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Article

Microalgal Lipid Profile and Their Dietary Impact on Drosophila melanogaster

by
Svitlana Miros
1,2,*,
Svitlana Bilokon
3,
Yiting Han
1,2 and
Ronald Halim
1,2
1
UCD Algae Group, School of Biosystems and Food Engineering, University College Dublin, Belfield, D04 V1W8 Dublin, Ireland
2
UCD Conway Institute, University College Dublin, Belfield, D04 V1W8 Dublin, Ireland
3
Department of Molecular Biology, Biochemistry and Genetics, Odesa I. I. Mechnikov National University, 65058 Odesa, Ukraine
*
Author to whom correspondence should be addressed.
Lipidology 2026, 3(2), 17; https://doi.org/10.3390/lipidology3020017
Submission received: 14 January 2026 / Revised: 4 March 2026 / Accepted: 30 April 2026 / Published: 9 May 2026
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Abstract

Background/Objectives: Microalgae are gaining increasing attention as sustainable sources of dietary lipids and other bioactive compounds; however, the relationship between microalgae lipid composition and physiological outcomes in vivo remains insufficiently understood. This study aimed to characterize antioxidant activity, total lipid content and fatty acid (FA) profiles of selected freshwater microalgae and to evaluate their dietary impact using Drosophila melanogaster as a whole-organism model. Methods: Four freshwater microalgal species (Chlorella vulgaris, Nannochloris limnetica, Scenedesmus communis, and Tetradesmus obliquus) were cultivated separately in 3N-BBM+V medium under controlled laboratory conditions. DPPH, FRAP and TPC were measured in microalgae methanolic extracts. Total lipids were extracted using a modified Breuer method and quantified gravimetrically. FA profiles were determined as fatty acid methyl esters by GC-FID. Freeze-dried microalgal biomass (3 mg/mL) was incorporated into standard D. melanogaster diet. Lifespan and body mass were assessed separately in females and males, as well as fecundity in general. Results: Total lipid content ranged from 17.3% to 28.1% of dry weight, with FA profiles dominated by C16 saturated, monounsaturated (omega-9), and omega-6 polyunsaturated fatty acids. Correlation analysis indicates that antioxidant properties of the studied microalgae are more closely linked to lipid fractions than to phenolic content. Dietary supplementation with microalgal biomass of three out of four microalgal species significantly extended median lifespan, particularly in males, without adverse effects on body mass or fecundity. Conclusions: These findings indicate that freshwater microalgae can serve as a physiologically safe dietary lipid source. D. melanogaster represents a suitable in vivo model for screening the nutritional potential of microalgal lipids.

Graphical Abstract

1. Introduction

Microalgae have attracted increasing attention as a natural source of valuable biomolecules and biomass for human and animal nutrition, as they provide lipids, proteins, polysaccharides, pigments, vitamins, and minerals, while simultaneously contributing to greenhouse gas mitigation via photosynthesis [1,2]. Lipid-rich microalgal species are of particular interest as they are considered a sustainable vegan source of essential lipids for dietary applications [3,4,5,6,7]. The FA composition of microalgal biomass is influenced by several factors, including strain selection, nutrient availability, cultivation conditions, and bioreactor configuration, by dictating key environmental factors such as light penetration, nutrient availability, temperature stability, and shear stress [8,9,10]. Continuous efforts are therefore focused on identifying suitable microalgal strains, optimizing cultivation conditions and characterizing their FA profiles [5,11,12]. In the present work, the microalgal species were selected based on their relevance as freshwater oleaginous microalgae with documented biotechnological potential and availability from recognized culture collections. One of the green algae (Chlorophyta), Chlorella vulgaris, is already widely used in human nutrition and as a feed ingredient [13,14], whereas other members, such as Tetradesmus obliquus and Scenedesmus communis, are well-studied candidates for animal feed and biorefinery applications due to their robust growth and favourable biochemical composition [15,16]. The Ochrophyta algae, Nannochloropsis limnetica, were included as a representative of eustigmatophytes, a group known for high lipid productivity and industrial relevance but still underexplored in dietary studies using whole-organism models [5]. To evaluate the effects of microalgae at the organismal level, rapid and cost-efficient screening models are required. Drosophila melanogaster is increasingly recognized as a valuable model organism in food and nutrition research due to its relatively short lifespan and generation time, as well as its genetic homology with humans, which makes it particularly suitable for such studies [17,18]. Although widely employed in nutritional research, only a limited number of studies have investigated the impact of microalgal supplementation on D. melanogaster [19,20].
The present study aims to analyze the lipid content and FA profiles of four freshwater microalgal strains and to assess their effects on key health parameters of D. melanogaster, including lifespan, body mass, and fecundity.

