Chitosan derived from crab shell waste: A natural coating against anthracnose disease in Capsicum annuum L. (Chilli)
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Abstract
Chitosan, a polysaccharide derived from crustacean shells, serves as a promising natural coating for horticultural products such as chili (Capsicum annuum L.) to control postharvest diseases like anthracnose, caused by Colletotrichum species. This research aimed to extract chitosan from crab shell waste and evaluate its physicochemical properties and efficacy against anthracnose in chili fruit. Chitosan was produced from crab shell waste through demineralization, deproteinization, and deacetylation. The process yielded 32.29% chitosan. Key characteristics of the extracted chitosan included a violet result in the biuret test (indicating protein presence), partial solubility in acetic acid, 0.02% ash content, a melting point of 193.2 °C, 6.87% moisture content, and a low viscosity of 192.9 centipoise (cP). In vitro assays demonstrated that chitosan at a concentration of 2 mg/mL significantly inhibited the growth of Colletotrichum gloeosporioides. In in vivo trials, pure chitosan (2 mg/mL) and shell-derived chitosan (6 mg/mL) slightly prolonged the disease incubation period to 3.67 days and reduced disease severity by 33.33% compared to the control. However, the chitosan coating did not significantly affect fruit shrinkage. These results suggest that while crab shell-derived chitosan has strong antifungal properties, its formulation may need modification to improve physical barrier properties such as moisture retention.
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References
Aberoumand A & Hoseinian M. 2025. Extraction of chitosan from shells of crab (Liocarcinus vernalis). Appl. Food Res. 5(1): 100964. https://doi.org/10.1016/j.afres.2025.100964
Abirami S, Ramachandran ER, Samrot AV, Sakthikavitha M, Revathi P, Varsini AM, Sundar D, Saigeetha S, Shobana N, & Prakash P. 2021. Extraction of chitosan from crab shell and fungi and its antibacterial activity against urinary tract infection causing pathogens. J. Pure Appl. Microbiol. 15(2): 968–975. https://doi.org/10.22207/jpam.15.2.55
Ahmad F, Kusumiyati K, Soleh MA, Khan MR, & Sundari RS. 2024. Chili cultivars vulnerability: A multi-factorial examination of disease and pest-induced yield decline across different growing microclimates and watering regimens. BMC Plant Biol. 24(1): 979. https://doi.org/10.1186/s12870-024-05541-3
Ahmed T, Noman M, Jiang H, Shahid M, Ma C, Wu Z, Nazir MM, Ali MdA, White JC, Chen J, & Li B. 2022. Bioengineered chitosan-iron nanocomposite controls bacterial leaf blight disease by modulating plant defense response and nutritional status of rice (Oryza sativa L.). Nano Today. 45: 101547. https://doi.org/10.1016/j.nantod.2022.101547
Akin HM, Anggraini D, Wibowo L, Prasetyo J, & Suharjo R. 2024. Antifungal evaluation of turmeric rhizome extract against Colletotrichum capsici, the causal agent of anthracnose on red-chili peppers (Capsicum annuum L.). J. Trop. Plant Pests Dis. 24(1): 75–81. https://doi.org/10.23960/jhptt.12475-81
Amalia KP, Ekayani M, & Nurjanah N. 2021. Pemetaan dan alternatif pemanfaatan limbah cangkang rajungan di Indonesia [Mapping and alternative utilization of shell crab waste in Indonesia]. JPHPI. 24(3): 310–318. https://doi.org/10.17844/jphpi.v24i3.37436
