Effects of Bacillus coagulans on haematological parameters of Cyprinus carpio

J. Bio. & Env. Sci. 20 2 5 34 | Jagadish and Mallikarjun RE RERE RESEARCH SEARCHSEARCH SEARCH PAPER PAPERPAPER PAPER OPEN ACCESS OPEN ACCESSOPEN ACCESS OPEN ACCESS Effects of Bacillus coagulans on haematological parameters of Cyprinus carpio Pruthvi Kargunda Jagadish, Ashashree Hassan Mallikarjun * Department of Zoology, Sahyadri college, Kuvempu University, Shimoga, Karnataka, India Article published on May 06, 2025 Key words: Probiotics, Bacillus coagulans , Cyprinus carpio , Haematological parameters, Recirculating aquaculture system Abstract This research assessed the impact of dietary Bacillus coagulans supplementation on hematological parameters and immunological state in common carp (Cyprinus carpio) cultivated in a recirculating aquaculture system. Fish were given diets enriched with different concentrations of B. coagulans for durations of 20, 40, and 60 days. Essential hematological parameters—such as red blood cell (RBC) count, hemoglobin (Hb), packed cell volume (PCV), white blood cell (WBC) count, mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH)—were evaluated to ascertain physiological and immunological responses. Results indicated substantial elevations in RBC, Hb, PCV, and WBC levels in the probiotic-treated groups, with the peak values seen in the T3 group at day 60. The MCV and MCH levels reached their peak early and stabilized by day 60, suggesting adaptive hematological control. The observations indicate that B. coagulans promotes hematopoiesis and immunological function in C. carpio, presenting a possible probiotic alternative to antibiotics for enhancing fish health and performance in sustainable aquaculture systems. * Corresponding Author: Ashashree Hassan Mallikarjun  ashashree20[email protected]om Journal of Biodiversity and Environmental Sciences (JBES) ISSN: 2220-6663 (Print) 2222-3045 (Online) Vol. 26, No. 5, p. 34-42, 2025 http://www.innspub.net J. Bio. & Env. Sci. 20 2 5 35 | Jagadish and Mallikarjun Introduction Aquaculture plays an important role in global food security and economic development, but sustainable techniques are essential to solve difficulties relating to fish health, growth performance, and environmental effect. Probiotics have emerged as viable alternatives to antibiotics in aquaculture, delivering advantages such as higher immunity, better digestion, and disease resistance (Troell et al., 2023). Cyprinus carpio, often known as common carp, faces ecological threats from habitat loss, overexploitation, and migratory obstacles (Humphries and Winemiller, 2009), leading to heightened dependence on aquaculture. Intensive agricultural techniques may adversely affect aquatic ecosystems and biodiversity (Saeedi et al., 2024). Sustainable management measures, including protected areas and community-based conservation, are vital for balancing aquaculture expansion with ecological preservation (Zebua et al., 2024). Concurrent progress in sturgeon aquaculture, particularly with Acipenser baerii, illustrates the advantages of enhanced breeding and nutritional methodologies, while also underscoring analogous difficulties in disease management and ecological sustainability (Pashko et al., 2024; Yurin et al., 2024). Probiotic supplementation, particularly with strains such as Bacillus and Lactobacillus, markedly enhances specified growth rates (SGR), feed conversion ratios (FCR), and immunological responses (Fachri et al., 2024; Calcagnile et al., 2024). Species-specific advantages have been recorded in grey mullet, striped catfish, and others, demonstrating resistance to diseases such as Nocardia seriolae and Edwardsiella ictaluri (Chan et al., 2024). Bacillus coagulans, a resilient spore-forming probiotic, improves gastrointestinal health, nutritional assimilation, and immune function in aquaculture species. In Cyprinus carpio, it shows potential for enhancing hematological indices—total erythrocyte count (TEC), hemoglobin (Hb), and packed cell