EUROPEAN JOURNAL OF ENTOMOLOGYENTOMOLOGY ISSN (online): 1802-8829 Eur. J. Entomol. 114: 222–229, 2017 http://www.eje.cz doi: 10.14411/eje.2017.027 ORIGINAL ARTICLE

Fitness traits of melanogaster (Diptera: Drosophilidae) after long-term laboratory rearing on different diets

JELENA TRAJKOVIĆ 1, VUKICA VUJIĆ 1, DRAGANA MILIČIĆ 1, GORDANA GOJGIĆ-CVIJOVIĆ 2, SOFIJA PAVKOVIĆ-LUČIĆ 1 and TATJANA SAVIĆ 3

1 University of Belgrade, Faculty of Biology, 16 Studentski trg, 11000 Belgrade, Serbia; e-mails: [email protected], [email protected], [email protected], sofi [email protected] 2 University of Belgrade, Institute of Chemistry, Technology and Metallurgy, 12 Njegoševa, 11000 Belgrade, Serbia; e-mail: [email protected] 3 University of Belgrade, Institute for Biological Research “Siniša Stanković”, 142 Despot Stefan Blvd, 11000 Belgrade, Serbia; e-mail: [email protected]

Key words. Diptera, Drosophilidae, , nutrition, fi tness components, protein content, C/N ratio

Abstract. Nutrition is one of the most important environmental factors that infl uence the development and growth in Drosophila. The food composition strongly affects their reproduction, welfare and survival, so it is necessary for fl ies to search for a mixture of macronutrients that maximizes their fi tness. We have fi ve D. melanogaster strains, which were reared for 13 years on fi ve differ- ent substrates: standard cornmeal-agar-sugar-yeast medium and four substrates modifi ed by adding tomato, banana, carrot and apple. This study was aimed at determining how such long-term rearing of fl ies on substrates with different protein content affects fi tness traits (dynamics of eclosion, developmental time and egg-to-adult survival). Further, we determined how transferring fl ies reared on fruit/vegetable substrates to a standard laboratory diet affected their fi tness. Results indicate that strains reared on the diet with the lowest content of protein and the highest C/N ratio had the slowest eclosion and developmental time, and lowest egg-to-adult survival (apple diet). The fl ies reared on the diet with the highest protein content and the lowest C/N ratio had the highest survival (tomato diet). reared on the carrot diet, which is quite similar in protein content and C/N ratio to the standard cornmeal diet, had the fastest development. Transferring fl ies to the standard cornmeal diet accelerate eclosion and developmen- tal time, but did not affect survival.

INTRODUCTION investigations (Kristensen et al., 2011; Güler et al., 2015) Food is essential for the survival of all organisms, as it including fl uctuating asymmetry studies (Vijendravarma et provides energy for different biological functions. During al., 2011), learning and memory (Kolss & Kawecki, 2008; the course of a lifetime there is a requirement for specifi c Wright, 2011), cuticular chemistry research (Fedina et nutrients for optimal body growth. However, in nature al., 2012; Pavković-Lučić et al., 2016) and sexual selec- are often exposed to changes in the quality and tion studies (Fricke et al., 2008; Abed-Vieilliard & Cortot, quantity of food, and in its availability. Consequently, there 2016). So far, nutrigenomic studies on D. melanogaster are changes in their resistance to stress, life-history traits have been used to further our understanding of human and reproduction (Djawdan et al., 1998; Bross et al., 2005; nutrigenomics, as metabolism is evolutionarily conserved Broughton et al., 2005; Carsten et al., 2005; Burger et al., (Ruden & Lu, 2006), and a promising way of developing 2007; Sisodia & Singh, 2012; Reddiex et al., 2013; Abed- strategies for dealing with metabolic diseases (Matzkin et Vieillard et al., 2014; Rodrigues et al., 2015; Kristensen et al., 2011). al., 2016). Most of the nutritional studies involving D. melanogaster D. melanogaster is one of the most frequently used model focus on adults, in spite of the fact that larvae could be a organisms in a variety of nutritional studies. Consuming better choice for studying this species nutritional require- food of different qualities is widely evaluated in ethanol- ments (Scherer et al., 2003; Schwarz et al., 2014). De- tolerance studies (McKechnie & Geer, 1993), mobility and pending on the nutritional composition of the diet, larvae cardiac (Bazzell et al., 2013), developmental increase in body mass (Sang, 1956, 1978), but are sensi- and metabolic studies (Kolss et al., 2009; Matzkin et al., tive to the amount of simple sugars and yeast in the diet 2011), ageing (Piper & Partridge, 2007), morphological (Durisko & Dukas, 2013; Neuser et al., 2005).

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222 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

