Reconsideration of phenoloxidase activity determination in white shrimp Litopenaeus vannamei |
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| Authors: | Junwa Huang Yang Yang Anli Wang |
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| Affiliation: | 1. Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-901, Brazil;2. Instituto Federal da Paraíba, Cabedelo, Paraíba, 58.015-020, Brazil;3. Instituto Agronômico de Pernambuco, Recife, Pernambuco, 50761-000, Brazil;4. Departamento de Pesca e Aquicultura, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, 52171-900, Brazil;5. Universidade Federal Rural de Pernambuco, Unidade Acadêmica de Serra Talhada, Pernambuco, 55608-680, Brazil;1. School of Ocean Science and Technology, Kerala University of Fisheries and Ocean Studies, Panangad, Kerala, India;2. Department of Processing Technology (Biochemistry), Kerala University of Fisheries and Ocean Studies, Panangad, Kerala, India;1. Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phyathai Road, Bangkok 10330, Thailand;2. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Paholyothin Road, Klong1, Klong Luang, Pathumthani 12120, Thailand;1. Laboratory of Immunology Applied to Aquaculture, Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, 88040-900 Florianópolis, SC, Brazil;2. Laboratory of Marine Shrimp, Department of Aquaculture, Federal University of Santa Catarina, 88040-900 Florianópolis, SC, Brazil |
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| Abstract: | ![]()
Though phenoloxidase (PO) activity has been used as an important index in immunological research of crustaceans, methods for the determination of PO activity are not consistent even for the same species. Plasma, the major location of PO activity, should be the most reasonable sample, instead of hemocytes or serum, for the determination of PO activity of shrimp. The current study provided a thorough characterization and reconsideration for PO activity assay in the plasma of Litopenaeus vannamei. Results show that the final concentration of l-dihydroxyphenylalanine (l-DOPA) for PO activity assay should be no less than 1.5 mg ml?1, and pH 6.6 should be used to maintain the stability of l-DOPA solution. This study provides direct evidence that PO activity is significantly inhibited by EDTA, and it is suggested to use EDTA-free anticoagulant in separating plasma for PO activity assay in future studies. Repeated measurements indicated that the assayed PO activities are significantly affected by preservation conditions, and plasma is quite unstable with spontaneous activation when put in ice or stored at ?20 °C. Thus samples need to be measured immediately or preserved at ?80 °C with assay as soon as possible after it is thawed, and should not be preserved for a second time for measuring PO activity. |
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