Supplementation of fish meal with moringa oleifera seed meal (MOSM) and its effect on growth performance and survival in ROHU, Labeo rohita (Hamilton, 1822) fry

Introduction

Global fisheries and aquaculture are progressive sectors whose production plays a crucial role in providing food and nutrition to the growing population in the present as well as in the future (FAO, 2022) [9,10] . Besides food production, this sector contributes to economic growth, employment opportunities, and livelihood generation among the local to literate populates (Ngasotter et al, 2020) [18] . Commercially important wild stock is declining day by day due to numerous reasons. One of the main reasons remains unsustainable fishing practices, which affected approximately 35.4% of the global marine fisheries stocks in 2019 (FAO, 2022) [9,10] . Therefore, from the past few years, the supply of seafood has remained constant and limited, to rectify these problems aquaculture is the foremost solution to increase production and to meet the growing world's demand for seafood (Das et al, 2022) [7] . Indian major carp are widely cultured farm fishes due to their fast growth, taste and consumer demand, thus forming the backbone of freshwater aquaculture in India (Bais, 2018). Labeo rohita is the principal species among the other two IMCs, the reason behind its popularity is its delicious taste and public preference. The Rohu is a species which is high in nutrients, including omega-3 fatty acids, vitamins A, B, and D. (Kitchen, 2022) [15] . For successful culture operations, it is important to provide a well-balanced diet and proper feeding as the cost of feed in an intensive culture system is about 40-60% (Bamidele et al, 1995) [6] . Fish meal is commonly employed in the formulation of aquaculture diets because it is a major source of protein and lipids (Miles and Chapman, 2006) [16] . The expensive rate of fish meal and the continuous reduction in the availability of dietary fish meal over the last decade, it is required to find alternative protein sources for aquafeed (Abdel-Latif et al, 2022) [1] . Nowadays, Plant-based protein sources are being utilized to replace or supplement proteins in fish diets because they are constantly available, relatively cheap, eco-friendly, environmentally safe, and sustainable (Gatlin et al, 2007) [11] . Moringa oleifera (also known as a tree of life, drumstick tree, or miracle tree) is a widely distributed plant in the Moringaceae family (Falowo et al, 2018) [8] . The moringa plant has various components, including roots, leaves, flowers, fruits, and seeds. These parts are rich in phytochemicals, especially tannins, anthraquinones, carotenoids, flavonoids, and proanthocyanidins. (Abdel-Latif et al, 2022) [1] . Moringa oleifera seeds are spherical or triangular in shape when completely grown and the kernel is covered by a thin woody shell with three paper wings (Bamidele et al, 1995) [6] . The seeds are high in protein, vitamins, and minerals, as well as high in essential amino acids (EAA), such as cysteine, methionine, and tryptophan (Yuangsoi et al, 2014) [22] . The major macromolecule in moringa seeds is protein and it contains more protein than leaves i.e., 33.53 % in seeds and 18.63 % in leaves (Mune et al, 2016) [17] . Therefore, M. oleifera seeds powder is suitable to use in aquaculture diets to improve the growth, survival, and immunity of culture organisms. Due to its coagulation capabilities, M. oleifera seed powder is utilized successfully in many nations for water purification in aquaculture farms (Kavitha et al, 2012) [14] .

Materials and Methods

Experimental laboratory

The research titled "supplementation of fish meal with Moringa oleifera seed meal (MOSM) and its effect on growth performance and survival in rohu, Labeo rohita (Hamilton, 1822) fry" took place at the College of Fisheries Science, KU, Veraval. The experiment lasted for 60 days, commencing on October 20, 2022, and concluding on December 20, 2022.

Experimental animal

For the experiment, fry of L. rohita (1.264±0.02 g) were procured from a commercial fish hatchery located in Gujarat. Fry were carefully packed in oxygen-filled polythene bags and brought to the College of Fisheries Science, KU, Veraval. The fry was acclimatized in the cleaned and disinfected FRP tanks (500 L) with continuous aeration and fed three times a day. The fish were provided with a basal diet containing 30% crude protein during this period.