D. melanogaster is a generalist that uses different are a rich source of antioxidants, extend the lifespan of D. fruits and vegetables for both feeding and reproduction melanogaster (Peng et al., 2011). (Shorrocks, 1972; Markow, 2015). Certain amounts of Using the aforementioned D. melanogaster strains, we protein, carbohydrate, lipid, vitamins and minerals are es- set out to determine (1) the extent to which the long-term sential for growth and survival of juveniles (Simpson & rearing of fl ies on different fruit/vegetable foods affects Raubenheimer, 1993; Simpson et al., 2004). The amount their fi tness traits and (2) whether these traits are affected of protein and carbohydrate in the larval food of D. mela- by transferring eggs to a standard laboratory diet, assum- nogaster affects their developmental time, egg-to-adult ing that plastic adaptation to the new nutritive environment survival and lifespan (Chippindale et al., 1998; Heilbronn may have fi tness consequences. & Ravussin, 2005; Fanson et al., 2009; Andersen et al., In nature, D. melanogaster lives, feeds, and breeds in the 2010; Kristensen et al., 2011; Merkey et al., 2011; Rodri- same place (Reaume & Sokolowski, 2006), so the ability to gues et al., 2015; Reis, 2016). However, it is only protein adapt to a new diet is important. D. melanogaster can de- that determines body and tissue growth in larvae (Britton tect the nutritional quality of a particular food and induce & Edgar, 1998; Colombani et al., 2003). Larval nutrition an adaptive plastic response (Partridge et al., 2005). Phe- further affects resistance of adults to heat and cold, starva- notypic plasticity, defi ned as “the ability of individual gen- tion and desiccation (Andersen et al., 2010; Kristensen et otypes to produce different phenotypes when exposed to al., 2016). Effects of nutrition can be sex specifi c (Lee & different environmental conditions” (Pigliucci et al., 2006) Micchelli, 2013; Reddiex et al., 2013; Nazario-Yepiz et al., can infl uence fi tness directly, if the ability to be plastic is 2017). Thus, for females of D. melanogaster the protein to adaptive (Sultan & Spencer, 2002; Crispo, 2008; Stomp et carbohydrate (P : C) ratio affects longevity, egg laying rate al., 2008), or indirectly, if the plastic response results in the and lifetime egg production (Lee et al., 2008; Fanson et al., development of an adaptive phenotype (Via et al., 1995). 2009; Rodrigues et al., 2015). During their lifetime, individuals may adapt by means of To our knowledge, there are insuffi cient studies on the developmental plasticity, since they may experience envi- fi tness consequences of long-term culturing of ronments in early life that are associated with particular (over decades), including D. melanogaster, on diets of conditions they will experience later in life (Monaghan, different qualities. In most nutritional studies on D. mela- 2008; Pilakouta et al., 2015). Selection for feeding on dif- nogaster, the quality of the diet is manipulated by alter- ferent foods can result in trade-offs associated with the ing the concentrations and ratios of yeast and sugar (Kris- adaptation, which could be manifested in terms of larval tensen et al., 2011; Matzkin et al., 2011; Fanson et al., development and survival (Kolss et al., 2009). If there is 2012; Güller et al., 2015), or by modifying the food by a diet-induced developmental plasticity then our strains using different species of yeast (Anagnostou et al., 2010), should differ in their effi ciency to utilize the carbohydrates dietary carbohydrates (Lushchak et al., 2014), lipids, vita- and proteins in the diet. The shift to a standard laboratory mins (Reis, 2016) and food additives (Neethu et al., 2014). food, prepared with yeast and sugar as an important protein However, this study involved using diets that are modifi ca- and energy sources, could affect their fi tness. tions of this fl ies’ natural food (tomato, banana, carrot and apple), prepared without adding sugar and yeast. Tomatoes MATERIAL AND METHODS are hosts for many insects: D. melanogaster is frequently Chemical analysis of substrates recorded infesting tomato crops, which in the past resulted Total dry weight of samples of substrate was determined by in a great deal of damage (De Camargo & Phaff, 1957; oven drying to constant weight at 105°C to (6 h + 2 h, depending Lange & Bronson, 1981). One of the most attractive blends on the sample) (Bradley, 2010). For the analysis of carbon (C), for D. melanogaster is produced by banana (Schubert et hydrogen (H), nitrogen (N) and sulphur (S) content a Vario EL III al., 2014), which was often used as the diet for maintain- CHNS/O Elemental Analyzer was used. Content of crude protein ing cultures of fl ies and in different experiments (Jaenike, was calculated from the nitrogen content by multiplying it by the factor 6.25 (AOAC, 1995). 1983; Demerc & Kaufman, 1996; Svilpe & Matjuškova, 2010; Stamps et al., 2012; Chhabra et al., 2013; Ho et al., strains and substrates 2013; Prakash & Krishna, 2015). Carrot is included in D. D. melanogaster fl ies were collected from a natural population melanogaster diets as it is a rich source of carotenoids. It and reared over 13 years (more than 300 generations) on fi ve dif- is reported that D. melanogaster larvae need large amounts ferent diets, in 20 replicated lines for each diet group. Flies were of dietary carotenoids for the biosynthesis of visual pig- reared on the standard cornmeal-sugar-agar-yeast diet (St), and ments (Giovannucci & Stephenson, 1999). Further, carot- four diets modifi ed by adding tomato (T), banana (B), carrot (C) and apple (A) (Fig. 1). Only the standard diet was prepared with enoids infl uence multitrophic interactions and affect additional sugar and yeast, which were not added to the fruit/veg- the evolutionary outcomes (Heath et al., 2013). Apples etable diets (Table 1, Kekić & Pavković-Lučić, 2003). Flies were are also highly attractive and commonly used for trap- reared in 250 ml glass bottles fi lled with 50 ml of food (20 bottles ping fruit fl ies, and experiments on habitat selection and per substrate), under standard laboratory conditions (temperature life-history traits (Jaenike, 1983; Hoffmann et al., 1984; of ~ 25°C, relative humidity of 60%, 300 lux of illumination, and Hoffmann, 1985; Pavković-Lučić et al., 2012; Kristensen 12L : 12D cycle). Large population, of about 2000 individuals per et al., 2016). It is reported that apple polyphenols, which strain, were maintained from generation to generation in density controlled, low competition conditions (about 100 individuals per bottle) in order to reduce genetic drift.

223 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

Fig. 1. Bottles containing D. melanogaster fl ies reared on standard, tomato, banana, carrot and apple diets.