2. Materials and Methods

2.1. Microalgae Species and Growth Conditions

Nannochloropsis limnetica (SAG 18.99) was purchased from the Culture Collection of Algae at the University of Gottingen, Göttingen, Germany (SAG). Chlorella vulgaris (CCAP 211/19), Scenedesmus communis (CCAP 276/4B), and Tetradesmus obliquus (CCAP 276/10) were obtained from the UK Culture Collection of Algae and Protozoa (CCAP, Scottish Association for Marine Science, Oban, Scotland, UK). Stock cultures and further cultivation were maintained on Bold Basal Modified (3N-BBM+V) cultivation medium [21]. Each species was cultured in 1 L working volume reactors with inline 0.22 µm filters to prevent contamination and aeration of 0.15 vvm. Cultures were grown in triplicate under fluorescent light (100 µmol photons m−2s−1 at the vessel’s surface), 16/8 h light/dark in batch mode until the onset of the stationary phase (21 days). At the end of cultivation, cultures reached an optical density of approximately 5 at 750 nm, corresponding to cell densities ranging from 13 × 107 to 14 × 107 cells/mL. Biomass was harvested by centrifugation at 3200× g, 10 min, RT. Freshly harvested microalgae biomass was frozen at −80 °C, solidifying the water inside the cells, then freeze-dried by a vacuum evaporator (Alpha 2–4 LD plus, Martin Christ, Osterode am Harz, Germany) at −70 °C for 48 h, until achieving a final low moisture content (usually <5%).

2.2. Total Lipid Content and Fatty Acid Analysis

Total lipids were extracted from freeze-dried biomass using a modified multi-step Breuer et al., 2013 [22] for microalgae biomass. The samples were weighed and transferred into a plastic tube (around 50 mg) to which 1 mL chloroform/methanol (4:5 v/v) solvent extraction mixture was added. The samples were bit-beaten 8 times with 6.5 m/s for 60 s each time, then transferred to the glass ‘extraction tube’, washing the plastic tube with the remains of the sample twice with 1 mL chloroform/methanol. Thus, the total volume of added chloroform/methanol consisted of 3 mL. Sonicated for 10 min in a water bath. Pipetted 2.5 mL of MilliQ water, containing 50 mM 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris) and 1 M NaCl to the ‘extraction tube’, which is set to pH 7 using a HCl solution, that caused a phase separation between chloroform and methanol-water. After centrifugation for 10 min the chloroform phases containing the extracted lipid were collected, pooled, filtered (0.2 μm syringe filter), and dried under flowing N2 gas. The total lipid concentration in biomass was calculated gravimetrically. For the next step, which included fatty acid methyl esters (FAME) preparation, the dried lipid extracts were re-dissolved in 2 mL of n-hexane, with tripentadecanoin (C15:0) (100 μg mL−1) added as an internal standard. Residual moisture was removed using anhydrous sodium sulphate. Fatty acids were converted to fatty acid methyl esters by direct (in situ) transesterification using methanolic HCl and heat, following the methods of Lepage and Roy (1986) [23] and Lewis et al. (2000) [24], as commonly applied to microalgal biomass. Specifically, 4 mL of 2 M L−1 potassium methoxide and 2 mL of 20% HCl in methanol were sequentially added, followed by heating at 70 °C for 30 min and centrifugation at 3200× g for 10 min at room temperature. The upper n-hexane layer containing FAMEs was filtered through a 0.2 μm PTFE membrane and transferred into GC vials with simultaneous dilution by hexane 1:9. FAME concentrations were determined by gas chromatography with flame ionization detection (GC-FID) using a 30 m HP-5 capillary column (Agilent Technologies, Santa Clara, CA, USA) and helium as the carrier gas at a split ratio of 10:1 and a flow rate of 2 mL min−1. The FAME separation method was employed based on their boiling points, and a four-step temperature profile was used, including an initial temperature of 50 °C for 2 min, a ramping rate of 10 °C min−1 to 150 °C, then at 17.5 °C min−1 to 185 °C, and finally to 300 °C. The injector and detector temperatures were set at 280 °C and 320 °C, respectively. Fatty acids (FA) were quantified using a 37-FAME standard mixture (Sigma Aldrich, Arklow, Ireland) and linear calibration curves, with identification based on retention times (Supplementary Materials). Transesterification efficiency was calculated from the conversion rate of the C15:0 internal standard, and this value was applied to correct the fatty acid quantities in each sample [25]. It should be noted that the non-polar HP-5 capillary column that was selected allows reliable quantification of major fatty acids, whose volume can be measured for supplementation purposes. This column is not optimized for resolving complex mixtures of minor fatty acids or positional and geometric isomers.

