Supplementation of pumpkin Cucurbita Maxima seed powder as a feed additive on growth and survival of rohu, Labeo rohita (Hamilton, 1822)

This study evaluates the effects of the dietary inclusion of Pumpkin (Cucurbita maxima) seed powder (PSP) on the growth and survival of Labeo rohita fingerlings over 60 days. Pumpkin seeds are rich in antioxidants and vitamin E. It has antinutritional and toxic factors, such as polyphenols, flavonoids, tannins, and cyanogenic glycosides. PSP was incorporated at varying levels (T0 (0%), T1 (1.5%), T2 (3%), T3 (4.5%), and T4 (6%) Pumpkin seed powder) into fish diets. Growth parameters and survival were assessed, and it showed that 4.5% PSP diet (T3) significantly improved growth rates and feed conversion ratio. However, survival rates were unaffected across treatments. Thus, a 4.5% PSP diet optimally enhances fish growth and Survival.

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Introduction

The aquaculture sector is vital to maintaining fish supply, especially with the decline in capture fish industry. Total production of aquatic animals has increased over the decades, going from 19 million tonnes (Live weight equivalent) in 1950 to a record of over 185 million tonnes in 2022, with an average annual growth rate of 3.2% (FAO, 2024). In 2022, global aquaculture production increased to 130.9 million tonnes, up 8.1 million tonnes from the 122.8 million tonnes produced in 2020. The most significant of the three Indian major carp species employed in carp polyculture systems is Rohu (Labeo rohita). It is a wellknown component of aquaculture because of its quick growth rate, capacity to adapt to a wide range of environmental circumstances, and strong market demand. Fish growth at various stages is primarily influenced by the type of feed, feeding frequency, feed intake, and the fish's ability to absorb nutrients (Abid & Ahmed, 2009) [1] . Farmed rohu is omnivorous and has a diet containing 30-35% protein (Rasal & Sundaray, 2020) [11] . Medicinal plants have been proven as growth promoters. They Introduction have the properties of growthpromoting ability, a tonic to improve the antimicrobial capability, and stimulating appetite and anti-stress characteristics (Citarasu et al., 2010) [2] . Pumpkin is a well-known edible plant with substantial medicinal properties due to its unique natural edible substances (Yadav et al., 2010) [15] . Pumpkin seed has considerable economic importance, generating huge amounts of seeds. Pumpkin seeds have been used traditionally for herbal, medicinal, and clinical purposes. Pumpkin seeds are rich in essential fatty acids such as palmitic, oleic, linoleic, and steric acids as well as arginine and glutamic acid. However, it has antinutritional and toxic factors, such as polyphenols, flavonoids, tannins and cyanogenic glycosides (Del Vechio et al., 2005) [4] . Pumpkin seeds contain approximately 30% crude protein (Sharma et al. 1986; Esuoso et al. 1998) [5,12] . The amount and quality of protein in fish feed significantly impact their growth and survival rates (Hua et al., 2019) [7] . Formulating fish feed utilizing relatively cheap, easily available, and extremely high-nutrient terrestrial resources, including Cucurbita maxima seeds is necessary.

Materials and Methods

The experiment was conducted at the College of Fisheries Science, KU, Veraval for 60 days. The experiment was conducted in 20 rectangular plastic aquarium tanks (2 x 1 x 1 feet) with adequate aeration. The Labeo rohita seed was sourced from a commercial IMC and acclimatized in a 500-liter FRP tank for 7 days. Fish with an average weight of 1.310±0.009 g was selected for the experiment.

Experimenatal Diet Preparation

Five experimental diets (T 0 to T 4 ) were prepared. Treatment groups include T 0 (control, no Pumpkin seed powder (PSP), T 1 with (1.5% PSP), T 2 (3% PSP), T 3 (4.5% PSP) and T 4 (6% PSP). The feed mixture was then subjected to heat treatment at 121 °C under 15 lbs of pressure for 15 minutes. After the steam cooking process, vitamins and minerals were added and carefully blended into the mixture.

