Comparison of the composition and color characteristics of haploid and diploid rainbow trout (Oncorhynchus mykiss) eggs produced under different rearing conditions

AQUATIC ANIMAL REPORTS 4(1) (2026) : 21-36 DOI: 10.5281/zenodo.17977631 [email protected] AQUATIC ANIMAL REPORTS Journal homepage: https://scopesscience.com/index.php/aqar/ Received: 09 August 2025; Received in revised form: 11December 2025 Accepted: 16 December 2025; Available online: 20 December 2025 RESEARCH PAPER Citation: Küçük, N. & Yılmaz, E. (2026). Comparison of the composition and color characteristics of haploid and diploid rainbow trout (Oncorhynchus mykiss) eggs produced under different rearing conditions. Aquatic Animal Reports, 4(1), 21-36. https://doi.org/10.5281/zenodo.17977631 COMPARISON OF THE COMPOSITION AND COLOR CHARACTERISTICS OF HAPLOID AND DIPLOID RAINBOW TROUT (Oncorhynchus mykiss) EGGS PRODUCED UNDER DIFFERENT REARING CONDITIONS Necmettin KÜÇÜK1,2, Ebru YILMAZ3* 1Program of Fisheries Engineering, Graduate School of Natural and Applied Sciences, Aydın Adnan Menderes University, Aydın, Türkiye 2 State Hydraulic Works (DSI), XXIst Regional Directorate, Aydin, Türkiye 3Bozdoğan Vocational School, Aydın Adnan Menderes University, Aydın, Türkiye Necmettin Küçük 1,2: E-mail: [email protected], ORCID ID: https://orcid.org/0009-0002-85898202 Ebru Yılmaz 3: E-mail: [email protected], ORCID ID: https://orcid.org/0000-0003-1905-1265 *Corresponding author: Ebru YILMAZ, [email protected], phone, +90-256-2207705 Abstract This study investigated the biochemical composition and color characteristics of haploid (n) and diploid (2n) eggs from rainbow trout (Oncorhynchus mykiss) broodstock reared at two farms in Seydikemer, Muğla. Four-year-old females and three-year-old males were used, and eggs were analyzed for proximate composition, fatty acid profile, and colorimetric parameters. While no significant differences were observed in dry matter, crude protein, crude fat, or ash content (p>0.05), diploid eggs exhibited higher total saturated and monounsaturated fatty acids, greater omega-3 levels, and higher EPA+DHA content. Haploid eggs showed higher omega-6 levels, DHA/EPA ratio, and hypocholesterolemic/hypercholesterolemic (HH) ratio (p<0.05). Color analysis revealed that diploid eggs had higher L* (lightness) and b* (yellowness), whereas haploid eggs were redder (higher a*, p<0.05). These results indicate that ploidy affects egg fatty acid composition and color, which may influence embryonic development and selective breeding programs. Keywords: Egg quality, Fatty acid, Lipid composition, Rainbow trout, Trout egg Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 22 Introduction Aquaculture has become a rapidly growing global sector, playing a strategic role in meeting the food demands of the world's increasing population (FAO, 2020). Key drivers of this growth include the depletion of natural fish stocks, the need for sustainable food production, and the high productivity offered by aquaculture. In this