Seasonal changes in humidity impact drought resistance in tropical Drosophila leontia: testing developmental effects of thermal versus humidity changes.

Drosophila leontia is native to highly humid equatorial tropical habitats but its desiccation sensitivity (~10h) is not consistent with its abundance during the drier autumn season in the subtropical regions. We have tested the effects of developmental acclimation on desiccation resistance and water balance related traits of D. leontia collected during rainy and autumn seasons. The isofemale lines of seasonal populations were reared under ecologically relevant growth temperatures (18 or 26 °C) or humidity conditions (35 or 85% RH) but tested at different times under identical experimental conditions. The larvae as well as flies reared under two thermal conditions (18 or 26 °C) showed no effect on desiccation related traits as well as storage and utilization of energy metabolites. In contrast, for D. leontia reared under low humidity (35% RH), significant changes at larval as well adult stages include increase in the desiccation resistance as well as cuticular lipid quantity, reduced levels of rate of body water loss, higher storage of carbohydrates but lower rate of utilization of carbohydrates as compared with flies reared at high humidity (85% RH). D. leontia has responded to rearing under low humidity conditions by increasing its desiccation resistance but not due to changes in the growth temperatures. These laboratory observations on seasonal populations highlight differences due to rearing conditions but not due to seasons. Further, direct analysis of wild-caught seasonal populations has shown trends similar to developmental acclimation effects. For wild caught flies, there are significant seasonal differences i.e. higher desiccation resistance as well as cuticular lipid quantity but reduced rate of water loss for autumn than rainy season flies. Thus, our laboratory observations are relevant for understanding seasonal adaptations of natural populations of tropical D. leontia to wet-dry conditions in the wild.

Developmental acclimation to low or high humidity conditions affect starvation and heat resistance of Drosophila melanogaster.

Several Drosophila species originating from tropical humid localities are more resistant to starvation and heat stress than populations from high latitudes but mechanistic bases of such physiological changes are largely unknown. In order to test whether humidity levels affect starvation and heat resistance, we investigated developmental acclimation effects of low to high humidity conditions on the storage and utilization of energy resources, body mass, starvation survival, heat knockdown and heat survival of D. melanogaster. Isofemale lines reared under higher humidity (85% RH) stored significantly higher level of lipids and showed greater starvation survival hours but smaller in body size. In contrast, lines reared at low humidity evidenced reduced levels of body lipids and starvation resistance. Starvation resistance and lipid storage level were higher in females than males. However, the rate of utilization of lipids under starvation stress was lower for lines reared under higher humidity. Adult flies of lines reared at 65% RH and acclimated under high or low humidity condition for 200 hours also showed changes in resistance to starvation and heat but such effects were significantly lower as compared with developmental acclimation. Isofemale lines reared under higher humidity showed greater heat knockdown time and heat-shock survival. These laboratory observations on developmental and adult acclimation effects of low versus high humidity conditions have helped in explaining seasonal changes in resistance to starvation and heat of the wild-caught flies of D. melanogaster. Thus, we may suggest that wet versus drier conditions significantly affect starvation and heat resistance of D. melanogaster.

Sex-specific divergence for adaptations to dehydration stress in Drosophila kikkawai.

Several studies on diverse Drosophila species have reported higher desiccation resistance of females, but the physiological basis of such sex-specific differences has received less attention. We tested whether sex-specific differences in cuticular traits (melanic females and non-melanic males) of Drosophila kikkawai correspond with divergence in their water balance mechanisms. Our results are interesting in several respects. First, positive clinal variation in desiccation resistance was correlated with cuticular melanisation in females but with changes in cuticular lipid mass in males, despite a lack of differences between the sexes for the rate of water loss. Second, a comparative analysis of water budget showed that females of the northern population stored more body water as well as hemolymph content and exhibited greater dehydration tolerance than flies from the southern tropics. In contrast, we found no geographical variation in the males for water content and dehydration tolerance. Third, an ~10-fold increase in the rate of water loss after organic solvent treatment of male D. kikkawai suggested a role of cuticular lipids in cuticular transpiration, but had no effect in the females. Fourth, geographical differences in the storage of carbohydrate content (metabolic fuel) were observed in females but not in males. Interestingly, in females, the rate of utilization of carbohydrates did not vary geographically, but males from drier localities showed a 50% reduction compared with wetter localities. Thus, body melanisation, increased body water, hemolymph, carbohydrate content and greater dehydration tolerance confer greater desiccation resistance in females, but a reduced rate of water loss is the only possible mechanism to cope with drought stress in males. Finally, acclimated females showed a significant increase in drought resistance associated with higher trehalose content as well as dehydration tolerance, while males showed no acclimation response. Thus, sex-specific differences in desiccation resistance of D. kikkawai are associated with divergence in some water balance strategies, despite a lack of differences in the rate of water loss between the two sexes.