Proximate Analysis of Ingredients

The proximate composition of the ingredients was analysed using the standard method of AOAC (2019). The composition of the experimental diet was formulated using Pearson's square method at a 30% protein level. The formula for the experimental diets, including their percentage composition, is detailed in Table 2 based on the treatment specifications. Completely Randomised Design (CRD) experimental design will be used in the experiment and estimate the arithmetic mean, standard deviation, and coefficient of variation using standard statistical Methods (Snedecor & Cochran, 2014) [13] .

Ingredients	Diet (30% Protein)	—	—	—	—
	T0 (Control)	T1 (1.5% PSP)	T2 (3.0% PSP)	T3 (4.5% PSP)	T4 (6% PSP)
Pumpkin seed powder	0	15	30	45	60
Sterilized fish meal	25	24	24	24	24
Wheat flour	15	14	14	13	13
Tapioca powder	13	13	12	12	12
Gnoc	41	41.5	41	40.5	39
Plant oil	2	2	2	2	2
Fish oil	2	2	2	2	2
Vitamin and minerals	2	2	2	2	2
Total	100	100	100	100	100

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Composition of Experimental Diet (in gms)

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The collected data was analyzed using a one-way analysis of variance (ANOVA) to identify significant differences between the means. The statistical analysis was performed using IBM SPSS version 22 software.

Results

The study examines Body weight gain, feed conversion ratio, specific growth rate, Protein efficiency ratio, survival rate, Average weight gain and Water quality parameters in L. rohita-fed diets with varying levels of Pumpkin seed powder.

Discussion

The growth of Labeo rohita was assessed by sampling 15 fish per tank at 15-day intervals. After 60 days, final weights were highest in T 3 (4.297±0.012 g), indicating pumpkin seed powder significantly enhanced growth. Over 60 days, Labeo rohita fry showed significant differences (p<0.05) in mean weight gain across treatments, with T 3 achieving the highest (2.982±0.007 g). Similar findings were reported by Teves et al. (2014) [14] for Nile tilapia (Oreochromis niloticus) fed squash (Cucurbita maxima) seed meal, where a 5% inclusion level resulted in the highest weight gain and specific growth rate. Musthafa et al. (2017) [8] also demonstrated that pumpkin (C. mixta) seed meal at 4 g kg⁻¹ and 6 g kg⁻¹ significantly (p< 0.05) improved weight gain and survival rates in fish. Its high protein and vitamin E content as key factors for improved growth and survival. The specific growth rate (SGR) data (Table 3) showed that T 3 and T 4 had the highest SGR, significantly exceeding T 0 , T 1 , and T 2 , while T 0 and T 1 had the lowest SGR with no significant difference (p>0.05) between them. These findings align with Dada et al. (2015) [3] for African catfish (Clarias gariepinus) fed fluted pumpkin leaf meal, where 5 g/kg feed led to improved growth, feed conversion ratio, and SGR. Similarly, Yanti et al. (2022) [16] observed that Barramundi (Lates calcarifer) fingerlings fed a diet with 5% pumpkin flour showed significant improvements in weight gain, daily growth, and SGR.

Parameters	Treatments	—	—	—	—
	T0 (Control)	T1 (PSP 1.5%)	T2 (PSP 3%)	T3 (PSP 4.5%)	T4 (PSP 6%)
IW (g)	1.310±0.009a	1.317±0.006a	1.325±0.006a	1.315±0.006a	1.310±0.010a
FW (g)	3.407±0.042e	3.535±0.104d	3.745±0.011c	4.297±0.012a	4.097±0.008b
MWG (g)	2.097±0.033e	2.217±0. 007d	2.420±0. 012c	2.982±0. 007a	2.787±0. 016b
SGR (%)	0.775±0.003e	0.803±0.006d	0.851±0.008c	1.001±0.005a	0.960±0.014b
Survival (%)	91.75± 2.39a	92.50±3.22a	93.00±3.53a	95.25±2.05a	95.75±2.05a
AWG (%)	160.08±1.581e	168.32±0.920d	182.66±1.569c	226.81±0.895a	212.85±2.908b