volume (PCV)—which are critical indications of health and stress (Mary and Raj, 2023; Naveenkumar et al., 2017; Pradhan et al., 2014). This research examines the impact of Bacillus coagulans on the growth and hematological parameters of C. carpio cultivated in a recirculating aquaculture system, with the objective of formulating probiotic-based approaches for sustainable and efficient fish farming. Materials and methods Collection and acclimatization of fish Fingerlings of Cyprinus carpio (L.) were purchased from the Fishery Department at B.R. Project, situated around 5 km from the Jnana Sahyadri campus, Shankaraghatta. Upon arriving at the laboratory, the fingerlings were acclimatized under controlled environmental conditions for a predetermined length before to the initiation of the experiment. The chosen fish had an average body weight of 15 ± 1 g. During the acclimatization phase, they were given a commercially available pelleted floating carp feed, which also served as the usual baseline diet for the research. Feeding was performed twice daily to achieve optimum adaptation to the laboratory circumstances and to reduce handling-induced stress. Commercial feed probiotic supplementation The probiotic strain Bacillus coagulans (10⁹ CFU/g), a gram-positive, spore-forming, lactic acid-producing bacterium recognized for its endurance in gastrointestinal settings, was received from Sanzyme Biologics Private Limited, Hyderabad, India. This commercially available formulation was added into the experimental diet provided for Cyprinus carpio fingerlings. The major purpose of its inclusion was to test its impact on important physiological parameters, including growth performance, biochemical indices, and digestive enzyme activities. The selection of B. coagulans was based on its proven stability under feed processing conditions and its established efficiency in altering gut microbiota, hence giving prospective benefits in aquaculture nutrition and health management. Experimental design A 60-day feeding study was undertaken using Cyprinus carpio fingerlings of uniform beginning weight to examine the physiological effects of probiotic administration. Fish were given a designed J. Bio. & Env. Sci. 20 2 5 36 | Jagadish and Mallikarjun meal comprising 32% crude protein at 3% of their body weight daily. The experiment comprised of four groups: a control (C) with no probiotics, and three treatment groups—T1, T2, and T3—supplemented with Bacillus coagulans at 0.05 × 10⁹, 0.1 × 10⁹, and 0.15 × 10⁹ CFU g⁻¹, respectively. Sampling occurred on days 20, 40, and 60, during which fish were killed, and blood samples were obtained for haematological investigation. Blood was taken by cardiac puncture using 2 mL syringes prepared with 150–200 μL of EDTA to avoid coagulation. The puncture site was placed ventrally, between the anterior bases of the pectoral fins. Samples were kept in 1.5 mL sterile Eppendorf tubes and refrigerated until analysis. This strategy enabled regular and sterile sample for the study of hematological responses to probiotic administration. Hematological examination Hematological parameters were evaluated using established protocols to assess the physiological responses of Cyprinus carpio to various dietary regimens. Total red blood cell (RBC) and white blood cell (WBC) counts were obtained using an advanced Neubauer hemocytometer, following the technique published by Hesser (1960). Hemoglobin (Hb) concentration was determined using the cyanmethemoglobin technique according to Blaxhall and Daisley (1973), while hematocrit (Hct) values were measured using micro-hematocrit capillary tubes based on the methods of Satheeshkumar et al. (2012). Differential leukocyte counts were determined using Giemsa staining, following the methodology published by Shah et al. (2009). Furthermore, erythrocyte indices including mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), and mean corpuscular volume (MCV) were determined utilizing formulas provided by Dacie and Lewis (2001), where