It was also reported previously that females live longer and are more resistant to different stresses than male flies. As a next step, we checked ‘life-quality’ indices, conventionally measured for Drosophila and other animal models of aging. Since R. rosea is known mainly by its adaptogenic properties, we also checked age-related changes in stress resistance of the treated flies.Climbing activity is considered as a marker of healthspan in Drosophila. The association of a decline in climbing activity with age has been established in D. melanogaster. We observed that R. rosea improved climbing activity of the flies over a wide concentration range. Rhodiola-fed flies showed significantly higher climbing activity from day 16 to day 40 compared with the controls. On day 33, 36-55% of females supplemented with 2.5 mg/ml, 5.0 mg/ml, and 10.0 mg/ml of R. rosea rhizome were able to pass 5 cm distance from food surface within 20 seconds, while among control flies only 10% could fulfill this task . Males showed similar results, with the strongest effect at 10.0 mg/ml R. rosea rhizome . In this study, R. rosea rhizome conferred resistance to heat shock and menadione-induced stress. The newly enclosed and older flies of both sexes did not demonstrate any difference between control and Rhodiola-fed groups in heat-induced coma onset . However, 34-day-old males,hydroponic nft channel fed with the food supplemented with 5.0 mg/ml and 10.0 mg/ml R. rosea rhizome had a 1.8- and 2.5-fold longer recovery time, than controls, respectively .

Compared with controls, 34-day-old females supplemented with 2.5 mg/ml, 5.0 mg/ml, and 10.0 mg/ml of R. rosea rhizome recovered from heat coma 1.4-, 2.1-, and 1.6-fold faster, respectively . Among males, only those receiving the 5.0 mg/ml R. rosea rhizome, recovered faster from heat coma than the controls. Recent studies have shown that heat-shock proteins are involved in lifespan extension. In particular, Rhodiola preparations increased levels of HSP-16 in C.elegans . Lifespan extension in D. melanogaster was also associated with enhanced Hsp22 levels. It is known that Hsp22 is synthesized after a rapid heat shock and may be important for cell survival under this stress. Kurapati and colleagues showed that the level of Hsp22 mRNA was increased in both long- and shortlived lines between days 40 and 50, but later the expression of this gene was decreased. In this study, we used a rapid exposure to high temperature and observed that the flies fed the diet supplemented with R. rosea rhizome powder were more resistant to heat shock at the middle of the cohort lifetime. Menadione is a compound that produces superoxide-anion in a redox-cycling way. In this study, an age-dependent change in menadione sensitivity was observed for both males and females . Among young flies , no significant difference in sensitivity was observed between control and Rhodiola-fed groups exposed to 20 mM menadione, whereas older flies fed the herb preparation were more resistant. A 2.3-foldhigher resistance was observed in 31-day-old males and a 2.4-fold higher resistance was observed in 20-day-old females . The difference among treated groups was not statistically significant. It was previously shown that treatment of pond snail Limnaea stagnalis larvae by R. rosea extract conferred resistance to both 600 μM menadione and heat shock under 43°C. Later, it was shown on C. elegans that adaptogens, including R. rosea, activate DAF-16/FOXO transcription factor, promoting change of its intracellular localization.

In another study, performed on bladder cancer cell lines, R. rosea extract and salidroside itself increased phosphorylation of AMP-activated protein kinase, leading, most probably, to the activation of mTOR pathway. It is known that both FOXO and TOR pathways are referred to stress resistance. In particular, one of the FOXO downstream targets is manganesecontaining superoxide dismutase, which is responsible for elimination of mitochondrial superoxide and probably capable of providing a defense against redoxcycling agents, such as menadione or paraquat. However, it was found that R. rosea did not activate antioxidant responses in human osteosarcoma-derived diploid fibroblast and neuroblastoma cell lines. In general, the findings on activation of HSP or antioxidant defenses by R. rosea are controversial. They imply that influence of R. rosea on signaling pathways is either transient, or that it does not operate directly with reactive oxygen species, but rather promotes more effective elimination of oxidized molecules .Many conditions, mutations, and compounds that prolong lifespan may reduce reproduction. From a medical and ethical point of view, lifespan extension at a cost of reduced reproduction would not be acceptable for the human population. In this context, there is a clear contemporary trend in searching for anti-aging remedies that do not affect reproduction. This point seemed to us reasonable in the actual experiments. In previous studies, high doses of R. rosea powder or extract decreased egg-laying ability of fruit flies. We checked the same for the preparation of R. rosea, collected in the Ukrainian Carpathian Mountains. A gradual decrease in fecundity with age in all treatment groups was observed, but R. rosea rhizome enhanced the fecundity in most age groups .

