Resistant Flies Research Articles

Functional Characterization of Carboxylesterases in Insecticide Resistant House Flies, Musca Domestica.

Carboxylesterase-mediated metabolism is thought to play a major role in insecticide resistance in various insects. Several carboxylesterase genes were found up-regulated in the resistant house fly strain, whereas their roles in conferring insecticide resistance remained to be explored. Here, we designed a protocol for the functional characterization of carboxylesterases. Three example experiments are presented: (1) expression and isolation of carboxylesterase proteins through a baculovirus-mediated insect Spodoptera frugiperda (Sf9) cell expression system; (2) a cell-based MTT (3-[4, 5-dimethykthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) cytotoxicity assay to measure the tolerance of insect cells to different permethrin treatments; and (3) in vitro metabolic studies to explore the metabolic capabilities of carboxylesterases toward permethrin. The carboxylesterase gene MdαE7 was cloned from a resistant house fly strain ALHF and used to construct a recombinant baculovirus for Sf9 cells infection. The cell viabilities against different permethrin treatments were measured with the MTT assay. The enhanced cell tolerance of the experimental group (MdαE7-recombinant baculovirus infected cells) compared with those of the control groups (CAT-recombinant baculovirus infected cells and GFP-recombinant baculovirus infected cells) to permethrin treatments suggested the capabilities of MdαE7 in metabolizing insecticides, thereby protecting cells from chemical damages. Besides that, carboxylesterase proteins were expressed in insect Sf9 cells and isolated to conduct an in vitro metabolic study. Our results indicated a significant in vitro metabolic efficiency of MdαE7 toward permethrin, directly indicating the involvement of carboxylesterases in metabolizing insecticides and thus conferring insecticide resistance in house flies.

Modulation of caveolae by insulin/IGF-1 signaling regulates aging of Caenorhabditis elegans.

Reducing insulin/IGF-1 signaling (IIS) extends lifespan, promotes protein homeostasis (proteostasis), and elevates stress resistance of worms, flies, and mammals. How these functions are orchestrated across the organism is only partially understood. Here, we report that in the nematode Caenorhabditis elegans, the IIS positively regulates the expression of caveolin-1 (cav-1), a gene which is primarily expressed in neurons of the adult worm and underlies the formation of caveolae, a subtype of lipid microdomains that serve as platforms for signaling complexes. Accordingly, IIS reduction lowers cav-1 expression and lessens the quantity of neuronal caveolae. Reduced cav-1 expression extends lifespan and mitigates toxic protein aggregation by modulating the expression of aging-regulating and signaling-promoting genes. Our findings define caveolae as aging-governing signaling centers and underscore the potential for cav-1 as a novel therapeutic target for the promotion of healthy aging.

Stress resistance and lifespan enhanced by downregulation of antimicrobial peptide genes in the Imd pathway.

Biological behaviors and longevity of ectothermic animals are remarkably influenced by ambient temperature. Development at 18°C significantly enhances the stress resistance of adult flies with more accumulation of nutrients (especially fat) in the body than development at 25°C. Gene expression analysis between the flies developed at 18°C and 25°C revealed that the Immune deficiency (Imd) pathway, including the downstream antimicrobial peptides (AMPs), is downregulated in the flies developed at 18°C. When hypomorphic imd mutant flies with reduced AMP expressions were developed at 25°C, they showed induced stress resistance with higher fat content in the body similar to the wild-type flies developed at 18°C. However, severe hypomorphic imd mutants could not enhance stress resistance due to the downregulation of another downstream JNK pathway that expresses stress tolerance genes. Interestingly, the downregulation of AMP genes, itself, extended lifespan with increased stress resistance. Especially, fat body-specific downregulation of Imd AMP genes exhibited a longer lifespan with higher heat resistance. The fat body is known to function in metabolic homeostasis, stress tolerance, growth, and longevity in Drosophila. Here, we provide the first evidence that mild downregulation of the Imd pathway with AMP genes increases fat content, stress resistance, and lifespan in adult flies.