context, rainbow trout (Oncorhynchus mykiss) stands out as one of the most widely farmed cold-water fish species worldwide due to its rapid growth performance, adaptability to environmental conditions, economic return, and ease of production (Escamilla-Rosales et al., 2024). Furthermore, this species possesses a rich nutritional profile with high-quality protein, omega-3 fatty acids, and various vitamins and minerals, making it highly valued for its nutritional value and significantly contributing to the continuously increasing consumer demand (Xu et al., 2022; Mahato et al., 2023). Its widespread acceptance in the consumer market is further bolstered by its soft texture, desirable white-topink flesh color, and mild flavor, enhancing its appeal as a high-quality animal protein source (Janampa-Sarmiento et al., 2020). In trout farming, production success largely depends on egg quality, as the chemical composition of the eggs directly determines embryonic development, fry survival rates, and overall productivity (Izquierdo et al., 2001; Brooks et al., 1997). In this context, the amino acid and fatty acid profiles of trout eggs are critically important and are generally at levels that support embryonic development. In particular, omega-3 polyunsaturated fatty acids such as EPA and DHA are essential components for fry growth, immune function, and physiological development (Tocher, 2010; Baki et al., 2021). The levels of these fatty acids in eggs largely depend on the broodstock diet. Differences in lipid sources used in broodstock diets can directly affect the fatty acid composition of the eggs; some vegetable oil-enriched feeds have been reported to increase EPA, ARA, and DHA levels, positively impacting fry development and survival rates (Mazorra et al., 2003; Yıldız et al., 2020). Grčević et al. (2019) emphasized that egg color should be considered as one of the important quality criteria in fish breeding studies. Since fish cannot synthesize carotenoids, these pigments must be obtained entirely from external sources via feed (Bjerkeng, 2008). In addition to giving the egg yolk a bright and vibrant color, carotenoids contribute to healthy embryonic development by protecting the embryo against oxidative damage thanks to their strong antioxidant properties. Therefore, sufficient carotenoid availability plays a critical role in improving embryonic development and increasing the survival rate of juvenile fish. Furthermore, the attractive color appearance of the egg significantly contributes to the marketability of the product by enhancing the perception of quality in both aquaculture facilities and consumer markets (Nakano & Wiegertjes, 2020; Shastak & Pelletier, 2023). Due to the influence of feed composition, environmental conditions, and genetic factors, significant differences arise in the chemical and biochemical properties of eggs obtained from different production centers (Bobe, 2015). Therefore, comparative analyses of rainbow trout eggs raised under various conditions are crucial for ensuring quality control and conducting improvement efforts in the industry (Baki et al., 2019; Baki et al., 2021). The aim of this study was to comparatively investigate the basic chemical composition, fatty acid profile and color parameters of haploid and diploid rainbow trout (Oncorhynchus mykiss) broodstock eggs obtained from different trout fish farms. Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 23 Material and Method Broodstock Spawning and Fertilization Process This study was conducted in two separate rainbow trout (Oncorhynchus mykiss) hatcheries located in the Seydikemer district of Muğla province. Four female and two male broodstock were used in the study. Dry spawning and fertilization were performed. The weight and length of the fish, which were stunned with phenoxyethanol, were measured. Haploid (n) eggs obtained from two four-year-old female broodstock were weighed, measured, labeled, and transported to the laboratory in styrofoam boxes covered with ice trays. Sperm from a threeyear-old male broodstock, checked for motility under a microscope, was added to the eggs and mixed. For the analysis of rainbow trout sperm, a Nikon Eclipse microscope equipped with a 20x objective and phase contrast was used. Sperm motility was examined using D532 buffer (1 mM CaCl₂, 20 mM Tris, 30 mM glycine, 125 mM NaCl, pH 9.0). During the examination, 2 µL of milt was mixed with 398 µL of activation solution at 6°C, and 0.7 µL of this mixture was placed on a glass slide, covered with a coverslip, and motility was observed from activation until cessation (Billard, 1977, Dietrich et al., 2008). Fertilization was achieved by contacting the mixture with water, and the eggs were allowed to rest for 30 minutes. The same procedures were followed for diploid (2n) eggs. All samples were stored at -80°C, and care was taken to ensure cleanliness and labeling of the materials. Similar procedures were performed at the other facility (Emre & Kürüm, 2007). Fecundity (Egg Production) Fecundity was estimated by dividing the number of eggs counted in the subsample by the total weight of the gonad (Le Cren, 1951; Avşar, 2005; Serezli, 2017). F = Fecundity (number), n = Number of eggs in the subsample (number), Wg = Gonad weight (g), g = Weight of the subsample (g). The equation F = n x Wg/g was used. Determination of Chemical Composition of Egg Samples Approximately 10-15 grams of egg sample were used for the analyses. Moisture analysis was performed by drying at 105°C. Crude protein content was determined by the Kjeldahl method using an EFLAB device equipped with an infrared combustion system. Following this, distillation (EFLAB) and titration with 0.1 mol HCl were performed. Ash analysis was performed by combustion at 600°C, and crude fat was determined using a Soxhlet apparatus (VELP SCIENTIFICA SER 148 model). After extraction with ether, the fat-ether mixture was distilled in the apparatus to separate the solvent from the fat (AOAC, 1998). Lipid Extraction and Fatty Acid Compositions Total lipids were