Experimental diet preparation

"Sterile-A" grade fish meal was obtained from a local fish meal plant located in Veraval. Other feed ingredients such as wheat flour, tapioca flour, ground nut oil cake, plant oil and vitamin minerals premix were obtained from the local market of Veraval. Fish oil was obtained from the nearby fish processing plant at Veraval. The moringa seeds were collected from the dry drumsticks of the moringa tree which is locally available. They were sun-dried for 6-7 days, ground into a powder using an electronic mixer grinder, and then sieved to remove any impurities. After processing, the obtained moringa seed powder was stored in a sealed plastic container at room temperature until further use. The proximate composition of ingredients was analysed by using the standard methods (AOAC, 2000) [13] . The Pearson square method was used to produce the experimental feeds. Based on the treatment details, five experimental diets were formulated and labeled as T 0 , T 1 , T 2 , T 3 , and T 4 . According to the feed formula, the required quantities of the ingredients were collected and then carefully weighed. The ingredients were combined in a plastic tray along with the necessary amount of water, and mixed thoroughly to create a dough. The prepared dough was placed in the autoclave at 121 °C and 15 lbs. pressure for 15-20 minutes. After the steaming process, vitamins and minerals premix was incorporated into the feed mixture. Feed pellets were prepared using a mechanical pelletizer. The prepared feed pellets were spread out on steel trays and dried under the direct sunlight. Then, the pelleted feed was stored in clearly labeled air-tight plastic bottles.

Experimental arrangement

The experiment was carried out in 15 plastic tanks. Prior to use, thorough washing and disinfection of all plastic tanks were performed using chlorinated water, followed by flushing and drying, to ensure cleanliness and sterility. The tanks were then arranged on stands within the Department of Aquaculture and filled with freshwater. The experiment encompassed five treatments and three replications, and it was executed utilizing a completely randomized design (CRD).

Experimental tank management L. rohita fry was stocked at the density of 20 nos. per tank in all 15 experimental tanks. During the experimental period, L. rohita fry were fed twice daily at 5% of their body weight, with morning and evening feeding sessions. The required amount of feed was weighed before feeding it to L. rohita fry. The diet contained 30% protein, and feed amounts were adjusted based on fortnightly sampling of the fish weight.

Freshwater that had been filtered and disinfected was used to partially replace the water in the experimental units every other day. To ensure appropriate amounts of dissolved oxygen in each tank, continuous aeration was supplied throughout the entire study period. Additionally, to prevent the accumulation of extra feed pellets and any residual feces, the experimental tanks were diligently cleaned and siphoned each day prior to both the morning and evening feedings. These cleaning measures helped maintain optimal water quality and hygiene within the tanks.

Growth, survival and feed utilization analysis

At the 15-days interval, all the fish in each treatment were counted with their body weight for analysis of the growth performance, feed utilization and survival. Before taking the weight, fish were starved overnight. To calculate the growth parameters and feed utilization in the experiment, the following formulas were used:

Body composition analysis

After the completion of the experiment, the proximate compositions of L. rohita were determined. For the proximate analysis, fish were taken from each experimental treatment and ice-killed. The experimental fish were subjected to determine the crude protein, moisture, crude fat, and ash contents using standard analytical methods (AOAC, 2000) [13] . Moisture was determined by taking a known weight of samples in a clean dried petri dish and drying them in a hot air oven at 100±1 °C overnight until a constant weight was attained. The crude protein of the samples was determined as total nitrogen by the micro Kjeldahl method after acid digestion. The Soxhlet extraction method was used to determine the crude fat content of the sample. The ash content was calculated by placing a known weight of the sample in a silica crucible, heating it in a muffle furnace at 600 °C for 6-8 hours.

Analysis of water quality parameters

Throughout the experimental period, water quality parameters including temperature, pH, dissolved oxygen (DO), total alkalinity, ammonia, and total hardness were examined weekly using the standard methods of APHA (2017) [3] .

Statistical analysis

Statistical analysis was performed by using standard statistical methods (Snedecor and Cochran, 2014) [19] .

Results

Growth performance and survival rate of L. rohita

The experimental results for the 60-day study on the supplementation of fishmeal with Moringa oleifera seed meal (MOSM) are provided in the Table 2. The outcomes include mean weight gain (MWG), percentage weight gain (PWG), specific growth rate (SGR), and survival rate (SR) for all five treatment groups were calculated. The higher growth performance was reported in T 1 (5% MOSM) treatment at the end of the experiment, followed by in T 2 , T 0 , T 3 and lowest in T 4 . Based on the statistical analysis, there were significant difference (p<0.05) observed in MWG, PWG and SGR among the treatment groups.

There was no significant difference (p>0.05) in the survival rate of L. rohita when given different amounts of moringa seed meal.

Feed utilization by L. rohita

In the present research feed utilization of L. rohita fry in the various treatments was analyzed through parameters such as feed conversion ratio (FCR) and protein efficiency ratio (PER), which are displayed in Table 3. Lowest (best) FCR was observed in T 1 treatment group followed by in T 2 . The highest PER was observed in T 1 treatment group followed by in T 2 . The statistical analysis of the FCR and PER revealed significant differences (p<0.05) between the T 1 and T 2 treatment groups compared to the others. However, no significant differences (p>0.05) were observed among the T 0 , T 3 , and T 4 treatment groups.