Experimental groups ber of adults that emerged per each day, and d is day of hatching. Two experimental groups (Fig. 2) were set up and scored for Since there were no differences between the replicates the results fi tness components. In experimental group I, fi tness components for each strain were pooled. Egg-to-adult survival was expressed were scored for fl ies reared on the diet they had been reared on for as the ratio of the number of fl ies that emerged and the number of 13 years (“native” diet). In experimental group II, fl ies laid eggs eggs placed in a vial. on their native diet, which were then transferred to the St diet, on Statistical analysis which their fi tness components were determined. The assumption of normality of variances and homoscedastic- Experimental procedure ity was confi rmed using Kolmogorov-Smirnov and Levene’s tests To estimate the fi tness components, thirty to fi fty 4–5 days old for both developmental time and survival. One-Way ANOVA was fertilized females were transferred to egg laying vials. They were used to analyse developmental time and egg-to-adult survival, de- left to oviposit in 60 ø mm Petri dishes for 12 h. Petri dishes were pending on the diets. Mean developmental time and mean egg-to- fi lled with the native substrate for every strain. Eggs were col- adult survival were calculated for each vial, and these values were lected and transferred in groups of 60 to new vials fi lled with 30 used as “units” in ANOVA. Further, a Post hoc Fisher’s LSD test ml of the native diet for every strain (for experimental group I) was used. Spearman’s rank test was used to analyse correlations or fi lled with 30 ml of standard diet (for experimental group II). between protein content and developmental time and egg-to-adult There were 5–7 replicates per strain and per experimental group. survival. All statistical analyses were performed in STATISTI- ® The number of fl ies that emerged was counted daily until no fur- CA , ver. 5.0 (StatSoft). ther fl ies emerged. RESULTS Assessment of fi tness components Chemical analysis of substrates We measured the following fi tness components: dynamics of eclosion, developmental time, and egg-to-adult survival. Dynam- Percentage of nitrogen (N), carbon (C), hydrogen (H) ics of eclosion is the percentage of fl ies the emerged per day. De- and sulphur (S) in the fi ve diets are presented in Table 2. velopmental time (Dt) was calculated as the average time weight- Considering the nature of these diets, the expected amount ed by the number of adults that emerged. It was determined using of lipid in these substrates is unlikely to exceed a few per- the following formula: Dt = (Σnd × d) / Σnd, where nd is the num- cent (USDA Food Composition Databases). C/N ratio indi- cates the proportion of protein relative to the total content Table 1. Composition of the fi ve diets used for long-term culturing of organic carbon, which in this case accurately refl ects of D. melanogaster strains (according to Kekić & Pavković-Lučić, 2003). Abbreviations: N – Nipagin, E – ethanol. Standard Tomato Banana, Carrot, Ingredients diet diet Apple diets Distilled water 1100 ml 200 ml 680 ml Quantity of fruits/vegetables / 900 g 600 g Cornmeal 104 g 60 g 20 g Sugar 94 g / / Yeast 20 g / / Agar 7 g 12 g 10 g 2.5g N/ 2g N/ 2g N/ Fungicide 30ml E 20ml E 20ml E *Amount is suffi cient for 20 (250 ml) glass bottles. Fig. 2. Scheme of the experimental design.

224 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

Fig. 3. Dynamics of eclosion (a), developmental time (b) and egg-to-adult survival (c) of D. melanogaster strains reared on fi ve different diets for 13 years. the protein: carbohydrate (P : C) ratio. Protein content was that of A fl ies the lowest (53.71% ± 3.48; p < 0.001). The highest in the tomato diet and lowest in the apple diet. Con- egg-to-adult viabilities of St, B and C fl ies were 84.44% ± sequently, the C/N ratio was the highest in the apple diet, 3.82, 82.67% ± 2.70 and 82.22% ± 3.71, respectively. The and the lowest in the tomato diet (Table 2). Spearmen’s rank coeffi cient revealed a signifi cant correla- tion between protein content and egg-to-adult survival (r Experimental group I s = 0.900, p < 0.05). Dynamics of eclosion and developmental time Dynamics of eclosion and mean developmental time (± Experimental group II S.E.) of fl ies reared on their native diets are presented in Dynamics of eclosion and developmental time Fig. 3a and b, respectively. St fl ies emerged from the 11th to Dynamics of eclosion and mean developmental time 19th day, with the largest number emerging on day 14. Their (± S.E.) in Experimental group II is presented in Fig. 4a mean developmental time was 13.82 ± 0.07 days. Eclosion and b, respectively. After transferring to the St diet, fl ies of both T and B fl ies started on the 13th day and that of T of all strains started to emerge on day 10, except T-to-St fl ies ended on the 17th day and of B fl ies on the 18th day. The fl ies, which started on day 12. The largest number of fl ies largest number of T fl ies emerged on day 14 and of B fl ies emerged on days 12 and 13. The duration of emergence on day 15. For the T and B fl ies development lasted, on varied among diets: that of C-to-St fl ies lasted 14 days average, 14.25 ± 0.05 and 14.97 ± 0.05 days, respectively. and of A-to-St fl ies 20 days. One-Way ANOVA detected Eclosion of C fl ies started on day 10 and ceased on day 14. signifi cant difference in developmental time among fl ies The highest percentage of adults of C strain emerged on the transferred from their native to the St diet (F = 3.734, df = 11th day, and mean developmental time was 11.08 ± 0.04 4, error df = 20, p < 0.05). LSD test revealed that both the days. On the other hand, A fl ies started emerging on day C-to-St fl ies (in days: 11.94 ± 0.06; p < 0.01) and B-to-St 14 and the last emerged on day 29, and the largest number fl ies (in days: 11.61 ± 0.08; p < 0.01) developed signifi - of fl ies emerged on day 20. Mean developmental time of A cantly faster than the St fl ies (in days: 13.82 ± 0.07; p < fl ies was 20.82 ± 0.22 days. One-Way ANOVA indicates 0.01) and B-to-St fl ies than the A-to-St fl ies (in days: 12.64 that the strains signifi cantly differed in developmental time ± 0.11; p < 0.05). Mean developmental time of T-to-St fl ies (F = 66.240, df = 4, error df = 25, p < 0.001). Post hoc LSD was 12.83 ± 0.07 days. test revealed that C fl ies developed the fastest (p < 0.001) Egg-to-adult survival and A fl ies the slowest (p < 0.001). Spearman’s rank test re- vealed no signifi cant correlations between developmental Mean egg-to-adult survival (± S.E.) of strains reared on time and protein content (r = –0.600, p > 0.05). fruit and vegetable diets and transferred to the St diet is s presented in Fig. 4c. One-Way ANOVA confi rmed signifi - Egg-to-adult survival cant differences in egg-to-adult survival among the strains Mean egg-to-adult survival (± S.E.) in Experimental transferred to the St diet (F = 4.082, df = 4, error df = 20, group I is presented in Fig. 3c. One-Way ANOVA revealed p < 0.05). LSD test indicated that the egg-to-adult survival signifi cant difference in egg-to-adult survival among of A-to-St fl ies (65.15% ± 5.52) was signifi cantly lower strains (F = 22.342, df = 4, error df = 25, p < 0.001). LSD than that of the C-to-St fl ies (84.17% ± 3.22, p < 0.05), T- Post hoc analysis indicates that the egg-to-adult survival to-St fl ies (86.94% ± 0.96, p < 0.01) and St fl ies (84.44% ± of T fl ies was the highest (88.61% ± 1.41; p < 0.001) and 3.82, p < 0.01). Egg-to-adult survival of B-to-St fl ies was 79.17% ± 7.77. Table 2. Content of macronutrients (g/100g of dry matter of sub- Experimental group I vs. Experimental group II strate). Abbreviations: N – Nitrogen, C – Carbon, H – Hydrogen, S – Sulphur. Dynamics of eclosion and developmental time Crude protein C/N The beginning of hatching and the largest number of Diet N % C % H % S % %, N × 6.25 ratio T fl ies emerging was recorded 1 day earlier, when T fl ies Standard diet 1.29 42.41 6.35 0.65 8.06 32.87 were transferred to the St diet. Also, the hatching of B Tomato diet 2.21 46.30 6.48 0.53 13.81 20.95 Banana diet 0.72 42.61 6.00 0.50 4.47 59.59 fl ies started 2 days earlier and the largest number of fl ies Carrot diet 1.22 41.91 5.92 0.48 7.63 34.35 emerged 3 days earlier on the St diet than on their native Apple diet 0.25 41.34 5.94 0.44 1.53 168.73 diet. The largest change in dynamics of eclosion was re- 225 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