extracted according to the method of Bligh & Dyer (1959). Fatty acid composition was determined after lipid extraction and methylation. Fatty acid methyl esters (FAMEs) were prepared by mixing 0.25 g of extracted lipid with 4 mL of heptane and 0.4 mL of 2 N KOH in methanol. The mixture was vortexed for 2 minutes and centrifuged at 5000 rpm for 5 minutes, and the clear supernatant was transferred into GC vials. Fatty acid compositions of the samples were analyzed using a GC/MS (Thermo Scientific ISQ LT) instrument equipped Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 24 with an automated sampler. The capillary column used was a Trace Gold TG-WaxMS (60 m) column with an inner diameter of 0.25 µm and a film thickness of 0.25 µm. The column temperature was initially held at 100°C for 3 minutes, then increased to 240°C at a rate of 4°C/min after an initial hold time of 6 minutes. Helium was used as the carrier gas at a flow rate of 1 mL/min, with a split ratio of 1:20. The injection block temperature was set to 240°C, and the MS transfer line and ion source temperatures were set to 250°C and 240°C, respectively. The mass spectrometer was operated in electron impact ionization mode (70 eV). For identification and comparison, a standard FAME mixture (Supelco, 37 components, Bellefonte, PA, USA) was used (Çorapcı et al., 2021; Kocatepe et al., 2025). Total fatty acids were calculated using the following formulas: ƩSFA=C6:0 + C8:0 + C10:0 + C11:0 + C12:0 + C13:0 + C14:0 + C15:0 + C16:0 + C17:0 + C18:0 + C20:0 + C21:0 + C22:0 + C23:0 + C24:0 ƩMUFA=C14:1 + C15:1 + C16:1 + C17:1 + C18:1n‐9c + C18:1n‐9t + C20:1n‐9c + C22:1n‐9 + C24:1 ƩPUFA=C18:2n‐6t + C18:2n‐6c + C18:3n‐3 + C18:3n‐6 + C20:2 + C22:2 + C20:3n‐3 + C20:3n‐6 + C20:5n‐3 + C20:4n‐6 + C22:6n‐3 ƩOmega‐3 (ɷ3)=C18:3n‐3 + C20:3n‐3 + C20:5n‐3 + C22:6n‐3 ƩOmega‐6 (ɷ6)=C18:2n‐6t + C18:2n‐6c + C18:3n‐6 + C20:4n‐6 + C20:3n‐6 ƩOmega‐9 (ɷ9)=C18:1n‐9c + C18:1n‐9t + C20:1n‐9c + C22:1n‐9 Atherogenicity (AI) Index: [(C12:0 + (4 × C14:0) + C16:0)] / (MUFA + Omega‐3 + Omega‐6) Thrombogenicity (IT) Index: (C14:0 + C16:0 + C18:0) / [(0.5 × MUFA) + (0.5 × Omega‐6) + (3 × Omega‐3) + (Omega‐3 / Omega‐6)] Hypocholesterolemic/Hypercholesterolemic Ratio (H/H) = (C18:1n‐9 + C18:2n‐6 + C18:3n‐3 + C20:4n‐6 + C20:5n‐3 + C22:6n‐3) / (C14:0 + C16:0) Color Analysis Egg color was measured using a color spectrophotometer (ColorFlex EZ, HunterLab, USA). The L∗, a∗, and b∗ parameters are as follows: L* represents brightness (lightness-darkness), a* represents redness-greenness, and b* represents yellowness-blueness. Three replicate readings were taken for each sample. Statistical Analysis Data obtained in the experiment were analyzed using the IBM SPSS 21 statistical program. The fatty acid levels of broodstock feeds and the fatty acid composition of haploid and diploid eggs were evaluated using a t-test. The independent sample t-test was applied to the groups in the study. First, the Levene test was used to check whether the variances were equal. If the variances were