Antibacterial Activity and Preliminary Protein Profile of Rock Oyster (Saccostrea cuccullata) Hemolymph

Main Article Content

Thipphawan Chothonglang
Sonthaya Phuynoi
Suriyan Tunkijjanukij
Attawut Khantavong
Teerasak E-kobon
Jintana Salaenoi


Oysters have gained popularity among consumers worldwide. They are commonly consumed either fresh (with or without lime and herbs) or cooked. Many consumers believe that oyster hemolymph has the ability to stop growth of pathogens and inhibit infection. The aims of this study were to examine the bacterial inhibition of rock oyster Saccostrea cuccullata hemolymph, and study protein alteration in four treatments: fresh hemolymph (FH), hemolymph boiled at 100 °C for 15 s (H1), hemolymph boiled at 100 °C for 5 min (H2), and fresh hemolymph combined with lime juice (FL). Protein patterns were studied by SDS-PAGE technique. The results showed that the FL treatment produced measurable inhibition of three bacterial species: Vibrio parahaemolyticus (AHPND), Escherichia coli (ATCC25922), and Staphylococcus aureus (ATCC25923). No antimicrobial activity was found in treatments FH, H1 or H2. The SDS polyacrylamide gels showed that acidity and heat altered protein band patterns of the oyster hemolymph. FH and H1 groups revealed 17 protein bands, while H2 and FL groups presented 10 and 11 protein bands, respectively. The results suggest that consuming oyster with lime juice could have an inhibitory effect on the growth of pathogenic bacteria compared with fresh and cooked ones.

Article Details

How to Cite
Chothonglang, T. ., Phuynoi, S. ., Tunkijjanukij, S. ., Khantavong, A. ., E-kobon, T. ., & Salaenoi, J. . (2023). Antibacterial Activity and Preliminary Protein Profile of Rock Oyster (Saccostrea cuccullata) Hemolymph. Journal of Fisheries and Environment, 47(1), 53–62. Retrieved from
Research Article


Bax, M., L. Aubry, C. Ferreira, J. Daudin, P. Gatellier, D. Rémond and V. Santé-Lhoutellier. 2012. Cooking temperature is a key determinant of in vitro meat protein digestion rate: Investigation of underlying mechanisms. Journal of Agricultural and Food Chemistry 60(10): 2569–2576.

Cai, B., J. Pan, Y. Wu, P. Wan and H. Sun. 2013. Immune functional impacts of oyster peptide-based enteral nutrition formula (OPENF) on mice: A pilot study. Chinese Journal of Oceanology and Limnology 31(4): 813–820.

Carlson-Bremer, D., T.M. Norton, F.J. Sanders, B. Winn, M. Spinks, B.A. Glatt, L. Mazzaro, P. Jodice, T.C. Chen and E.S. Dierenfeld. 2014. Circulating fat-soluble vitamin concentrations and nutrient composition of aquatic prey eaten by American Oystercatchers (Haematopus palliatus palliatus) in the Southeastern United States. Journal of Avian Medicine and Surgery 28(3): 216–224.

Chothonglang, T. 2019. Effects of temperature and lime juice on blood cockle (Anadara granosa) and oyster (Saccostrea commercialis) hemolymph proteomes and antibacterial activity. Master Thesis, Kasetsart University, Bangkok, Thailand. 109 pp.

Drake, S.L., D. Elhanafi, W. Bang, M.A. Drake, D.P. Green and L.A. Jaykus. 2006. Validation of a green fluorescent protein-labeled strain of Vibrio vulnificus for use in the evaluation of postharvest strategies for handling of raw oysters. Applied and Environmental Microbiology 72(11): 7205–7211.

Dupont, S., A. Lokmer, E. Corre, J.C. Auguet, B. Petton, E. Toulza, C. Montagnani, G. Tanguy, D. Pecqueur, C. Salmeron, L. Guillou, C. Desnues, B. La Scola, J. Bou Khalil, J. de Lorgeril, G. Mitta, Y. Gueguen and J.M. Escoubas. 2020. Oyster hemolymph is a complex and dynamic ecosystem hosting bacteria, protists and viruses. Animal Microbiome 2(1): 12. DOI: 10.1186/s42523-020-00032-w.

Enejoh, O.S., I.O. Ogunyemi, M.S. Bala, I.S. Oruene, M.M. Suleiman and S.F. Ambali. 2015. Ethnomedical importance of Citrus Aurantifolia (Christm) swingle. The Pharma Innovation Journal 4(8): 1–6.

Froelich, B.A. and R.T. Noble. 2016. Vibrio bacteria in raw oysters: managing risks to human health. Philosophical Transactions of the Royal Society B 371: 20150209. DOI: 10.1098/rstb.2015.0209.

Hardoko, H. and F. Yuliana. 2014. Stability study of antibacterial activity of mixed lime juice and honey of heating temperature on Staphylococcus aureus and Streptococcus pyogenes. International Journal of Pure and Applied Sciences and Technology. 21(2): 1-7.

Jayana, B.L., T. Prasai, A. Singh and K.D. Yami. 2010. Study of antimicrobacterial activity of lime juice against Vibrio cholera. Scientific World 8: 8 DOI: 10.3126/sw.v8i8.3847.