Fig. 4. Dynamics of eclosion (a), developmental time (b) and egg-to-adult survival (c) of D. melanogaster strains reared on different diets for 13 years, after transfer to the standard diet. corded for A fl ies. Emergence of A fl ies started 4 days ear- diet with the highest percentage of protein, and the lowest lier and ended 9 days earlier on the St diet than on their C/N ratio (tomato diet, C/N ratio = 20.95), had the highest native diet. Further, the largest number of fl ies emerged egg-to-adult survival. On the diet with the lowest percent- 7 days earlier on the St diet. Flies of T, B and A strains age of protein and highest C/N ratio (apple diet, C/N ratio developed signifi cantly faster on the St diet than on their = 168.73) the emergence of adults was delayed and de- native diets (T strain: F = 10.48, df = 1, error df = 7, p < velopment prolonged, and the lowest egg-to-adult survival 0.05; B strain: F = 65.099, error df = 7, df = 1, p < 0.001; recorded. However, the banana diet also has a high C/N A strain: F = 66.030, df = 1, error df = 12, p < 0.001). On ratio (C/N ratio = 59.59), but the fi tness components did the other hand, dynamics of eclosion of C fl ies remained not differ signifi cantly from those recorded for the strain the same on the St and C diets, while the largest number reared on the standard diet, or on the tomato diet (which of fl ies emerged 1 day earlier on the St diet. Based on the had the smallest C/N ratio). As eggs and larvae are more One-Way ANOVA, there was no difference in the develop- sessile than adult fl ies, it is expected that their oviposition mental times C fl ies reared on their native diet and the St behaviour and selection of pupation sites has been strongly diet (F = 5.06, df = 1, error df = 9, p > 0.05). selected for (Markow, 2015). However, Jaenike (1983), re- ports that females lay more eggs on apple than on tomato, Egg-to-adult survival although our study indicates that the apple diet had “the Egg-to-adult survival of fl ies transferred to the St diet did lowest” quality in terms of both larval development and not differ signifi cantly from that recorded on their native egg-to-adult survival. In nature, a prolonged developmen- diets (T fl ies: F = 0.1390, df = 1, error df = 7, p > 0.05; B tal time increases the risk of running out of food and not fl ies: F = 0.1904, df = 1, error df = 7, p > 0.05; C fl ies: F = completing their development. In such condition, selec- 0.328, df = 1, error df = 9, p > 0.05; A fl ies: F = 3.1634, df tion against very slow development may be strong. Thus, = 1, error df = 12, p > 0.05). relatively fast larval development in nature is an important DISCUSSION aspect of the adaptation to larval nutrition (Kolss et al., 2009). Diet affects many biological processes, starting from cel- D. melanogaster larvae have a natural propensity to bal- lular metabolism up to behaviour. Quality and amount of ance their diet. Flies fed on a diet defi cient in proteins or nutritive resources, as well as the balance of macronutri- carbohydrates later preferred the diet containing the nutri- ents in food, have a strong effect on the life-history traits ents they required. Also, when larvae are offered a choice of D. melanogaster (Lee et al., 2008; Kolss et al., 2009; between a balanced, protein rich or carbohydrate rich diets, Kristensen et al., 2011; Schwarz et al., 2014; May et al., they chose the balanced diet (Schwarz et al., 2014). Re- 2015; Rodrigues et al., 2015; Simpson et al., 2015; Abed- cently, Rodrigues et al. (2015) report that the shortest de- Vieillard & Cortot, 2016). Nutritive demands may change velopmental time is recorded for fl ies fed on a diet with during the course of life and may be sex-specifi c, if the an intermediate P : C ratio (ratio of about 1 : 2–1 : 4). Also, sexes maximize fi tness in different ways (Lee et al., 2008; egg-laying rate and lifetime egg production is maximized Maklakov et al., 2008; Lihoreau et al., 2016). when fed on diets with a P : C ratio 1 : 2 and 1 : 4, respec- D. melanogaster larvae feed and live on rotting fruit, ac- tively (Lee et al., 2008). Krijger et al. (2001) report that quiring most of their protein from the yeasts present on among neotropical Drosophila species, those with a short fruit when it is decomposing (Begon, 1982). It is record- development had a competitive advantage over those with ed that D. melanogaster reared on a protein rich diet are a long development. In our study, fl ies reared on the carrot more viable, heavier and larger, and have more ovarioles diet had a shorter development and dynamics of eclosion (Rodrigues et al., 2015). On the other hand, a low level than the other four strains. Although the carrot diet and of protein in the diet results in prolonged development, standard diet did not differ in protein content and C/N ratio delay in emergence and decrease in egg-to-adult survival, (carrot diet, C/N ratio = 34.35; standard diet, C/N ratio = fecundity and growth in D. melanogaster (Wang & Clark, 32.87), it is possible that these two diets may differ in the 1995; Tu & Tatar, 2003; Kolss et al., 2009; Rodrigues et quality of the protein (i.e., in amino acid composition). al., 2015). Results obtained in this study confi rm the results of the above mentioned studies. The strain reared on the 226 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

