Growth and hematological effects of cellulolytic Bacillus spp. isolated from rabbit gut

Main Article Content

B. TURAEVA
G. J. KUTLIEVA
X. F. KAMOLOVA
M. A. NEMATOVA

Abstract

Abstract. Turaeva B, Kutlieva GJ, Kamolova XF, Nematofa MA. 2026. Growth and hematological effects of cellulolytic Bacillus spp. isolated from rabbit gut. Biodiversitas 27 (1): d270131. https://doi.org/10.13057/biodiv/d270131. This study aimed to isolate cellulolytic Bacillus strains from the rabbit gastrointestinal tract and evaluate their probiotic potential. Thirteen spore-forming isolates were obtained, of which Bacillus subtilis PBUZ-1 and PBUZ-3 exhibited the strongest cellulolytic activity. Both strains demonstrated high cellulase activity, with PBUZ-1 producing 1.69±0.16 U/mL in whey-based medium compared to 0.29±0.02 U/mL in Getchenson medium (p<0.001, Cohen’s d = 12.49). In vivo trials were conducted on Hikol rabbits supplemented with lyophilized strains (1 × 10⁶ CFU/g feed) for 30 days. Rabbits receiving probiotics showed superior growth performance, with body weights exceeding controls by 312-1050 g at 110-120 days (p<0.01, Cohen’s d = 2.45-3.83). Hematological analysis revealed significant increases in red and white blood cell counts, hemoglobin, and platelets (p<0.05). Biochemical profiles also improved, with higher total protein (65.17±2.24 g/L) and albumin (4.7±0.30 g/L), while bilirubin remained within physiological ranges. No adverse effects were observed, and physiological parameters such as temperature and heart rate stayed within normal limits. These findings demonstrate that rabbit gut-derived Bacillus subtilis strains enhance growth, hematological indices, and systemic health in rabbits. Importantly, this study provides the first evidence that such strains possess dual cellulolytic and probiotic functions, offering a safe and sustainable alternative to synthetic feed additives in rabbit farming and potentially other herbivorous livestock.

Article Details

Section

Articles

References

Abdel-Aziz SH, Ibrahim AM, Guirgis AA, Dawwam GE, Elsababty ZE. 2021. Isolation and screening of cellulase producing bacteria isolated from soil. Benha J Appl Sci 6 (3): 207-213. https://doi.org/10.21608/bjas.2021.188849.

Abdelsalam M, Fathi M, El-Raffa A, Abd El-latif G, Abou-Emera O, Abd El-Fatah M, Rayan G. 2025. Influence of probiotic supplementation and rabbit line on growth performance, carcass yield, blood biochemistry and immune response under hot weather. Animal Biosci 38 (9): 2033. https://doi.org/10.5713/ab.24.0904.

Borah M, Mandal M, Konwar BK. 2025. Characterization of probiotic strains of Bacillus sp. from fermented palm wine (Nypa fructicans sp.) and exploration of cellulolytic potential for use as an addition in animal feed. Intl Microbiol 28: 965-977. https://doi.org/10.1007/s10123-024-00589-5.

Cai Z, Wang Y, You Y, Yang N, Lu S, Xue J, Xing X, Sha S, Zhao L. 2024. Introduction of cellulolytic bacterium Bacillus velezensis Z2. 6 and its cellulase production optimization. Microorganisms 12 (5): 979. https://doi.org/10.3390/microorganisms12050979.

Chen B, Zeng Y, Wang J, Lei M, Gan B, Wan Z, Wu L, Luo G, Cao S, An T, Zhang Q, Pan K, Jing B, Ni X, Zeng D. 2025. Targeted screening of fiber degrading bacteria with probiotic function in herbivore feces. Probiotics Antimicrob Prot 17: 1473-1497. https://doi.org/10.1007/s12602-024-10215-5.

Dicks LMT, Botha M, Loos B, Smith C. 2015. Adhesion of Lactobacillus reuteri strain Lr1 to equine epithelial cells and competitive exclusion of Clostridium difficile from the gastrointestinal tract of horses. Ann Microbiol 65: 1087-1096. https://doi.org/10.1007/s13213-014-0954-4.

Dobrzyński J, Wróbel B, Górska EB. 2023. Taxonomy, ecology, and cellulolytic properties of the genus Bacillus and related genera. Agriculture 13: 1979. https://doi.org/10.3390/agriculture13101979.

Fu Y, Luo X-D, Li J-Z et al. 2024. Host-derived Lactobacillus plantarum alleviates hyperuricemia by improving gut microbial community and hydrolase-mediated degradation of purine nucleosides. eLife 13: e100068. https://doi.org/10.7554/eLife.100068.

Ghiasi SE, Gheibipour M, Motamedi H, Dar MA. 2024. Screening and evaluation of the ruminal cellulolytic bacteria and their potential application as probiotics. Iran J Microbiol 16 (3): 389. https://doi.org/10.18502/ijm.v16i3.15796

Guo M, Wu F, Hao G, Qi Q, Li R, Li N, Wei L, Chai T. 2017. Bacillus subtilis improves immunity and disease resistance in rabbits. Front Immunol 8: 354. https://doi.org/10.3389/fimmu.2017.00354.

