow dung fly Scathophaga stercoraria L. (Diptera: Scathophagidae) is an established model system for studies of sexual selection and sexual conflict, but karyotypical data are lacking to date. Here, the karyotype of S. stercoraria was characterized using conventional Giemsa-staining and C-banding techniques. The diploid chromosome set consists of 6 pairs of bi-armed meta- or submetacentric chromosomes. The sex chromosomes are the largest chromosomes and constitute 30% of the total length of the diploid set in females and about 25% in males. Males are the heterogametic sex, and the length of the Y chromosome is about three-quarters of that of the X chromosome. C-banding revealed that both sex chromosomes are largely heterochromatic. In contrast, in the five autosome pairs, heterochromatin is limited to narrow bands in the centromeric regions. This karyotypic information will help provide a more profound understanding of the inheritance of phenotypic variation in reproductive traits and the chromosomal locations of underlying genes.

Sperm competition and ejaculate economics

Sperm competition was identified in 1970 as a pervasive selective force in post-copulatory sexual selection that occurs when the ejaculates of different males compete to fertilise a given set of ova. Since then, sperm competition has been much studied both empirically and theoretically. Because sperm competition often favours large ejaculates, an important challenge has been to understand the evolution of strategies through which males invest in sperm production and economise sperm allocation to maximise reproductive success under competitive conditions. Sperm competition mechanisms vary greatly, depending on many factors including the level of sperm competition, space constraints in the sperm competition arena, male mating roles, and female influences on sperm utilisation. Consequently, theoretical models of ejaculate economics are complex and varied, often with apparently conflicting predictions. The goal of this review is to synthesise the theoretical basis of ejaculate economics under sperm competition, aiming to provide empiricists with categorised model assumptions and predictions. We show that apparent contradictions between older and newer models can often be reconciled and there is considerable consensus in the predictions generated by different models. We also discuss qualitative empirical support for some of these predictions, and detail quantitative matches between predictions and observations that exist in the yellow dung fly. We argue that ejaculate economic theory represents a powerful heuristic to explain the diversity in ejaculate traits at multiple levels: across species, across males and within individual males. Future progress requires greater understanding of sperm competition mechanisms, quantification of trade-offs between ejaculate allocation and numbers of matings gained, further knowledge of mechanisms of female sperm selection and their associated costs, further investigation of non-sperm ejaculate effects, and theoretical integration of pre- and post-copulatory episodes of sexual selection.

Effects of the parasiticide ivermectin on the structure and function of dung and soil invertebrate communities in the field (Madrid, Spain)

The effects of the parasiticide ivermectin on dung and soil fauna as well as dung decomposition were assessed in a field study conducted near Madrid (Spain). Groups of cattle were treated with an injection of Ivomec ® at a dosage of 200 μg ivermectin kg −1 body weight at four different time points before collecting the fresh dung of all cattle groups on day 0. Dung pats prepared from the dung of the ivermectin-treated cattle were exposed in the field from day 0 onwards in parallel with dung pats that were prepared from the dung of untreated cattle and that was either spiked with a high concentration of ivermectin (positive control) or not spiked (negative control). Ivermectin concentrations in freshly excreted dung ranged from 0.31 to 0.81 mg ivermectin kg −1 dung dry weight. Ivermectin soil concentrations were highest below ivermectin-spiked dung in the uppermost soil layer with up to 0.085 mg ivermectin kg −1 soil dw. No ivermectin-related effects on collembolans and mites were found. A significant decrease in the abundance of adult dung beetles was observed at 0.81 mg ivermectin kg −1 dung dw, but the two most abundant species showed contrasting responses. For the dung beetle species, Volinus distinctus, a no observed effect concentration (NOEC) and a median effect concentration (EC 50) of 0.50 and 0.62 mg ivermectin kg −1 dung dw, respectively, were determined. Dung fly larvae were the most sensitive dung fauna group as their abundance was significantly reduced in all ivermectin treatments, resulting in a NOEC of <0.31 mg ivermectin kg −1 dw. Staphylinid beetles were abundant in the dung, but apparently not affected by ivermectin. Dung from ivermectin-treated cattle degraded more slowly than dung from untreated cattle, resulting in a NOEC of <0.78 mg ivermectin kg −1 dung dw for this functional endpoint. Adult dung beetles of both dominant species were attracted to dung pats spiked with ivermectin. The mobility of ivermectin appeared to differ between dung from ivermectin-treated cattle and dung spiked with ivermectin. This finding challenges approaches to assess the fate and effect of veterinary pharmaceuticals in higher tier tests by using spiked dung. This field study detected consistent results regarding effects of ivermectin on structural and functional endpoints in the dung compartment but effects on soil microarthropods are of little relevance in comparison. This study further advances guidance on higher tier tests that is required to make the results more applicable to the environmental risk assessment of veterinary pharmaceuticals.

