Effect of leaf powder of giloy (Tinospora cordifolia) in fish feed on survival and growth of post larvae of Cyprinus carpio fingerlings

Introduction

Aquaculture continues to drive growth, with global production soaring to 130.9 million tons in 2022, representing a significant 6.6% increase from 122.8 million tons in 2020. (FAO, 2024) [6] . With the wild fisheries resources continuing to decline aquaculture have become a primary source of global food fish production. To address the rising demand for protein driven by population growth, the aquaculture sector has witnessed substantial expansion. However, this growth has been accompanied by a rising use of synthetic chemicals-such as pesticides to treat parasites, fungal infections, and unwanted organisms in culture ponds, aiming to prevent disease outbreaks, enhance fish health, and improve environmental conditions, ultimately impacting production and farmer incomes. Fish is key provider of inexpensive protein and essential fatty acids and maintaining fish health is paramount to human nutrition and food security. Global aquatic and fisheries resources, spanning from inland waters (lakes, reservoirs, ponds, and rivers) to marine ecosystems (seas and oceans), yield numerous long-term benefits for human well-being, including, sustainable food sources, economic growth, environmental sustainability and social welfare. Despites its abundant provision, human induced-stressors, including pesticides disrupts the delicate balance of aquatic ecosystems harm fish populations, impair fish health and degrade environmental quality. This disruption threatens the long-term sustainability of aquatic resources, undermining the benefits they provide to human well-being. Consumption of pesticide-contaminated animals and their by-products can transfer toxicity to humans and wildlife, including carnivorous fish species that feed on contaminated aquatic insects, potentially propagating harmful effects up the food chain (Maurya & Singh (2021) [10] . T. cordifolia, also known as the queen of all herbs, is a climbing shrub with a number of coiling branches. The whole plant has been divided into different parts i.e., stem, leaves, flower, and fruits. Other parts, like aril roots, lamina, and seeds, are present as well. Tetra-to penta-arch structures are the characteristics of aerial roots. Other than that, the root cortex has an outer, thick wall and an inner parenchymatous zone. The lamina is ovate, 10-20 cm long and up to 15 cm broad; the base is deeply cordate, membranous, pubescent, and whitish tomentose with a prominent reticulum beneath. Seeds are curved in shape.

The embryo turned into acurve automatically. However, the endocarp is well ornamented and confers vital taxonomic characters. Another important part of the plant, its root possesses anti-ulcer and anti-activity. Both the stem and root of T. cordifolia, together with other medicinal drugs, are prescribed as an anti-dote against scorpion sting and snakebite. Dry barks of T. cordifolia have antiinflammatory, antiallergic, antipyretic, antispasmodic, and anti-leprotic properties.Herbal immune stimulants are promising in aquaculture since they boost growth performance, fish immunity, and antimicrobial properties, making them viable alternatives to chemicaltreatments and antibiotics (Elumalai et al., 2021) [5] . Tinospora cordifoliais a well-knownmedicinal plant in the Menispermaceae family and is a significant supplier of newmedications. It has a wide range of therapeutic applications and phytochemicals responsible for its pharmacological effects (Sajith & Farhan, 2022) [11] . Antibiotics and therapies (Elumalai et al., 2021) [5] , one well-known medicinal plant in the Menispermaceae family, Tinosporacordifolia, is a major source of novel drugs. Its pharmacological effects are caused by phytochemicals and a variety of medicinal uses (Sajith & Farhan, 2022) [11] . Tinospora leaves are heart-shaped, 5-10 cm long, and high in protein, calcium, and phosphorus (Sinha et al., 2004; Kushwaha, (2020) [12,8] . In catfish Pangasius sutchi fingerlings, it has been documented that diets enriched with T. cordifolia leaf extract enhanced performance, survival, and haemato-biochemical profiles (Latha et al., 2020) [9] . Likewise, El Basuini et al., (2022) [4] discovered that dietary T.cordifolia improves growth performance, antioxidative capacity, immune response, and resistance against hypoxia stress in the Nile tilapia. The current study aimed to evaluate the effects of T. cordifolia stem powder on growth performance, and antioxidant-enzyme activities in C. carpio, an economically important farmed fish species in India.

Materials and Methods

Experiment Design

The feeding trial was conducted at the aquaculture research facility of the, Sinha fish farm village-Jargaon Block-Chhura District-Gariabandh The experiment was carried out for a total duration of 150 days.