Body proximate composition analysis of L. rohita

The proximate composition of L. rohita fry was estimated shortly after the experiment was completed and the results obtained from the study were presented in Table 4. The proximate composition of the experimental fish L. rohita fry indicated that the protein and fat content was higher in the treated groups.

Discussion

The availability of low-cost protein sources for the formulation of aquafeed lowers the production cost in aquaculture systems (FAO, 2020) [9,10] . Among the various feed ingredients available, fish meal is commonly employed in the formulation of aquaculture diets because it is a major source of protein and lipids (Miles and Chapman, 2006) [16] .

As a result, due to the expensive rate of fish meal and the continuous reduction in the availability of dietary fish meal over the last decade, it is required to find alternative protein sources for aquafeed (Abdel-Latif et al, 2022) [1] . Moringa oleifera seeds (MOS) are high in protein, vitamins, and minerals, as well as high in essential amino acids (EAA), such as cysteine, methionine, and tryptophan (Yuangsoi et al, 2014) [22] . However, there is less information available on the use of moringa seed powder in the diet of rohu. The findings obtained demonstrate that, the of 5% MOSM supplementation yielded significantly superior outcomes in terms of mean weight gain (MWG), percent weight gain (PWG), specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER) when compared to the other treatments. Possibly, this could be due to the high crude protein content and sufficient levels of essential amino acids in MOSM, meeting the specific requirements of L. rohita.

The present study revealed that increasing the level of MOSM more than 10% led to a decline in the growth performance and feed utilization of L. rohita. A similar result was observed in a study by Tabassum et al. (2021) [20] on Cirrhinus mrigala fingerlings, replacing fish meal with M. oleifera leaf meal (MOLM) in their diets showed positive effects on growth parameters The researchers came to the conclusion that adding MOLM at a level of 10% enhanced growth performance without having a negative effect on fish health. However, it was noted that higher levels of replacement had negative effects on the observed parameters. Likewise, Al-Dubakel and Taher (2020) [2] conducted a study where they supplemented M. oleifera leaf meal (MOLM) into the diet of common carp at various levels (0%, 5%, 10%, and 15%). In comparison to other supplementation doses, their research showed that feeding animals 5% MOLM increased growth performance and feed utilization. Feeding L. rohita with varying levels of MSM in the present study did not significantly influence the survival rate.

The proximate analysis of the fish samples revealed variations in their composition across different treatment groups. The highest moisture content was observed in T 0 , T 1 exhibited the highest protein content, T 2 had the highest fat content and Ash content was highest in T 3 treatment group. The similar result was obtained by the Arsalan et al. (2016) [4] , who investigated the effect of Moringa oleifera leaf meal (MOLM) diets on the carcass composition of L. rohita fingerlings. Their investigation led them to the conclusion that MOLM can substitute for fish meal in diets up to 10%, enhancing the nutritional value of L. rohita fingerlings.

To conclude, the research findings provide evidence that expensive fishmeal can be effectively supplemented with affordable plant products, such as Moringa oleifera seed meal (MOSM) at a rate of 10%. This supplementation has demonstrated positive outcomes in terms of growth, body composition, and feed utilization, all without any adverse effects.

Conclusion

The present study demonstrates that Moringa oleifera seed meal (MOSM) can effectively replace fishmeal in the diet of Labeo rohita fry, improving growth performance, feed utilization, and body composition. The optimal supplementation level was found to be 5% MOSM, which yielded the best results for mean weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio. Higher supplementation levels led to diminished growth, while no significant differences were observed in survival rates across treatments. Overall, MOSM proves to be a cost-effective and sustainable alternative protein source for aquaculture feed, reducing reliance on fishmeal without compromising fish health or performance.

Acknowledgement

AcknowledgementsFirst and foremost, I extend my deepest appreciation to my supervisor, (Dr. V. C. Bajaniya), for their unwavering guidance, valuable insights, and continuous support throughout the research process. I also want to acknowledge the [College of Fisheries Science, Kamdhenu University, Veraval] for providing the necessary resources and research facilities that facilitated the smooth progress of this study.