equal, the significance value in the "Equal variance assumed" line was used. If the variances were not equal, the t-test result was determined by examining the significance value in the "Unequal variance assumed" line. If the sig (2-tailed) value was less than 0.05, a significant difference was concluded between the two groups. If this value was greater than 0.05, there was no significant difference between the groups (Yazıcıoğlu & Erdoğan, 2014). Nutrient content, fatty acid composition, and color analyses of haploid and diploid eggs were analyzed using the Tukey test. Data were analyzed using one-way analysis of variance (ANOVA) using the SPSS 21 statistical program and subjected to Tukey's multiple comparison test. Differences between groups were evaluated as p<0.05 (Logan, 2010). Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 25 Results Water quality parameters were measured during the winter months at two different trout farms in Seydikemer. Temperature, pH, and dissolved oxygen levels were measured as 12.25±0.53°C, 7.75±0.18, and 8.15±0.25 mg/L in facility 1. Temperature, pH, and dissolved oxygen levels were measured as 11.50±0.35°C, 7.40±0.15, and 8.50±0.30 mg/L in facility 2. Table 1. Metric measurements of the trial fish. Weight (g) Length (cm) Spawned Eggs (g) Post-spawned weight (g) Gonad Weight (g) Station 1 1. fish 3.010 64 408 2.602 602 2. fish 3.470 64 572 2.898 694 Station 2 1. fish 2.912 55 527 2.385 582.4 2. fish 2.900 57 411 2.489 580 According to the metric measurements, the trial fish at Station 1 exhibited body weights of 3.010-3.470 g, a length of 64 cm, and gonad weights ranging from 602 to 694 g. At Station 2, the fish showed body weights of 2.900-2.912 g, lengths of 55-57 cm, and gonad weights between 580 and 582.4 g (Table 1). Table 2. Haploid and diploid trout egg composition and estimated fertility at two stations Egg Composition Sample (count) Sample (gram) Diameter (mm) Fecundity (count) Station 1 Haploid 63 5.668±0.32 4.5±0.15 6.691±0.27 Diploid 78 7.634±0.45 5.5±0.20 7.713±0.35 Total 14.404 Station 2 Haploid 58 4.490±0.28 5±0.18 7.523±0.31 Diploid 57 6.060±0.38 5.5±0.22 7.492±0.33 Total 15.015 In the comparison between haploid and diploid eggs, the sample weights of diploid eggs were measured as 7.634±0.45 g and 6.060±0.38 g, while those of haploid eggs were 5.668±0.32 g and 4.490±0.28 g, respectively. Diameter measurements showed that diploid eggs had values of 5.5±0.20 mm and 5.5±0.22 mm, whereas haploid eggs measured 4.5±0.15 mm and 5±0.18 mm. Regarding fecundity, diploid eggs exhibited values of 7.713±0.35 and 7.492±0.33, compared to haploid eggs, which were 6.691±0.27 and 7.523±0.31. These results indicate that diploid eggs surpass haploid eggs in terms of weight, diameter, and fecundity (Table 2). Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 26 Table 3. Chemical composition of broodstock feeds Chemical Composition of Feeds Station 1 Station 2 Crude Protein (%) 50 45.3 Crude Fat (%) 14 19.4 Crude Fiber (%) 2.5 0.8 Crude Ash (%) 8.5 8.9 Gross Energy (GE; kJ/g) 21.58 22.71 The chemical composition of the feeds given to broodstock fish varied among the stations. The feed used at the first station contained 50% crude protein, 14% crude fat, 2.5% crude fiber, and 8.5% crude ash, yielding a gross energy value of 21.58 kJ/g. The feed used at the second station contained 45.3% crude protein, 19.4% crude fat, 0.8% crude fiber, 8.9% crude ash, and a gross energy value of 22.71 kJ/g (Table 3). Table 4. Analysis of chemical composition of diploid and haploid rainbow trout eggs from various location Chemical Composition of Eggs Haploid (1st station) Haploid (2st station) Diploid (1st station) Diploid (2st station) Dry matter (%) 29.75±1.30 28.76±1.64 30.3±2.35 29.72±3.56 Crude Protein (%) 22.09±1.63 21.02±1.37 22.37±2.48 20.98±0.67 Crude Fat (%) 3.69±0.56 3.43±0.32 3.65±0.08 4.26±1.48 Crude Ash (%) 3.96±0.62 4.30±0.24 4.29±0.35 4.47±1.41 Significant differences (p<0.05) exist between values in the same rows that are indicated by different letters. The values (n=3) are presented as mean ± SEM. Analysis of the chemical composition of diploid and haploid rainbow trout eggs revealed no statistically significant differences in dry matter, crude protein, crude fat or crude ash contents (p>0.05). Dry matter ranged from 28.76±1.64% to 29.75±1.30% in haploid eggs and from 29.72±3.56% to 30.3±2.35% in diploid eggs. Crude protein was similar between groups; haploids ranged from 21.02±1.37% to 22.09±1.63% and diploids ranged from 20.98±0.67% to 22.37±2.48%. Crude fat content ranged from 3.43±0.32% to 3.69±0.56% in haploids and from 3.65±0.08% to 4.26±1.48% in diploids. Raw ash content also did not show any significant difference, varying between 3.96±0.62% and 4.30±0.24% in haploids and 4.29±0.35% and 4.47±1.41% in diploids (Table 4). Fatty acid analysis of trout broodstock feeds from the first and second stations revealed significant differences in several saturated, monounsaturated, and polyunsaturated fatty acids. Specifically, C14:0, C16:0, C17:0, C18:0, C20:0, C22:0, C23:0, ΣSFA, C17:1, C18:1n9t, C20:1, ΣMUFA, C18:2n6t, C18:2n6c, C18:3n3, C20:2, C20:3n-3, C20:5n-3, C20:3n-6, C20:4n-6, C22:6n-3, C22:2, C22:1n9, ΣPUFA, ƩOmega-3, ƩOmega-6, ƩOmega-9, ɷ3/ɷ6, ɷ6/ɷ3, EPA/DHA, and DHA/EPA showed significant differences (p<0.05) between the two Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 27 stations. In contrast, C15:0, C15:1, C16:1, C21:0, C24:0, C24:1, C14:1, and C18:3n6 did not differ significantly (p>0.05) (Table 5). Table 5. Comparison of fatty acid levels of different broodstock feed groups (Independent Sample T-Test) Fatty Acids Groups N X ss t-test t sd p C14:0 (%) 1st Stn. 3 2.87 0.05 -10.161 4 0.001 2nd Stn. 3 4.60 0.29 -10.161 2.127 0.008 C15:0 (%) 1st Stn. 3 0.48 0.02 1.006 4 .371 2nd Stn. 3 0.45 0.04 1.006 2.748 .395 C16:0 (%) 1st Stn. 3 15.19 0.14 11.345 4 .000 2nd Stn. 3 12.25 0.42 11.345 2.476 .003 C17:0 (%) 1st Stn. 3 0.78 0.01 17.482 4 .000 2nd Stn. 3 0.50 0.02 17.482 2.616 .001 C18:0 (%) 1st Stn. 3 2.87 0.05 -10.161 