Procurement of Experimental Fishes

The healthy common carp fingerlings were collected from Sinha fish farm with average body weighing 10.5 g. Then these fishes were transfer in acclimatization tank for 15 days.

Procurement of Experimental Plant and Collection of Ingredients

Giloy (Tinospora cordifolia) stem powders were collected from the Sinha fish farm village-Jargaon, Gariabandh, Chhattisgarh, India. The collected giloy were washed with tap water to remove debris and air dried under shade. After complete drying, the giloy was powdered using a warring blender. A plastic container was use for storing the leaves powder for further use. Remain other ingredients (Rice bran, MOC, wheat flour, fish meal) and the vitamin premixes were purchased from the local market of Gariabandh, Chhattisgarh. The feeding ingredient and their composition were mentioned in Table no1.

Experimental Feed Preparation

For the preparation of experimental diets all the ingredients were (Rice bran, MOC, wheat flour, soybean meal, fish meal) finely powdered and it were mixed all together according to the percentage amount calculation of the ingredient with water, vitamin was added in all the three feeds including the control tank. The feed was formulated with 35 per cent protein content that was 3, 5 and 8 percent of Giloy (Tinospora cordifolia) stem powders added. After complete mixing the dough the different treatment feed was put in an autoclave for 30 minutes to kill the microorganism.

The autoclaved dough was added with vitamin premix and after complete mixing the dough feed were put in a feed pelletizer and extruded according to the required size for the fingerling fish mouth. Extruded pelleted feed are then dried at 40 °C for four hours to remove the moisture from the feed. The control feed used contains 0 per cent Giloy powders, for the three treatment tanks, three different percentages of Giloy powders were used to prepare the other three experimental diets, for the control experiment, feed containing fish meal, wheat flour, mustard oil cake (MOC) and rice barn as the ingredients was used. The feeding was done at the rate 3 per cent of their body weight.

Water quality analysis

Various water quality parameters viz. water temperature (°C), pH, dissolved oxygen (mg/l), nitrite, nitrate and ammonia (APHA, 2005) [3] .

Growth Parameters

The following formulas were used to calculate the fish's growth performance.

Statistical Data Analysis

The data obtained from this study were analysed using the Statistical Package for the Social Sciences (SPSS) 2019. The data were presented as Mean ± SE. Results will be considered statistically significant at the 5 per cent level (p< 0.05).

Results and Discussion

The growth performance and feed utilization parameters of common carp (Cyprinus carpio) fingerlings fed diets supplemented with different levels of Tinospora cordifolia stem powder for 150 days are presented in Table 1.

Growth Parameters

The present study investigated the effect of dietary supplementation with varying levels of Tinospora cordifolia (Giloy) stem powder on the growth performance and feed utilization of Cyprinus carpio fingerlings. The growth performance of Cyprinus carpio fed diets supplemented with varying levels of Tinospora cordifolia (Giloy) was summarized in Table 2. The initial body weights of fish were comparable across all treatments, ranging from 10.48 g (T 1 ) to 10.55 g (T 2 ), and confirming uniformity at the start of the experiment. A significant improvement in growth parameters was observed in fish fed diets containing Giloy. The highest final body weight (40.80 g) was recorded in the T 2 group (5% Giloy), followed by T 3 (8%) with 38.50 g, T 1 (3%) with 36.80 g, and the lowest in the control group T 0 (31.25 g). Similarly, weight gain was significantly higher in T 2 (30.25 g), demonstrating the most effective growth promotion, while the control group showed the lowest gain (20.73 g). The percent weight gain followed a similar trend: T 2 (286.73%) > T 3 (266.67%) > T 1 (251.14%) > T 0 (197.05%). This pattern indicates that the incorporation of Giloy in the diet enhances growth, particularly at a 5% inclusion level. The Specific Growth Rate (SGR) was also highest in the T 2 group (2.05%/day), indicating faster biomass accumulation compared to other treatments. The SGR in the control group was the lowest at 1.81%/day. This result supports the conclusion that Giloy supplementation positively affects metabolic and anabolic processes, possibly due to its bioactive compounds like alkaloids and diterpenoid lactones known for immunostimulant and growth-promoting effects. Feed intake across the treatments was relatively consistent, ranging from 37.31 g (T 0 ) to 38.70 g (T 2 ). The Feed Conversion Ratio (FCR), an important index of feed efficiency, improved with increasing Giloy supplementation up to 5%. The lowest FCR was observed in T 2 (1.52), indicating better feed utilization efficiency. In contrast, the control group had the highest FCR (1.80), suggesting less efficient feed conversion into body mass. FCR slightly increased in T 3 (1.58), suggesting that a further increase beyond 5% may not yield additional benefits and could be less efficient. The survival rate remained high across all groups, ranging from 95.56% (T 0 ) to 97.78% (T 1 , T 2 , and T 3 ). The lack of significant differences in survival rate among treatments indicates that Giloy supplementation up to 8% does not adversely affect the health or survivability of C. carpio. Elumalai et al., 2021; Sajith & Farhan, 2022 [5,11] they suggested that the Giloy is safe and well-tolerated by fish even at higher concentrations. The results clearly demonstrate that 5% Giloy supplementation (T 2 ) provides optimal enhancement of growth performance and feed conversion in common carp. The improved weight gain, SGR, and FCR in this group may be attributed to the bioactive phytochemicals present in Tinospora cordifolia, which are known to enhance nutrient absorption, digestive enzyme activity, and immune response in fish. Elumalai et al., 2021 [5] ; Sinha et al., 2004 [12] and Sajith & Farhan, 2022 [11] they were suggested that the consistent with earlier studies where herbal additives improved fish performance by enhancing gut health and boosting immunity. The lowest FCR was observed in T5 (1%) which was significantly different with T 1 , T 2 , T 3 and T 4 (p<0.05). The mean value of different treatments shown that best FCR was recorded in T5 (2.41) followed by T 4 (2.74), T 3 (5.35), T 2 (6.87) and control T 1 (6.51). Kour et al. (2004) [7] lead a feeding trial on C. mrigala to measure the impact of Bala (Sida cordifolia) herb diverse with ground nut oil cake and rice bran with ratio 1:1. Though the 8% Giloy group (T 3 ) too presented better growth comparative to the control, its act was slightly lower than T 2 , suggesting a threshold effect beyond which no additional benefits occur, and excessive inclusion may even hinder feed efficiency. Therefore, inclusion levels beyond 5% may not be economically justified.