Rapid effects of humidity acclimation on stress resistance in Drosophila melanogaster.

We tested the hypothesis whether developmental acclimation at ecologically relevant humidity regimes (40% and 75% RH) affects desiccation resistance of pre-adults (3rd instar larvae) and adults of Drosophila melanogaster Meigen (Diptera: Drosophilidae). Additionally, we untangled whether drought (40% RH) acclimation affects cold-tolerance in the adults of D. melanogaster. We observed that low humidity (40% RH) acclimated individuals survived significantly longer (1.6-fold) under lethal levels of desiccation stress (0-5% RH) than their counter-replicates acclimated at 75% RH. In contrast to a faster duration of development of 1st and 2nd instar larvae, 3rd instar larvae showed a delayed development at 40% RH as compared to their counterparts grown at 75% RH. Rearing to low humidity conferred an increase in bulk water, hemolymph content and dehydration tolerance, consistent with increase in desiccation resistance for replicates grown at 40% as compared to their counterparts at 75% RH. Further, we found a trade-off between the levels of carbohydrates and body lipid reserves at 40% and 75% RH. Higher levels of carbohydrates sustained longer survival under desiccation stress for individuals developed at 40% RH than their congeners at 75% RH. However, the rate of carbohydrate utilization did not differ between the individuals reared at these contrasting humidity regimes. Interestingly, our results of accelerated failure time (AFT) models showed substantial decreased death rates at a series of low temperatures (0, -2, or -4°C) for replicates acclimated at 40% RH as compared to their counter-parts at 75% RH. Therefore, our findings indicate that development to low humidity conditions constrained on multiple physiological mechanisms of water-balance, and conferred cross-tolerance towards desiccation and cold stress in D. melanogaster. Finally, we suggest that the ability of generalist Drosophila species to tolerate fluctuations in humidity might aid in their existence and abundance under expected changes in moisture level in course of global climate change.

Biofouling in Membrane Bioreactors: Mechanism, Interactions and Possible Mitigation Using Biosurfactants.

Biofouling roots damage to membrane bioreactors (MBRs), such as physical, functional and organisational changes and even therefore clogging of the membrane pores and successive microbial degradation. Further, it blocks the pores, results into a biomass cake and in due course reduces the membrane flux and leads to an increase in the operational costs. MBR fouling contributed to the rise in transmembrane pressure (TMP) and decrease in permeate flux (in case of constant pressure operation mode). Chemical surfactants adopted for the cleaning of membrane surfaces have certain disadvantages such as toxicity manifestations, damage to the membranes and high CMC concentrations. Biosurfactant surfactants have attained increasing interest due to their low toxicity, biodegradability, stability to extreme environmental conditions such as temperatures, pH and tolerance to salinity. The biosurfactants trapped the foulants via micelle formation, which distresses hydrophobic interactions amongst bacteria and the surface. Rhamnolipids as an anionic biosurfactant pose a significant interfacial potential and have affinity to bind organic matter. The present review discusses the problem of biofouling in MBRs, type and interactions of foulants involved and also highlights the mechanisms of biosurfactant cleaning, effect of different parameters, effect of concentration, TMP, flux recovery, permeability, mitigation practices and challenges.

Biostimulation of Anaerobic Digestion Using Iron Oxide Nanoparticles (IONPs) for Increasing Biogas Production from Cattle Manure.

