Bromelain-Enriched Diet Improves Morphology and Spawning Performance in Clarias macrocephalus
Keywords:
Bromelain, Clarias macrocephalus, external morphology, functional feed, reproductive performanceAbstract
This study aimed to investigate the effects of a bromelain-supplemented diet on the external morphological and reproductive characteristics of Clarias macrocephalus, and to assess how these changes influence spawning performance. Functional food enzymes are attracting increasing interest in aquaculture due to their potential to improve digestion and physiological performance. Bromelain, a proteolytic enzyme derived from pineapple, was evaluated for its effects on external morphology and reproductive characteristics. The experiment comprised two dietary treatments: a control diet and a bromelain-supplemented diet. External morphology was assessed by examining skin pigmentation, tail curvature, mucus secretion, muscle tone and reproductive traits such as genital papillae development and abdominal distension. External morphologies were scored using a standardised scale of 1–5, and statistical analyses were performed using independent t-tests and chi-square tests. Fish fed the bromelain diet showed significantly greater skin pigmentation (3.65 ± 0.49 vs. 1.15 ± 0.37), increased tail curvature (3.40 ± 0.60 vs. 1.60 ± 0.50), higher mucus secretion (3.80 ± 0.89 vs. 1.50 ± 0.51) and reduced muscle tone (3.40 ± 0.60 vs. 1.25 ± 0.55) compared to the control group (p<0.001). In contrast, reproductive characteristics including genital papilla structure and abdominal distension showed no significant differences (p>0.05), although successful spawning and a high fertilisation rate (87.4 ± 5.2%) were observed in the bromelain-fed pairs. The findings suggest that bromelain supplementation has a marked effect on the external morphology and physiological condition of catfish, which could enhance spawning readiness despite limited visible changes in reproductive anatomy. This indicates that the effects of bromelain have an internal or metabolic potential. The study highlights the potential of bromelain as a feed additive in aquaculture and recommends further research to investigate its effects on reproductive hormones, gonadal histology and synergistic interactions with bioactive compounds to optimise reproductive outcomes in C. macrocephalus culture.
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Alves, R.N. & Agustí, S. 2022. Transcriptional changes in the gilthead seabream (Sparus aurata) skin in response to ultraviolet b radiation exposure. Frontiers in Marine Science, 9. DOI: https://doi.org/10.3389/fmars.2022.966654
AOAC. 2003. Official Methods of Analysis of AOAC International. 17th Ed. Association of the Official Analytical Chemists (AOAC) International, Gaithersburg.
Aripin, S., Jintasataporn, O. & Yoonpundh, R. 2015. Effects of zinc amino acid in walking catfish (Clarias macrocephalus) female broodstock first sexual maturation. Journal of Aquaculture Research and Development, 6: 1-6.
Badriyya, E., S, S.S., Pratiwi, A.R., Dillasamola, D., Aldi, Y. & Husni, E. 2020. Topical anti-inflammatory activity of bromelain. Pharmacognosy Journal, 12(6s): 1586-1593. DOI: https://doi.org/10.5530/pj.2020.12.217
Bao, Y., Wang, K., Yang, H., Regenstein, J.M., Ertbjerg, P. & Zhou, P. 2020. Protein degradation of black carp (Mylopharyngodon piceus) muscle during cold storage. Food Chemistry, 308: 125576. DOI: https://doi.org/10.1016/j.foodchem.2019.125576
Barua, A., Rana, S., Shawon, F.I., Muntaha, M.K., Ahmed, S.I., Shimul, S.A. & Nahid, S.A.A. 2021. Effect of background color on the growth pattern and coloration of guppy (Poecilia reticulata). Bangladesh Journal of Fisheries, 33(1): 13-20. DOI: https://doi.org/10.52168/bjf.2021.33.02
Bautista, L.M., Tumbokon, B.L., Plasus, M.M. & Serrano, A.E. 2022. Benfotiamine in high carbohydrate diet enhances growth and reproductive performance of the Asian native catfish Clarias macrocephalus subadults. The Israeli Journal of Aquaculture Bamidgeh, 74: 11. DOI: https://doi.org/10.46989/001c.36448