Water Quality Parameters Analysis

The mean values of water quality parameters monitored in the experimental tanks throughout the 150-day culture period were presented in Table 3.

. for temperature and 6.5-8.5 for pH) for optimal carp growth and physiological functions. No significant fluctuation in temperature or pH was observed due to Giloy supplementation, suggesting that dietary treatments did not alter these basic water conditions. Dissolved oxygen concentrations were slightly higher in treatment groups receiving Giloy, with the maximum DO recorded in T 2 (6.4 mg/L) and the minimum in the control group T 0 (6.2 mg/L). These differences, although minor, may be attributed to improved water quality as a result of enhanced feed digestion and reduced metabolic waste, particularly in T 2 . All values remained above 5.0 mg/L, indicating a welloxygenated environment suitable for carp culture. The levels of nitrogenous waste products-TAN, NO₂-N, and NO₃-N showed a decreasing trend with increasing Giloy supplementation up to T 2 . The lowest TAN (0.22 mg/L) and NO₂-N (0.040 mg/L) were recorded in T 2 , while the control group showed the highest levels (TAN: 0.28 mg/L, NO₂-N: 0.055 mg/L). Nitrate levels also followed this trend, with the lowest NO₃-N in T 2 (1.65 mg/L). These findings suggest that Giloy supplementation, particularly at 5%, may have contributed to better nitrogen metabolism or reduced nitrogenous excretion. The lower concentrations of TAN and NO₂-N in the Giloy-supplemented groups could be attributed to improved nutrient assimilation, potentially reducing the amount of nitrogenous waste excreted into the water. This may also indicate that Giloy has a role in enhancing gut health or supporting beneficial microbial activity, which could contribute to the detoxification of ammonia and nitrite in the culture system. Maintaining optimal water quality was critical in aquaculture, as deviations can adversely affect fish health and growth. The results indicate that the inclusion of Giloy stem powder in the diet did not negatively impact water quality. In fact, there was a slight improvement in parameters such as DO and nitrogen compounds in the T 2 treatment, which also corresponded with the best growth performance observed in fish. This correlation supports the idea that 5% Giloy supplementation not only enhances fish growth but may also contribute indirectly to better environmental conditions in the culture system. These findings align with studies reporting that the use of herbal feed additives can lead to improved nutrient utilization and reduced organic and nitrogenous waste output in aquaculture systems. The stabilization of water parameters across all treatments also indicates good management practices and experimental reliability. (Anita et al. 2016) [2] reported significant improvements in SGR and FCR in Amur carp (C. carpio) fed diets with 0.75% T. cordifolia leaf powder.25 similarly, (Yadav et al., 2023; Al-Turaihi et al. 2023) [13,1] found that 6 g/kg (0.6%) T. cordifolia leaf powder enhanced growth in common carp. The present study, using stem powder, identified 1.0% as optimal, which falls within a comparable range of effective dosages. Also found that 6 g/kg (0.6%) of T. cordifolia stem powder (as well as root powder) significantly enhanced growth performance in common carp 9, further supporting the efficacy of the stem part.