MCHC (g/dL) = (Hb / Hct) × 100, MCH (pg) = (Hb / RBC) × 10, and MCV (fL) = (Hct/ RBC) × 10. These indicators supplied useful insights into the erythrocytic state and general health condition of the fish treated to probioticsupplemented diets. Statistical analysis The results were represented as mean ± standard deviation (SD), and differences between groups were examined using one-way analysis of variance (ANOVA). Statistical significance was judged at levels of p < 0.05 and p < 0.001. All statistical analyses were done using SPSS software. Results The haematological examination of Cyprinus carpio fed with probiotic-supplemented diets over 20, 40, and 60 days demonstrated consistent, doseand time-dependent improvements across all measured parameters compared to the control group (Tables 1– 3). Red Blood Cell (RBC) counts substantially increased in the high-dose T3 group, increasing from 1.61 ± 0.02 × 10⁶/µL at day 20 to 2.50 ± 0.02 × 10⁶/µL at day 60, suggesting greater erythropoiesis and oxygen-carrying ability. Haemoglobin (Hb) concentrations showed a similar pattern, reaching 9.11 ± 0.06 g/dL in T3 by day 60, compared 5.69 ± 0.05 g/dL in the control, showing enhanced metabolic support. Packed Cell Volume (PCV) significantly increased in the probiotic groups, with T3 reaching a high of 27.85 ± 0.14% at day 60, compared to 21.77 ± 0.53% in the control. Early rises in Mean Corpuscular Volume (MCV) and Mean Corpuscular Haemoglobin (MCH) were seen, notably in T3, but values progressively stabilized, indicating a shift toward erythrocyte homeostasis. While Mean Corpuscular Haemoglobin Concentration (MCHC) changed somewhat, it remained within physiological limits, suggesting no deleterious impact on red cell integrity. White Blood Cell (WBC) counts exhibited the most dramatic elevation, with T3 reaching 217.26 ± 1.87/µL by day 60 vs 176.47 ± 0.17/µL in the control, showing immune system activation. Overall, the findings demonstrate that Bacillus coagulans supplementation considerably improves hematological health, oxygen transport, and immunological function in C. carpio, indicating its potential as a useful probiotic in aquaculture. J. Bio. & Env. Sci. 20 2 5 37 | Jagadish and Mallikarjun Table 1. Hematological parameters of C. carpio treated with various amounts of probiotics throughout 20 days of trial Parameter s Control T1 T2 T3 RBC (×10 ⁶/µL) 1.28 ± 0.02 1.38 ± 0.02 1.47 ± 0.01 1.61 ± 0.02 Hb (g/dL) 5.69 ± 0.05 6.80 ± 0.08 7.82 ± 0.02 8.77 ± 0.02 PCV (%) 14.63 ± 0.12 17.35 ± 0.25 22.65 ± 0.31 25.14 ± 0.12 MCV (fL) 114.78 ± 1.65 125.73 ± 1.77 153.99 ± 2.95 155.90 ± 1.85 MCH (pg) 44.64 ± 0.74 49.27 ± 0.87 53.15 ± 0.39 54.36 ± 0.57 MCHC (g/dL) 38.85 ± 0.33 39.17 ± 0.52 34.54 ± 0.43 34.84 ± 0.31 WBC (/µL) 154.67 ± 1.86 167.65 ± 11.50 204.80 ± 2.26 212.43 ± 2.68 Table 2. Hematological parameters of C. carpio treated with various amounts of probiotics throughout 40 days of trial Parameter s Control T1 T2 T3 RBC (×10 ⁶/µL) 1.62 ± 0.02 1.85 ± 0.02 2.18 ± 0.02 2.34 ± 0.07 Hb (g/dL) 7.08 ± 0.03 8.70 ± 0.04 8.80 ± 0.02 9.13 ± 0.16 PCV (%) 16.55 ± 0.20 19.47 ± 0.14 24.70 ± 0.09 26.39 ± 0.24 MCV (fL) 102.15 ± 1.77 105.06 ± 1.49 113.55 ± 1.08 113.08 ± 3.22 MCH (pg) 43.75 ± 0.39 46.93 ± 0.49 40.47 ± 0.43 39.21 ± 1.15 MCHC (g/dL) 42.80 ± 0.43 44.68 ± 0.42 35.64 ± 0.15 34.61 ± 0.59 WBC (/µL) 174.03 ± 0.94 181.57 ± 1.46 204.30 ± 2.06 214.48 ± 1.63 Table 3. Hematological parameters of C. carpio treated with various amounts of probiotics throughout 60 days of trial Parameters Control T1 T2 T3 RBC (×10 ⁶/µL) 2.12 ± 0.08 2.22 ± 0.01 2.33 ± 0.02 2.50 ± 0.02 Hb (g/dL) 7.23 ± 0.06 7.63 ± 0.03 8.94 ± 0.02 9.11 ± 0.06 PCV (%) 21.77 ± 0.53 23.82 ± 0.15 25.64 ± 0.12 27.85 ± 0.14 MCV (fL) 103.23 ± 3.72 107.52 ± 0.47 110.11 ± 1.26 111.08 ± 0.40 MCH (pg) 34.21 ± 1.04 34.46 ± 0.24 38.38 ± 0.46 36.40 ± 0.31 MCHC (g/dL) 33.16 ± 0.87 32.05 ± 0.20 34.87 ± 0.16 32.73 ± 0.18 WBC (/µL) 176.47 ± 0.17 183.07 ± 1.96 206.44 ± 0.56 217.26 ± 1.87 Discussion Probiotic supplementation has emerged as a