Bond, E.R. & Forsgren, K.L. 2021. External reproductive structures of male surfperches (Embiotocidae): Urogenital papilla. The Anatomical Record, 305(7): 1712-1723. DOI: https://doi.org/10.1002/ar.24836
Bottega, R., Persico, I., De Seta, F., Romano, F. & Di Lorenzo, G. 2021. Anti-inflammatory properties of a proprietary bromelain extract (Bromeyal™) after in vitro simulated gastrointestinal digestion. International Journal of Immunopathology and Pharmacology, 35: 1-9. DOI: https://doi.org/10.1177/20587384211034686
Butler, J.M., Whitlow, S.M., Gwan, A., Chakrabarty, P. & Maruska, K.P. 2017. Swim bladder morphology changes with female reproductive state in the mouth-brooding African cichlid Astatotilapia burtoni. Journal of Experimental Biology, 220(23): 4463-4470. DOI: https://doi.org/10.1242/jeb.163832
Byrnes, M.A., Darnell, K.M. & Darnell, M.Z. 2022. Changes in the morphology of widgeon grass (Ruppia maritima) with the onset of reproduction and impacts on fish assemblages at the Chandeleur Islands, LA. Frontiers in Environmental Science, 10: 978772. DOI: https://doi.org/10.3389/fenvs.2022.978772
Chakraborty, A.J., Mitra, S., Tallei, T.E., Tareq, A.M., Nainu, F., Cicia, D. et al. 2021. Bromelain a potential bioactive compound: A comprehensive overview from a pharmacological perspective. Life, 11(4): 317. DOI: https://doi.org/10.3390/life11040317
Chen, S., Maulu, S., Wang, J., Xie, X., Liang, X., Wang, H., Wang, J. & Xue, M. 2024. The application of protease in aquaculture: Prospects for enhancing the aquafeed industry. Animal Nutrition, 16: 105-121. DOI: https://doi.org/10.1016/j.aninu.2023.11.001
Da, C.T., Xuyen, B.T.K., Nguyen, T.K.O., Tang, V.T., Ha, P.T.T., Pham, M.T. & Berg, H. 2024. Vitamin solutions effects on reproduction of broodstock, growth performance, and survival rate of Pangasius catfish fingerlings. Animals, 14(15): 2203. DOI: https://doi.org/10.3390/ani14152203
Das Neves, J.L., Coetzee, H., Barnhoorn, I. & Wagenaar, I. 2019. The urogenital papillae as an indicator of sexual maturity in male African catfish, Clarias gariepinus (Burchell, 1822). Aquaculture Research, 50(12): 3519-3527. DOI: https://doi.org/10.1111/are.14283
Department of Statistics Malaysia (DOSM). 2025. Agriculture Census 2024: Aquaculture highlights. Department of Statistics Malaysia, Putrajaya.
Díaz-Puertas, R., Adamek, M., Mallavia, R. & Falcó, A. 2023. Fish skin mucus extracts: An underexplored source of antimicrobial agents. Marine Drugs, 21(6): 350. DOI: https://doi.org/10.3390/md21060350
Dong, X., Wu, L., Zhu, X., Han, D., Yang, Y. & Xie, S. 2011. Effects of dietary oxidized fish oil on growth performance and skin colour of Chinese longsnout catfish (Leiocassis longirostris Günther). Aquaculture Nutrition, 17(4): e861-e868. DOI: https://doi.org/10.1111/j.1365-2095.2011.00854.x
Duong, T.Y., Thi Nguyen, N.T. & Le, N.S. 2022. Strain-specific effects of broodstock size on fecundity and offspring performance in bighead catfish (Clarias macrocephalus Günther, 1864). Aquaculture Research, 53(11): 3996-4006. DOI: https://doi.org/10.1111/are.15902
El-Sayed, A.F.M. (Ed.). 2006. Tilapia Culture. CABI Publishing, Wallingford. DOI: https://doi.org/10.1079/9780851990149.0000
Feng, X., Hang, S., Zhou, Y., Liu, Q. & Yang, H. 2018. Bromelain kinetics and mechanism on myofibril from golden pomfret (Trachinotus blochii). Journal of Food Science, 83(8): 2148-2158. DOI: https://doi.org/10.1111/1750-3841.14212
Fragkoulis, S., Kourkouta, C., Geladakis, G., Printzi, A., Glaropoulos, A. & Koumoundouros, G. 2022. Recovery of haemal lordosis in European seabass Dicentrarchus labrax (Linnaeus 1758). Aquaculture Journal, 2(1): 1-12. DOI: https://doi.org/10.3390/aquacj2010001
Geladakis, G., Kourkouta, C., Somarakis, S. & Koumoundouros, G. 2022. Developmental temperature shapes the otolith morphology of metamorphosing and juvenile gilthead seabream (Sparus aurata Linnaeus, 1758). Fishes, 7(2): 82. DOI: https://doi.org/10.3390/fishes7020082
Gopalraaj, J., Velayudhannair, K., Arockiasamy, J.P. & Radhakrishnan, D.K. 2024. The effect of dietary supplementation of proteases on growth, digestive enzymes, oxidative stress, and intestinal morphology in fishes - A review. Aquaculture International, 32(1): 745-765. DOI: https://doi.org/10.1007/s10499-023-01191-8