Braga SDP, Lundgren GA, Macedo SA, Tavares JF, Vieira WAS, Câmara MPS, & de Souza EL. 2019. Application of coatings formed by chitosan and Mentha essential oils to control anthracnose caused by Colletotrichum gloesporioides and C. brevisporum in papaya (Carica papaya L.) fruit. Int. J. Biol. Macromol. 139: 631–639. https://doi.org/10.1016/j.ijbiomac.2019.08.010
Carmona SL, Villarreal-Navarrete A, Burbano-David D, Gómez-Marroquín M, Torres-Rojas E, & Soto-Suárez M. 2021. Protection of tomato plants against Fusarium oxysporum f.sp. lycopersici induced by chitosan. Revista Colombiana De Ciencias Hortícolas. 15(3): e1282. https://doi.org/10.17584/rcch.2021v15i3.12822
Chen K, Tian R, Jiang J, Xiao M, Wu K, Kuang Y, Deng P, Zhao X, & Jiang F. 2024. Moisture loss inhibition with biopolymer films for preservation of fruits and vegetables: a review. Int. J. Biol. Macromol. 263(1): 130337. https://doi.org/10.1016/j.ijbiomac.2024.130337
Cong H, Wu Q, Zhang Z, & Kan J. 2023. Improvement of functional characteristics of Hypophthalmichthys molitrix protein by modification with chitosan oligosaccharide. Front. Nutr. 10: 1140191. https://doi.org/10.3389/fnut.2023.1140191
De Bona GS, Vincenzi S, De Marchi F, Angelini E, & Bertazzon N. 2021. Chitosan induces delayed grapevine defense mechanisms and protects grapevine against Botrytis cinerea. J. Plant Dis. Prot. 128(3): 715–724. https://doi.org/10.1007/s41348-021-00432-3
Du DX & Vuong BX. 2019. Study on preparation of water-soluble chitosan with varying molecular weights and its antioxidant activity. Adv. Mater. Sci. Eng. 2019(1): 1–8. https://doi.org/10.1155/2019/8781013
Eddya M, Tbib B, & El-Hami K. 2020. A comparison of chitosan properties after extraction from shrimp shells by diluted and concentrated acids. Heliyon. 6(2): e03486. https://doi.org/10.1016/j.heliyon.2020.e03486
Fachry ME & Alpiani. 2021. Literature review: Economic value of utilization of crab shell waste (case study of PT. Toba Surimi Industri in Tanjungpinang City, Riau Island Province). Akuatikisle: J. Aqua. Coast. & Isle. 5(2): 49–52. https://doi.org/10.29239/j.akuatikisle.5.2.49-52
Hendricks KE, Christman MC, & Roberts PD. 2017. A statistical evaluation of methods of in-vitro growth assessment for Phyllosticta citricarpa: Average colony diameter vs. area. PLoS ONE. 12(1): e0170755. https://doi.org/10.1371/journal.pone.0170755
Hodiyah I, Suryaman M, Hartini E, Juhaeni AH, Laksana BY, Aisyah A, & Benatar GV. 2024. Diversity of morphology, pathogenicity, and host range of Colletotrichum spp. associated with chili anthracnose in East Priangan, Indonesia. Biodiversitas. 25(2): e250212. https://doi.org/10.13057/biodiv/d250212
Hu X, Saravanakumar K, Sathiyaseelan A, & Wang M. 2020. Chitosan nanoparticles as edible surface coating agent to preserve the fresh-cut bell pepper (Capsicum annuum L. var. grossum). Int. J. Biol. Macromol. 165(A): 948–957. https://doi.org/10.1016/j.ijbiomac.2020.09.176
Ibram A, Ionescu AM, & Cadar E. 2019. Comparison of extraction methods of chitin and chitosan from different sources. EJNM. 2(2): 23–36. https://doi.org/10.26417/688wvv48e
Ismail AM, Elshewy ES, Ali IH, Muhanna NAES, & Khafagi EY. 2024. Encapsulation of clove oil nanoemulsion in chitosan-based nanocomposite: In vitro and in vivo antifungal activity against Rhizoctonia solani and Sclerotium rolfsii. Phyton-Int. J. Exp. Bot. 93(11): 2787–2811. https://doi.org/10.32604/phyton.2024.057518
Jieying S, Shuangfeng G, Tingting L, Zhicheng Y, Lei W, Caie W, Dandan Z, Gongjian F, & Xiaojing L. 2025. The molecular mechanism of chitosan-based OEO nanoemulsion edible film in controlling Alternaria alternata and in application for apricot preservation. Food Control. 176: 111354. https://doi.org/10.1016/j.foodcont.2025.111354
