Molecular Characteristics of Indonesian Isolate Enterocytozoon hepatopenaei Based on Sequence Analysis of 18S rRNA Genes

Yohanes Kristiawan Artanto

Abstract


ABSTRACT

 

Enterocytozoon hepatopenaei (EHP) infection have been reported as an obstacle of whiteleg shrimp (Penaeus vannamei) culture in Indonesia. However, understanding of the molecular characteristics of EHP species in Indonesia is not widely known. The aims of this study were to determine the identity and characters of DNA, and their phylogeny of EHP species from several different locations in Indonesia with specific references to 18S rRNA gene. The EHPs were collected from cultured P.vannamei from Lampung, Pangandaran, Sidoarjo, Banyuwangi, Probolinggo, Blitar, Makassar, and Lombok. Thirteen (13) samples were randomly selected to explore their gene characters through 18S rRNA gene sequencing. The primers used were EHP_F and EHP_1R. Parameter observed were DNA sequencing, nucleotide sequence homology with related available genes in the Gen Bank database, multiple sequence alignment, and reconstruction of genetic relationship trees. DNA sequence homology analysis showed that all samples had 99.89-100% similarity to Indian Enterocytozoon hepatopenaei (Accession Number MH259890.1 and MH260592.1). The alignment results illustrated that all EHP sequences of Indonesian isolates were 100% identical each other. The phylogenetic tree topology provided information that all sample accessions were in the same clade and spread evenly. The conclusion were that the Indonesian EHP species were identical (100%) and it could be said that they were genetically homogeneous.

 

Keywords: Enterocytozoon hepatopenaei, 18S rRNA gene, Indonesia.


References


References

Campbell, N.A., J.B. Reece, L.G. Mitchell. 2010. Biologi. Edisi ke-8. Terj. Dari: Biology. 8th ed. oleh Manulu, W. Jakarta: Erlangga.

Efron, B. 1979. Bootstrap Methods: Another Look at the Jackknife. Ann. Statist. 7(1): 1 – 26.

Felsenstein, J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783-791.

Finkeldey R. 2005. Pengantar Genetika Hutan Tropis. Djamhuri E, Siregar IZ, Siregar UJ, Kertadikara AW, penerjemah. Bogor: Fakultas Kehutanan, Institut Pertanian Bogor.

Ha, N.T., H, D.T., Thuy, N.T., Lien, V.T.K. 2010. Enterocytozoon hepatopenaei parasitizing on tiger shrimp (Penaeus monodon) infected by white feces culture in Vietnam, has been detected (In Vietnamese with English abstract). Agriculture and Rural Development : Science and Technology (translation from Vietnamese), 12 : 45–50.

Hall, B.G. 2001. Phylogenetic trees made easy : A how-to manual for molecular biologists. Sinauer Associates, Inc., Sunderland.

Hillis, D.M., Marble and C. Moritz. 1996. Applications Of Molecular Systematics: The State Of The Field And A Look To The Future. In: Hillis, D.M.C. Moritz And B.K. Mable (Eds.). 1996. Molecular systematics. 2nd Ed. Sinauer Associates, Inc., Sunderland. pp. 515-543.

Kesavan, K., Mani, R., Toshiaki, I., Sudhakaran, R. 2016. Quick report on prevalence of shrimp microsporidian parasite Enterocytozoon hepatopenaei in India. Aquaculture Research, 1-5, doi: 10.1111/are.13078.

Kolondam, B.J., Lengkong, E., Mandang, J.P., Pinaria, P., Runtunuwu, S. 2012. Barcode DNA berdasarkan gen rbcL dan matK Anggrek Payus Limondok (Phius tancarvilleae), J. Bioslogos 2(2):1–8.

Nei, M and Kumar, S. 2000. Molecular Evolution and Phylogenetics. Oxford University Press, New York.

Osawa, S., S. Zhi-Hui and Y. Imura. 2004. Molecular Phylogeny and Evolution of Carabid Graound Beetles. Springer-Verlag Tokyo: SNP Best-set Typesetter Ltd.