Molecular characterization of pyrethroid resistance in the olive fruit fly Bactrocera oleae

Α reduction of pyrethroid efficacy has been recently recorded in Bactrocera oleae, the most destructive insect of olives. The resistance levels of field populations collected from Crete-Greece scaled up to 22-folds, compared to reference laboratory strains. Sequence analysis of the IIS4–IIS6 region of para sodium channel gene in a large number of resistant flies indicated that resistance may not be associated with target site mutations, in line with previous studies in other Tephritidae species. We analyzed the transcriptomic differences between two resistant populations versus an almost susceptible field population and two laboratory strains. A large number of genes was found to be significantly differentially transcribed across the pairwise comparisons. Interestingly, gene set analysis revealed that genes of the ‘electron carrier activity’ GO group were enriched in one specific comparison, which might suggest a P450-mediated resistance mechanism. The up-regulation of several transcripts encoding detoxification enzymes was qPCR validated, focusing on transcripts coding for P450s. Of note, the expression of contig00436 and contig02103, encoding CYP6 P450s, was significantly higher in all resistant populations, compared to susceptible ones. These results suggest that an increase in the amount of the CYP6 P450s might be an important mechanism of pyrethroid resistance in B. oleae.

Geographic variation in responses of European yellow dung flies to thermal stress

Climatic conditions can be very heterogeneous even over small geographic scales, and are believed to be major determinants of the abundance and distribution of species and populations. Organisms are expected to evolve in response to the frequency and magnitude of local thermal extremes, resulting in local adaptation. Using replicate yellow dung fly (Scathophaga stercoraria; Diptera: Scathophagidae) populations from cold (northern Europe) and warm climates (southern Europe), we compared 1) responses to short-term heat and cold shocks in both sexes, 2) heat shock protein (Hsp70) expression in adults and eggs, and 3) female reproductive traits when facing short-term heat stress during egg maturation. Contrary to expectations, thermal traits showed minor geographic differentiation, with weak evidence for greater heat resistance of southern flies but no differentiation in cold resistance. Hsp70 protein expression was little affected by heat stress, indicating systemic rather than induced regulation of the heat stress response, possibly related to this fly group's preference for cold climes. In contrast, sex differences were pronounced: males (which are larger) endured hot temperatures longer, while females featured higher Hsp70 expression. Heat stress negatively affected various female reproductive traits, reducing first clutch size, overall reproductive investment, egg lipid content, and subsequent larval hatching. These responses varied little across latitude but somewhat among populations in terms of egg size, protein content, and larval hatching success. Several reproductive parameters, but not Hsp70 expression, exhibited heritable variation among full-sib families. Rather than large-scale clinal geographic variation, our study suggests some local geographic population differentiation in the ability of yellow dung flies to buffer the impact of heat stress on reproductive performance.

Oxidation of SQSTM1/p62 mediates the link between redox state and protein homeostasis

Cellular homoeostatic pathways such as macroautophagy (hereinafter autophagy) are regulated by basic mechanisms that are conserved throughout the eukaryotic kingdom. However, it remains poorly understood how these mechanisms further evolved in higher organisms. Here we describe a modification in the autophagy pathway in vertebrates, which promotes its activity in response to oxidative stress. We have identified two oxidation-sensitive cysteine residues in a prototypic autophagy receptor SQSTM1/p62, which allow activation of pro-survival autophagy in stress conditions. The Drosophila p62 homologue, Ref(2)P, lacks these oxidation-sensitive cysteine residues and their introduction into the protein increases protein turnover and stress resistance of flies, whereas perturbation of p62 oxidation in humans may result in age-related pathology. We propose that the redox-sensitivity of p62 may have evolved in vertebrates as a mechanism that allows activation of autophagy in response to oxidative stress to maintain cellular homoeostasis and increase cell survival.