Hassaan, M.S., El-Sayed, A.I.M., Soltan, M., Iraqi, M.M., Goda, A., Davies, S.J. & Ramadan, H. 2019. Partial dietary fish meal replacement with cotton seed meal and supplementation with exogenous protease alters growth, feed performance, hematological indices and associated gene expression markers (GH, IGF-I) for Nile tilapia, Oreochromis niloticus. Aquaculture, 503: 282-292. DOI: https://doi.org/10.1016/j.aquaculture.2019.01.009
Hung, T., Ellison, L., Stevenson, T., Sandford, M., Schultz, A.A. & Eads, A.R. 2022. Early weaning in endangered delta smelt: Effect of weaning time on growth and survival. North American Journal of Aquaculture, 84(2): 249-260. DOI: https://doi.org/10.1002/naaq.10230
Kerniske, F.F., Castro, J.P., De la Ossa-Guerra, L.E., Mayer, B.A., Abilhoa, V., de Paiva Affonso, I. & Artoni, R.F. 2021. Spinal malformations in a naturally isolated Neotropical fish population. PeerJ, 9: e12239. DOI: https://doi.org/10.7717/peerj.12239
Kwan, G.T., Walsh, K.A., Thompson, A.R., Ben-Aderet, N., Fennie, H.W., Semmens, B.X. & Swalethorp, R. 2024. Trophic level influences larval shortbelly rockfish development. Marine and Coastal Fisheries, 16(6). DOI: https://doi.org/10.1002/mcf2.10319
Liu, X., Ding, C., Bond, N., Sun, J., Ding, L., Chen, J. & Tao, J. 2023. Spawning cohort trade-offs of reproductive time and output in cyprinid fish along an elevation gradient. Ecology of Freshwater Fish, 33(2). DOI: https://doi.org/10.1111/eff.12763
Luis, A.I., Campos, E.V., Oliveira, J.L., Vallim, J.H., Proença, P.L., Castanha, R.F. & Fraceto, L.F. 2021. Ecotoxicity evaluation of polymeric nanoparticles loaded with ascorbic acid for fish nutrition in aquaculture. Journal of Nanobiotechnology, 19(1): 163. DOI: https://doi.org/10.1186/s12951-021-00910-8
Marín, O.M. 2016. Permanent germinal epithelium and reproductive cycle of tropical gar Atractosteus tropicus males (Lepisosteiformes: Lepisosteidae), Tabasco, Mexico. Revista de Biología Tropical, 64(4). DOI: https://doi.org/10.15517/rbt.v64i4.21123
Molinari, G.S., McCracken, V.J., Wojno, M., Rimoldi, S., Terova, G. & Kwasek, K. 2020. Can intestinal absorption of dietary protein be improved through early exposure to plant-based diet? DOI: https://doi.org/10.1101/2020.01.24.917856
Morioka, S., Vongvichith, B., Phommachan, P. & Chantasone, P. 2013. Growth and morphological development of laboratory-reared larval and juvenile bighead catfish Clarias macrocephalus (Siluriformes: Clariidae). Ichthyological Research, 60(1): 16-25. DOI: https://doi.org/10.1007/s10228-012-0301-3
Mugwanya, M., Dawood, M.A., Kimera, F. & Sewilam, H. 2023. Replacement of fish meal with fermented plant proteins in the aquafeed industry: A systematic review and meta-analysis. Reviews in Aquaculture, 15(1): 62-88. DOI: https://doi.org/10.1111/raq.12701
Na-Nakorn, U. & Brummett, R.E. 2009. Use and exchange of aquatic genetic resources for food and aquaculture: Clarias catfish. Reviews in Aquaculture, 1(3-4): 214-223. DOI: https://doi.org/10.1111/j.1753-5131.2009.01010.x
Ng, W.K. & Romano, N. 2013. A review of the nutrition and feeding management of farmed tilapia throughout the culture cycle. Reviews in Aquaculture, 5(4): 220-254. DOI: https://doi.org/10.1111/raq.12014
Öngün, O., Koca, S.B. & Koca, H.U. 2021. The effects of supplemented diets with turmeric powder on pigmentation and growth of yellow tail cichlid Pseudotropheus acei (Regan 1922). Acta Aquatica Turcica, 16(3): 386-394. DOI: https://doi.org/10.22392/actaquatr.865465
Ordóñez-Grande, B., Fernández-Alacid, L., Sanahuja, I., Sánchez-Nuño, S., Fernández-Borrás, J., Blasco, J. & Ibarz, A. 2020. Evaluating mucus exudation dynamics through isotopic enrichment and turnover of skin mucus fractions in a marine fish model. Conservation Physiology, 8(1): coaa095. DOI: https://doi.org/10.1093/conphys/coaa095
Oyedokun, S., Isioma, K. & Babangida, K.J. 2023. Comparative fertilization and hatchability rate of black and albino pigmented Clarias gariepinus (Burchell, 1822) and their hybridization. International Research Journal of Modernization in Engineering Technology and Science.