2.3. Determination of Antioxidant Activity and Total Phenolic Content of Microalgae

The antioxidant potential of the investigated microalgal biomass was evaluated using DPPH radical scavenging activity, ferric reducing antioxidant power (FRAP), and total phenolic content (TPC) assays on the methanolic microalgae extracts. To obtain the extracts, freeze-dried biomass was ground into a fine powder (30 mg), mixed with 1 mL of methanol, and vortexed for 30 s before being placed in a sonication bath (Ultrawave Ltd., Cardiff, UK) in the dark at room temperature for 15 min. The extraction mixture was stored at 4 °C for 24 h before being centrifuged at 7500× g for 10 min. The extraction process was repeated twice using 1 mL of methanol for each cycle to achieve colourless biomass. All supernatants were pooled and stored at −20 °C until further use.

2.3.1. DPPH Radical Scavenging Assay

The DPPH radical scavenging assay was adapted from [26] for a 96-well plate format. Trolox (Sigma-Aldrich, Cat. No. 238813) served as the reference standard. Methanolic extracts were diluted four-fold prior to analysis. A volume of 100 μL of diluted extract was combined with 200 μL of 72 μM DPPH solution prepared in methanol and incubated in darkness at room temperature for 30 min. Absorbance was then measured at 515 nm. Results were expressed as mg Trolox equivalents per gram of dry biomass (TE/g DW) using the equation: DPPH value = Trolox concentration (mg/L) × extraction volume (L)/dry biomass (g). The Trolox concentration in each sample was obtained from the calibration curve. The extraction volume corresponded to the methanol volume multiplied by the dilution factor, and dry biomass represented the mass of the extracted sample.

2.3.2. Ferric Reducing Antioxidant Power (FRAP)

The FRAP reagent was prepared according to [27] by mixing 300 mM acetate buffer (pH 3.6), 10 mM TPTZ dissolved in 40 mM HCl, and 20 mM FeCl3 in a 10:1:1 (v/v) ratio and pre-warming the mixture to 37 °C. For analysis, 900 μL of FRAP reagent was mixed with 100 μL of methanolic extract and 50 μL of Trolox standard. After incubation in the dark at 37 °C for 40 min, absorbance was recorded at 593 nm using a SpectraMax M3 microplate reader (Molecular Devices, San Jose, CA, USA) against a methanol blank. FRAP values were expressed as mg Trolox equivalents per gram dry biomass (TE/g DW) according to the equation: FRAP value = Trolox concentration (mg/L) × extraction volume (L)/dry biomass (g). Trolox concentration was derived from the calibration curve, the extraction volume corresponded to the methanol extraction volume multiplied by the dilution factor, and dry biomass represented the sample weight.

2.3.3. Total Phenolic Content (TPC)

Total phenolic content was quantified using the Folin–Ciocalteu method [28] adapted for a 96-well plate. Methanolic extracts were diluted four-fold before analysis. A 100 μL aliquot of diluted extract was mixed with 100 μL Folin–Ciocalteu reagent and allowed to react for 5 min. Subsequently, 700 μL of 20% Na2CO3 was added and the mixture was incubated for 20 min at room temperature in darkness. Samples were centrifuged at 18,500× g for 3 min, and 250 μL of supernatant was transferred to a microplate for absorbance measurement at 765 nm (SpectraMax M3, Molecular Devices, USA). Gallic acid (Thermo Scientific, Waltham, MA, USA, Cat. No. 220765000) was used as the calibration standard. Results were expressed as mg gallic acid equivalents per gram dry biomass (mg GAE/g DW) using the equation: TPC value = Gallic acid concentration (mg/L) × extraction volume (L)/dry biomass (g). The gallic acid concentration was calculated from the standard curve, the extraction volume corresponded to the methanol volume multiplied by the dilution factor, and dry biomass represented the weight of the extracted sample.

2.4. D. melanogaster Assay

Parental flies of wildtype Canton-S (CS) strain (Bloomington Drosophila stock centre at Indiana University, Bloomington, IN, USA) were kept at 25 °C and held on standard fly media (agar 0.25 g, yeast 0.375 g, sugar 1.25 g and semolina 2.0 g per 25 mL of distilled water) in a climate room with a LD 12 h:12 h cycle and 50% relative humidity. For experimental variants, 3 mg of freeze-dried microalgae biomass was added per 1 mL (0.3%) of fly standard media. The selected dose was based on the information from two published studies [19,20]. The lifespan and body weight were measured separately for females and males. For that, 100 flies of each sex were raised in 250 mL bottles containing 20 mL of medium (20 flies per vial) under the same conditions as noted above. Flies were transferred into fresh food vials with the same component every 4 days. Day 3 post-ecdysis, and the deaths of the flies were recorded. Flies were considered dead when neither voluntary movement nor responses to external stimulation could be observed. The average body weights of flies from the 5 replicated vials (20 flies per vial) were measured using a Denver Instrument SI-234 balance (accuracy 0.0001 g). The experiments stopped on day 45 when most flies died.
To assess fecundity, 5 pairs of D. melanogaster (5 males and 5 females) were placed in a vial containing 5 mL of standard nutritional medium (control) or the same medium supplemented with microalgal powder (experimental variants). Each treatment consisted of 5 vials, resulting in a total of 25 pairs per condition. All vials were maintained for 72 h. After this period, the adult flies were removed, and the number of emerging adult offspring (imago) was recorded. The total number of offspring per pair was used as a measure of reproductive output (fecundity).