The effect of synthesised IONPs employing a nontoxic leaf extract of Azadirachta indica as a reducing, capping, and stabilizing agent for increasing biogas and methane output from cattle manure during anaerobic digestion (AD) was investigated in this study. Furthermore, the UV-visible spectra examination of the synthesized nanoparticles revealed a high peak at 432 nm. Using a transmission electron microscope, the average particle size of IONPs observed was 30-80 nm, with irregular, ultra-small, semi-spherical shapes that were slightly aggregated and well-distributed. IONPs had a polydisparity index (PDI) of 219 nm and a zeta potential of -27.0 mV. A set of six bio-digesters were fabricated and tested to see how varying concentrations of IONPs (9, 12, 15, 18, and 21 mg/L) influenced biogas, methane output, and effluent chemical composition from AD at mesophilic temperatures (35 ± 2 °C). With 18 mg/L IONPs, the maximum specific biogas and methane production were 136.74 L/g of volatile solids (VS) and 64.5%, respectively, compared to the control (p < 0.05), which provided only 107.09 L/g and 51.4%, respectively. Biogas and methane production increased by 27.6% and 25.4%, respectively using 18 mg/L IONPs as compared to control. In all treatments, the pH of the effluent was increased, while total volatile fatty acids, total solids, volatile solids, organic carbon content, and dehydrogenase activity decreased. Total solid degradation was highest (43.1%) in cattle manure + 18 mg/L IONPs (T5). According to the results, the IONPs enhanced the yield of biogas and methane when compared with controls.

Divergence of water balance mechanisms in two melanic Drosophila species from the western Himalayas.

Drosophila busckii is more abundant under colder and drier montane habitats in the western Himalayas as compared to Drosophila melanogaster but the mechanistic basis of such climatic adaptations is largely unknown. We tested the hypothesis whether genetic variation or phenotypic plasticity of cuticular traits confer adaptive protection against desiccation stress in two melanic Drosophila species living under drier montane localities. For D. melanogaster, changes in melanisation are known to be associated with reduced water loss but there are no data on D. busckii. We investigated changes in body melanisation, cuticular lipids, desiccation resistance, water loss, extractable hemolymph volume (%), and dehydration tolerance in six sympatric populations of D. busckii and D. melanogaster over an altitudinal range of 640-2236 m. D. busckii is a melanic species but changes in cuticular water loss are negatively correlated with cuticular lipid mass and not with body melanisation. In D. melanogaster, there are no plastic effects (14-28 °C) for cuticular lipid mass but variation in body melanisation is associated with desiccation-related traits. Effects of organic solvents (hexane or chloroform: methanol), developmental plasticity and seasonal variation in cuticular lipids affect body water loss in D. busckii but no such changes occur in D. melanogaster. Thus, sympatric populations of D. busckii and D. melanogaster have evolved different water balance mechanisms under shared environmental conditions in the western Himalayas. Multiple measures of desiccation resistance in these species show clinal variation with altitude, consistent with adaptation to increased desiccation stress.

Divergence of water balance mechanisms in two sibling species (Drosophila simulans and D. melanogaster): effects of growth temperatures.

Drosophila simulans is more abundant under colder and drier montane habitats in the western Himalayas as compared to its sibling D. melanogaster but the mechanistic bases of such climatic adaptations are largely unknown. Previous studies have described D. simulans as a desiccation sensitive species which is inconsistent with its occurrence in temperate regions. We tested the hypothesis whether developmental plasticity of cuticular traits confers adaptive changes in water balance-related traits in the sibling species D. simulans and D. melanogaster. Our results are interesting in several respects. First, D. simulans grown at 15 °C possesses a high level of desiccation resistance in larvae (~39 h) and in adults (~86 h) whereas the corresponding values are quite low at 25 °C (larvae ~7 h; adults ~13 h). Interestingly, cuticular lipid mass was threefold higher in D. simulans grown at 15 °C as compared with 25 °C while there was no change in cuticular lipid mass in D. melanogaster. Second, developmental plasticity of body melanisation was evident in both species. Drosophila simulans showed higher melanisation at 15 °C as compared with D. melanogaster while the reverse trend was observed at 25 °C. Third, changes in water balance-related traits (bulk water, hemolymph and dehydration tolerance) showed superiority of D. simulans at 15 °C but of D. melanogaster at 25 °C growth temperature. Rate of carbohydrate utilization under desiccation stress did not differ at 15 °C in both the species. Fourth, effects of developmental plasticity on cuticular traits correspond with changes in the cuticular water loss i.e. water loss rates were higher at 25 °C as compared with 15 °C. Thus, D. simulans grown under cooler temperature was more desiccation tolerant than D. melanogaster. Finally, desiccation acclimation capacity of larvae and adults is higher for D. simulans reared at 15 °C but quite low at 25 °C. Thus, D. simulans and D. melanogaster have evolved different strategies of water conservation consistent with their adaptations to dry and wet habitats in the western Himalayas. Our results suggest that D. simulans from lowland localities seems vulnerable due to limited acclimation potential in the context of global climatic change in the western Himalayas. Finally, this is the first report on higher desiccation resistance of D. simulans due to developmental plasticity of both the cuticular traits (body melanisation and epicuticular lipid mass) when grown at 15 °C, which is consistent with its abundance in temperate regions.