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Growth performance and survival of L. rohita fed a diet with 0% (Control), 1.5%, 3%, 4.5%, and 6% supplementation of pumpkin

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At the end of the 60-day experimental period, the final mean percentage weight gain for Treatment T 3 , with 4.5% pumpkin seed powder, showed the highest weight gain, followed by T 4 with 6% inclusion of PSP. Similarly, Teves et al. (2014) [14] found that Oreochromis niloticus fed squash (Cucurbita maxima) seed meal at a 5% inclusion level achieved the highest weight gain and SGR. Naiel et al. (2024) [9] reported that Nile tilapia-fed ochratoxin Acontaminated diets supplemented with pumpkin seed extract exhibited significantly improved final biomass, daily weight gain, SGR, and percentage weight gain. The survival rates of L. rohita are represented in Table 3. Statistical analysis revealed no significant difference (p>0.05) among treatments, indicating consistent survival rates across all groups. Similarly, Yanti et al. (2022) [16] reported that a 5% pumpkin flour diet significantly improved survival, in Barramundi (Lates calcarifer). Musthafa et al. (2017) [8] demonstrated that diets with 4 g kg⁻¹ and 6 g kg⁻¹ pumpkin (C. mixta) seed meal significantly (p<0.05) enhanced survival rates.

Feed utilization by Labeo rohita

Feed utilization by Labeo rohita

Discussion

After 60 days, the feed conversion ratio (FCR) for T 3 was 2.248±0.011 (Table 4). Statistical analysis revealed significant differences (p>0.05) among treatments, with T 3 showing the lowest FCR, indicating superior feed efficiency. Mounes et al. (2024) demonstrated that replacing soybean meal with pumpkin seed cake in Nile tilapia diets significantly improved FCR. Similarly, Omosowone and Ogunrinde (2018) [10] found that a diet with a 25% inclusion level of pumpkin seed meal resulted in the best FCR for Heteroclarias. These findings highlight the effectiveness of pumpkin seed as a dietary supplement for improving FCR, enhancing growth performance, and potentially reducing feed costs in aquaculture. Analysis showed significant variation in protein efficiency ratio (PER) among treatments, with T 3 achieving the highest PER (1.482±0.007), followed by T 4 (1.402±0.009), and T 0 the lowest. High PER reflects efficient feed protein conversion, consistent with studies by Musthafa et al. (2017) [8] and Teves et al. (2014) [14] , highlighting the benefits of pumpkin seed meal in enhancing growth efficiency.

Parameters	Treatments	—	—	—	—
	T0 (Control)	T1 (PSP 1.5%)	T2 (PSP 3%)	T3 (PSP 4.5%)	T4 (PSP 6%)
FCR	2.895±0.033a	2.802±0.010b	2.631±0.018c	2.248±0.011e	2.376±0.015d
PER	1.151±0.013e	1.189±0.004d	1.266±0.009c	1.482±0.007a	1.402±0.009b

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Evolution of Feed Conversion ratio (FCR) and Protein efficiency ratio (PER) of L. Rohita recorded in different treatments at the

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Physico-chemical water parameters

Water quality was monitored weekly for temperature, pH, dissolved oxygen, and total hardness throughout the 60-day study. It was in the ideal range throughout the experiment.

Conclusion

The study concluded that supplementing the Labeo rohita diet with pumpkin seed powder (PSP) positively affected growth, feed conversion, and immune response. The best growth was observed in fish fed 4.5% PSP (T 3 ), showing the highest mean weight gain, specific growth rate (SGR), and favorable feed conversion ratio (FCR). Survival rates remained high, and no significant difference in mortality was noted. The study concluded that 4.5% PSP supplementation improved both growth and Survival in Labeo rohita, yielding the best outcomes.