2.5. Statistical Analysis

The data related to total lipids and FAs concentration presented in this study are reported as mean ± standard deviation.
Drosophila survival days estimates were expressed as the median ± percentage difference from the control group, as performed in [29]. The body weight of flies and fecundity were expressed as mean ± standard deviation.
GraphPad Prism 10.4 (Boston, MA, USA) was used for statistical analysis. A one-way analysis of variance (ANOVA) with post hoc Tukey’s test was applied for comparison of total lipid and total FA content, while Dunnett’s test was used for body weight and fecundity. Microalgae biomass antioxidant activity and D. melanogaster lifespan comparisons were conducted using the nonparametric Kruskal–Wallis criterion with post hoc Mann–Whitney tests. Differences were considered statistically significant at p-value < 0.05.
A Pearson correlation analysis via GraphPad Prism 10.4 was performed to evaluate the relationships between lipid-related parameters and antioxidant properties of microalgal biomass. The variables included total lipid content, total fatty acids (TFA), FRAP, DPPH, and TPC. Correlation coefficients (r) were calculated using normalized mean values of each parameter. The strength of correlations was interpreted as weak (|r| < 0.3), moderate (0.3 ≤ |r| < 0.7), and strong (|r| ≥ 0.7).

3. Results

3.1. Total Lipid and Fatty Acid Profile

The total lipid content and fatty acid (FA) profile of four freshwater microalgae species are shown in Table 1.
The total lipid concentration varied among the analyzed microalgae strains, ranging from 17.3 ± 2.1% in S. communis to 28.1 ± 1.7% in N. limnetica. Lipid content of C. vulgaris and T. obliquus was 27.3 ± 1.9% and 21.9 ± 1.1%, respectively. The total fatty acid content in C. vulgaris and N. limnetica was 1.7 times lower than the content of total lipids, and 3.7 and 3-fold lower in S. communis and T. obliquus. All microalgal species produced saturated FA C16:0, as well as monounsaturated C18:1n-9 (omega-9), and polyunsaturated C18:2n-6c (omega-6) FAs, though in varying proportions. Only small amounts of additional fatty acids were observed: C14:1 (1.7 ± 0.1%), C18:0 (3.4 ± 0.3%), and C16:1 (4.3 ± 0.2%) in N. limnetica, and C18:0 (4.5 ± 0.2%) and C16:1 (5.7 ± 0.1%) in C. vulgaris. Thus, the most relevant FAs for dietary application identified in microalgae strains were Linoleic acid (C18:2n-6c) and Oleic acid (C18:1n-9).

3.2. Antioxidant Activity and Total Phenolic Content of Microalgae

Figure 1 demonstrates the antioxidant activity along with the microalgae strains.
Both C. vulgaris and N. limnetica demonstrated the strongest DPPH radical scavenging activity (≈2.1 ± 0.1 mg TE g−1), forming a statistically distinct group (p < 0.05) from S. communis and T. obliquus, which exhibited significantly lower activity (≈1.7 ± 0.1 mg TE g−1). A similar trend was observed in the FRAP assay, where C. vulgaris showed the highest reducing power (25.5 ± 0.7 mg TE g−1), followed by N. limnetica (22.0 ± 0.7 mg TE g−1), T. obliquus (19.1 ± 0.7 mg TE g−1), and S. communis displaying the lowest value (7.6 ± 0.7 mg TE g−1).
In contrast, the pattern of total phenolic content differed from the antioxidant assays. S. communis contained the highest TPC (1.44 ± 0.01 mg GAE g−1), whereas C. vulgaris and N. limnetica showed intermediate levels of 1.23 ± 0.01 and 1.13 ± 0.7 mg GAE g−1, respectively. T. obliquus had significantly lower phenolic content (0.34 ± 0.04 mg GAE g−1).

3.3. Correlation Between Antioxidant Activity and Lipid Content

The correlation matrix was visualized as a heatmap (Figure 2).
Pearson correlation analysis revealed strong positive relationships between lipid-related variables and antioxidant capacity measured by FRAP and DPPH assays. Total lipid content showed strong positive correlations with TFA (r = 0.84), FRAP (r = 0.87), and DPPH (r = 0.87). Similarly, TFA strongly correlated with FRAP (r = 0.84) and DPPH (r = 0.78). A strong positive association was also observed between FRAP and DPPH (r = 0.73).
In contrast, total phenolic content (TPC) exhibited weak or moderate correlations with the other variables. TPC showed a weak positive correlation with TFA (r = 0.17) and DPPH (r = 0.30), and a weak negative correlation with total lipids (r = −0.03) and FRAP (r = −0.27).