Perelló-Amorós, M., García-Pérez, I., Sánchez-Moya, A., Innamorati, A., Vélez, E.J., Achaerandio, I. & Gutiérrez, J. 2021. Diet and exercise modulate GH-IGFs axis, proteolytic markers and myogenic regulatory factors in juveniles of gilthead sea bream (Sparus aurata). Animals, 11(8): 2182. DOI: https://doi.org/10.3390/ani11082182
Péter, G., Lukić, J., Milla, S., Sándor, Z.J., Brlás-Molnár, Z., Ardó, L. et al. 2023. Reproduction of hatchery-reared pike-perch (Sander lucioperca) fed diet with low-marine-ingredients: Role of dietary fatty acids. Fishes, 8(5): 219. DOI: https://doi.org/10.3390/fishes8050219
Qin, J., Deng, X., Lei, Y., Liu, P., Lu, S. & Zhang, J. 2020. Effects of $mu$-calpain oxidation on Coregonus peled myofibrillar protein degradation in vitro. Journal of Food Science, 85(3): 682-688. DOI: https://doi.org/10.1111/1750-3841.15048
Rehman, A., Ahmad, N., Mussarat, S., Majid, A., Alnomasy, S.F. & Khan, S.N. 2021. Nanoparticles as alternatives for the control of Haemonchus contortus: A systematic approach to unveil new anti-haemonchiasis agents. Frontiers in Veterinary Science, 8. DOI: https://doi.org/10.3389/fvets.2021.789977
Saleh, E.S., Tawfeek, S.S., Abdel-Fadeel, A.A., Abdel-Daim, A.S., Abdel-Razik, A.R.H. & Youssef, I.M. 2022. Effect of dietary protease supplementation on growth performance, water quality, blood parameters and intestinal morphology of Nile tilapia (Oreochromis niloticus). Journal of Animal Physiology and Animal Nutrition, 106(2): 419-428. DOI: https://doi.org/10.1111/jpn.13591
Serrano, A.E., Bautista, L.M. & Tumbokon, B.L. 2022. Effects of vitamin E on growth and maturation in the Asian catfish (Clarias macrocephalus) at puberty. Israeli Journal of Aquaculture - Bamidgeh, 74: 1–12. DOI: https://doi.org/10.46989/001c.57186
Sharma, S.A., Surveswaran, S., Arulraj, J. & Velayudhannair, K. 2021. Bromelain enhances digestibility of Spirulina-based fish feed. Journal of Applied Phycology, 33(2): 967-977. DOI: https://doi.org/10.1007/s10811-020-02337-4
Shoemaker, C.A. & LaFrentz, B.R. 2015. Growth and survival of the fish pathogenic bacterium, Flavobacterium columnare, in tilapia mucus and porcine gastric mucin. FEMS Microbiology Letters, 362(4): 1-5. DOI: https://doi.org/10.1093/femsle/fnu060
Tahiluddin, A.B., Bornales, J.C., Limbaro, G.R.A., Paudac, M.A.T.U., Amarille, R.K., Sirad, N.R. et al. 2025. Environmental impacts of aquaculture in the Philippines. Israeli Journal of Aquaculture-Bamidgeh, 77(2): 51-81. DOI: https://doi.org/10.46989/001c.133778
Taft, N.K., Taft, B.N., Henck, H. & Mehner, T. 2018. Variation in flexural stiffness of the lepidotrichia within and among the soft fins of yellow perch under different preservation techniques. Journal of Morphology, 279(8): 1045-1057. DOI: https://doi.org/10.1002/jmor.20831
Thaiso, K., Niamphithak, P., Charoenwattanasak, S., Tola, S. & Yuangsoi, B. 2024. Effect of dietary exogenous protease supplementation on growth performance and flesh quality of Nile tilapia (Oreochromis niloticus) reared at different stocking densities. Aquaculture, Aquarium, Conservation & Legislation, 17(5): 2074-2085.