The standard cornmeal-sugar-agar-yeast medium is com- choice and parasitoid resistance in Drosophila. — Oikos 119: monly used for maintaining cultures of D. melanogaster 533–541. fl ies under laboratory conditions. In that sense, fi tness ANDERSEN L.H., KRISTENSEN T.N., LOESCHCKE V., TOFT S. & components of the fl ies reared on the standard diet could be MAYNTZ D. 2010: Protein and carbohydrate composition of larval food affect tolerance to thermal stress and desiccation the control for all other diets (Kolss et al., 2009). Changes in adult Drosophila melanogaster. — J. Insect Physiol. 56: in fi tness components after transferring eggs from their na- 336–340. tive fruit/vegetable diets to the standard diet could be the AOAC 1995: Offi cial Methods of Analysis of AOAC Interna- result of plastic responses to the different nutrition. After tional, 16th ed. Association of Offi cial Analytical Chemists transferring eggs to the standard diet, the developmental (AOAC), Gaithersburg, MD, USA. time of almost all the strains changed, but egg-to-adult sur- BAZZELL B., GINZBERG S., HEALY L. & WESSELLS R.J. 2013: Di- vival was not affected. Thus, it is possible that regulation etary composition regulates Drosophila mobility and cardiac of the developmental time buffered survival, because the physiology. — J. Exp. Biol. 21: 859–868. developmental time was not correlated with protein con- BEGON M. 1982: Yeast and Drosophila. In Ashburner M., Carson H.L. & Thompson J.N. Jr. (eds): The and Biology of tent, while egg-to-adult survival was. Drosophila. 3b. Academic Press, London, pp. 345–384. D. melanogaster has a genetic potential to adapt to BELDADE P., MATEUS A.R. & KELLER R.A. 2011: Evolution and different nutritional conditions based on transcriptional molecular mechanisms of adaptive developmental plasticity. dynamics (Kolss et al., 2009; Whitaker et al., 2014). De- — Mol. Ecol. 20: 1347–1363. velopmental plasticity provides developing individuals BRADLEY R.L. JR. 2010: Moisture and total solids analysis in food with multiple phenotypes each expressed under different analysis. In Nielsen S.S. (ed.): Food Analysis, 4th ed. Springer nutritional regimes (Xie et al., 2015). This is an adaptive Science + Business Media, New York, pp. 88–91. process, which may result in changes manifested at physio- BRITTON J.S. & EDGAR B.A. 1998: Environmental control of the logical, morphological and behavioural levels (Monaghan, cell cycle in Drosophila: nutrition activates mitotic and en- doreplicative cells by distinct mechanisms. — Development 2008; Kolss et al., 2009; Shingleton et al., 2009; Beldade 125: 2149–2158. et al., 2011; Kristensen et al., 2011; Trajković et al., 2013; BROSS T.G., ROGINA B. & HELFAND S.L. 2005: Behavioral, physi- Güller et al., 2015; Rodrigues et al., 2015). Phenotypic cal, and demographic changes in Drosophila populations plasticity might allow individuals to “jump” from one fi t- through dietary restriction. — Aging Cell 4: 309–317. ness peak to another, without crossing fi tness valleys (Price BROUGHTON S.J., PIPER M.D., IKEYA T.M., JACOBSON J., DRIEGE Y., et al., 2003; Crispo, 2007). This study reveals that such a MARTINEZ P., HAFEN E., WITHERS D.J., LEEVERS S.J. & PARTRIDGE scenario is possible in just one generation. L. 2005: Longer lifespan, altered metabolism, and stress resist- Results of this study demonstrate that different fi tness ance in Drosophila from ablation of cells making insulin-like components do not respond similarly to the different pro- ligands. — Proc. Nat. Acad. Sci. USA 102: 3105–3110. BURGER J.M., HWANGBO D.S., CORBY-HARRISV. & PROMISLOW D.E. tein compositions and C/N ratios of the larval diet, and in- 2007: The functional costs and benefi ts of dietary restriction in dicate a plastic response when D. melanogaster is exposed Drosophila. — Aging Cell 6: 63–71. to different nutritional environments. In that sense, differ- CARSTEN L.D., WATTS T. & MARKOW T.A. 2005: Gene expression ences in fi tness traits recorded in the strains used is a chal- patterns accompanying a dietary shift in Drosophila mela- lenge for future studies, which could include establishing nogaster. — Mol. Ecol. 14: 3203–3208. relationships with morphological characteristics (e.g. body CHIPPINDALE A.K., GIBBS A.G., SHEIK M., YEE K.J., DJWADAN M., size), physiological properties (e.g. tolerance of stress) and BRADLEY T.J. & ROSE M.R. 1998: Resource acquisition and the different behavioural traits. Adaptive signifi cance of phe- evolution of stress resistance in Drosophila melanogaster. — notypic plasticity, possible trade-offs between larval and Evolution 52: 1342–1352. CHHABRA R., KOLLI S. & BAUER J.H. 2013: Organically grown adult traits, as well as underlying molecular mechanisms food provides health benefi ts to Drosophila melanogaster. — should also be further investigated. PLoS ONE 8: e52988, 8 pp. COLOMBANI J., RAISIN S., PANTALACCI S., RADIMERSKI T., MONTAGNE ACKNOWLEDGMENTS. This work was funded by The Serbian Ministry of Education, Science and Technological Development J. & LEOPOLD P. 2003: A nutrient sensor mechanism controls (Grant No. 173012). We would also like to thank to L. Filipović Drosophila growth. — Cell 114: 739–749. for laboratory assistance. The clarity of this article was greatly CRISPO E. 2007: The Baldwin effect and genetic assimilation: re- improved by constructive and useful comments from two anony- visiting two mechanisms of evolutionary change mediated by mous reviewers. phenotypic plasticity. — Evolution 61: 2469–2479. CRISPO E. 2008: Modifying effects of phenotypic plasticity on in- REFERENCES teractions among natural selection, adaptation and gene fl ow. — J. Evol. Biol. 21: 1460–1469. ABED-VIEILLARD D. & CORTOT J. 2016: When choice makes sense: DE CAMARGO R. & PHAFF H.J. 1957: Yeasts occurring in Droso- Menthol infl uence on mating, oviposition and fecundity in phila fl ies and in fermenting tomato fruits in Northern Califor- Drosophila melanogaster. — Front. Integr. Neurosci. 10: 5, 11 nia. — J. Food Sci. 22: 367–372. pp. DEMERC M. & KAUFMAN B.P. 1996: Drosophila Guide. Carnegie ABED-VIEILLARD D., CORTOT J., EVERAERTS C. & FERVEUR J.-F. Institution, Washington, 46 pp. 2014: Choice alters Drosophila oviposition site preference on DJAWDAN M., CHIPPINDALE A.K., ROSE M.R. & BRADLEY T.J. 1998: menthol. — Biol. Open 3: 22–28. Metabolic reserves and evolved stress resistance in Drosophila ANAGNOSTOU C., LEGRAND E.A. & ROHLFS M. 2010: Friendly food melanogaster. — Physiol. Zool. 71: 584–594. for fi tter fl ies? Infl uence of dietary microbial species on food