Kehila S, Alkalai-Tuvia S, Chalupowicz D, Poverenov E, & Fallik E. 2021. Can edible coatings maintain sweet pepper quality after prolonged storage at sub-optimal temperatures?. Horticulturae. 7(10): 387. https://doi.org/10.3390/horticulturae7100387
Koirala P, Bhandari Y, Khadka A, Kumar SR, & Nirmal NP. 2024. Nanochitosan from crustacean and mollusk byproduct: Extraction, characterization, and applications in the food industry. Int. J. Biol. Macromol. 262: 130008. https://doi.org/10.1016/j.ijbiomac.2024.130008
Kurniasih M, Kartika D, & Riyanti R. 2016. Optimasi kondisi adsorpsi kolesterol menggunakan karboksimetil kitosan [Optimizing conditions to cholesterol adsorbed with carboxymethyl chitosan]. Molekul. 11(1): 112–124. https://doi.org/10.20884/1.jm.2016.11.1.200
Ma M, Liu Y, Zhang S, & Yuan Y. 2024. Edible coating for fresh-cut fruit and vegetable preservation: Biomaterials, functional ingredients, and joint non-thermal technology. Foods. 13(23): 3937. https://doi.org/10.3390/foods13233937
Martínez-Blay V, Pérez-Gago MB, de la Fuente B, Carbó R, & Palou L. 2020. Edible coatings formulated with antifungal GRAS salts to control citrus anthracnose caused by Colletotrichum gloeosporioides and preserve postharvest fruit quality. Coatings. 10(8): 730. https://doi.org/10.3390/coatings10080730
Megahed AA, Masoud HM, Helmy MSE, Ibrahim MAA, El-Mougy NS, & Abdel-Kader MM. 2023. Efficiency of some abiotic and biotic agents on Vicia faba L. rust and chocolate spot diseases. Plant Prot. 7(3): 449–463. https://doi.org/10.33804/pp.007.03.4798
Morachis-Valdez AG, Gómez-Oliván LM, García-Argueta I, Hernández-Navarro MD, Díaz-Bandera D, & Dublán-García O. 2017. Effect of chitosan edible coating on the biochemical and physical characteristics of carp fillet (Cyprinus carpio) stored at −18 ºC. Int. J. Food Sci. 2017(1): 1–10. https://doi.org/10.1155/2017/2812483
Muflikh YN & Kiloes AM. 2024. Insight into the buying behaviour of consumers for chilli in Indonesia: Households and food businesses in selected cities. Appl. Food Res. 4(1): 100413. https://doi.org/10.1016/j.afres.2024.100413
Musmade A & Mahatma L. 2021. Extraction and characterization of chitosan by simple technique from mud crabs. Int. J. Curr. Microbiol. App. Sci. 10(6): 513–518. https://doi.org/10.20546/ijcmas.2021.1006.055
Rungjindamai N. 2016. Isolation and evaluation of biocontrol agents in controlling anthracnose disease of mango in Thailand. J. Plant Protect. Res. 56(3): 306–311. https://doi.org/10.1515/jppr-2016-0034
Noor NM & Zakaria L. 2024. Pathogenic variations in Colletotrichum spp. causing chilli anthracnose in Peninsular Malaysia. Plant Pathol. 73(7): 1788–1793. https://doi.org/10.1111/ppa.13925
Oo MM, Yoon H, Jang HA, & Oh S. 2018. Identification and characterization of Colletotrichum species associated with bitter rot disease of apple in South Korea. Plant Pathol. J. 34(6): 480–489. https://doi.org/10.5423/ppj.ft.10.2018.0201
Paul SK, Sarkar S, Sethi LN, & Ghosh SK. 2018. Development of chitosan-based optimized edible coating for tomato (Solanum lycopersicum) and its characterization. J. Food Sci. Technol. 55(7): 2446–2456. https://doi.org/10.1007/s13197-018-3162-6
Pellis A, Guebitz GM, & Nyanhongo GS. 2022. Chitosan: Sources, processing and modification techniques. Gels. 8(7): 393. https://doi.org/10.3390/gels8070393