promising method in aquaculture, giving many advantages such as better growth, improved immunity, improved nutrient absorption, and increased economic efficiency. Several studies have proven that probiotics significantly affect specific growth rate (SGR) and feed conversion ratio (FCR), notably in freshwater fish such as Nile tilapia and African catfish (Ariyanto and Anika, 2024; Omar et al., 2024; Hadijah et al., 2024). These effects are mostly related to the synthesis of digestive enzymes and enhanced gut shape, which promote effective feed consumption (Ringø et al., 2020; Mahmoodian et al., 2024). In addition to digestive efficiency, probiotics also strengthen the immune system by raising immunoglobulin M (IgM) levels and upregulating immune-related genes such as TNF and IL-6 (Choi et al., 2024; Ferdous et al., 2023). They improve gut health by altering the microbiota, promoting beneficial bacterial populations, and reducing harmful microorganisms (Cerezuela et al., 2011; Mahmoodian et al., 2024). Furthermore, enzymes like as amylase, protease, and lipase generated by probiotics increase nutrient digestion and reduce anti-nutritional components prevalent in plant-based diets (Amenyogbe et al., 2024). These steps not only decrease mortality and antibiotic consumption but also increase sustainability and profitability in aquaculture operations (Fachri et al., 2024; Vulla et al., 2024; Han et al., 2024). The efficiency of probiotic genera such as Bacillus and Lactobacillus has been widely confirmed in diverse aquaculture species. In grass carp (Ctenopharyngodon idella), dietary Bacillus subtilis increased SGR, FCR, and gut enzyme activity, while modifying intestinal microbiota (Wu et al., 2012). Similarly, Lactobacillus plantarum generated from grass carp greatly boosted growth performance, immunological responses, and digestive enzyme activity in Labeo rohita (Yasmin et al., 2024). J. Bio. & Env. Sci. 20 2 5 38 | Jagadish and Mallikarjun Synbiotic combinations such Bacillus licheniformis with fructooligosaccharide (FOS) further enhanced immune responses and survival rates post-pathogen exposure (Sukul et al., 2023). The current research assessed the effects of dietary Bacillus coagulans supplementation on haematological markers of Cyprinus carpio across a 60-day trial period. Consistent improvements were reported across all haematological indicators in treated groups. RBC counts in the high-dose T3 group increased considerably from 1.28 ± 0.02 ×10⁶/µL (control) to 2.50 ± 0.02 ×10⁶/µL by day 60, while Hb concentration jumped from 5.69 ± 0.05 g/dL to 9.11 ± 0.06 g/dL. Similarly, PCV levels rose from 14.63 ± 0.12% to 27.85 ± 0.14%, suggesting better erythropoiesis and oxygen delivery. MCV and MCH peaked at day 20, followed by stability by day 60, indicating a return to erythrocyte homeostasis. The most noticeable change was in WBC count, which climbed from 154.67 ± 1.86/µL (control) to 217.26 ± 1.87/µL (T3, day 60), showing immunological enhancement. These findings are consistent with earlier research revealing that B. coagulans enhances RBC, Hb, and hematocrit (HCT) levels, consequently boosting oxygen-carrying capacity and general physiological performance (Zhang et al., 2023; Xu et al., 2014). The rise in WBCs suggests an active immune system, which is critical for disease resistance in intensive aquaculture systems. Other studies have also underlined the importance of WBCs as indicators of immunological state, stress, and environmental circumstances (Sayed-Lafi et al., 2023; Satkar et al., 2024; Ahmed et al., 2020; Haghparast et al., 2020; Lataretu et al., 2013). Despite these hopeful results, heterogeneity in metrics like MCV, MCH, and MCHC across various research shows that the benefits of probiotics may rely on individual strains, dose, fish species, and culture conditions (Ayala et al., 2008; Adriani et al., 2013). Additionally, the immunomodulatory effects identified in the current research may also be ascribed to cytokine modulation, resulting to decreased