227 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

DURISKO Z. & DUKAS R. 2013: Attraction to and learning from so- LANGE W.H. & BRONSON L. 1981: Insect pests of tomatoes. — cial cues in fruit fl y larvae. — Proc. R. Soc. (B) 280: 20131398, Annu. Rev. Entomol. 26: 345–371. 7 pp. LEE W.-C. & MICCHELLI C.A. 2013: Development and characteri- FANSON B.G., WELDON C.W., PÉREZ-STAPLES D., SIMPSON S.J. & zation of a chemically defi ned food for Drosophila. — PLoS TAYLOR P.W. 2009: Nutrients, not caloric restriction, extend ONE 8(7): e67308, 10 pp. lifespan in Queensland fruit fl ies (Bactroceratryoni). — Aging LEE K.P., SIMPSON S.J., CLISSOLD F.J., BROOKS R., BALLARD J.W.O., Cell 8: 514–523. TAYLOR P.W., SORAN N. & RAUBENHEIMER D. 2008: Lifespan FANSON B.G., YAP S. & TAYLOR P.W. 2012: Geometry of com- and reproduction in Drosophila: New insights from nutritional pensatory feeding and water consumption in Drosophila mela- geometry. — Proc. Natl. Acad. Sci. USA 105: 2498–2503. nogaster. — J. Exp. Biol. 215: 766–773. LIHOREAU M., POISSONNIER L.-A., ISABEL G. & DUSSUTOUR A. FEDINA T.Y., KUO T.H., DREISEWERD K., DIERICK H.A., YEW J.Y. 2016: Drosophila females trade off good nutrition with high- & PLETCHER S.D. 2012: Dietary effects on cuticular hydrocar- quality oviposition sites when choosing foods. — J. Exp. Biol. bons and sexual attractiveness in Drosophila. — PLoS ONE 7: 219: 2514–2524. e49799, 11 pp. LUSHCHAK O.V., GOSPODARYOV D.V., ROVENKO B.M., YURKEVYCH FRICKE C., BRETMAN A. & CHAPMAN T. 2008: Adult male nutrition I.S., PERKHULYN N.V. & LUSHCHAK V.I. 2014: Specifi c dietary and reproductive success in Drosophila melanogaster. — Evo- carbohydrates differentially infl uence the life span and fecun- lution 62: 3170–3177. dity of D. melanogaster. — Gerontol. (A, Biol. Sci. Med. Sci.) GIOVANNUCCI D.R. & STEPHENSON R.S. 1999: Identifi cation and 69: 3–12. distribution of dietary precursors of the Drosophila visual pig- MAKLAKOV A.A., SIMPSON S.J., ZAJITSCHEK F., HALL M.D., DESS- ment chromophore: analysis of carotenoids in wild type and MANN J., CLISSOLD F., RAUBENHEIMER D., BONDURIANSKY R. & ninaD mutants by HPLC. — Vision Res. 39: 219–229. BROOKS R.C. 2008: Sex-specifi c fi tness effects of nutrient in- GÜLER P., AYHAN N., KOŞUKCU C. & ÖNDER B.Ş. 2015: The effects take on reproduction and lifespan. — Curr. Biol. 18: 1062– of larval diet restriction on developmental time, preadult sur- 1066. vival, and wing length in Drosophila melanogaster. — Turk. J. MARKOW T.A. 2015: The secret lives of Drosophila fl ies. — eLife Zool. 39: 395–403. 4: e06793, 9 pp. HEATH J.J., CIPOLLINI D.F. & STIREMAN J.O. 2013: The role of MATZKIN L.M., JOHNSON S., PAIGHT C., BOZINOVIC G. & MARKOW carotenoids and their derivatives in mediating interactions be- T.A. 2011: Dietary protein and sugar differentially affect de- tween insects and their environment. — Arthr. Plant Interact. velopment and metabolic pools in ecologically diverse Droso- 7: 1–20. phila. — J. Nutr. 141: 1127–1133. HEILBRONN L.K. & RAVUSSIN E. 2005: Calorie restriction extends MAY C.M., DOROSZUK A. & ZWAAN B.J. 2015: The effect of devel- life span – but which calories? — PLoS Med. 2(8): e231, 3 pp. opmental nutrition on life span and fecundity depends on the HO T., REJTO D., SIU E. & SORATHIA S. 2013: Effects of nutrients adult reproductive environment in Drosophila melanogaster. from bananas on maturation