Torrecillas, S., Montero, D., Dominguez, D., Robaina, L. & Izquierdo, M. 2019. Skin mucus fatty acid composition of gilthead sea bream (Sparus aurata): A descriptive study in fish fed low and high fish meal diets. Fishes, 4(1): 15. DOI: https://doi.org/10.3390/fishes4010015
Vercauteren, M., De Swaef, E., Declercq, A., Polet, H., Aerts, J., Ampe, B. et al. 2019. Scrutinizing the triad of Vibrio tapetis, the skin barrier and pigmentation as determining factors in the development of skin ulcerations in wild common dab (Limanda limanda). Veterinary Research, 50(1). DOI: https://doi.org/10.1186/s13567-019-0659-6
Wainwright, D.K., Lauder, G.V. & Gemmell, B.J. 2024. Hydrodynamic function of the slimy and scaly surfaces of teleost fishes. Integrative and Comparative Biology, 64(2): 480-495. DOI: https://doi.org/10.1093/icb/icae066
Wiboonsirikul, J., Klahan, R. & Khuwijitjaru, P. 2024. Extraction of crude bromelain from pineapple (Ananas comosus L.) fruit waste and its in vitro protein digestibility. Journal of Agricultural Sciences Sri Lanka, 19(1). DOI: https://doi.org/10.4038/jas.v19i1.9798
Wiszniewski, G., Jarmołowicz, S., Hassaan, M.S., Mohammady, E.Y., Soaudy, M.R., Łuczyńska, J. & Siwicki, A. 2019. The use of bromelain as a feed additive in fish diets: Growth performance, intestinal morphology, digestive enzyme and immune response of juvenile sterlet (Acipenser ruthenus). Aquaculture Nutrition, 25(6): 1289-1299. DOI: https://doi.org/10.1111/anu.12949
Wu, J., Yu, T., Wang, Q., Zhang, C., Fu, D., Liu, W. & Wu, J. 2024. Effects of dietary microbial protease on growth performance, nutrient apparent digestibility, hepatic antioxidant capacity, protease activities and intestinal microflora in juvenile genetically improved farmed tilapia, Oreochromis niloticus. Aquaculture Reports, 34: 101894. DOI: https://doi.org/10.1016/j.aqrep.2023.101894
Yaseen, T., Fatima, M., Shah, S.Z.H., Ali, W. & Qudratullah, S. 2023. Effect of bromelain-fermented diets on digestive enzyme activities and muscle proximate composition of Labeo rohita. Journal of Zoology and Systematics, 24-28. DOI: https://doi.org/10.56946/jzs.v1i1.191
Zhao, H., Feng, L., Jiang, W., Liu, Y., Jiang, J., Wu, P. et al. 2015. Flesh shear force, cooking loss, muscle antioxidant status and relative expression of signaling molecules (Nrf2, Keap1, TOR, and CK2) and their target genes in young grass carp (Ctenopharyngodon idella) muscle fed with graded levels of choline. PLoS ONE, 10(11): e0142915. DOI: https://doi.org/10.1371/journal.pone.0142915
Zheng, C.C., Wu, J.W., Jin, Z.H., Ye, Z.F., Yang, S., Sun, Y.Q. & Fei, H. 2019. Exogenous enzymes as functional additives in finfish aquaculture. Aquaculture Nutrition, 26(2): 213-224. DOI: https://doi.org/10.1111/anu.12995
Zheng, S., Shi, Y., Zhang, J., Dai, J., Hu, Y. & Zhong, L. 2023. Effects of replacing fish meal with stickwater hydrolysate and meal on the growth, serum biochemical indexes, and muscle quality of yellow catfish (Tachysurus fulvidraco). Fishes, 8(12): 566. DOI: https://doi.org/10.3390/fishes8120566
Zheng, T., Song, Z., Qiang, J., Tao, Y., Zhu, H., Ma, J. & Xu, P. 2021. Transport stress induces skin innate immunity response in hybrid yellow catfish (Tachysurus fulvidraco♀ × P. vachellii♂) through TLR/NLR signaling pathways and regulation of mucus secretion. Frontiers in Immunology, 12. DOI: https://doi.org/10.3389/fimmu.2021.740359
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