stress and enhanced hematological stability (Zhang et al., 2023; Xu et al., 2014). Given the difficulties associated with antibiotic usage in aquaculture—such as environmental pollution, antimicrobial resistance, and food safety concerns— probiotics provide a sustainable option for boosting fish health and production (Nagesh et al., 2025). The outcomes of this research underline the potential of Bacillus coagulans as an effective dietary supplement for enhancing haematological parameters and increasing immunological function in C. carpio, contributing to sustainable aquaculture techniques. Conclusion The current research reveals that nutritional supplementation with Bacillus coagulans considerably boosts hematological parameters and immunological responses in Cyprinus carpio under a recirculating aquaculture system. Prolonged dosing for 60 days resulted in considerable improvements in RBC count, hemoglobin concentration, hematocrit value, and white blood cell levels, suggesting increased oxygen-carrying ability, erythropoiesis, and immunological state. The early increases in MCV and MCH followed by normalization imply adaptive physiological responses and hematological stability with ongoing probiotic consumption. These data demonstrate the potential of B. coagulans as a sustainable alternative to antibiotics, leading to enhanced fish health, survival, and production in aquaculture. Incorporating probiotics such as B. coagulans into aquafeeds might therefore play a crucial role in promoting eco-friendly and healthoriented aquaculture practices. Acknowledgments The authors extend their heartfelt thanks to Dr. Ashashree for her essential advice and assistance during the duration of this study. Sincere thanks are expressed to the Department of Zoology, Sahyadri College, Kuvempu University, Shivamogga, Karnataka, India, for providing vital research facilities. The authors are also thanks to Stellixir J. Bio. & Env. Sci. 20 2 5 39 | Jagadish and Mallikarjun Biotech Pvt. Ltd., Bengaluru, for giving vital laboratory facilities that greatly helped to the successful completion of this work. References Ahmed I, Reshi QM, Fazio F. 2020. The influence of the endogenous and exogenous factors on hematological parameters in different fish species: A review. Aquaculture International 28, 869–899. Al-Saeedi N. 2024. Carp fish and effects on the aquatic environment. University of Thi-Qar Journal of Agricultural Research 13(2), 327–334. Amenyogbe E, Droepenu EK, Ayisi CL, Boamah GA, Duker RQ, Abarike ED, Huang JS. 2024. Impact of probiotics, prebiotics, and synbiotics on digestive enzymes, oxidative stress, and antioxidant defense in fish farming: Current insights and future perspectives. Frontiers in Marine Science 11, 1368436. Ariyanto YS, Anika M. 2024. Probiotics on commercial fish growth: A meta-analysis. Jurnal Sumberdaya Hayati 10(4), 205–216. Ariyanto YS, Anika M. 2024. Probiotics on commercial fish growth: A meta-analysis. Jurnal Sumberdaya Hayati 10(4), 205–216. Ariyanto YS, Anika M. 2024. Probiotics on commercial fish growth: A meta-analysis. Jurnal Sumberdaya Hayati 10(4), 205–216. Arthanari M, Dhanapalan S. 2016. Assessment of the haematological and serum biochemical parameters of three commercially important freshwater fishes in river Cauvery Velur, Namakkal district, Tamil Nadu, India. International Journal of Fisheries and Aquatic Studies 4(1), 155–159. Askwith DAJ, Raj A. 2023. Effects of dietary probiotic Lactobacillus acidophilus and prebiotic Spirulina platensis on haemotological profile of Labeo rohita (Hamilton, 1822) fingerlings. Blaxhall PC, Daisley KW. 1973. Routine haematological methods for use with fish blood. Journal of Fish Biology 5(6), 771–781. Calcagnile M, Tredici SM, Alifano P. 2024. A comprehensive review on probiotics and their use in aquaculture: Biological control, efficacy, and safety through the genomics and wet methods. Heliyon 10(24). Cerezuela R, Meseguer J, Esteban MA. 2011. Current knowledge in synbiotic use for fish aquaculture: A