Kohn, M., T. Farley, T. Ando, M. Curtis, S.A. Wilson, Q. Jin, S.S. Monroe, R.C. Baron, L.M. McFarland and R.I. Glass. 1995. An outbreak of Norwalk virus gastroenteritis associated with eating raw oysters: Implications for maintaining safe oyster beds. JAMA 273(6): 466–471.

Morse, D.L., J.J. Guzewich, J.P. Hanrahan, R. Stricof, M. Shayegani, R. Deibel, J.C. Grabau, N.A. Nowak, J.E. Herrmann, G. Cukor and N.R. Blacklow. 1986. Widespread outbreaks of clam- and oyster-associated gastroenteritis. North England Journal of Medicine (USA) 314(11): 678–681.

Nuchchanart, W., S. Poompuang, S. Tunkijjanukij and S. Nakasathien. 2007. Characterization of antibacterial protein from hemolymph of oyster Crassostrea belcheri. Kasetsart Journal-Natural Science 41(1): 169–179.

Oiken, E.I., E.S. Omoregie, F.E. Oviasogie and K. Oriakhi. 2016. Phytochemical, antimicrobial, and antioxidant activities of different citrus juice concentrates. Food Science and Nutrition 4(1): 103–109.

Okeke, M.I., A.S. Okoli and C.U. Iroegbu. 2015. Antibacterial activity of Citrus limonum fruit juice extract. Pakistan Journal of Pharmaceutical Sciences 28(5): 1567–1571.

Palka, K. and H. Daun. 1999. Changes in texture, cooking losses, and myofibrillar structure of bovine M. semitendinosus during heating. Meat Science 51: 237–243.

Pathirana, S., S.H.M.P. Wimalasena, C.J.D.S. Benthotage and S. Hossain. 2018. Antibacterial activity of lime (Citrus aurantifolia) essential oil and limonene against fish pathogenic bacteria isolated from cultured olive flounder (Paralichthys olivaceus). Archives of Polish Fisheries 26(2): 131–139.

Pila, E., J.T. Sullivan, X.Z. Wu, J. Fang, S.P. Rudko, M.A. Gordy and P.C. Hanington. 2016. Haematopoiesis in molluscs: a review of haemocyte development and function in gastropods, cephalopods and bivalves. Developmental and Comparative Immunology 58: 119–128.

Prangtong, O. and S. Tunkijjanukij. 2004. Isolation of Antibacterial Substance from the Haemolymphs of Crassostrea belcheri (Sowerby, 1871) and Crassostrea iredalei (Faustino, 1932). Kamphaengsaen Academic Journal 2(3): 126–136.

Qin, X., X. Fan, L. Zhang, H. Zheng, C. Zhang and J. Yuan. 2018. Extraction, purification, and structure characterization of polysaccharides from Crassostrea rivularis. Food Science and Nutrition Published 6: 1621–1628.

Schmitt, P., M. Duperthuy, C. Montagnani, E. Bachère and D. Destoumieux-Garzón. 2011. Immune responses in the Pacific oyster Crassostrea gigas: An overview with focus on summer mortalities. In: Oysters: Physiology, Ecological Distribution and Mortality (ed. J.G. Qin), pp. 227–273. Nova Science Publishers, New York, USA.

Scotti, P.D., S.C. Dearing and D.R. Greenwood. 2007. Characterisation of cavortin, the major haemolymph protein of the Pacific oyster (Crassostrea gigas), New Zealand Journal of Marine and Freshwater Research 41(1): 91–101.

Shakya, A., L. Bhawana, K. Pragati, D. Ritu, D. Puspa and C. Richa. 2019. Comparative study of antibacterial activity of juice and peel extract of citrus fruits. Tribhuvan University Journal of Microbiology 6: 82–88.

Singh, T.K., S.K Øiseth, L. Lundina and L. Day. 2014. Influence of heat and shear induced protein aggregation on the in vitro digestion rate of whey proteins. Food and Function Journal 5: 2686–2698.

Tomotake, H., T. Koga, M. Yamato, A. Kassu and F. Ota. 2006. Antibacterial activity of citrus fruit juices against Vibrio species. Journal of Nutritional Science and Vitaminology (Tokyo) 52(2): 157–160.

Watanachote, J., N. Khongchareonporn, N. Littirut and S. Leethochavalit. 2013. Parasites and immunity of marine shellfishes along the East Coast of Thailand. Burapha University, Chonburi, Thailand. 73 pp.

Xue, Q., L.K. Schey, K.A. Volety, E.F. Chu and L.F.J. Peyre. 2004. Purification and characterization of lysozyme from plasma of the eastern oyster (Crassostrea virginica). Comparative Biochemistry and Physiology 139: 11–25.

Xue, Q., J. Gauthier, K. Schey, Y. Li, R. Cooper, R. Anderson and J.L. Peyre. 2012. Identification of a novel metal binding protein, segon, in plasma of the eastern oyster, Crassostrea virginica. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 163(1): 74–85.

Ziegler, G., K. Paynter and D. Fisher. 2002. Maxtrix metalloproteinase-link activity from hemocytes of the eastern oyster (Crassostrea virginica). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 131(3): 361–337.