Heat- and humidity-induced plastic changes in body lipids and starvation resistance in the tropical fly Zaprionus indianus during wet and dry seasons.

Insects in tropical wet or dry seasons are likely to cope with starvation stress through plastic changes (developmental as well as adult acclimation) in energy metabolites. Control and experimental groups of Zaprionus indianus flies were reared under wet or dry conditions, but adults were acclimated at different thermal or humidity conditions. Adult flies of the control group were acclimated at 27°C and low (50%) or high (60%) relative humidity (RH). For experimental groups, adult flies were acclimated at 32°C for 1 to 6 days and under low (40%) or high (70%) RH. For humidity acclimation, adult flies were acclimated at 27°C but under low (40%) or high (70%) RH for 1 to 6 days. Plastic changes in experimental groups as compared with the control group (developmental as well as adult acclimation) revealed significant accumulation of body lipids owing to thermal or humidity acclimation of wet season flies, but low humidity acclimation did not change the level of body lipids in dry season flies. Starvation resistance and body lipids were higher in the males of dry season flies but in the females of wet season flies. Adults acclimated under different thermal or humidity conditions exhibited changes in the rate of utilization of body lipids, carbohydrates and proteins. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity; and a reduction in fecundity under starvation. Thus, thermal or humidity acclimation of adults revealed plastic changes in energy metabolites to support starvation resistance of wet or dry season flies.

Host plant accessions determine bottom-up effect of snapmelon (Cucumis melo var. momordica) against melon fly (Bactrocera cucurbitae (Coquillett)).

The melon fly, Bactrocera cucurbitae (Tephritidae: Diptera) is an important pest of snapmelon (Cucumis melo var. momordica), leading to significant losses in yield in the hot arid agro-climate of India. The accessions IC- 430190 (11.21%), DKS-AHS 2011/4 (14.97%) and DKS-AHS 2011/3 (18.57%) were found to be novel resistant accessions against melon fly, B. cucurbitae infestation. Free amino acid and total soluble solid (TSS) were in positive correlation with percent fruit infestation whereas phenols, tannin, total alkaloids and flavonoid contents had significant negative correlation with percent fruit infestation. The percent fruit infestation had significant positive correlation with fruit length, fruit diameter and flesh thickness and negative correlation with length of ovary pubescence, rind hardness at immature stage, rind hardness at mature stage and pericarp thickness. Based on Kaiser Normalization method, two principal components (PCs) were extracted explaining cumulative variation of 82.80% in melon fly infestation. PC1 explained 53.41% of the variation while PC2 explained 29.39% of variation. The flavonoid, total alkaloid, tannins, phenols content, length of ovary pubescence and rind hardness were the novel antibiosis and antixenotic characters found in snapmelon resistant melon fly, B. cucurbitae and therefore, could be used as marker traits in plant breeding programs to select resistant accessions.

Carboxylesterase genes in pyrethroid resistant house flies, Musca domestica

Carboxylesterases are one of the major enzyme families involved in the detoxification of pyrethroids. Up-regulation of carboxylesterase genes is thought to be a major component of insecticide resistant mechanisms in insects. Based on the house fly transcriptome and genome database, a total of 39 carboxylesterase genes of different functional clades have been identified in house flies. In this study, eleven of these genes were found to be significantly overexpressed in the resistant ALHF house fly strain compared with susceptible aabys and wild-type CS strains. Eight up-regulated carboxylesterase genes with their expression levels were further induced to a higher level in response to permethrin treatments, indicating that constitutive and inductive overexpression of carboxylesterases are co-responsible for the enhanced detoxification of insecticides. Spatial expression studies revealed these up-regulated genes to be abundantly distributed in fat bodies and genetically mapped on autosome 2 or 3 of house flies, and their expression could be regulated by factors on autosome 1, 2 and 5. Taken together, these results demonstrate that multiple carboxylesterase genes are co-upregulated in resistant house flies, providing further evidence for their involvement in the detoxification of insecticides and development of insecticide resistance.