time of Drosophila melanogaster: — Ecol. Evol. 5: 1156–1168. preference of Drosophila melanogaster for bananas from dif- MCKECHNIE S.W. & GEER B.W. 1993: Long-chain dietary fatty ferent sources. — Expedition 3: 17 pp. acids affect capacity of Drosophila melanogaster to tolerate HOFFMANN A.A. 1985: Effects of experience on oviposition and ethanol. — J. Nutr. 123: 106–116. attraction in Drosophila: comparing apples and oranges. — MERKEY A.B., WONG C.K., HOSHIZAKI D.K. & GIBBS A.G. 2011: Am. Nat. 126: 41–51. Energetics of metamorphosis in Drosophila melanogaster. — HOFFMANN A.A., PARSONS P.A. & NIELSEN K.M. 1984: Habitat J. Insect Physiol. 57: 1437–1445. selection: olfactory response of Drosophila melanogaster de- MESHRIF W.S. & ELKHOLY S.E. 2015: Genotype and environment pends on resources. — Heredity 53: 139–143. shape the fi tness of Drosophila melanogaster. — J. Basic Appl. JAENIKE J. 1983: Induction of host preference in Drosophila mela- Zool. 68: 1–9. nogaster. — Oecologia 58: 320– 325. MONAGHAN P. 2008: Early growth conditions, phenotypic de- KEKIĆ V. & PAVKOVIĆ-LUČIĆ S. 2003: Fruit and vegetable food velopment and environmental change. — Phil. Trans. R. Soc. media suitable for maintaining D. melanogaster fl ies. — Dros- Lond. (B) 363: 1635–1645. oph. Inf. Serv. 86: 147. NAZARIO-YAPIZ N.O., RAMIREZ LOUSTALOT-LACLETTE M. & KOLSS M. & KAWECKI T.J. 2008: Reduced learning ability as a MARKOW T.A. 2017: Drosophila species as models for nutri- consequence of evolutionary adaptation to nutritional stress in tional studies: Development, metabolic pools on diets with Drosophila melanogaster. — Ecol. Entomol. 33: 583–588. contrasting relative sugar: protein ratios. — J. Nutr. Biol. 1: KOLSS M., VIJENDRAVARMA R.K., SCHWALLER G. & KAWECKI T.J. 101–107. 2009: Life-history consequences of adaptation to larval nutri- NEETHU B.K., BABU Y.R. & HARINI B.P. 2014: Flavors supple- tional stress in Drosophila. — Evolution 63: 2389–2401. mented in diet regulate the hatchability and viability in Droso- KRIJGER C.L., PETERS Y.C. & SEVENSTER J.G. 2001: Competitive phila. — Drosoph. Inf. Serv. 97: 24–28. ability of neotropical Drosophila predicted from larval devel- NEUSER K., HUSSE J., STOCK P. & GERBER B. 2005: Appetitive opment times. — Oikos 92: 325–332. olfactory learning in Drosophila larvae: effects of repetition, KRISTENSEN T.N., OVERGAARD J., LOESCHKE V. & MAYNTZ D. 2011: reward strength, age, gender, assay type and memory span. — Dietary protein content affects evolution for body size, body Anim. Behav. 69: 891–898. fat and viability in Drosophila melanogaster. — Biol. Lett. 7: PAVKOVIĆ-LUČIĆ S., SAVIĆ T., JELIĆ M., KENIG B., TANASKOVIĆ M., 269–272. STAMENKOVIĆ-RADAK M. & ANĐELKOVIĆ M. 2012: Note on the KRISTENSEN T.N., HENNINGSEN A.K., AASTRUP C., BECH-HANSEN fauna of Drosophila (Diptera: Drosophilidae) and the fi rst re- M., HOBERG BJERRE L.B., CARLSEN B., HAGSTRUP M., GRAARUP cord of Opomyza fl orum (Diptera: ) from Moun- JENSEN S., KARLSEN P., KRISTENSEN L. ET AL. 2016: Fitness com- tain Goč, Serbia. — Acta Entomol. Serb. 17: 45‒53. ponents of Drosophila melanogaster developed on a standard PAVKOVIĆ-LUČIĆ S., TODOSIJEVIĆ M., SAVIĆ T., VAJS V., TRAJKOVIĆ laboratory diet or a typical natural food source. — Insect Sci. J., ANĐELKOVIĆ B., LUČIĆ L., KRSTIĆ G., MAKAROV S., TOMIĆ V. 23: 771–779. ET AL. 2016: “Does my diet affect my perfume?” Identifi cation