Rajendran, K.V., Shivam, Saloni, Ezhil, Praveena. P., Joseph, Sahaya, Rajan, J., Sathish, Kumar. T., Avunje, Satheesha, Jagadeesan, V., Prasad, Babu S., Pande, Ashish, Navaneeth, Krishnan, A., Alavandi, S.V., Vijayan, K.K. 2016. Emergence of Enterocytozoon hepatopenaei (EHP) in farmed Penaeus (Litopenaeus) vannamei in India. Aquaculture 454: 272-280, doi: 10.1016/j.aquaculture.2015.12.034.

Schmitt. T. dan K. Haubrich. 2008. The Genetic Structure of the Mountain Forest Butterfly Erebia euryale Unravels the Late Pleistocene and Postglacial History of The Mountain Coniferous Forest Biome in Europe. Molecular ecology 17(9): 2194-2207 (doi: 10.1111/j.1365-294x.2007.03687.x).

Sritunyalucksana, K., Sanguanrut, P., Salachan, P.V., Thitamadee, S., Flegel, T.W. 2014. Urgent appeal to control spread of the shrimp microsporidian parasite Enterocytozoon hepatopenaei (EHP). Network of Aquaculture Centres in Asia-Pacific (NACA) http://www.enaca.org/ modules/news/article.php?article_id=2039.

Stackebrandt, E., Goebel, B.M. 1994. Taxonomic note: a place for DNA–DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol. 44, 846–849.

Suebsing, R., Prombun, P., Srisala, J., Kiatpathomchai, W. 2013. Loop-mediated isothermal amplification combined with colorimetric nanogold for detection of the microsporidian Enterocytozoon hepatopenaei in penaeid shrimp. Journal of Applied Microbiology 114 : 1254-1263.

Swofford, D.L., Olsen, G.J., Waddell, P.J. and Hillis, D.M. 1996. Phylogenetic Inference. In: Hillis, D.M., Moritz, C. and Mable, B.K., Eds., Molecular Systematics, 2nd Edition, Sinauer Associates, Sunderland (MA), 407-514.

Tang, K.F.J., Aranguren, L.F., Piamsomboon, P., Han, J.E., Maskaykina, I.Y., and Schmidt, M.M. 2017. Detection of the microsporidian Enterocytozoon hepatopenaei (EHP) and Taura syndrome virus in Penaeus vannamei cultured in Venezuela. Aquaculture, 480, 17-21.

Tang, K.F.J., Han, J.E., Aranguren, L.F., White-Noble, B., Schmidt, M.M., Piamsomboon, P., Risdiana, E., Hanggono, B. 2016. Dense populations of the microsporidian Enterocytozoon hepatopenaei (EHP) in feces of Penaeus vannamei exhibiting white feces syndrome and pathways of their transmission to healthy shrimp. Journal of Invertebrate Pathology 140, 1-7, doi: 10.1016/j.jip.2016.08.004.

Tang, K.F.J., Pantoja, C.R., Redman, R.M., Han, J.E., Tran, L.H., Lightner, D.V. 2015. Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. Journal of Invertebrate Pathology 130 : 37-41, doi: 10.1016/j.jip.2015.06.009.

Tangprasittipap, A., Srisala, J., Chouwdee, S., Somboon, M., Chuchird, N., Limsuwan, C., Srisuvan, T., Flegel, T.W., Sritunyalucksana, K. 2013. The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei. BMC Veterinary Research 9, 139-148.

Thitamadee, S., Prachumwat, A., Srisala, J., Jaroenlak, P., Salachanb, P.V., Sritunyalucksana, K., Flegel, T.W., Itsathitphaisarn, O. 2016. Review of current disease threats for cultivated penaeid shrimp in Asia. Aquaculture 452 : 69–87.

Tourtip, S., Wongtripop, S., Stentiford, G.D., Bateman, K.S., Sriurairatana, S., Chavadej, J., Sritunyalucksana, K., Withyachumnarnkul, B. 2009. Enterocytozoon hepatopenaei sp.nov. (Microsporida : Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae) : Fine structure and phylogenetic relationships. Journal of Invertebrate Pathology 102 : 21–29.




DOI: http://dx.doi.org/10.20884/1.oa.2019.15.1.694

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