3.4. Effect of Microalgae-Treated Food on D. melanogaster Physiological Parameters

3.4.1. Lifespan

Survival days estimates were expressed separately for females and males, as the percentage of median difference from the control group (Table 2).
In the control group, the median lifespan differed between females and males, at 18 and 12 days, respectively. Inclusion of microalgal freeze-dried biomass at 3 mg/mL extended the female median lifespan by 45.9% for C. vulgaris, 8.1% for S. communis, and 27.0% for T. obliquus. A more significant extension was observed in males, where supplementation with C. vulgaris, N. limnetica, S. communis, and T. obliquus biomass increased their median lifespan by up to 75.0%, 25%, 29.2% and 50.0%, respectively.

3.4.2. Body Mass and Fertility

The fly’s body mass, obtained separately for females and males (Figure 3A) and fertility (Figure 3B), are shown below.
The average body mass of D. melanogaster females was 1.23 ± 0.03 mg, and that of males was 0.95 ± 0.09 mg in the control variant. Supplementation with microalgal biomass had no significant effect on their body mass, except in females treated by N. limnetica, where the imago weight decreased by 21% (p < 0.05).
The average progeny from control D. melanogaster couples was equal to 60.07 ± 2.8. A statistically significant increase of 20% and 18% in fertility was observed only with the addition of T. obliquus and S. communis, respectively, to the nutritional medium.