Conclusion

In light of these findings, it is recommended that the incorporated into the diets of common carp fingerlings at a 5 percent inclusion level of T. cordifolia. This quantities suggestions optimal growth improvement, feed efficiency, and water quality maintenance, making it a valuable and natural alternative to synthetic feed additives in sustainable aquaculture systems. This research also help in making the strategy for fish culture in mainly grow out stage hence we suggesting some filed trial also.

Acknowledgement

AcknowledgementsThe authors gratefully acknowledge the Department of Aquaculture, School of School, Sanjeev Agrawal Global Educational (SAGE) University Bhopal, India for providing the necessary facilities for conducting this research. Disclaimer (Artificial Intelligence)Author(s) hereby declare that NO generative AI technologies such as Large Language Models (ChatGPT, COPILOT, etc.) and text-to-image generators have been used during the writing or editing of this manuscript.

References

1. Al-Turaihi AS, Khawaja AM, Shaker IH, Al-Zubaydi JH. Hydrochemical study of artesian wells in Babylon Governorate-Middle of Iraq. IOP Conference Series: Earth and Environmental Science. 2023;1222(1):012044.
2. Anita, Dubey MK, Khati A, Chauhan RS. Immunostimulatory and growth promoting potential of Tinospora cordifolia (Thunb.) Miers on fingerlings of Amur carp. Indian Journal of Experimental Biology. 2016;54:659-663.
3. APHA. Standard methods for the examination of water and wastewater. 21st ed. Washington D.C.: American Public Health Association; 2005.
4. El Basuini MF, Teiba II, Shahin SA, Mourad MM, Zaki MA, Labib EM, Dawood MA. Dietary Guduchi (Tinospora cordifolia) enhanced the growth performance, antioxidative capacity, immune response and ameliorated stress-related markers induced by hypoxia stress in Nile tilapia (Oreochromis niloticus). Fish & Shellfish Immunology. 2022;120:337-344.
5. Elumalai K, Kumar SS, Sundaram PS, Elumalai M, Eluri K, Keerthana M, Sivannan S. Cellulitis occurred due to the Streptococcus and Staphylococcus species: an efficient antibiotics therapy. SN Comprehensive Clinical Medicine. 2021;3(4):979-981.
6. FAO. The State of World Fisheries and Aquaculture 2024-Blue transformation in action. Rome: Food and Agriculture Organization of the United Nations; 2024. Available from: https://doi.org/10.4060/cd0683en
7. Kour D, Sharma LL, Sharma BK. Use of herb, Bala (Sida cordifolia Linn.) as growth promoter in the supplementary feed of Cirrhinus mrigala (Ham.). Indian Journal of Fisheries. 2004;51(4):501-504.
8. Kushwaha PDS. Skoog (MS) medium [dissertation]. Varanasi: Banaras Hindu University; 2020.
9. Latha C, Krishnakumar V, Munuswamy N. Growth, survival and haemato-biochemical profiles of the freshwater catfish, Pangasius sutchi (Fowler, 1937) fingerlings fed on Tinospora cordifolia leaf extract supplemented diet. Egyptian Journal of Aquatic Biology and Fisheries. 2020;24(1):249-262.
10. Maurya SP, Singh R. Sustainable water resources. In: Sustainable resource management. Elsevier; 2021. p. 147-162.
11. Sajith KS, Farhan H. Pharmacological effects of Tinospora cordifolia (Giloy) in human body. The Pharma Innovation Journal. 2022;11:7-10.
12. Sinha K, Mishra N, Singh J, Khanuja S. Tinospora cordifolia (Guduchi), a reservoir plant for therapeutic applications: a review. Indian Journal of Traditional Knowledge. 2004;3(3):257-270.
13. Yadav MK, Ojha ML, Keer NR. Effect of vegetable oils on growth, feed utilization and digestive enzyme activities of Labeo rohita (Hamilton, 1822). Indian Journal of Ecology. 2023;50(5):1772-1775.