Divergence of larval resource acquisition for water conservation and starvation resistance in Drosophila melanogaster.

Laboratory selection experiments have evidenced storage of energy metabolites in adult flies of desiccation and starvation resistant strains of D. melanogaster but resource acquisition during larval stages has received lesser attention. For wild populations of D. melanogaster, it is not clear whether larvae acquire similar or different energy metabolites for desiccation and starvation resistance. We tested the hypothesis whether larval acquisition of energy metabolites is consistent with divergence of desiccation and starvation resistance in darker and lighter isofemale lines of D. melanogaster. Our results are interesting in several respects. First, we found contrasting patterns of larval resource acquisition, i.e., accumulation of higher carbohydrates during 3rd instar larval stage of darker flies versus higher levels of triglycerides in 1st and 2nd larval instars of lighter flies. Second, 3rd instar larvae of darker flies showed ~40 h longer duration of development at 21°C; and greater accumulation of carbohydrates (trehalose and glycogen) in fed larvae as compared with larvae non-fed after 150 h of egg laying. Third, darker isofemale lines have shown significant increase in total water content (18%); hemolymph (86%) and dehydration tolerance (11%) as compared to lighter isofemale lines. Loss of hemolymph water under desiccation stress until death was significantly higher in darker as compared to lighter isofemale lines but tissue water loss was similar. Fourth, for larvae of darker flies, about 65% energy content is contributed by carbohydrates for conferring greater desiccation resistance while the larvae of lighter flies acquire 2/3 energy from lipids for sustaining starvation resistance; and such energy differences persist in the newly eclosed flies. Thus, larval stages of wild-caught darker and lighter flies have evolved independent physiological processes for the accumulation of energy metabolites to cope with desiccation or starvation stress.

Divergent strategies for adaptation to desiccation stress in two Drosophila species of immigrans group.

Water balance mechanisms have been investigated in desert Drosophila species of the subgenus Drosophila from North America, but changes in mesic species of subgenus Drosophila from other continents have received lesser attention. We found divergent strategies for coping with desiccation stress in two species of immigrans group--D. immigrans and D. nasuta. In contrast to clinal variation for body melanization in D. immigrans, cuticular lipid mass showed a positive cline in D. nasuta across a latitudinal transect (10°46'-31°43'N). Based on isofemale lines variability, body melanization showed positive correlation with desiccation resistance in D. immigrans but not in D. nasuta. The use of organic solvents has supported water proofing role of cuticular lipids in D. nasuta but not in D. immigrans. A comparative analysis of water budget of these two species showed that higher water content, reduced rate of water loss and greater dehydration tolerance confer higher desiccation resistance in D. immigrans while the reduced rate of water loss is the only possible mechanism to enhance desiccation tolerance in D. nasuta. We found that carbohydrates act as metabolic fuel during desiccation stress in both the species, whereas their rates of utilization differ significantly between these two species. Further, acclimation to dehydration stress improved desiccation resistance due to increase in the level of carbohydrates in D. immigrans but not in D. nasuta. Thus, populations of D. immigrans and D. nasuta have evolved different water balance mechanisms under shared environmental conditions. Multiple measures of desiccation resistance in D. immigrans but reduction in water loss in D. nasuta are consistent with their different levels of adaptive responses to wet and dry conditions on the Indian subcontinent.