Resistance to pyrethroid insecticides in house flies, Musca domestica L., (Diptera: Muscidae) collected from urban areas in Punjab, Pakistan.

House flies are one of the major public health pests in urban settings. People usually use insecticides containing pyrethroids for the management of house flies; however, there is a lack of information on pyrethroid resistance in house flies from urban areas. In the present study, resistance to four pyrethroids (beta-cyfluthrin, deltamethrin, permethrin, transfluthrin) was assessed in house flies collected from urban areas of Punjab, Pakistan. Significant levels of resistance to all the pyrethroids were found in different strains of house flies. The resistance ratios (RRs) at the median lethal dose (LD50) level were in the range of 5.25- to 11.02-fold for beta-cyfluthrin, 7.22- to 19.31-fold for deltamethrin, 5.36- to 16.04-fold for permethrin, and 9.05- to 35.50-fold for transfluthrin. Pairwise comparison of the log LD50s revealed a highly significant correlation (p<0.01) between deltamethrin and permethrin, suggesting the possibility of a cross-resistance mechanism. The results revealed the occurrence of pyrethroid resistance in house flies from urban areas of Punjab. Regular resistance monitoring surveys and integrated approaches for the management of house flies are needed to retain the efficacy of these insecticides for a longer period of time.

Two novel house fly Vssc mutations, D600N and T929I, give rise to new insecticide resistance alleles

The house fly, Musca domestica, is a serious pest because it transmits a large diversity of human and veterinary diseases. Insecticides, particularly pyrethroids, are commonly used to control house flies. However, the evolution of pyrethroid resistance has reduced the effectiveness of these insecticides. A major mechanism of resistance to pyrethroids is target site insensitivity caused by the mutations in the voltage-sensitive sodium channel (Vssc) gene (e.g. kdr [L1014F] and super-kdr [M918T+L1014F]). Recently, two novel Vssc alleles, super-kdr+D600N and kdr+T929I were detected in a field collected resistant house fly population in Kansas, USA in 2013. To determine the levels of resistance that these new alleles confer to pyrethroids, we isolated strains having the unique Vssc alleles, but being otherwise congenic to the susceptible strain, aabys. We compared levels of resistance conferred to 14 pyrethroids and determined the inheritance of resistance to 8 pyrethroids. Our results revealed that super-kdr+D600N conferred higher levels of resistance to seven pyrethroids relative to super-kdr, and kdr+T929I showed super-kdr-like levels of resistance in house flies. Our results are compared with previous studies and reveal that addition of T929I to the kdr mutation (L1014F) increased resistance to all pyrethroids (except etofenprox), and enhanced resistance by ~1000-fold to acrinathrin and flumethrin. The implications of these results on the evolution of resistance are discussed.

Correlation between desiccation stress response and epigenetic modifications of genes in Drosophila melanogaster: An example of environment-epigenome interaction

Animals from different phyla including arthropods tolerate water stress to different extent. This tolerance is accompanied by biochemical changes which in turn are due to transcriptional alteration. The changes in transcription can be an indirect effect on some of the genes, ensuing from the effect of stress on the regulators of transcription including epigenetic regulators. Within this paradigm, we investigated the correlation between stress response and epigenetic modification underlying gene expression modulation during desiccation stress in Canton-S. We report altered resistance of flies in desiccation stress for heterozygote mutants of PcG and TrxG members. Pc/+ mutant shows lower survival, while ash1/+ mutants show higher survival under desiccation stress as compared to Canton-S. We detect expression alteration in stress related genes as well the genes of the Polycomb and trithorax complex in Canton-S subjected to desiccation stress. Concomitant with this, there is an altered enrichment of H3K27me3 and H3K4me3 at the upstream regions of the stress responsive genes. The enrichment of activating mark, H3K4me3, is higher in non-stress condition. H3K27me3, the repressive mark, is more pronounced under stress condition, which in turn, can be correlated with the binding of Pc. Our results show that desiccation stress induces dynamic switching in expression and enrichment of PcG and TrxG in the upstream region of genes, which correlates with histone modifications. We provide evidence that epigenetic modulation could be one of the mechanisms to adapt to the desiccation stress in Drosophila. Thus, our study proposes the interaction of epigenome and environmental factors.