References

1. Abdel-Latif HM, Abdel-Daim MM, Shukry M, Nowosad J, Kucharczyk D. Benefits and applications of Moringa oleifera as a plant protein source in Aquafeed: A review. Aquaculture. 2022;547: 737369. https://doi.org/10.1016/j.aquaculture.2021.737369
2. Al-Dubakel AY, Taher MM. Use of moringa (Moringa oleifera) leaf meal as partial replacement of soybean meal in common carp feed. Iraqi Journal of Aquaculture. 2020;17(2):103-115.
3. Arsalan MZH, Hussain SM, Asrar M, Anwar H, Rehan MMH, Shahzad MM, Wahab N. Effects of Moringa oleifera leaf meal (MOLM)-based diets on carcass composition and hematology of Labeo rohita fingerlings. Journal of Biodiversity and Environmental Sciences. 2016;9(1):214-223.
4. Bais B. Fish scenario in India with emphasis on Indian major carps. International Journal of Avian & Wildlife Biology. 2018;3(6):409-411.
5. Bamidele NA, Obasa SO, Ikeiwenwe NB, Abdulraheem I, Adeoye AA, Odebiyi OC. Effect of dried moringa (Moringa oleifera) seed meal-based diets on the growth, hematological, biochemical parameters and histopathology of the African catfish (Clarias gariepinus) fingerlings. International Journal of Fisheries and Aquatic Studies. 1995;2(4S):27-34.
6. Das BK, Behera BK, Sajina AM, Mishal P, Swain HS, Meena DK, et al., editors. Souvenir of 1st Indian Fisheries Outlook 2022. Director, ICAR-CIFRI;
7. Falowo AB, Mukumbo FE, Idamokoro EM, Lorenzo JM, Afolayan AJ, Muchenje V. Multi-functional application of Moringa oleifera Lam. in nutrition and animal food products: A review. Food Research International. 2018;106:317-334. https://doi.org/10.1016/j.foodres.2017.12.079
8. FAO. The State of World Fisheries and Aquaculture 2020: Sustainability in action. Rome: FAO; c2020. https://doi.org/10.4060/cc0461en
9. FAO. The State of World Fisheries and Aquaculture 2022: Towards Blue Transformation. Rome: FAO; c2022. https://doi.org/10.4060/ca9229en
10. Gatlin DM, Barrows FT, Brown P, Dabrowski K, Gaylord TG, Hardy RW, et al. Expanding the utilization of sustainable plant products in aquafeeds: A review. Aquaculture Research. 2007;38(6):551-579. https://doi.org/10.1111/j.1365-2109.2007.01704.x
11. Kavitha C, Ramesh M, Kumaran SS, Lakshmi SA. Toxicity of Moringa oleifera seed extract on some hematological and biochemical profiles in a freshwater fish, Cyprinus carpio. Experimental and Toxicologic Pathology. 2012;64(7-8):681-687. https://doi.org/10.1016/j.etp.2011.01.001
12. Kitchen C. Labeo rohita nutrition facts -Cully's Kitchen. Cully's Kitchen. 2022 Jan 8. Available from: https://cullyskitchen.com/labeo-rohita-nutrition-facts/
13. Miles RD, Chapman FA. Benefits of fish meal in aquaculture diets. EDIS. 2006;2006(12). https://doi.org/10.32473/edis-fa122-2006
14. Mune MAM, Nyobe EN, Bassogog CBB, Minka SR. A comparison on the nutritional quality of proteins from Moringa oleifera leaves and seeds. Cogent Food & Agriculture. 2016;2(1):1213618. https://doi.org/10.1080/23311932.2016.1213618
15. Ngasotter S, Panda SP, Mohanty U, Akter S, Mukherjee S, Waikhom D, et al. Current scenario of fisheries and aquaculture in India with special reference to Odisha: A review on its status, issues and prospects for sustainable development. International Journal of Bio-resource and Stress Management. 2020;11(1):370-380. https://doi.org/10.23910/1.2020.2126a
16. Tabassum S, Hussain SM, Ali S, Arsalan M, Ahmad B, Asrar M, et al. Partial replacement of fish meal with Moringa oleifera leaf meal in practical diets of Cirrhinus mrigala fingerlings. Brazilian Journal of Biology. 2021;83:246333. https://doi.org/10.1590/1519-6984.246333
17. Yuangsoi B, Masumoto T. Replacing moringa leaf (Moringa oleifera) partially by protein replacement in soybean meal of fancy carp (Cyprinus carpio). Songklanakarin Journal of Science & Technology. 2012;34(5):479-485.
18. Yuangsoi B, Klahan R, Charoenwattanasak S. Partial replacement of protein in soybean meal by moringa seed cake (Moringa oleifera) in bocourti's catfish (Pangasius bocourti). Songklanakarin Journal of Science & Technology. 2014;36(2):125-135.