228 Trajković et al., Eur. J. Entomol. 114: 222–229, 2017 doi: 10.14411/eje.2017.027

and quantifi cation of cuticular compounds in fi ve Drosophila SHORROCKS B. 1972: Drosophila. Ginn, London, 144 pp. melanogaster strains maintained over 300 generations on dif- SIMPSON S.J. & RAUBENHEIMER D. 1993: A multilevel analysis of ferent diets. — Chem. Biodivers. 13: 224–232. feeding-behaviour – the geometry of nutritional decisions. — PARTRIDGE L., PIPER M.D.W. & MAIR W. 2005: Dietary restriction Phil. Trans. R. Soc. Lond. (B) 342: 381–402. in Drosophila. — Mech. Ageing Dev. 126: 938–950. SIMPSON S.J., SIBLY R.M., LEE K.P., BEHMER S.T. & RAUBENHEIMER PENG C., CHAN H.Y.E., HUANG Y., YU H. & CHEN Z.-Y. 2011: D. 2004: Optimal foraging when regulating intake of multiple Apple polyphenols extend the mean lifespan of Drosophila nutrients. — Anim. Behav. 68: 1299–1311. melanogaster. — J. Agric. Food Chem. 59: 2097–2106. SIMPSON S.J., LE COUTEUR D.G. & RAUBENHEIMER D. 2015: Putting PIGLIUCCI M., MURREN C.J. & SCHLICHTING C.D. 2006: Phenotypic the balance back in diet. — Cell 161: 18–23. plasticity and evolution by genetic assimilation. — J. Exp. STAMPS J.A., YANG L.H., MORALES V.M. & BOUNDY-MILLS K.L. Biol. 209: 2362–2367. 2012: Drosophila regulate yeast density and increase yeast PILAKOUTA N., JAMIESON S., MOORAD J.A. & SMISETH P.T. 2015: community similarity in a natural substrate. — PLoS ONE Parental care buffers against inbreeding depression in burying 7(7): e42238, 11 pp. beetles. — Proc. Natl. Acad. Sci. USA 112: 8031–8035. STOMP M. VAN DIJK M.A., VAN OVERZEE H.M.J., WORTEL M.T., PIPER M.D. & PARTRIDGE L. 2007: Dietary restriction in Droso- SIGON C.A.M., EGAS M.H., HOOGVELD H., GONS H.J. & HUIS- phila: delayed aging or experimental artefact? — PLoS Genet. MAN J. 2008: The timescale of phenotypic plasticity and its im- 3(4): e57, 6 pp. pact on competition in fl uctuating environments. — Am. Nat. PRAKASH V.S. & KRISHNA M.S. 2015: Effect of organic fruits (ba- 172: E169–E185. nana and papaya) on locomotor ability and resistance to eitheri- SULTAN S.E. & SPENCER H.G. 2002: Metapopulation structure fa- zatoin in Drosophila melanogaster. — Int. J. Recent Sci. Res. vors plasticity over local adaptation. — Am. Nat. 160: 271–283. 6: 4656–4660. SVILPE E. & MATJUŠKOVA N. 2010: Infl uence of shiitake mush- PRICE T.D., QVARNSTRÖM A. & IRWIN D.E. 2003: The role of phe- room Lentinula edodes on reproduction of Drosophila mela- notypic plasticity in driving genetic evolution. — Proc. R. Soc. nogaster. — Proc. Latv. Acad. Sci. (B) 64: 223–228. Lond. (B) 270: 1433–1440. TRAJKOVIĆ J., PAVKOVIĆ-LUČIĆ S. & SAVIĆ T. 2013: Mating success REAUME C.J. & SOKOLOWSKI M.B. 2006: The nature of Drosophila and wing morphometry in Drosophila melanogaster after long- melanogaster. — Curr. Biol. 16: R623–R638. term rearing on different diets. — Behaviour 150: 1431–1448. REDDIEX A.J., GOSDEN T.P., BONDURIANSKY R. & CHENOWETH S.F. TU M.-P. & TATAR M. 2003: Juvenile diet restriction and the aging 2013: Sex-specifi c fi tness consequences of nutrient intake and and reproduction of adult Drosophila melanogaster. — Aging the evolvability of diet preferences. — Am. Natur. 182: 91–102. Cell 2: 327–333. REIS T. 2016: Effects of synthetic diets enriched in specifi c nu- UNITED STATES DEPARTMENT OF AGRICULTURE. AGRICULTURAL RE- trients on Drosophila development, body fat, and lifespan. — SEARCH SERVICE. USDA Food Composition Databases. URL: PLoS ONE 11(1): e0146758, 12 pp. https://ndb.nal.usda.gov/ndb/. RODRIGUES M.A., MARTINS N.E., BALANCE L.F., BROOM L.N., DIAS UNCKLESS R.L., ROTTSCHAEFER S.M. & LAZZARO B.P. 2015: A A.J.S., FERNANDES A.S.D., RODRIGUES F., SUCENA E. & MIRTH genome-wide association study for nutritional indices in Dro- C.K. 2015: Drosophila melanogaster larvae make nutritional sophila. — G3 (Bethesda) 5: 417–425. choices that minimize developmental time. — J. Insect Phy- VIA S., GOMULKIEWICZ R., DE JONG G., SCHEINER S.M., SCHLICHTING siol. 81: 69–80. C.D. & VAN TIENDEREN P.H. 1995: Adaptive phenotypic plas- RUDEN D.M. & LU X. 2006: Evolutionary conservation of me- ticity: consensus and controversy. — Trends Ecol. Evol. 10: tabolism explains how Drosophila nutrigenomics can help us 212–217. understand human nutrigenomics. — Genes Nutr. 1: 75–83. VIJENDRAVARMA R.K., NARASIMHA S. & KAWECKI T.J. 2011: Adap- SANG J.H. 1956: The quantitative nutritional requirements of tation to larval malnutrition does not affect fl uctuating assym- Drosophila melanogaster. — J. Exp. Biol. 33: 45–72. metry in Drosophila melanogaster. — Biol. J. Linn. Soc. 104: SANG J.H. 1978: The nutritional requirements of Drosophila. In 19–28. Ashburner M. & Wright T.R.F. (eds): The Genetics and Biology WANG L. & CLARK A.G. 1995: Physiological genetics of the re- of Drosophila. Vol. 2b. Academic Press, London, pp. 159–192. sponse to a high-sucrose diet by Drosophila melanogaster. — SCHERER S., STOCKER R.F. & GERBER B. 2003: Olfactory learning Biochem. Genet. 33: 149–165. in individually assayed Drosophila larvae. — Learn. Mem. 10: WHITAKER R., GIL M.P., DING F., TATAR M., HELFAND S.L. & NER- 217–225. ETTI N. 2014: Dietary switch reveals fast coordinated gene SCHUBERT M., HANSSON B.S. & SACHSE S. 2014: The banana code expression changes in Drosophila melanogaster. — Aging 6: – natural blend processing in the olfactory circuitry of Droso- 355–368. phila melanogaster. — Front. Physiol. 5(59): 13 pp. WRIGHT G.A. 2011: Appetitive learning: memories need calories. SCHWARZ S., DURISKO Z. & DUKAS R. 2014: Food selection in lar- — Curr. Biol. 21: R301–302. val fruit fl ies: dynamics and effects on larval development. — XIE J., DE CLERCQ P., PAN C., LI H., ZHANG Y. & PANG H. 2015: Naturwissenschaften 101: 61–68. Larval nutrition-induced plasticity affects reproduction and SHINGLETON A.W., ESTEP C.M., DRISCOL M.V. & DWORKIN I. 2009: gene expression of the ladybeetle, Cryptolaemus montrouzieri. Many ways to be small: different environmental regulators of — BMC Evol. Biol. 15: 276, 9 pp. size generate distinct scaling relationships in Drosophila mela- Received December 5, 2016; revised and accepted March 23, 2017 nogaster. — Proc. R. Soc. Lond. (B) 276: 2625–2633. Published online April 18, 2017

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