4. Discussion

Among the tested strains (Table 1), C. vulgaris and N. limnetica demonstrated the highest total lipid contents (27.3–28.1% DW), significantly exceeding that of S. communis and comparable to T. obliquus. These values fall within the upper range reported for lipid-producing freshwater microalgae grown under nutrient-replete conditions, where lipid contents typically range from 15 to 30% DW [7,10]. The relatively high lipid accumulation in C. vulgaris and N. limnetica supports their frequent consideration as promising candidates for nutritional and biotechnological applications.
The proportion of total fatty acids (TFA) relative to dry weight showed a strong correlation (0.84) with total lipid content and varies between species. Although TFA constituted a major fraction of total lipids, the observed differences between gravimetrically determined total lipids and the quantified fatty acids indicate the presence of non-saponifiable components, such as pigments, sterols, and other neutral lipids, particularly in S. communis [30,31], as well as minor analytical losses.
It should be noted that a quite limited diversity of fatty acids was detected in the present study compared with numerous reports on the phylum Chlorophyta (Chlorella, Scenedesmus, Tetradesmus) [32,33] and the phylum Ochrophyta (Nannochloropsis species) [34,35]. This difference is most likely explained by the specific cultivation conditions applied. All strains were grown in nitrogen-replete 3N-BBM medium and harvested at the onset of the stationary phase. It is known that nitrogen availability is a key regulator of lipid metabolism in microalgae and strongly influences fatty acid composition [36,37,38]. Under nitrogen-sufficient conditions, microalgae prioritize rapid growth and protein synthesis, and lipid metabolism is typically dominated by saturated and C18 fatty acids, particularly C16:0, C18:1 and C18:2 [39]. In contrast, nitrogen limitation and other environmental stresses promote membrane remodelling and stimulate the biosynthesis of highly unsaturated fatty acids, including C18:3, C16 polyunsaturated fatty acids, and long-chain PUFA such as EPA and ARA [40,41]. Therefore, the simplified fatty acid profile observed here is consistent with nutrient-replete cultivation rather than contradictory to the known lipid potential of these taxa. In addition, numerous studies demonstrate that the high variability of FA content depends on the cultivation conditions and simplification of the FA profile during the non-stress cultivation [1,8,41,42,43]. In addition, the GC-FID analysis performed using a non-polar HP-5 column was intended for general screening of major fatty acids; minor components present at very low abundance may have remained below the detection threshold [44]. However, considering the relatively low supplementation level of microalgae biomass (0.3%) in the present study, such minor components are unlikely to substantially influence the overall dietary effect. Several studies have also demonstrated that analytical methodology can significantly affect lipidomic outcomes [30,45]. In our case, the chloroform/methanol extraction procedure was selected and optimized for the analysis of small amounts of microalgal biomass.
Overall, our findings are consistent with previous reports [46] demonstrating the dominance of C16:0 (palmitic acid), C18:1n-9c (oleic acid), and C18:2n-6c (linoleic acid) as 16- and 18-carbon precursors during the entire cultivation process. A relatively high proportion of omega-6 fatty acids is characteristic of many freshwater green microalgae grown under non-stress conditions [39]. We did not observe elongation to long-chain FAs (C20–C22), which is consistent with non-stress cultivation, as such conversion processes are often associated with stress-induced enzymatic activity [39]. Moreover, desaturation process may be enhanced under low-light and temperature conditions [47]. Stearic acid (C18:0) and palmitoleic acid (C16:1) were synthesized from C16:0 only in C. vulgaris and N. limnetica, which have the highest lipid concentration, while myristic acid (C14:0) was detected exclusively in N. limnetica, where it was synthesized de novo, highlighting species-specific differences in elongation and desaturation processes [43].
The correlation analysis performed between total lipids, total fatty acids, and antioxidant capacity (FRAP and DPPH) revealed strong positive correlations (Figure 2), which suggest that fatty acids and lipid-associated components, such as carotenoids, tocopherols, or other lipophilic compounds, contribute substantially to the antioxidant potential of the studied microalgal biomass. The close relationship between FRAP and DPPH indicates that both assays consistently reflect the antioxidant capacity of the samples, despite being based on different reaction mechanisms. In contrast, TPC showed weak correlations with most parameters, suggesting that phenolic compounds were not the main contributors to antioxidant activity in these microalgae. Similar findings were reported in a previous study on C. vulgaris and Scenedesmus obliquus, where antioxidant properties differed between lipid and phenolic extracts of the same species and varied depending on the growth medium [48]. This observation aligns with the well-established fact that microalgal lipid composition is influenced by cultivation conditions.
Dietary supplementation with freeze-dried microalgal biomass at 0.3% of daily food intake significantly affected the lifespan of D. melanogaster, with pronounced sex-specific responses (Table 2). Overall, microalgal feeding resulted in lifespan extension in most experimental variants, particularly in males, suggesting a beneficial physiological effect even at relatively low supplementation levels.
Among females, supplementation with C. vulgaris and T. obliquus led to a substantial increase in median lifespan (+45.9% and +27.0%, respectively), whereas S. communis induced only a modest effect, and N. limnetica showed no measurable improvement compared with the control. In contrast, male flies exhibited a more uniform and pronounced response, with all microalgal treatments significantly extending median lifespan (+25.0% to +75.0%). The strongest effect was observed in males fed C. vulgaris, consistent with this strain’s relatively high total lipid and fatty acid content.
Sex-dependent differences in lifespan responses are well documented in D. melanogaster and are often attributed to differences in metabolic rate, nutrient allocation, reproductive investment, and stress sensitivity between females and males [49]. Male flies generally exhibit shorter lifespans and higher sensitivity to dietary interventions, which may explain the greater relative lifespan extension observed in males across all treatments. Similar sex-specific effects have been reported for dietary lipid manipulation, antioxidant supplementation, and caloric modulation in Drosophila [50,51,52].
The observed lifespan extension did not correlate directly with total lipid or total fatty acid content of the microalgal biomass. For example, N. limnetica, despite exhibiting high lipid levels, failed to enhance female lifespan, whereas T. obliquus, with intermediate lipid content, significantly prolonged lifespan in both sexes. This suggests that lifespan modulation is likely influenced by a combination of factors beyond total lipid quantity, including fatty acid composition, lipid class distribution, antioxidant compounds, pigments, sterols, and other bioactive metabolites present in microalgal cells.
In contrast to the pronounced effects observed on lifespan, dietary supplementation with freeze-dried microalgal biomass exerted comparatively modest effects on body mass and fecundity in D. melanogaster (Figure 1). This suggests that low-level microalgal supplementation primarily influences longevity-related physiological pathways rather than gross growth or reproductive output. Body mass responses were largely sex- and strain-dependent. Female flies fed N. limnetica biomass exhibited a significant reduction in body mass compared with controls, whereas no substantial changes were detected in males or in flies fed other microalgal strains. Body mass in Drosophila is tightly regulated by nutrient-sensing pathways, particularly insulin/IGF and TOR signalling, which respond sensitively to dietary lipid composition rather than total caloric input alone [53,54]. The observed reduction in female body mass upon N. limnetica feeding may therefore reflect altered lipid utilization or allocation toward maintenance rather than somatic growth.
Importantly, reduced body mass in Drosophila is not necessarily indicative of compromised health. Previous studies have shown that moderate reductions in body mass or lipid storage can accompany lifespan extension and improved metabolic efficiency, particularly under dietary interventions that modulate lipid quality rather than quantity [55,56]. In this context, the absence of widespread body mass reduction across treatments supports the conclusion that microalgal supplementation did not induce nutritional stress or malnutrition.
Fecundity responses were similarly moderate but revealed strain-specific effects. Flies fed S. communis and T. obliquus biomass demonstrated a significant increase in fertility, whereas no significant changes were observed for C. vulgaris or N. limnetica. Reproductive output in D. melanogaster is highly sensitive to dietary lipid composition, particularly the balance between saturated, monounsaturated, and polyunsaturated fatty acids, which influence oogenesis, membrane synthesis, and hormone signalling [57,58].
Interestingly, the enhancement of fecundity did not correlate with total lipid or total fatty acid content of the microalgal biomass, reinforcing the notion that specific lipid classes or associated bioactive compounds may play a more important role than overall lipid abundance. Microalgae are known to contain sterols, glycolipids, carotenoids, and antioxidants that can influence reproductive physiology either directly or indirectly by reducing oxidative stress [31,39]. Such components may contribute to the observed fertility enhancement in S. communis and T. obliquus treatments.
Taken together, the body mass and fecundity data indicate that microalgal supplementation at low dietary levels does not negatively affect growth or reproductive performance in D. melanogaster. Instead, the largely neutral or positive effects on these parameters, combined with significant lifespan extension, suggest an overall improvement in physiological robustness rather than a trade-off between longevity and reproduction. This finding is particularly relevant for nutritional lipid research, as it supports the potential use of microalgae-derived lipids as functional dietary components without adverse effects on reproductive fitness.
Only limited literature exists on microalgae-based diets in D. melanogaster. Qiu et al. reported that very high supplementation levels (up to 40%) of C. vulgaris and S. obliquus reduced lifespan and body mass, likely due to dietary imbalance or reduced food palatability [20]. In contrast, lower concentrations of wastewater-grown microalgae were shown to extend lifespan [19]. The present study supports these findings by demonstrating that low-dose microalgal supplementation can exert beneficial effects on longevity without negative physiological consequences.