The flies fed a diet supplemented with R. rosea rhizome demonstrated a higher fecundity on days 3 , 7 , 9 , 11 , 15, and 19 . It seems that R. rosea rhizome at 2.5 and 5.0 mg/ml provided a higher fecundity in young flies until approximately day 11, while 10.0 mg/ml of R. rosea rhizome enhanced fecundity from day 7 until day 19. The maximum effect was found at 10.0 mg/ml. There was found to be a positive relationship between fly fecundity and supplementation with 10.0 mg/ml R. rosea . To investigate the possible positive effect of R. rosea preparation on the egg-laying ability of female Drosophila, we measured the mating speed and duration of copulation in the control and Rhodiola-treated flies. These parameters, as well as the percentage of mated females and copulated flies, were not significantly different at any concentration of Rhodiola rhizome used . The number of eclosed offspring was also not affected. A decrease in fecundity is often considered to be due to caloric restriction. Current data on fecundity suggest that R. rosea preparation might not mimic calorie restriction. All our observations on fecundity partially support the assumption that R. rosea may preserve bio-genic amines fromoxidation by inhibition of monoamine oxidase. In particular, it has recently been shown that octopamine is required for egg release from ovary, while serotonin is needed for male ejaculation.Recent studies have shown that bio-active compounds of R. rosea, particularly salidroside, may affect the mTOR pathway. It is known,nft hydroponic system that dietary protein-to-carbohydrate ratio influences TOR signaling in D. melanogaster and hence, lifespan, stress resistance, and food consumption. In this way, a specific diet could modulate the lifespan prolonging effect of R. rosea. In this study, we conducted a pilot experiment assessing lifespan extension by R. rosea on eight diets with different P:C ratios. The lifespan assays were conducted using mortality cages. Yeast was used as the protein source, while sucrose was used as the carbohydrate source. We obtained similar results on the diet with composition close to the one described previously. Diet-response surfaces show that the highest median lifespan was reached on diets with a P:C ratio around 0.36 for the diet with 5 g/l yeast and sucrose . These data converge with those described by Skorupa et al.. However, the environment, namely the vials or mortality cage, as well as peculiarities of protein and micro-nutrient source, might also influence lifespan parameters. In particular, in our previous study, fruit flies lived approximately 80 days in vials on the diet with 10% sucrose with only 0.25% yeast extract, whereas in this study, fruit flies lived around 30 days in cages on a similar diet, . Rhizome powder of R. rosea added to the diet at a concentration of 5 mg/ml extended mean lifespan by up to 11% to 15% on diets with P:C ratios of 0.044, 0.2, and 0.36, except the diet with 15% yeast and sucrose , and up to 21% on the diet with P:C ratio 0.011 . However, there was no significant extension on the diets with P:C ratios 1 and 0.36 , while on the diet with a P:C ratio 1.5 R. rosea supplementation decreased lifespan by about 10%.

These data lead us to suggest that the macro-nutrients, being at high concentration in the food, may cancel out the beneficial effect of R. rosea on Drosophila lifespan. This effect seems to be caused by the increased amount of protein in the diet. Additionally, the P:C ratio along with food consumption should also be taken into account.An “ecological or environmental edge” can be defined as the transition between two landscape elements , and it can be permanent , or it may be more temporary Fig. 1. Generally, the effects of ecological and environmental edges on spatial distributions of animals and plants have been described and discussed mostly from the perspective of how ecosystem fragmentation affects biodiversity . “Habitat islands,” originating from the process of landscape fragmentation, are characterized by differences in physical environments between edges and interior, leading to bio-geographic changes in original wildlife communities . For example, a study of five groups of carabid species in the Aggtelek National Park, Hungary, revealed greater carabid species diversity near forest edges as compared to forest interiors due to increase in the number of edge-preferring carabid species and higher herb cover near forest edges . Gall-forming insects in natural forest patches in the Brazilian Pantanal also showed different species composition between edge and interior with one third of 113 species only sampled along forest edges and with another third sampled exclusively in the interior . Similarly, Altamirano et al. showed that the spatial distribution of galling insects sampled at transects in nine forests in Central Argentina were most predominant along forest edges and that many species benefitted from unspecified forest edge conditions. In broader terms, it has been reported that there is an increase in richness and abundance of insect generalist species near ecological or environmental edges, while the interior of fragmented habitats tend to favor specialized and competitor insects . The topic of biographic changes in insect communities as the result of ecosystem fragmentations and edge-biased distributions has been discussed and reviewed by several ecologists . In addition to a rich body of literature on relationships between biodiversity and ecological or environmental edges in natural and semi-natural ecosystems, there are numerous reports of similar edge-biased distributions of insects in agricultural systems. However, this specific topic has never been reviewed, and it has not been emphasized how this unique aspect of spatial distribution patterns is highly relevant to both insect monitoring and pest management in agricultural systems. Here, we describe published reports of insect distributions with edge-biased distributions across a wide range of agricultural systems. We discuss the following two questions: What are the potential mechanisms responsible for an edge-biased distribution of insect populations in agricultural systems? How can increased knowledge about edgebiased distributions and their governing mechanisms help to enhance the efficiency of agricultural pest management practices?An edge-biased distribution of insects in agricultural systems was first reported in 1950 in a study on black bean aphids, Aphis fabae Scop. on twigs of Euonymus europaeus L. and Viburnum opulus L. . The author noted that the onset of infestations by black bean aphids was near field edges and eventually spread throughout the field, although field edges would be the areas with the highest aphid density. Winder et al. conducted a large-scale study of the effect of grid size on the analysis of the spatial distribution of English grain aphids, Sitobion avenae Fabricius and found that significant edge effect was detected for large grid size with higher count of aphids along field edges. However, the edge-biased distribution was not evident at small grid sizes. Thus, sampling unit size was highlighted as a critical aspect of the ability to detect and quantify spatial trends, such as edge-biased distributions. Another large-scale spatial distribution study of common farmland arthropods in winter wheat fields also demonstrated higher density of species along edges . The studies by Winder et al. and Holland et al. noted the effervescent occurrence of edge-biased distributions. These findings agree with Wilson and Morton who showed that edgebiased distribution of two-spotted spider mite, Tetranychus urticae Koch , in cotton was only prominent early in the growing season but eventually diminished as the season progressed.