Antibacterial and antivirulence factor activities of protein hydrolysates from Phatthalung Sangyod rice (Oryza sativa L.) seeds against zoonotic and foodborne pathogens
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Published:2023-10
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Volume:
Page:2002-2015
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ISSN:2231-0916
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Container-title:Veterinary World
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language:en
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Short-container-title:Vet World
Author:
Rodjan Prawit1ORCID, Sangkanu Suthinee2ORCID, Mitsuwan Watcharapong3ORCID, Pongpom Monsicha4ORCID, Saengsawang Phirabhat5ORCID, Tedja Irma6ORCID, Lamai Jarunet7ORCID, Pruksaphon Kritsada8ORCID, Jeenkeawpieam Juthatip3ORCID
Affiliation:
1. School of Agricultural Technology and Food Industry, Walailak University, Nakhon Si Thammarat, 80160, Thailand; Center of Excellence in Innovation of Essential Oil and Bio-active Compound, Walailak University, Nakhon Si Thammarat, 80160, Thailand. 2. Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai 90112, Songkhla, Thailand. 3. Center of Excellence in Innovation of Essential Oil and Bio-active Compound, Walailak University, Nakhon Si Thammarat, 80160, Thailand; Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand; One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand. 4. Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand. 5. Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand; One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand. 6. Department of Biochemistry and Molecular Biology, Monash Biomedicine Discovery Institute, Monash University, Victoria 3800, Australia. 7. Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand. 8. Department of Medical Technology, School of Allied Health Sciences, Walailak University, Nakhon Si Thammarat, 80160, Thailand.
Abstract
Background and Aim: Antimicrobial resistance is an emerging public health threat. Foodborne illnesses are typically caused by bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus, which are frequently resistant to common antimicrobial agents. Rice is a staple grain in most parts of the world. Our previous work showed that Phatthalung Sangyod rice seed protein hydrolysates (SYPs), especially SYP4, exhibit antifungal activity against several fungal species that are pathogenic for both humans and animals and are non-cytotoxic to animal red blood cells. In this study, we aimed to determine the effects of the bioactive peptides in SYPs against several pathogenic bacteria in humans and animals.
Materials and Methods: After isolating SYP1, it was treated as follows: heated (SYP2), and hydrolyzed using pepsin (SYP3), and proteinase K (SYP4). Then, we used 500 μg of protein to evaluate the antibacterial effects on four pathogenic bacteria, including E. coli, P. aeruginosa, B. cereus, and S. aureus, using agar well diffusion. Using a broth microdilution assay, we determined the minimum inhibitory and bactericidal concentration (MIC and MBC, respectively) values of active SYPs. Using the agar well diffusion and microtube incubation methods, we also assessed the inhibitory effects of SYPs on the bacterial quorum sensing (QS) activity of Chromobacterium violaceum. Sangyod rice seed protein hydrolysates were evaluated for their ability to inhibit the biofilm formation of bacterial cells by a crytal violet assay. Furthermore, using the dropping method, we tested the inhibitory effects of SYPs on the bacterial pigments pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus.
Results: Our results showed that the crude protein lysate (SYP1) did not exhibit antibacterial activity against any of the test bacteria. Intriguingly, after boiling (SYP2) and enzymatic hydrolysis (SYP3 and SYP4), the protein hydrolysates were transformed into bioactive peptides and displayed antibacterial properties against all of the test bacteria at a concentration of 500 μg as determined by agar well diffusion. SYP4 demonstrated the highest antibacterial activity as it completely inhibited all test strains, with inhibition zones ranging from 16.88 ± 0.25 to 21.25 ± 0.5 mm, and also yielded the highest MIC/MBC values against P. aeruginosa, B. cereus, and E. coli, at 256 and >256 μg/mL, respectively. We observed that at least 256 μg/mL of SYP4 is required to exhibit optimal antibacterial activity. At 16–128 μg/mL, it exhibited antibiofilm activity against S. aureus. Furthermore, at 256 μg/mL, SYP4 inhibited pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Although SYP2 and SYP3 displayed weak antibacterial activity and their MIC values could not be obtained for all bacteria, they showed strong QS inhibition in C. violaceum at 256 μg protein. Moreover, SYP2 and SYP3, at a minimum concentration of 32 μg/mL, significantly reduced violacein production. SYP3 also showed biofilm reduction activity on S. aureus at least 16-512 μg/mL.
Conclusion: Sangyod Phatthalung protein hydrolysates exerted excellent inhibitory effects against the growth of bacteria and their virulence factors, such as QS, biofilm formation, and/or pigment production. These factors include zoonotic and foodborne pathogens. Therefore, daily consumption of Sangyod Phatthalung rice might reduce the risk of bacterial pathogenesis and foodborne diseases. In conclusion, functional foods or alternate methods of treating bacterial illnesses may be developed in humans and animals.
Keywords: antibacterial peptide, foodborne pathogens, Phatthalung Sangyod rice, protein hydrolysate, zoonotic.
Funder
Walailak University
Publisher
Veterinary World
Subject
General Veterinary
Reference53 articles.
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