5. Conclusions

This study demonstrates that freshwater microalgae represent a promising and physiologically safe dietary source of lipids with high antioxidant activity and measurable biological effects in vivo. The four investigated microalgal strains cultured in BBM with three-fold nitrogen concentration exhibited fatty acid profiles dominated by C16, omega-9 monounsaturated, and omega-6 polyunsaturated fatty acids.
Correlation analysis demonstrated strong positive relationships between total lipids, total fatty acids, and antioxidant capacity (FRAP and DPPH), while total phenolic content showed weak associations with these parameters. These findings suggest that lipid-associated compounds are likely the major contributors to the antioxidant potential of the studied microalgal biomasses, highlighting the importance of lipid fractions in determining their nutritional and functional value.
Dietary supplementation of Drosophila melanogaster with low levels (0.3%) of freeze-dried microalgal biomass resulted in significant lifespan extension, particularly in males, without detrimental effects on body mass or reproductive performance.
The modest and strain-specific effects on body mass and fecundity, together with the absence of adverse trade-offs between longevity and reproduction, indicate an overall improvement in physiological robustness rather than nutritional stress. These findings support the suitability of D. melanogaster as a rapid and cost-effective whole-organism model for screening the nutritional and functional properties of microalgal lipids.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/lipidology3020017/s1.

Author Contributions

Conceptualization, S.M. and R.H.; methodology, S.M., S.B. and Y.H.; investigation, S.M., S.B. and Y.H.; data curation, S.M.; writing—original draft preparation, S.M.; writing—review and editing, S.M., S.B. and R.H.; visualization, S.M. and Y.H.; supervision, R.H.; project administration, R.H.; funding acquisition, R.H. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Irish Research Council’s Ukrainian Researchers Scheme (URS/2023/3L) “Cell-Wall Autolysis for the Scalable Fractionation of Microalgae into Biofuels and Novel Food Products”.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Acknowledgments

The authors would like to thank Hans-Georg Eckhardt for technical assistance with GC-FID.