Petriccione M, Mastrobuoni F, Pasquariello MS, Zampella L, Nobis E, Capriolo G, & Scortichini M. 2015. Effect of chitosan coating on the postharvest quality and antioxidant enzyme system response of strawberry fruit during cold storage. Foods. 4(4): 501–523. https://doi.org/10.3390/foods4040501
Pham TT, Nguyen LLP, Dam MS, & Baranyai L. 2023. Application of edible coating in extension of fruit shelf life: Review. AgriEngineering. 5(1): 520–536. https://doi.org/10.3390/agriengineering5010034
Popović NT, Lorencin V, Strunjak-Perović I, & Čož-Rakovac R. 2023. Shell waste management and utilization: Mitigating organic pollution and enhancing sustainability. Appl. Sci. 13(1): 623. https://doi.org/10.3390/app13010623
Riseh RS, Hassanisaadi M, Vatankhah M, Babaki SA, & Barka EA. 2022. Chitosan as a potential natural compound to manage plant diseases. Int. J. Biol. Macromol. 220: 998–1009. https://doi.org/10.1016/j.ijbiomac.2022.08.109
Salgado-Cruz MD, Salgado-Cruz J, García-Hernández AB, Calderón-Domínguez G, Gómez-Viquez H, Oliver-Espinoza R, Fernández-Martínez MC, & Yáñez-Fernández J. 2021. Chitosan as a coating for biocontrol in postharvest products: A bibliometric review. Membranes. 11(6): 421. https://doi.org/10.3390/membranes11060421
Silva WB, Silva GMC, Santana DB, Salvador AR, Medeiros DB, Belghith I, da Silva NM, Cordeiro MHM, & Misobutsi GP. 2017. Chitosan delays ripening and ROS production in guava (Psidium guajava L.) fruit. Food Chem. 242: 232–238. https://doi.org/10.1016/j.foodchem.2017.09.052
Soesanto L, Prastyani N, Utami DS, & Manan A. 2020. Application of raw secondary metabolites from four entomopathogenic fungi against chilli disease caused by viruses. J. Trop. Plant Pests Dis. 20(2): 100–107. https://doi.org/10.23960/j.hptt.220100-107
Song W, Zhang Q, Guan Y, Li W, Xie S, Tong J, Li M, & Ren L. 2022. Synthesis and characterization of porous chitosan/Saccharomycetes adsorption microspheres. Polymers. 14(11): 2292. https://doi.org/10.3390/polym14112292
Than PP, Prihastuti H, Phoulivong S, Taylor PWJ, & Hyde KD. 2008. Chilli anthracnose disease caused by Colletotrichum species. J. Zhejiang Univ. Sci. B. 9(10): 764–778. https://doi.org/10.1631/jzus.b0860007
Tovar CDG, Delgado-Ospina J, Porras DPN, Peralta-Ruiz Y, Cordero AP, Castro JI, Valencia MNC, Mina JH, & López CC. 2019. Colletotrichum gloesporioides inhibition in situ by chitosan–Ruta graveolens essential oil coatings: Effect on microbiological, physicochemical, and organoleptic properties of guava (Psidium guajava L.) during room temperature storage. Biomolecules. 9(9): 399. https://doi.org/10.3390/biom9090399
Trung TS, Tram LH, Van Tan N, Van Hoa N, Minh NC, Loc PT, & Stevens WF. 2020. Improved method for production of chitin and chitosan from shrimp shells. Carbohydr. Res. 489: 107913. https://doi.org/10.1016/j.carres.2020.107913
Yan D, Li Y, Liu Y, Li N, Zhang X, & Yan C. 2021. Antimicrobial properties of chitosan and chitosan derivatives in the treatment of enteric infections. Molecules. 26(23): 7136. https://doi.org/10.3390/molecules26237136
Zhang X, Liang S, Wu Q, Charles TC, He R, Wu J, Zhao Y, Zhao Z, & Wang H. 2022. Mode of action of nanochitin whisker against Fusarium pseudograminearum. Int. J. Biol. Macromol. 217: 356–366. https://doi.org/10.1016/j.ijbiomac.2022.07.056
Zhang HM & Shen L. 2025. Chitosan as a potential biocontrol agent of plant pathogens and its multiple acting mechanisms: A review. Int. J. Biol. Macromol. 334(1): 148984. https://doi.org/10.1016/j.ijbiomac.2025.148984