Temperature-dependent melanism and phenoloxidase activity in the dimorphic sepsid fly Sepsis thoracica.

Climate is changing towards both higher average temperatures and more frequent and severe heat waves. Whereas numerous studies have investigated temperature effects on animal life histories, assessments of their immune function are limited. In the size- and colour-dimorphic black scavenger (or dung) fly Sepsis thoracica (Diptera: Sepsidae), we experimentally studied how developmental temperature and larval density influence phenoloxidase (PO) activity, a key enzyme in insect pigmentation, thermoregulation, and immunity. Flies from five latitudinal European populations were raised at three developmental temperatures (18, 24, 30 °C). PO activity increased with developmental temperature differently in the sexes and the two male morphs (black and orange), altering the sigmoid relationship between melanism, i.e. colouration and fly size. PO activity further positively correlated with larval rearing density, potentially because of higher risks of pathogen infection or greater developmental stress following stronger resource competition. Populations varied somewhat in PO activity, body size and colouration, however with no clear latitudinal pattern. Overall our results indicate that morph- and sex-specific PO activity, and thus likely immune function, in S. thoracica depends on temperature and larval density, modifying the underlying putative trade-off between immunity and body size. The strong dampening of the immune system of all morphs at cool temperatures suggests low-temperature stress in this warm-adapted species common in southern Europe. Our results also support the population density dependent prophylaxis hypothesis, which predicts higher investment in immunity when facing limited resource availability and increased pathogen infection probability.

Prediction and enrichment analyses of the Homo sapiens-Drosophila melanogaster COPD-related orthologs: potential for modeling of human COPD genomic responses with the fruit fly.

The significant similarities in airway epithelial cells between mammals and the fruit fly Drosophila melanogaster have rendered the latter an important model organism for studies of chronic inflammatory lung diseases. Focusing on the chronic obstructive pulmonary disease (COPD), we here mapped human gene orthologs associated with this disease in D. melanogaster to identify functionally equivalent genes for immediate, further screening with the fruit fly model. The DIOPT-DIST tool was accessed for the prediction of the COPD-associated orthologs between humans and Drosophila. Enrichment analyses with respect to pathways of the retrieved functional homologs were performed using the ToppFun and FlyMine tools, identifying 73 unique human genes as well as 438 fruit fly genes. The ToppFun analysis verified that the human gene list is associated with COPD phenotypes. Furthermore, the FlyMine investigation highlighted that the Drosophila genes are functionally connected mainly with the "ABC-family proteins mediated transport" and the "ß-catenin-independent WNT signaling pathway." These results suggest an evolutionarily conserved role toward responses to inhaled toxicants and CO2 in both species. We reason that the predicted orthologous genes should be further studied in the Drosophila models of cigarette smoke-induced COPD.

Comparative reproductive dormancy differentiation in European black scavenger flies (Diptera: Sepsidae).

Seasonality is a key environmental factor that regularly promotes life history adaptation. Insects invading cold-temperate climates need to overwinter in a dormant state. We compared the role of temperature and photoperiod in dormancy induction in the laboratory, as well as winter survival and reproduction in the field and the laboratory, of 5 widespread European dung fly species (Diptera: Sepsidae) to investigate their extent of ecological differentiation and thermal adaptation. Unexpectedly, cold temperature is the primary environmental factor inducing winter dormancy, with short photoperiod playing an additional role mainly in species common at high altitudes and latitudes (Sepsis cynipsea, neocynipsea, fulgens), but not in those species also thriving in southern Europe (thoracica, punctum). All species hibernate as adults rather than juveniles. S. thoracica had very low adult winter survivorship under both (benign) laboratory and (harsh) field conditions, suggesting flexible quiescence rather than genetically fixed winter diapause, restricting their distribution towards the pole. All other species appear well suited for surviving cold, Nordic winters. Females born early in the season reproduce before winter while late-born females reproduce after winter, fulgens transitioning earliest before winter and thoracica and punctum latest; a bet-hedging strategy of reproduction during both seasons occurs rarely but is possible physiologically. Fertility patterns indicate that females can store sperm over winter. Winter dormancy induction mechanisms of European sepsids are congruent with their geographic distribution, co-defining their thermal niches. Flexible adult winter quiescence appears the easiest route for insects spreading towards the poles to evolve the necessary overwinter survival.

Interplay between male quality and male-female compatibility across episodes of sexual selection.

The processes underlying mate choice profoundly influence the dynamics of sexual selection and the evolution of male sexual traits. Consistent preference for certain phenotypes may erode genetic variation in populations through directional selection, whereas divergent preferences (e.g., genetically compatible mates) provide one mechanism to maintain such variation. However, the relative contributions of these processes across episodes of selection remain unknown. Using Drosophila melanogaster, we followed the fate of male genotypes, previously scored for their overall reproductive value and their compatibility with different female genotypes, across pre- and postmating episodes of selection. When pairs of competitor males differed in their intrinsic quality and their compatibility with the female, both factors influenced outcomes from mating success to paternity but to a varying degree between stages. These results add further dimensions to our understanding of how the interactions between genotypes and forms of selection shape reproductive outcomes and ultimately reproductive trait evolution.

The effect of cigarette smoke extract exposure on the size and sexual behaviour of Drosophila melanogaster.

Drosophila melanogaster is a widely used animal model in human diseases and to date it has not been applied to the study of the impact of tobacco use on human sexual function. Hence, this report examines the effects of different concentrations of cigarette smoke extract (CSE) exposure on the size and sexual behavior of D. melanogaster. Wild-type flies were held in vials containing CSE-infused culture media at concentrations of 10%, 25%, and 50% for three days, and their offspring were reared under the same conditions before measuring their body size and mating behavior. CSE exposure during development reduced the tibia length and body mass of emerging adult flies and prolonged the time required for successful courtship copulation success, while courtship behaviors (wing extension, tapping, abdomen bending, attempted copulation) remained largely unchanged. Our findings indicate that CSE exposure negatively affects the development of flies and their subsequent reproductive success. Future experiments should investigate the CSE effect on male female fertility.

The Drosophila septate junctions beyond barrier function: Review of the literature, prediction of human orthologs of the SJ-related proteins and identification of protein domain families.

The involvement of Septate Junctions (SJs) in critical cellular functions that extend beyond their role as diffusion barriers in the epithelia and the nervous system has made the fruit fly an ideal model for the study of human diseases associated with impaired Tight Junction (TJ) function. In this study, we summarized current knowledge of the Drosophila melanogaster SJ-related proteins, focusing on their unconventional functions. Additionally, we sought to identify human orthologs of the corresponding genes as well as protein domain families. The systematic literature search was performed in PubMed and Scopus databases using relevant key terms. Orthologs were predicted using the DIOPT tool and aligned protein regions were determined from the Pfam database. 3-D models of the smooth SJ proteins were built on the Phyre2 and DMPFold protein structure prediction servers. A total of 30 proteins were identified as relatives to the SJ cellular structure. Key roles of these proteins, mainly in the regulation of morphogenetic events and cellular signalling, were highlighted. The investigation of protein domain families revealed that the SJ-related proteins contain conserved domains that are required not only for cell-cell interactions and cell polarity but also for cellular signalling and immunity. DIOPT analysis of orthologs identified novel human genes as putative functional homologs of the fruit fly SJ genes. A gap in our knowledge was identified regarding the domains that occur in the proteins encoded by eight SJ-associated genes. Future investigation of these domains is needed to provide functional information.

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.