Conflicts of Interest

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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Lipidology 03 00017 g001
Figure 1. Microalgae biomass antioxidant activity: (A) DPPH radical-scavenging activity (mg TE/g); (B) ferric reducing antioxidant power (mg TE/g); (C) total polyphenol content (mg GAE/g). Ch (C. vulgaris), Nl (N. limnetica), Sc (S. communis), To (T. obliquus). Different letters above the bars indicate significant differences between groups (p < 0.05). Comparisons for parameters between microalgae strains were performed by nonparametric Kruskal–Wallis criterion with a post hoc Mann–Whitney test (p < 0.05) (n = 3).
Figure 1. Microalgae biomass antioxidant activity: (A) DPPH radical-scavenging activity (mg TE/g); (B) ferric reducing antioxidant power (mg TE/g); (C) total polyphenol content (mg GAE/g). Ch (C. vulgaris), Nl (N. limnetica), Sc (S. communis), To (T. obliquus). Different letters above the bars indicate significant differences between groups (p < 0.05). Comparisons for parameters between microalgae strains were performed by nonparametric Kruskal–Wallis criterion with a post hoc Mann–Whitney test (p < 0.05) (n = 3).
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Figure 2. Pearson correlation matrix showing relationships between total lipids, total fatty acids (TFA), antioxidant activities (FRAP and DPPH), and total phenolic content (TPC) in microalgal biomass. ***—p-value < 0.0001, **—p-value < 0.0025 (n = 12).
Figure 2. Pearson correlation matrix showing relationships between total lipids, total fatty acids (TFA), antioxidant activities (FRAP and DPPH), and total phenolic content (TPC) in microalgal biomass. ***—p-value < 0.0001, **—p-value < 0.0025 (n = 12).
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Figure 3. D. melanogaster body mass (A) (n = 100) and fertility (B) (n = 25) measurement after feeding with microalgae freeze-dried biomass (3 mg/mL). Ch (C. vulgaris), Nl (N. limnetica), Sc (S. communis), To (T. obliquus). Comparisons of parameters between microalgae-treated flies against control flies were performed by one-way ANOVA with post hoc Dunnett’s test. Values are means ± SD. ****—p-value < 0.0001, ***—p-value < 0.01, **—p-value < 0.05.
Figure 3. D. melanogaster body mass (A) (n = 100) and fertility (B) (n = 25) measurement after feeding with microalgae freeze-dried biomass (3 mg/mL). Ch (C. vulgaris), Nl (N. limnetica), Sc (S. communis), To (T. obliquus). Comparisons of parameters between microalgae-treated flies against control flies were performed by one-way ANOVA with post hoc Dunnett’s test. Values are means ± SD. ****—p-value < 0.0001, ***—p-value < 0.01, **—p-value < 0.05.
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Table 1. The total lipid content and fatty acid profile of microalgae grown on 3N-BBM+V medium.
Table 1. The total lipid content and fatty acid profile of microalgae grown on 3N-BBM+V medium.
Component, % DWC. vulgarisN. limneticaS. communisT. obliquus
Total Lipids27.3 ± 1.9 A28.1 ± 1.7 A17.3 ± 2.1 B21.9 ± 1.1 B
Total Fatty Acids15.7 ± 0.5 A25.6 ± 0.5 AB4.7 ± 0.4 B,C7.3 ± 0.2 C
Fatty acids, (% TFA)
Saturated
C14:0–1.7 ± 0.1––
C16:020.4 ± 0.217.6 ± 0.322.2 ± 4.216.7 ± 2.5
C18:04.5 ± 0.23.4 ± 0.3––
Monounsaturated
C16:15.7 ± 0.14.3 ± 0.2––
C18:1n-9
Polyunsaturated		55.4 ± 4.152.5 ± 3.450.8 ± 3.259.2 ± 1.9
C18:2n-6c19.1 ± 0.825.4 ± 0.226.9 ± 1.024.1 ± 4.5
A, B, C letters indicate statistical grouping based on a one-way ANOVA with post hoc multiple comparisons using Tukey’s test (p < 0.05). Groups that do not share the same letter are significantly different. Values are means ± SD (n = 3).
Table 2. Lifespan of D. melanogaster following dietary supplementation with freeze-dried microalgal biomass.
Table 2. Lifespan of D. melanogaster following dietary supplementation with freeze-dried microalgal biomass.
VariantFemalesMales
Survival DaysMedian Lifespan Change
(% vs. Control)		Survival DaysMedian Lifespan Change
(% vs. Control)
MedianMeanMaxMedianMeanMax
Control18.517.2350 A12.013.8300 A
C. vulgaris27.024.541+45.9 B21.018.932+75.0 B
N. limnetica18.518.9320.0 A15.015.330+25.0 B
S. communis20.020.036+8.1 B15.515.029+29.2 B
T. obliquus23.524.144+27.0 B18.016.831+50.0 B
A, B letters indicate statistical grouping based on nonparametric Kruskal–Wallis criterion with a post hoc Mann–Whitney test (p < 0.05) within females and males separately. Groups that do not share the same letter as the control are significantly different (n = 100).
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Miros, S.; Bilokon, S.; Han, Y.; Halim, R. Microalgal Lipid Profile and Their Dietary Impact on Drosophila melanogaster. Lipidology 2026, 3, 17. https://doi.org/10.3390/lipidology3020017

AMA Style

Miros S, Bilokon S, Han Y, Halim R. Microalgal Lipid Profile and Their Dietary Impact on Drosophila melanogaster. Lipidology. 2026; 3(2):17. https://doi.org/10.3390/lipidology3020017

Chicago/Turabian Style

Miros, Svitlana, Svitlana Bilokon, Yiting Han, and Ronald Halim. 2026. "Microalgal Lipid Profile and Their Dietary Impact on Drosophila melanogaster" Lipidology 3, no. 2: 17. https://doi.org/10.3390/lipidology3020017

APA Style

Miros, S., Bilokon, S., Han, Y., & Halim, R. (2026). Microalgal Lipid Profile and Their Dietary Impact on Drosophila melanogaster. Lipidology, 3(2), 17. https://doi.org/10.3390/lipidology3020017

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