Results of an international ring test with the dung fly Musca autumnalis in support of a new OECD test guideline

A standardized bioassay using the face fly, Musca autumnalis L. (Diptera: Muscidae), was developed to test the lethal and sublethal toxicity of parasiticide residues in livestock dung. The repeatability of this test was assessed for the parasiticide ivermectin in seven tests performed in four laboratories in Germany and France. Additional results of limit tests were provided by two laboratories from the UK. Test results had an acceptable range of heterogeneity. The calculated effect concentration at which 50% emergence was observed (EC50) averaged 4.65 ± 2.17 (Standard Deviation (SD) µg ivermectin/kg fresh dung (range: 1.20–7.7)). Effects on emergence were, with one exception, not observed below the No Observed Effect Concentration (NOEC) ranging between 1.11 and 3.33 µg ivermectin/kg. No effect on development time was observed. We conclude that the face fly is suitably sensitive, and the methods sufficiently repeatable, to support use of this standardized bioassay by the international community in the registration of new veterinary pharmaceuticals. Following these considerations, this species was accepted as a possible test organism in a recently published OECD Guideline (No. 228).

Systematic ambiguity in the well-established model system insect Scathophaga stercoraria (Diptera: Scathophagidae): sister species S. soror revealed by molecular evidence

Even for well-established insect model systems, such as the yellow dung fly Scathophaga stercoraria (Linnaeus) (Diptera: Scathophagidae), there may be hidden systematic ambiguities that require clarification. Dung flies from the Afrotropical Region have been considered (i) as con-specific and not different from all the other Holarctic Scathophaga stercoraria; (ii) as a local and peculiar African subspecies of S. stercoraria (Scathophaga stercoraria soror Wiedemann), or (iii) as a separate valid species (Scathophaga soror Wiedemann). Our study represents an attempt, based on mitochondrial (COI, 12S, and 16S), nuclear (ITS2) as well as microsatellite markers, to clarify this problem. Results strongly suggest that S. soror is a separate taxon from S. stercoraria. Due to the importance of S. stercoraria as a model system for studies in ecology, behaviour and evolution, the systematic position of S. soror (relative to S. stercoraria) is not solely of interest for systematists, but for evolutionary ecologists as well.

A review of the natural history and laboratory culture methods for the yellow dung fly, Scathophaga stercoraria.

The yellow dung fly Scathophaga stercoraria (L.) (Diptera: Scathophagidae) is a widespread and locally abundant fly associated with the dung of large mammals, especially farm animals. This species has recently become a standard test organism for evaluating toxic effects of veterinary pharmaceuticals in livestock dung. In this context, a review of its natural history and a general description of the field and laboratory rearing methods of this species are provided here to benefit the scientific community as well as government regulators and applicants of eco-toxicological studies. For guidance, means and ranges are included for all relevant standard life history traits stemming from previously published data on Swiss populations.

The assessment of insemination success in yellow dung flies using competitive PCR

In spite of considerable interest in postcopulatory sexual selection, separating the effects of sperm competition from cryptic female choice remains difficult because mechanisms underlying postcopulatory processes are poorly understood. One methodological challenge is to quantify insemination success for individual males within the sperm stores of multiply mated females to discover how insemination translates into eventual paternity. Any proposed method must be applicable in organisms without extensive DNA sequence information (which include the majority of model species for sexual selection). Here, we describe the development and application of microsatellite competitive-multiplex-PCR for quantifying relative contributions to a small number of sperm in storage. We studied how DNA template characteristics affect PCR amplification of known concentrations of mixed DNA and generated regressions for correcting observations of allelic signal strength based on such characteristics. We used these methods to examine patterns of sperm storage in twice-mated female yellow dung flies, Scathophaga stercoraria. We confirm previous findings supporting sperm displacement and demonstrate that average paternity for the last mate accords with the mean proportion of sperm stored. We further find consistent skew in storage across spermathecae, with more last male sperm stored in the singlet spermatheca on one side of the body than in the doublet on the opposite side. We also show that the time between copulations may be important for effectively sorting sperm. Finally, we demonstrate that male size may influence the opportunity for sperm choice, suggesting future work to disentangle the roles of male competition and cryptic female choice.