Sand Flies and Their Control Methods

The main aim of managing arthropod vectors that carry the disease agents is interrupting the infection cycle. Therefore, the management of the disease implies that all precautions related to all elements (i.e., human, arthropod vector, and reservoir) in the infection cycle need to be taken. There are important points that need to be considered while dealing with sand flies (Diptera: Psychodidae: Phlebotominae), which in many regions worldwide, particularly in tropical and subtropical areas, are vectors of diseases such as leishmaniasis and sand fly fever and are the arthropods of the infection cycle. Because the larval control of the sand flies is very difficult and almost impossible, the management is mainly conducted for the adults. The most effective strategy for reducing both sand fly fever and leishmaniasis is managing sand flies, particularly in areas where humans are located. In this review, the morphology, biology, and taxonomy of sand flies; the integrated fighting and management methods such as insecticide-impregnated bed nets and use of curtains, zooprophylaxis, indoor and outdoor residual applications, larvicides, repellents, and insecticide-impregnated dog collars; and data regarding many issues such as insecticide resistance in sand flies have been emphasized on in the review.

Analysis of Differentially Expressed Genes Related to Resistance in Spinosad- and Neonicotinoid-Resistant Musca domestica L. (Diptera: Muscidae) Strains.

BackgroundThe housefly is a global pest that has developed resistance to most insecticides applied against it. Resistance of the spinosad-resistant strain 791spin and the neonicotinoid-resistant 766b strain is believed to be due to metabolism. We investigate differentially expressed genes in these two resistant strains related to metabolism in comparison with an insecticide-susceptible reference strain.ResultsGenes involved in metabolism of xenobiotics were primarily up-regulated in resistant flies with some differences between resistant strains. The cyp4g98 and cyp6g4 genes proved interesting in terms of neonicotinoid resistance, while cyp4d9 was overexpressed in 791spin compared to spinosad-susceptible strains. GSTs, ESTs and UGTs were mostly overexpressed, but not to the same degree as P450s. We present a comprehensive and comparative picture of gene expression in three housefly strains differing significantly in their response to insecticides. High differential expression of P450s and genes coding for cuticle protein indicates a combination of factors involved in metabolic neonicotinoid and spinosad resistance.ConclusionResistance in these strains is apparently not linked to the alteration of a single gene but is composed of several changes including differential expression of genes encoding metabolic detoxification enzymes.

Studying DDT Susceptibility at Discriminating Time Intervals Focusing on Maximum Limit of Exposure Time Survived by DDT Resistant Phlebotomus argentipes (Diptera: Psychodidae): an Investigative Report.

Extensive application of routine insecticide i.e., dichlorodiphenyltrichloroethane (DDT) to control Phlebotomus argentipes (Diptera: Psychodidae), the proven vector of visceral leishmaniasis in India, had evoked the problem of resistance/tolerance against DDT, eventually nullifying the DDT dependent strategies to control this vector. Because tolerating an hour-long exposure to DDT is not challenging enough for the resistant P. argentipes, estimating susceptibility by exposing sand flies to insecticide for just an hour becomes a trivial and futile task.Therefore, this bioassay study was carried out to investigate the maximum limit of exposure time to which DDT resistant P. argentipes can endure the effect of DDT for their survival. The mortality rate of laboratory-reared DDT resistant strain P. argentipes exposed to DDT was studied at discriminating time intervals of 60 min and it was concluded that highly resistant sand flies could withstand up to 420 min of exposure to this insecticide. Additionally, the lethal time for female P. argentipes was observed to be higher than for males suggesting that they are highly resistant to DDT's toxicity. Our results support the monitoring of tolerance limit with respect to time and hence points towards an urgent need to change the World Health Organization's protocol for susceptibility identification in resistant P. argentipes.

