Shrimp vibriosis , caused predominantly by virulent strains of Vibrio spp., continues to pose a serious threat to global aquaculture. Outbreaks of the disease are often associated with mass mortalities in shrimp farms, reduced productivity, and severe economic losses. Conventional disease management strategies, including the use of antibiotics and chemotherapeutics, have proven increasingly ineffective due to the emergence of multidrug-resistant bacterial strains, besides raising concerns about environmental impact and food safety. Consequently, the search for sustainable alternatives has turned attention toward antimicrobial peptides (AMPs), naturally occurring defense molecules that offer broad-spectrum activity, low propensity for resistance development, and eco-friendly applicability in aquaculture systems.
In the present study, we explored the dual functional potential of crustin , a recombinant AMP derived from the crab Scylla serrata , with particular focus on its antimicrobial activity against vibrio species associated with shrimp vibriosis , and its anticancer properties against human breast (MDA-MB-231) and prostate (PC-3) carcinoma cell lines.
Antimicrobial assays revealed that recombinant crustin exhibited potent inhibitory activity against virulent Vibrio isolates, confirming its capacity to serve as a preventive or therapeutic agent in shrimp culture. Beyond its antimicrobial effects, crustin demonstrated significant anticancer potential. Cytotoxicity studies using MTT assays revealed dose-dependent inhibition of proliferation in breast (MDA-MB-231) and prostate (PC-3) carcinoma cell lines, with IC₅₀ values of 250 µg/mL and 300 µg/mL, respectively . Multiple complementary assays including propidium iodide (PI) staining, Annexin V-FITC analysis, dual acridine orange/ethidium bromide (AO/EB) staining, and DNA fragmentation provided clear evidence of apoptosis induction. Further, Rhodamine 123 staining confirmed mitochondrial membrane depolarization, implicating activation of the intrinsic (mitochondria-mediated) apoptotic pathway. Additionally, f low cytometry analysis revealed G0/G1 phase cell cycle arrest, supporting the role of crustin in suppressing tumor cell proliferation through both apoptotic and cell cycle regulatory mechanisms.
Overall, the findings highlight recombinant crustin as a multifunctional bioactive molecule with significant translational relevance. Its strong anti-Vibrio activity positions it as a potential natural alternative to antibiotics for sustainable aquaculture, while its selective anticancer effects underscore its promise as a lead molecule for novel therapeutic strategies. This dual functionality not only broadens the application spectrum of AMPs but also emphasizes the untapped potential of marine-derived biomolecules in addressing challenges in both aquaculture and human health.