Ivannikova RF, Pimenov NV, Sotnikova LF, Kostilev VA, Gorlov IF, Knyazhechenko OA, Frolova MV. 2021. Adaptive and productive effects when using bacteria of the genus Bacillus as the basis of a feed additive on a rabbit model. IOP Conf Ser: Earth Environ Sci 677: 022098. https://doi.org/10.1088/1755-1315/677/2/022098.

Khongkool K, Taweechotipatr M, Payungporn S, Sawaswong V, Lertworapreecha M. 2025. Gut microbiota modulation and immunity enhancement by Bacillus amyloliquefaciens NL1.2: A fiber-degrading probiotic isolated from native Thai swine. Vet World 18 (6): 1487-1507. https://doi.org/10.14202/vetworld.2025.1487-1507.

Liao Y, Wu S, Zhou G, Mei S, Yang Z, Li S, Jin Z, Deng Y, Wen M, Yang Y. 2024. Cellulolytic Bacillus cereus produces a variety of short-chain fatty acids and has potential as a probiotic. Microbiol Spectr 12: e03267-23. https://doi.org/10.1128/spectrum.03267-23.

Liu B, Cui Y, Ali Q, Zhu X, Li D, Ma S, Wang Z, Wang C, Shi Y. 2022. Gut microbiota modulate rabbit meat quality in response to dietary fiber. Front Nutr 9: 849429. https://doi.org/10.3389/fnut.2022.849429.

Mancini S, Paci G. 2021. Probiotics in rabbit farming: Growth performance, health status, and meat quality. Animals 11 (12): 3388. https://doi.org/10.3390/ani11123388.

Manjunatha DB, Rajeshwari YB, Mahadevappa DG, Shree JS. 2016. Effect of cellulolytic enzymes and probiotics on growth performance of broiler rabbits. J Anim Res 6 (6): 1053-1056. https://doi.org/10.5958/2277-940X.2016.00153.4.

Mun D, Kyoung H, Kong M, Ryu S, Jang KB, Baek J, Park KI, Song M, Kim Y. 2021. Effects of Bacillus-based probiotics on growth performance, nutrient digestibility, and intestinal health of weaned pigs. J Anim Sci Technol 63 (6): 1314. https://doi.org/10.5187/jast.2021.e109.

Nematova MA, Murodova SS. 2024. The influence of rhizobacteria on some secondary metabolites of Ferula: A review. Sib J Life Sci Agric 16 (1): 356-379. https://doi.org/10.12731/2658-6649-2024-16-1-705.

Ngalimat MS, Yahaya RSR, Baharudin MMA-A, Yaminudin SM, Karim M, Ahmad SA, Sabri S. 2021. A review on the biotechnological applications of the operational group Bacillus amyloliquefaciens. Microorganisms 9: 614. https://doi.org/10.3390/microorganisms9030614.

Pu G, Hou L, Zhao Q, Liu G, Wang Z, Zhou W, Niu P, Wu C, Li P, Huang R. 2025. Interactions between gut microbes and host promote degradation of various fiber components in Meishan pigs. mSystems 10: e01500-24. https://doi.org/10.1128/msystems.01500-24.

Sun Q, Vega NM, Cervantes B, Mancuso CP, Mao N, Taylor MN, Collins JJ, Khalil AS, Gore J, Lu TK. 2022. Enhancing nutritional niche and host defenses by modifying the gut microbiome. Mol Syst Biol 18: msb20209933. https://doi.org/10.15252/msb.20209933.

Thomas L, Ram H, Singh VP. 2018. Inducible cellulase production from an organic solvent tolerant Bacillus sp. SV1 and evolutionary divergence of endoglucanase in different species of the genus Bacillus. Braz J Microbiol 49 (2): 429-442. https://doi.org/10.1016/j.bjm.2017.05.010.

Wang J, Ni X, Wen B, Zhou Y, Liu L, Zeng Y, Zhao W, Khalique A, Wang P, Pan K, Yu Z, Jing B, Liu H, Zeng D. 2020. Bacillus strains improve growth performance via enhancing digestive function and anti-disease ability in young and weaning rex rabbits. Appl Microbiol Biotechnol 104: 4493-4504. https://doi.org/10.1007/s00253-020-10536-9.

Wita A, Białas W, Wilk R, Szychowska K, Czaczyk K. 2019. The influence of temperature and nitrogen source on cellulolytic potential of microbiota isolated from natural environment. Pol J Microbiol 68 (1): 105-114. https://doi.org/10.21307/pjm-2019-012.

Zhang H, Liu J, Miao Z. 2020. The effect of probiotic Bacillus amyloliquefaciens on growth performance, immune organ index and immune function of weaned meat rabbits. Vet Archiv 90 (3): 271-277. https://doi.org/10.24099/vet.arhiv.0564.