Experimental mixtures of dung fauna affect dung decomposition through complex effects of species interactions

Dung fauna plays an important role in dung decomposition, a key ecosystem process in nutrient cycling in grazed grasslands. The diversity of a three-species community (dung beetles, dung flies and epigeic earthworms) was systematically manipulated to produce different relative abundance distributions (evenness levels) and the resulting communities were introduced to standardised dung pats in laboratory experiments. Dung decomposition was modelled using an analysis that disentangled species identity effects and the interactions among species that contribute to the diversity effect. This revealed that the net diversity effect was composed of positive (fly–earthworm), negative (beetle–earthworm) and neutral (fly–beetle) effects of species interaction. These pairwise interactions resulted in complex, but systematically varying and predictable effects on dung decomposition as the relative abundances of species changed. Decomposition was consistently greater in communities with higher decomposer biomass. The utility of the adopted analytical approach was emphasised by comparison with an ANOVA that found that dung decomposition did not differ among evenness levels. Thus, the averaging of decomposition across different community structures within evenness levels masked the different effects of species interactions. These results highlight methodological insights into the quantification of diversity–function relationships.

Longevity and Developmental Stability in the Dung FlySepsis cynipsea,as Affected by the Ectoparasitic Mite,Pediculoides mesembrinae

Fluctuating asymmetry (FA) is a widely employed measure of developmental stability. It has been found to increase with many stressors including parasite infection. Associations between parasites and FA may exist for several reasons in addition to parasites being the direct cause of increased FA. Developmentally stable individuals may have superior immune systems, and be less susceptible to parasite infection, and/or may be less exposed to parasites than developmentally unstable ones. Mites negatively impact host fitness in a number of insects, and if FA is a reflection of general genetic quality, as has been proposed, associations between mite number and FA are predicted. Potential relationships were investigated between an ectoparasitic mite, Pediculoides mesembrinae (Canestrini) (Phthiraptera: Menoponidae) and FA in the common dung fly Sepsis cynipsea (L.) (Diptera: Sepsidae). While it was found that mite infested flies died much faster than flies without mites, indicating that mites indeed stress their hosts, counter to expectations, no associations between mites and FA were found in any analyses. Additionally, FA in mite-infected flies generally did not differ from previously published FA data from uninfected S. cynipsea. Nevertheless, parasitized males tended to be somewhat less asymmetrical than non-parasitized males, but based on our data, it does not appear that mite infestation is generally associated with developmental stability in S. cynipsea.

Sperm release and use at fertilization by yellow dung fly females (Scathophaga stercoraria)

Females of many species mate multiple times and store transferred sperm in storage organs. The mechanisms underlying sperm release from the stores at fertilization remain poorly understood, although they are central to an understanding of the female influence on post-copulatory male competition. Using double-mated females of the yellow dung fly, we counted the sperm sticking to the surface of deposited eggs of two successive clutches to obtain insight into the physiological processes associated with fertilization. The number of sperm released to fertilize an egg decreased between the first and second clutches, as well as within clutches from early to late eggs. These results indicate that: (1) sperm are lost from the stores over time independent of egg laying and (2) the number of sperm released depends on the amount of sperm stored. The lower number of sperm on eggs of the second clutches was accompanied by a strong increase of the proportion of sperm adhering to the micropyle region, suggesting that sperm use is more efficient and sperm release better controlled when sperm supply is substantially reduced. Finally, our approach indicates that sperm storage capacity of the female is higher than assumed from counts of spermathecal sperm.