review. Journal of Aquaculture Research & Development S1, 1–7. Chan CH, Chen LH, Chen KY, Chen IH, Lee KT, Lai LC, Yan TR. 2024. Single-strain probiotics enhance growth, anti-pathogen immunity, and resistance to Nocardia seriolae in grey mullet (Mugil cephalus) via gut microbiota modulation. Animal Microbiome 6(1), 67. Choi W, Moniruzzaman M, Bae J, Lee S, Min T, Bai SC. 2024. Modulatory effect of three intestinally isolated dietary probiotic bacteria in low fish meal diets on growth, feed utilization, serum biochemistry, innate immunity, gut health and resistance against bacterial challenge in juvenile olive flounder Paralichthys olivaceus. Dacie JV, Lewis SM. 1975. Practical haematology, 5 th ed. The English Language Book Society and Churchill Livingstone. Fachri M, Amoah K, Huang Y, Cai J, Alfatat A, Ndandala CB, Chen H. 2024. Probiotics and paraprobiotics in aquaculture: A sustainable strategy for enhancing fish growth, health and disease prevention – a review. Frontiers in Marine Science 11, 1499228. Fachri M, Amoah K, Huang Y, Cai J, Alfatat A, Ndandala CB, Chen H. 2024. Probiotics and paraprobiotics in aquaculture: A sustainable strategy for enhancing fish growth, health and disease prevention—a review. Frontiers in Marine Science 11, 1499228. J. Bio. & Env. Sci. 20 2 5 40 | Jagadish and Mallikarjun Ferdous Z, Hossain MK, Hadiuzzaman M, Rafiquzzaman SM, Halim KA, Rahman T, Shahjahan M. 2024. Multi-species probiotics enhance survival, growth, intestinal microbiota and disease resistance of rohu (Labeo rohita) larvae. Water Biology and Security 3(1), 100234. Hadijah H, Loar L, Mardiana M, Kantun W, Zainuddin Z. 2023. Dietary probiotics and its effect on growth rate, survival rate, and feed conversion ratio of Clarias gariepinus. Jurnal Riset Akuakultur 18(4), 227–238. Haghparast MM, Alishahi M, Ghorbanpour M, Shahriari A. 2020. Evaluation of hematoimmunological parameters and stress indicators of common carp (Cyprinus carpio) in different C/N ratio of biofloc system. Aquaculture International 28(6), 2191–2206. Han C, Shi H, Cui C, Wang J, Li L, Bei W, Wang S. 2024. Strain-specific benefits of Bacillus on growth, intestinal health, immune modulation, and ammonia-nitrogen stress resilience in hybrid grouper. Antioxidants 13(3), 317. Hesser EF. 1960. Methods for routine fish hematology. The Progressive Fish-Culturist 22(4), 164–171. Humphries P, Winemiller KO. 2009. Historical impacts on river fauna, shifting baselines, and challenges for restoration. BioScience 59(8), 673–684. Kumar P, Kaur VI, Tyagi A, Khairnar S. 2023. Synergistic effect of prebiotic with gut isolated probiotic bacteria on survival, growth and carcass composition of Labeo rohita. Indian Journal of Ecology 50(4), 1163–1169. Lataretu A, Furnaris F, Mitranescu E. 2013. Hematologic profile as stress indicator in fish. Mahmoodian S, Meimandipour A, Faeed M, Shams Ara M, Fatemi SSA, Ghasemi M, Chaharmahali M. 2024. Dietary probiotic prototypes and their effects on growth performance, immune function, and gut microbiota of rainbow trout (Oncorhynchus mykiss). Mikhailov AN, Moruzi I, Pishchenko EV, Kalmykova GV, Mager SN. 2024. The effectiveness of probiotics in aquaculture (review). Rybovodstvo i Rybnoe Hozâjstvo 12, 876–885. https://doi.org/10.33920/sel-09-2412-04 Naveenkumar BT, Kaur Y, Tyagi A, Shanthanagouda AH. 2017. Baseline studies on selected hematological parameters of Indian major carps, exotic carps and catfishes. J Fish Life Sci 2(2), 60–67. Omar AA, Gado MS, Kandel HE, Farrag FA, Shukry M. 2024. Probiotic efficacy in aquaculture: The role of Technospore® (Bacillus coagulans) in improving Nile Tilapia (Oreochromis niloticus) performance and disease resistance: A study on gut health, immunological response, and gene expression. Probiotics and Antimicrobial Proteins, 1–18. Pashko S, Tretiak O, Pashko M, Kolos O. 2024. Peculiarities of cultivation of Siberian sturgeon (Acipenser baerii Brandt, 1869) breeders using floating cages in the climatic conditions of the forest steppe of Ukraine. Ribogospodarsʹka