Activation of the Tor/Myc signaling axis in intestinal stem and progenitor cells affects longevity, stress resistance and metabolism in drosophila.

The TOR (target of rapamycin) signaling pathway and the transcriptional factor Myc play important roles in growth control. Myc acts, in part, as a downstream target of TOR to regulate the activity and functioning of stem cells. Here we explore the role of TOR-Myc axis in stem and progenitor cells in the regulation of lifespan, stress resistance and metabolism in Drosophila. We found that both overexpression of rheb and myc-rheb in midgut stem and progenitor cells decreased the lifespan and starvation resistance of flies. TOR activation caused higher survival under malnutrition conditions. Furthermore, we demonstrate gut-specific activation of JAK/STAT and insulin signaling pathways to control gut integrity. Both genetic manipulations had an impact on carbohydrate metabolism and transcriptional levels of metabolic genes. Our findings indicate that activation of the TOR-Myc axis in midgut stem and progenitor cells influences a variety of traits in Drosophila.

Pyrethroid Insecticide Resistance Mechanisms in the Adult Phlebotomus papatasi (Diptera: Psychodidae).

Phlebotomus papatasi is one of the most medically important sand fly species in the Old World, serving as a vector of Leishmania parasites and phleboviruses. Chemical control is still considered the most effective method for rapidly reducing populations of flying insects involved in vector-borne disease transmission, but is increasingly threatened by insecticide resistance in the target insect posing significant problems for entomologists responsible for control programs. This study was conducted to determine pyrethroid resistance mechanisms and the biological, physiological, and molecular impacts of resistance in Ph. papatasi, and to compare their resistance mechanisms against those reported for mosquitoes and other intensely studied dipterans. Field-collected Ph. papatasi from Aswan, Egypt, were subjected to sublethal doses of permethrin and reared as a resistant strain under laboratory conditions through 16 generations. Biological parameter observations of resistant Ph. papatasi revealed an association of resistance with productivity cost. Physiological analysis revealed that concentrations of oxidase and esterase enzymes increased in early generations of the resistant colony, and then subsided through the F16 generation to levels similar to those in a susceptible colony. The activity levels of acetylcholinesterase were higher in field-collected Ph. papatasi than in susceptible colony flies, but decreased significantly despite subsequent exposure to permethrin. The molecular search for gene mutations in the resistant strain of Ph. papatasi failed to identify any mutations common in pyrethroid-resistant mosquitoes. Our study revealed that the mechanism of pyrethroid resistance in sand flies is different than that in mosquitoes, at least at the genetic level.

Nuclear hormone receptor DHR96 mediates the resistance to xenobiotics but not the increased lifespan of insulin-mutant Drosophila

Lifespan of laboratory animals can be increased by genetic, pharmacological, and dietary interventions. Increased expression of genes involved in xenobiotic metabolism, together with resistance to xenobiotics, are frequent correlates of lifespan extension in the nematode worm Caenorhabditis elegans, the fruit fly Drosophila, and mice. The Green Theory of Aging suggests that this association is causal, with the ability of cells to rid themselves of lipophilic toxins limiting normal lifespan. To test this idea, we experimentally increased resistance of Drosophila to the xenobiotic dichlordiphenyltrichlorethan (DDT), by artificial selection or by transgenic expression of a gene encoding a cytochrome P450. Although both interventions increased DDT resistance, neither increased lifespan. Furthermore, dietary restriction increased lifespan without increasing xenobiotic resistance, confirming that the two traits can be uncoupled. Reduced activity of the insulin/Igf signaling (IIS) pathway increases resistance to xenobiotics and extends lifespan in Drosophila, and can also increase longevity in C. elegans, mice, and possibly humans. We identified a nuclear hormone receptor, DHR96, as an essential mediator of the increased xenobiotic resistance of IIS mutant flies. However, the IIS mutants remained long-lived in the absence of DHR96 and the xenobiotic resistance that it conferred. Thus, in Drosophila IIS mutants, increased xenobiotic resistance and enhanced longevity are not causally connected. The frequent co-occurrence of the two traits may instead have evolved because, in nature, lowered IIS can signal the presence of pathogens. It will be important to determine whether enhanced xenobiotic metabolism is also a correlated, rather than a causal, trait in long-lived mice.