4 .001 2nd Stn. 3 4.60 0.29 -10.161 2.127 .008 C20:0 (%) 1st Stn. 3 1.40 0.02 19.400 4 .000 2nd Stn. 3 1.07 0.02 19.400 3.994 .000 C21:0 (%) 1st Stn. 3 0.03 0.02 -2.457 4 .070 2nd Stn. 3 0.07 0.01 -2.457 3.200 .086 C22:0 (%) 1st Stn. 3 1.31 0.05 17.717 4 .000 2nd Stn. 3 0.54 0.05 17.717 3.980 .000 C23:0 (%) 1st Stn. 3 0.14 0.02 -3.280 4 .031 2nd Stn. 3 0.21 0.02 -3.280 3.816 .033 C24:0 (%) 1st Stn. 3 0.58 0.01 .898 4 .420 2nd Stn. 3 0.57 0.03 .898 2.580 .445 ΣSFA (%) 1st Stn. 3 32.26 0.22 8.995 4 .001 2nd Stn. 3 27.07 0.97 8.995 2.219 .009 C14:1 (%) 1st Stn. 3 0.21 0.00 -.354 4 .742 2nd Stn. 3 0.22 0.03 -.354 2.129 .756 C15:1 (%) 1st Stn. 3 0.09 0.00 1.768 4 .152 2nd Stn. 3 0.07 0.01 1.768 2.560 .191 C16:1 (%) 1st Stn. 3 0.42 0.00 2.457 4 .070 2nd Stn. 3 0.38 0.03 2.457 2.148 .125 C17:1 (%) 1st Stn. 3 0.45 0.01 -9.865 4 .001 2nd Stn. 3 0.73 0.04 -9.865 2.528 .004 C18:1n9t (%) 1st Stn. 3 1.61 0.04 -9.099 4 .001 2nd Stn. 3 1.94 0.04 -9.099 3.906 .001 C20:1 (%) 1st Stn. 3 2.68 0.03 110.363 4 .000 2nd Stn. 3 0.35 0.00 110.363 2.102 .000 C24:1 (%) 1st Stn. 3 1.16 0.29 1.012 4 .369 2nd Stn. 3 0.98 0.03 1.012 2.057 .416 ΣMUFA (%) 1st Stn. 3 34.34 0.35 6.126 4 .004 2nd Stn. 3 30.01 1.17 6.126 2.367 .017 1st Stn. 3 0.22 0.00 -23.702 4 .000 Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 28 Fatty Acids Groups N X ss t-test t sd p C18:2n6t (%) 2nd Stn. 3 0.40 0.01 -23.702 2.941 .000 C18:2n6c (%) 1st Stn. 3 18.50 0.06 30.207 4 .000 2nd Stn. 3 14.41 0.22 30.207 2.287 .001 C18:3n3 (%) 1st Stn. 3 4.05 0.05 -39.588 4 .000 2nd Stn. 3 6.78 0.10 -39.588 3.152 .000 C18:3n6 (%) 1st Stn. 3 0.17 0.01 -2.055 4 .109 2nd Stn. 3 0.21 0.03 -2.055 2.322 .158 C20:2 (%) 1st Stn. 3 1.65 0.01 -20.348 4 .000 2nd Stn. 3 1.95 0.02 -20.348 2.941 .000 C20:3n-3 (%) 1st Stn. 3 1.98 0.02 -98.508 4 .000 2nd Stn. 3 4.88 0.04 -98.508 3.298 .000 C20:5n-3 (%) 1st Stn. 3 1.90 0.02 -82.575 4 .000 2nd Stn. 3 5.76 0.07 -82.575 2.569 .000 C20:3n-6 (%) 1st Stn. 3 0.49 0.00 7.155 4 .002 2nd Stn. 3 0.44 0.01 7.155 2.941 .006 C20:4n-6 (%) 1st Stn. 3 0.80 0.01 -23.888 4 .000 2nd Stn. 3 1.24 0.03 -23.888 2.424 .001 C22:6n-3 (%) 1st Stn. 3 3.39 0.16 -33.310 4 .000 2nd Stn. 3 6.68 0.05 -33.310 2.456 .000 C22:2 (%) 1st Stn. 3 0.20 0.00 5.892 4 .004 2nd Stn. 3 0.11 0.02 5.892 2.00 .028 C22:1n9 (%) 1st Stn. 3 0.11 0.00 6.364 4 .003 2nd Stn. 3 0.08 0.00 6.364 4.00 .003 ΣPUFA (%) 1st Stn. 3 33.38 0.18 -52.247 4 .000 2nd Stn. 3 42.90 0.25 -52.247 3.709 .000 ƩOmega-3 (%) 1st Stn. 3 11.33 0.15 -98.888 4 .000 2nd Stn. 3 24.12 0.16 -98.888 3.974 .000 ƩOmega-6 (%) 1st Stn. 3 20.19 0.05 25.970 4 .000 2nd Stn. 3 16.71 0.22 25.970 2.254 .001 ƩOmega-9 (%) 1st Stn. 3 31.99 0.15 5.917 4 .004 2nd Stn. 3 27.61 1.27 5.917 2.056 .026 ɷ3/ɷ6 1st Stn. 3 0.561 0.00 -67.569 4 .000 2nd Stn. 3 1.44 0.02 -67.569 2.421 .000 ɷ6/ɷ3 1st Stn. 3 4.56 0.05 81.798 4 .000 2nd Stn. 3 2.12 0.00 81.798 2.046 .000 EPA/DHA 1st Stn. 3 0.56 0.03 -15.605 4 .000 2nd Stn. 3 0.86 0.00 -15.605 2.100 .003 DHA/EPA 1st Stn. 3 1.78 0.10 10.537 4 .000 2nd Stn. 3 1.15 0.00 10.537 2.019 .009 