Lethal and sublethal toxic effects of a test chemical (ivermectin) on the yellow dung fly (Scathophaga stercoraria) based on a standardized international ring test

A standardized bioassay using the yellow dung fly, Scathophaga stercoraria L. (Diptera: Scathophagidae), was developed to test the lethal and sublethal toxicity of parasiticide residues in livestock dung. The repeatability of the bioassay was assessed for the parasiticide ivermectin in 13 tests performed by seven laboratories in Germany, the United Kingdom, Switzerland, and Canada. Test results had an acceptable range of heterogeneity. The calculated median effective concentration for 50% (EC50) egg-to-adult mortality was 20.9 +/- 19.1 microg ivermectin/kg dung fresh weight (FW) (mean +/- standard deviation; range, 6.33-67.5 microg/kg). Mortality was not observed below a calculated no-observable-effect concentration (NOEC) of 8.1 +/- 7.7 microg/kg FW. However, prolonged development time (and, in a subset of tests, reduced body size) was observed above a calculated NOEC of 0.8 +/- 0.8 microg/kg FW. An oviposition site choice test revealed that yellow dung fly females do not discriminate among dung of different ivermectin concentrations. Thus, the yellow dung fly is suitably sensitive, and the methods are sufficiently repeatable, to support use of this standardized bioassay by the international community in the registration of new veterinary pharmaceuticals.

Response to selection and realized heritability of sperm length in the yellow dung fly (Scathophaga stercoraria)

Sperm length shows considerable phenotypic variation both inter- and intra-specifically, but a general explanation for this variation is lacking. In addition, our understanding of the genetic variation underlying sperm length variation is also limited because there have been few studies on the genetics of sperm size. One factor that could explain the variation in sperm length is that length influences sperm competitiveness, and there is some evidence for this. However, in yellow dung flies (Scathophaga stercoraria), microevolutionary responses to experimental variation at levels of sperm competition indicate that sperm length does not influence sperm competitiveness, although this lack of response may simply indicate sperm length lacks evolutionary potential (that is, it is constrained in some way), in spite of evidence that sperm length is heritable. Here we report on a laboratory study, in which we artificially selected upwards and downwards on sperm length in S. stercoraria. We found that sperm length significantly diverged after four generations of selection, but the response to selection was asymmetrical: upward selection generated a rapid response, but downward did not. We estimated the realized heritability of sperm length to be approximately 50%, which is consistent with previous sire-son estimates. We also assessed the fertility of males from upward and downward lines and found they did not differ. Results are discussed in the context of sperm competition.

CLEPTOPARASITISM AND DETRITIVORY IN DUNG BEETLE FOSSIL BROOD BALLS FROM PATAGONIA, ARGENTINA

Abstract: Traces within traces is a new ichnological field that is meant to shed light on significative palaeoecological aspects. Dung beetle fossil brood balls (Coprinisphaera ispp.), from the Middle Eocene – Lower Miocene Sarmiento Formation of Patagonia, Argentina, show two different trace fossils excavated in its infillings and/or wall that reveal the presence and relationships among different components of past dung communities. Tombownichnus pepei n. isp. is represented by elongated pits, circular to elliptical in cross‐section, occurring in the centre or beside ovoid mounds in the internal surface of the Coprinisphaera wall. These traces record the activity of cleptoparasites, such as other dung beetles or flies, whose larvae were probably carried passively with the dung for provisions. Tombownichnus pepei would represent the pupation chambers excavated by full grown larvae in the Coprinisphaera wall after completing their development inside provisioned dung. The other trace fossil, Lazaichnus fistulosus is represented by circular to subcircular borings occurring in Coprinisphaera walls, in connection with an internal gallery in their infillings. Its connection also with meniscate burrows and chambers in the surrounding palaeosol attributable to aestivation chambers of earthworms revealed that these organisms would have been active cleptoparasites or detritivores in dung beetle fossil brood balls.

Size-dependent alternative male mating tactics in the yellow dung fly, Scathophaga stercoraria

Whenever males can monopolize females and/or resources used by females, the opportunity for sexual selection will be great. The greater the variation among males in reproductive success, the greater the intensity of selection on less competitive males to gain matings through alternative tactics. In the yellow dung fly, Scathophaga stercoraria, males aggressively compete for access to receptive, gravid females on fresh dung. Larger males are better able to acquire mates and to complete copulation successfully and guard the female throughout oviposition. Here we demonstrate that when an alternative resource is present where females aggregate (i.e. apple pomace, where both sexes come to feed), smaller males will redirect their searching for females from dung to the new substrate. In addition, we identify a class of particularly small males on the alternative substrate that appears never to be present searching for females on or around dung. Smaller males were found to have a mating 'advantage' on pomace, in striking contrast to the pattern observed on dung, providing further support for the existence of an alternative male reproductive tactic in this species.