Figures and tables from the paper

Ingredients (%)	Crude protein (%)	Crude fat (%)	Ash (%)	Moisture (%)
Fishmeal	55.03	4.20	15.20	7.91
Pumpkin seed powder (PSP)	24.12	22.17	3.31	6.184
Wheat flour	11.94	1.3	5.32	9.81
GNOC	35.56	4.92	8.07	7.19
Tapioca	4.81	0.61	0.14	8.13

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Proximate Analysis of Ingredients

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1. Abid M, Ahmed MS. Efficacy of feeding frequency on growth and survival of Labeo rohita (Ham.) fingerlings under intensive rearing. J Anim Plant Sci. 2009;19(2):111-113.
2. Citarasu T. Herbal biomedicines: a new opportunity for aquaculture industry. Aquac Int. 2010;18(3):403-414.
3. Dada AA. Use of fluted pumpkin (Telfairia occidentalis) leaf powder as a feed additive in African catfish (Clarias gariepinus) fingerlings. Int. J Biol Chem Sci. 2015;9(1):301-307.
4. Del-Vechio G, Correa AD, Abreu CMPD, Santos CDD. Effect of the thermal processing on pumpkin seeds (Cucurbita spp.) on the levels of antinutritional factors and/or toxics. Ciênc Agrotecnol. 2005;29(2):369-376.
5. Esuoso K, Lutz H, Kutubuddin M, Bayer E. Chemical composition and potential of some underutilized tropical biomass. I: fluted pumpkin (Telfairia occidentalis). Food Chem. 1998;61(4):487-492.
6. FAO. The State of World Fisheries and Aquaculture 2024. Blue Transformation in action. Rome: FAO; 2024. Available from: https://doi.org/10.4060/cd0683en
7. Hua K, Cobcroft JM, Cole A, Condon K, Jerry DR, Mangott A, et al. The future of aquatic protein: implications for protein sources in aquaculture diets. One Earth. 2019;1(3):316-329.
8. Musthafa MS, Ali ARJ, Kumar MS, Paray BA, AlSadoon MK, Balasundaram C, et al. Effect of Cucurbita mixta (L.) seed meal enrichment diet on growth, immune response and disease resistance in Oreochromis mossambicus. Fish Shellfish Immunol. 2017;68:509-515.
9. Naiel MA, Fawzy RM, Zaki EM, Abo-Elmatty AM, ElTarabily KA, Negm SS. Pumpkin seed extract attenuates toxic effects of ochratoxin-contaminated diets in terms of growth performance, and antioxidant and immunological indices of Nile tilapia. Aquac Rep. 2024;38:102300.
10. Omosowone OO, Ogunrinde AM. Effect of partial or total supplementation of soybean meal with fluted pumpkin (Telfairia occidentalis) seed meal in the diet of hybrid Catfish (Heteroclarias) fingerlings. Food Sci Nutr. 2018;6(5):1190-1195.
11. Rasal KD, Sundaray JK. Status of genetic and genomic approaches for delineating biological information and improving aquaculture production of farmed rohu, Labeo rohita (Ham, 1822). Rev Aquac. 2020;12(4):2466-2480.
12. Sharma PB, Lal BM, Madaan TR, Chatterjee SR. Studies on the nutritional quality of some cucurbit kernel proteins. J Sci Food Agric. 1986;37(4):418-420.
13. Snedecor GW, Cochran WG. Statistical Methods. 8th ed. Wiley Blackwell; 2014. p. 491.
14. Teves JFC, Fernandez MNM, Ragaza JA. Effects of replacing fishmeal with squash seed meal (f) on performance of juvenile Nile tilapia (Oreochromis niloticus). Aquac Aquar Conserv Legis. 2014;7(2):6875.
15. Yadav M, Jain S, Tomar R, Prasad GBKS, Yadav H. Medicinal and biological potential of pumpkin: an updated review. Nutr Res Rev. 2010;23(2):184-190.
16. Yanti RZ, Muchlisin ZA, Muhammadar AA, Fadli N. Application of pumpkin flour to the diet of Barramundi Lates calcarifer fingerling. Depik J Ilmu-Ilmu Perair Pesisir Perikan. 2022;11(2):176-181.