Anti Perkinsus olseni Monoclonal Antibody Generation Using Hypnospores as Antigens
Article Sidebar
Main Article Content
Abstract
Antibodies are valuable for the detection, identification, and diagnosis of pathogens in many fields, including aquaculture. In this study, we aimed to produce monoclonal antibodies (mAbs) against the parasite Perkinsus olseni, isolated from the clam Paphia undulata in Thailand. The mAbs were characterized using dot blotting and immunohistochemistry techniques in order to identify their class and their ability to detect the hypnospores and zoospores of P. olseni. Immunoreactions between the obtained mAbs and P. olseni produced black or grey spots according to the concentration of parasites detected by the mAbs. The quantitative detection thresholds of hypnospores for the two most strongly reactive mAbs were 105 cells/mL (producing grey spots) to 107 cells/mL (producing black spots), and more than 107 cells/mL (grey spots) for zoospores. Both mAbs showed immunoreactivity at the surface of trophozoite cells of P. olseni in infected clam tissues as assessed by immunohistochemistry analysis, indicating that they could be used for the detection and identification of P. olseni infection in commercial Pa. undulata production.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Copyright Transfer Statement
The copyright of this article is transferred to Current Applied Science and Technology journal with effect if and when the article is accepted for publication. The copyright transfer covers the exclusive right to reproduce and distribute the article, including reprints, translations, photographic reproductions, electronic form (offline, online) or any other reproductions of similar nature.
The author warrants that this contribution is original and that he/she has full power to make this grant. The author signs for and accepts responsibility for releasing this material on behalf of any and all co-authors.
Here is the link for download: Copyright transfer form.pdf
References
Choi, K.S. and Park, K.I., 2010. Review on protozoan parasite Perkinsus olseni (Lester and Davis 1981) infection in Asian waters. In: A. Ishimatsu and H.J. Lie, eds. Coastal Environmental and Ecosystem Issues of the East China Sea. Nagasaki: Terrapub and Nagasaki University, pp. 269-281.
Smolowitz, R., 2013. A review of current state of knowledge concerning Perkinsus marinus effects on Crassostrea virginica (Gmelin) (the Eastern Oyster). Veterinary Pathology, 50, 404-411, https://doi.org/10.1177/0300985813480806.
Ramilo, A. Pintado, J., Villalba, A. and Abollo, E., 2016. Perkinsus olseni and P. chesapeaki detected in a survey of perkinsosis of various clam species in Galicia (NW Spain) using PCR-DGGE as a screening tool. Journal of Invertebrate Pathology, 133, 50-58, https://doi.org/10.1016/j.jip.2015.11.012.
Park, K-.I., Ngo, T.T.T., Choi, S-.D., Cho, M. and Choi, K.-S., 2006. Occurrence of Perkinsus olseni in the Venus clam Protothaca jedoensis in Korean water. Journal of Invertebrate Pathology, 93, 81-87, https://doi.org/10.1016/j.jip.2006.04.007.
Waki, T., Takahashi, M., Eki, T., Hiasa, M., Umeda, K., Karakawa, N. and Yoshinaga, T., 2018. Impactof Perkinsus olseni infection on a wild population of Manila clam Ruditapes philippinarum in Ariake Bay, Japan. Journal of Invertebrate Pathology, 153, 134-144, https://doi.org/10.1016/j.jip.2018.03.001.
Leethochavalit, S., Chalermwat, K., Upatham, E.S., Choi, K.-S., Sawangwong, P. and Kruatrachue, M., 2004. Occurrence of Perkinsus sp. in undulate surf clams Paphia undulata from the Gulf of Thailand. Diseases of Aquatic Organisms, 60, 165-171, https://doi.org/10.3354/dao060165.
Manual of Diagnostic Tests for Aquatic Animals, 2019. Infection with Perkinsus marinus. [online] Avaliable at: https://www.woah.org/fileadmin/Home/eng/Health_standards/aahm/ current/chapitre_perkinsus_marinus.pdf.
Manual of Diagnostic Tests for Aquatic Animals, 2017. Infection with Perkinsus olseni. [online] Available at: http://www.animalhealthsurveillance.agriculture.gov.ie/individualdis easelistings/infectionwithperkinsusolseni/.
Villalba, A., Reece, K.S., Ordás, M.C., Casas, S.M. and Figueras, A., 2004. Perkinsosis in molluscs: A review. Aquatic Living Resources, 17, 411-432, https://doi.org/10.1051/alr:2004050.
Park, K.-I., Yang, H.-S., Kang, H.-S., Cho, M., Park, K.-J. and Choi, K.-S., 2010. Isolation and identification of Perkinsus olseni from feces and marine sediment using immunological and molecular techniques. Journal of Invertebrate Pathology, 105, 216-269, https://doi.org/10.1016/j.jip.2010.07.006.