Identification of Drosophila Zfh2 as a Mediator of Hypercapnic Immune Regulation by a Genome-Wide RNA Interference Screen.

Hypercapnia, elevated partial pressure of CO2 in blood and tissue, develops in many patients with chronic severe obstructive pulmonary disease and other advanced lung disorders. Patients with advanced disease frequently develop bacterial lung infections, and hypercapnia is a risk factor for mortality in such individuals. We previously demonstrated that hypercapnia suppresses induction of NF-κB-regulated innate immune response genes required for host defense in human, mouse, and Drosophila cells, and it increases mortality from bacterial infections in both mice and Drosophila. However, the molecular mediators of hypercapnic immune suppression are undefined. In this study, we report a genome-wide RNA interference screen in Drosophila S2* cells stimulated with bacterial peptidoglycan. The screen identified 16 genes with human orthologs whose knockdown reduced hypercapnic suppression of the gene encoding the antimicrobial peptide Diptericin (Dipt), but did not increase Dipt mRNA levels in air. In vivo tests of one of the strongest screen hits, zinc finger homeodomain 2 (Zfh2; mammalian orthologs ZFHX3/ATBF1 and ZFHX4), demonstrate that reducing zfh2 function using a mutation or RNA interference improves survival of flies exposed to elevated CO2 and infected with Staphylococcus aureus. Tissue-specific knockdown of zfh2 in the fat body, the major immune and metabolic organ of the fly, mitigates hypercapnia-induced reductions in Dipt and other antimicrobial peptides and improves resistance of CO2-exposed flies to infection. Zfh2 mutations also partially rescue hypercapnia-induced delays in egg hatching, suggesting that Zfh2's role in mediating responses to hypercapnia extends beyond the immune system. Taken together, to our knowledge, these results identify Zfh2 as the first in vivo mediator of hypercapnic immune suppression.

Effects of the isoflavone prunetin on gut health and stress response in male Drosophila melanogaster

The traditional Asian diet is rich in fruits, vegetables and soy, the latter representing a significant source of dietary isoflavones. The isoflavone prunetin was recently identified to improve intestinal epithelial barrier function in vitro and to ameliorate general survival and overall health state in vivo in male Drosophila melanogaster. However, the prunetin-mediated health benefits in the fruit fly were ascertained under standard living conditions. As the loss of intestinal integrity is closely related to a reduction in Drosophila lifespan and barrier dysfunction increases with age, effects on prunetin-modulated gut health under oxidative or pathogenic stress provocation remain to be elucidated. In this study, male adult D. melanogaster were administered either a prunetin or a control diet. Gut-derived junction protein expression and pathogen-induced antimicrobial peptide expressions as well as the stem cell proliferation in the gut were evaluated. Furthermore, survival following exposure to hydrogen peroxide was assessed. Prunetin ingestion did not attenuate bacterial infection and did not protect flies from oxidative stress. Intestinal mRNA expression levels of adherence and septate junction proteins as well as the stem cell proliferation were not altered by prunetin intake. Prunetin does not improve the resistance of flies against severe injuring, exogenous stress and therefore seems to function in a preventive rather than a therapeutic approach since the health-promoting benefits appear to be exclusively restricted to normal living circumstances.