Nauka Ukraïni 2(68), 40–57. https://doi.org/10.61976/fsu2024.02.040 Pradhan SC, Patra AK, Pal A. 2014. Hematological and plasma chemistry of Indian major carp, Labeo rohita (Hamilton, 1822). Journal of Applied Ichthyology 30(1), 48–54. J. Bio. & Env. Sci. 20 2 5 41 | Jagadish and Mallikarjun Rahayu S, Amoah K, Huang Y, Cai J, Wang B, Shija VM, Jiang M. 2024. Probiotics application in aquaculture: Its potential effects, current status in China and future prospects. Frontiers in Marine Science 11, 1455905. Rahayu S, Amoah K, Huang Y, Cai J, Wang B, Shija VM, Jiang M. 2024. Probiotics application in aquaculture: Its potential effects, current status in China and future prospects. Frontiers in Marine Science 11, 1455905. Ringø E, Van Doan H, Lee SH, Soltani M, Hoseinifar SH, Harikrishnan R, Song SK. 2020. Probiotics, lactic acid bacteria and bacilli: Interesting supplementation for aquaculture. Journal of Applied Microbiology 129(1), 116–136. Saini VP, Ojha ML, Gupta MC, Nair P, Sharma A, Luhar V. 2014. Effect of dietary probiotic on growth performance and disease resistance in Labeo rohita (Ham.) fingerlings. International Journal of Fisheries and Aquatic Studies 1(6), 7–11. Satheeshkumar P, Ananthan G, Kumar DS, Jagadeesan L. 2012. Haematology and biochemical parameters of different feeding behaviour of teleost fishes from Vellar estuary, India. Comparative Clinical Pathology 21, 1187–1191. Satkar SG, Bhusare S, Sahu A, Aitwar VS, Kunjir S, Girkar M. 2024. Stress-induced hematological and serum alterations as biomarkers in fish health assessment. International Journal of Advanced Biochemistry Research. https://doi.org/10.33545/26174693.2024.v8.i2sf.614 Sayed-Lafi RM, Sultan FA. 2023. Changes in hematological parameters of common carp (Cyprinus carpio) fingerlings fed on pomegranate (Punica granatum) peel supplement. Shah AW, Muni Parveen MP, Mir SH, Sarwar SG, Yousuf AR. 2009. Impact of helminth parasitism on fish haematology of Anchar Lake, Kashmir. Sukul T, Kari ZA, Téllez-Isaías G, Ghosh K. 2023. Autochthonous Bacilli and fructooligosaccharide as functional feed additives improve growth, feed utilisation, haematoimmunological parameters and disease resistance in rohu, Labeo rohita (Hamilton). Animals 13(16), 2631. Troell M, Costa‐Pierce B, Stead S, Cottrell RS, Brugere C, Farmery AK, Barg U. 2023. Perspectives on aquaculture's contribution to the Sustainable Development Goals for improved human and planetary health. Journal of the World Aquaculture Society 54(2), 251–342. Vulla KE, Mmanda FP, Nyangoko BP, Makule EE. 2024. Unlocking potential benefits on applications of probiotics in inland aquaculture industry: A review. Aquaculture, Fish and Fisheries 4(6), e70027. Wu ZX, Feng X, Xie LL, Peng XY, Yuan J, Chen XX. 2012. Effect of probiotic Bacillus subtilis Ch9 for grass carp, Ctenopharyngodon idella (Valenciennes, 1844), on growth performance, digestive enzyme activities and intestinal microflora. Journal of Applied Ichthyology 28(5), 721–727. Yasmin R, Saharia PK, Hussain IA, Talukdar A, Baishya S, Bhagawati K, Singh A. 2025. Probiotic strain Lactobacillus plantarum GCLP4 derived from the gut of Ctenopharyngodon idella modulates growth, digestive enzymes and immune‐ biochemical parameters of Labeo rohita fingerlings. Journal of Animal Physiology and Animal Nutrition 109(2), 268–279. Yurin DA, Данилова АА, Maksim E, Gneush A. 2024. Studying reproductive feed application for sturgeon. Vestnik Krasnoârskogo Gosudarstvennogo Agrarnogo Universiteta 4, 149– 154. https://doi.org/10.36718/1819-4036-2023-4-149154 J. Bio. & Env. Sci. 20 2 5 42 | Jagadish and Mallikarjun Zebua O, Zega A, Zebua RD, Laoli D, Dawolo J, Telaumbanua BV. 2024. Krisis biodiversitas perairan: Investigasi solusi berbasis komunitas untuk pemulihan ekosistem aquatik. Manfish: Jurnal Ilmiah Perikanan dan Peternakan 2(2), 69–79. Zhou Y, Zeng Z, Xu Y, Ying J, Wang B, Majeed M, Li W. 2020. Application of Bacillus coagulans in animal husbandry and its underlying mechanisms. Animals 10(3), 454.