Ruano, F., Batista, F.M. and Arcangeli, G., 2015. Perkinsosis in the clams Ruditapes decussatus and R. philippinarum in the Northeastern Atlantis and Mediterranean Sea: A review. Journal of Invertebrate Pathology, 131, 58-67, https://doi.org/10.1016/j.jip.2015.07.015.
Eto, S.F., Fernandes, D.C., Moraes, A.C., Prado, J.R., Baldassi, A.C., Manrique, W.G., Silva, I.C., Medeiros, A.S.R., Belo, M.A.A., Balbuena, T.S., Samara, S.I. and Pizauro, J.M., 2018. Validation of IgY for the diagnosis of Streptococcus agalactiae-caused endocarditis and bacterial meningitis in Nile tilapia (Oreochromis niloticus). Fish and Shellfish Immunology, 76, 153-160, https://doi.org/10.1016/j.fsi.2018.02.048.
Kumar, G., Rathore, G., Sengupta, U., Kapoor, D. and Lakra, W.S., 2010. Production of monoclonal antibodies specific to major outer membrane protein of Edwardsiella tarda. Comparative Immunology, Microbiology, and Infectious Diseases, 33, 133-14, https://doi.org/10.1016/j.cimid.2008.08.008.
Sithigorngul, P., Rukpratanporn, S., Longyant, S., Chaivisuthangkura, P., Sithigorngul, W. and Menasveta, P., 2002. Monoclonal antibodies specific to yellow head virus (YHV) of Penaeus monodon. Diseases of Aquatic Organisms, 49, 71-76, https://doi.org/10.3354/dao049071.
Wangman, P., Senapin, S., Chaivisuthangkura, P., Longyant, S., Rukpratanporn, S. and Sithigorngul, P., 2012. Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein. Diseases of Aquatic Organisms, 98, 121-131, https://doi.org/10.3354/dao02431.
Choi, K.S., Lewis D.H., Powell, E.C., Frelier, P.F. and Ray, S.M., 1991. A Polyclonal antibody developed from Perkinsus marinus hypnospores fails to cross react with other life stages of Perkinsus marinus in oyster (Crassostrea virginica) tissue. Journal of Shellfish Research, 10, 411-415.
Dungan, C.F. and Roberson, B.S., 1993. Binding specificities of mono-and polyclonal antibodies to the protozoan oyster pathogen Perkinsus marinus. Diseases of Aquatic Organisms, 15, 9-22, https://doi.org/10.3354/dao015009.
Earnhart, C.G., Gauthier, D.T., Vogelbein, W.K. and Kaattari, S.L., 2005. Monoclonal antibody analysis of Perkinsus marinus extracellular products. International Journal for Parasitology, 35, 171-184, https://doi.org/10.1016/j.ijpara.2004.10.025.
Romestand, B., Torreilles, J. and Roch, P., 2001. Production of monoclonal antibodies against the protozoa, Perkinsus marinus: estimation of parasite multiplication in vitro. Aquatic Living Resources, 14, 351-357, https://doi.org/10.1016/S0990-7440(01)01140-8.
Kaewsalubnin, S., Leethochavalit, S., Watanachote, J., Komolpis, K. and Khongchareonporn, N., 2015. Production and characterization of monoclonal antibody aganist Perkinsus olseni in undulated surf clams Paphia undulata. Food and Applied Bioscience Journal, 3, 231-238, https://doi.org/10.14456/fabj.2015.22.
Ellin, R. and Bushek, D., 2006. Adaptation of Ray’s fluid thioglycollate medium assay to detect quantify planktonic stages of Perkinsus spp. parasites. Journal of Shellfish Research, 25(3), 1037-1042, https://doi.org/10.2983/0730-8000(2006)25[1037:AORFTM]2.0.CO;2.
Ray, S.M., 1952. A culture technique for the diagnosis of infections with Dermocystidium marinum Mackin, Owen, and Collier in oysters. Science, 116, 360-361, https://doi.org/10.1126/science.116.3014.360.
Audemard, C., Carnegie, R.B. and Burreson, E.M., 2008. Shellfish tissues evaluated for Perkinsus spp. using the Ray's fluid thioglycollate medium culture assay can be used for downstream molecular assays. Diseases of Aquatic Organisms, 80, 235-239, https://doi.org/10.3354/dao01944.
Almeida, M., Berthe, F., Thebault, A. and Dinis, M.T., 1999. Whole clam culture as a quantitative diagnostic procedure of Perkinsus atlanticus (Apicomplexa, Perkinsea) in clams Ruditapes decussatus. Aquaculture, 177(1-4), 325-332, https://doi.org/10.1016/s0044-8486(99)00095-2.