N: Number of samples in each group, X : Mean. ss: Standard deviation, sd: Degrees of freedom, p: Significance level Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 29 Table 6. Fatty acid composition comparison between diploid and haploid rainbow trout eggs from various locations Fatty acids Haploid (1st station) Haploid (2st station) Diploid (1st station) Diploid (2st station) C14:0 (%) 1.25±0.02c 1.85±0.06a 1.42±0.01b 1.18±0.08c C15:0 (%) 0.28±0.02ab 0.26±0.00b 0.30±0.00a 0.16±0.01c C16:0 (%) 13.44±0.38ab 10.92±0.03c 13.88±0.07a 12.85±0.23b C17:0 (%) 0.64±0.01a 0.41±0.01c 0.57±0.01b 0.30±0.00d C18:0 (%) 10.11±0.20b 8.14±0.04c 10.60±0.10a 8.31±0.00c C20:0 (%) 0.27±0.01b 0.15±0.00c 0.29±0.00a 0.13±0.01d C21:0 (%) 0.07±0.13ab 0.02±0.04b 0.25±0.00a 0.02±0.01b C22:0 (%) 0.26±0.03a 0.02±0.00b 0.34±0.07a 0.13±0.06b C23:0 (%) 0.20±0.05b 0.21±0.01ab 0.29±0.02a 0.01±0.00c C24:0 (%) 1.04±0.03a 0.76±0.03b 1.04±0.11a 0.43±0.06c ΣSFA (%) 27.61±0.56b 22.80±0.12c 29.04±0.10a 23.24±0.18c C14:1 (%) 0.03±0.02a 0.04±0.00a 0.04±0.00a 0.02±0.00a C15:1 (%) 0.02±0.01a 0.02±0.00ab 0.02±0.00ab 0.01±0.00b C16:1 (%) 0.30±0.02a 0.24±0.00b 0.31±0.00a 0.16±0.02c C17:1 (%) 0.38±0.04a 0.25±0.02b 0.41±0.04a 0.12±0.00c C18:1n9c (%) 19.64±0.04c 20.04±0.16b 21.50±0.03a 15.93±0.11d C18:1n9t (%) 2.03±0.13a 1.70±1.19a 2.00±0.03a 2.33±0.04a C20:1 (%) 2.24±0.08b 0.18±0.00d 2.47±0.04a 0.44±0.02c C24:1 (%) 0.14±0.00b 0.07±0.01c 0.13±0.02b 1.07±0.01a ΣMUFA (%) 24.98±0.07b 22.85±1.08c 27.09±0.06a 20.17±0.16d C18:2n6t (%) 0.49±0.01b 0.32±0.01c 0.56±0.01a 0.18±0.00d C18:2n6c (%) 0.03±0.00b 13.31±0.08a 0.03±0.00b 13.17±0.06a C18:3n3 (%) 3.11±0.02c 3.27±0.01b 3.48±0.01a 2.32±0.09d C18:3n6 (%) 1.54±0.02b 0.58±0.00c 1.72±0.08a 0.41±0.03d C20:2 (%) 4.46±0.02c 4.81±0.02ab 4.85±0.01a 4.72±0.06b C20:3n-3 (%) 1.56±0.04b 1.30±0.00c 1.81±0.00a 0.01±0.00d C20:5n-3 (%) 6.30±0.06b 5.58±0.13d 6.79±0.03a 5.81±0.05c C20:3n-6 (%) 6.37±0.03a 3.47±0.04c * 4.28±0.04b C20:4n-6 (%) 7.94±0.11b 5.89±0.01c 8.35±0.03a 5.45±0.03d C22:6n-3 (%) 15.47±0.31c 15.28±0.11c 16.01±0.11b 19.81±0.14a C22:2 (%) 0.17±0.00b 0.04±0.00c 0.21±0.02a 0.02±0.00c C22:1n9 (%) 0.06±0.08b 0.12±0.00ab 0.20±0.01a 0.07±0.02b ΣPUFA (%) 45.28±0.47c 53.87±0.08b 43.85±0.12d 56.21±0.11a ƩOmega-3 (%) 26.45±0.40b 25.43±0.13c 28.10±0.11a 27.95±0.09a ƩOmega-6 (%) 16.40±0.07b 23.58±0.12a 10.68±0.10c 23.51±0.02a ƩOmega-9 (%) 23.98±0.21b 22.05±1.10c 26.17±0.03a 18.78±0.15d ɷ3/ɷ6 1.61±0.01b 1.07±0.01d 2.63±0.03a 1.19±0.00c ɷ6/ɷ3 0.62±0.00c 0.92±0.00a 0.38±0.00d 0.84±0.00b EPA/DHA 0.40±0.00a 0.36±0.01b 0.42±0.00a 0.29±0.00c DHA/EPA 2.45±0.03c 2.74±0.07c 2.35±0.01b 3.41±0.05a AI 0.31±0.00b 0.35±0.01a 0.35±0.00a 0.33±0.00ab TI 0.24±0.00a 0.20±0.00b 0.24±0.00a 0.20±0.00b HH 3.74±0.13c 5.12±0.12a 3.60±0.02c 4.46±0.05b EPA+DHA 21.78±0.36c 20.86±0.15d 22.80±0.11b 25.62±0.11a Küçük&Yılmaz, Egg Composition &Color in Haploid & Diploid Trout Aquatic Animal Reports 4(1)(2026)21-36 36 Meng, Y., Liu, X., Guan, L., Bao, S., Zhuo, L., Tian, H., ... & Ma, R. 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