Aim: The objective of this research was to determine the species and strains of Leucocytozoon caulleryi and study the phylogenetics of L. caulleryi of broiler chickens in endemic areas in Indonesia.

Materials and Methods: Blood samples were collected from broiler chickens originated from endemic area in Indonesia, i.e., Pasuruan, Lamongan, Blitar, Lumajang, Boyolali, Purwokerto, and Banjarmasin in 2017. Collected blood was used for microscopic examination, sequencing using BLAST method to identify the nucleotide structure of cytochrome b (cyt b) gene that determines the species, and the phylogenetics analysis of L. caulleryi that infected broiler chickens in endemic areas in Indonesia, using Mega 5 software.

Results: The results showed that Plasmodium sp. and L. caulleryi were infected broiler chickens in endemic areas in Indonesia. L. caulleryi in one area had very close phylogenetic relations with those in other areas. The genetic distance between L. caulleryi taxa from various endemic areas is very close (<5%).

Conclusion: There is a very close phylogenetics among strains of L. caulleryi that infected broiler chickens in various endemic areas in Indonesia.

Keywords: broiler chickens, cytochrome b gene, endemic, Indonesia, Leucocytozoon caulleryi.

1. Zhao, W., Liu, J., Xu, R., Zhang, C., Pang, Q., Chen, X., Liu, S., Hong, L., Yuan, J., Li, X., Chen, Y., Li, J. and Su, Z. (2015) The gametocytes of Leucocytozoon sabrazesi infect chicken thrombocytes, not other blood cells. PLos ONE, 10(7): 1-17. [Crossref]

2. Murdock, C.C., Foufopoulos, J. and Simon, C.P. (2013) A transmission model for the ecology of an avian blood parasite in a temperate ecosystem. PLos ONE, 8(9): 1-14. [Crossref]

3. Purwanto B. (2009) If the Chicken Got Malaria. Trobos Magazine, July Edition.

4. Choupina, A.B. and Martin, I.M. (2014) Molecular marker for genetic diversity, gene flow and genetic population structure of freshwater mussel species. Braz. J. Biol., 74(3): 167-170. [Crossref] [PubMed]

5. Valkiunas, G., Sehgal, R.N.M., Iezhova, T.A. and Hull, H.C. (2010) Identification of Leucocytozoon toddy group (Haemosporida: Leucocytizoidae), with remark on the species taxonomy of leucocytozoids. J. Parasitol., 96(1): 170-177. [Crossref] [PubMed]

6. Dimitrov, K.M., Lee, D.H., Williams-Coplin, D., Olivier, T.L. and Miller, P.J. (2016) Newcastle disease causing recent outbreaks worldwide show unexpectedly high genetic similarity with historical virulent isolates from the 1940s. J. Clin. Microbiol., 54(5): 1228-1235. [Crossref] [PubMed] [PMC]

7. Schaer, J., Perkin, S.L., Decher, J., Leendertz, F.H., Fahr, J. and Weber, N. (2013) High diversity of west african bat malaria parasites and a tight link with rodent Plasmodium taxa. Proc. Natl. Acad. Sci., 10(43): 17451-17419. [Crossref]

8. Zagalska-Neubauer, M. and Bensch, S. (2016) High prevalence of Leucocytozoon parasites in fresh water breeding gulls. J. Ornithol., 157: 525-532. [Crossref]

9. Xu, S., Schaack, S., Seyfert, A., Choi, E., Lynch, M. and Cristescu, M.E. (2012) High mutation rates in the mitochondrial genom of Daphnia pulex. Mol. Biol. Evol., 29(2): 763-769. [Crossref] [PubMed] [PMC]

10. Amaral, M.R.X., Albrecht, M., McKinley, A.S., Ferreira, C.A.M., Cavalcante, S.J.S. and Diniz, F.M. (2015) Mitochondrial DNA variation reveals a sharp genetic break within the distribution of the blue land crab Cardisoma guanhumi in the western central Atlantic. Molecules, 20: 15158-15174. [Crossref] [PubMed]

11. Silva-Neto, A.A., Ferreira, P.B., Torres, R.A., Texeira, R.H.F., Duarte, D.M.B., Barbosa, A.C., Vargas, R.C. and Garcia, C.E. (2016) Diagnostic Cytochrome b gene profiles for the identification of Paca (Cuniculus paca) bushmeat: Implications for the monitoring of illegal hunting and wildlife trade. Braz. J. Biol., 76(1): 55-58. [Crossref]

12. Hellgren, O.J., Waldenstrom, J. and Bensch, S. (2004) A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. J. Parasitol., 90: 797-802. [Crossref] [PubMed]

13. Omori, S., Sato, Y., Hirakawa, S., Isobe, T., Yukawa, M. and Murata, K. (2008) Two extra chromosomal genomes of Leucocytozoon caulleryi: Complete nucleotide sequences of the mitochondrial genome and existence of the apicoplast genome. Parasitol. Res., 103: 953-957. [Crossref]

14. Tamura, K., Dudley, J., Nei, M. and Kumar, S. (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol., 24: 1596-1599. [Crossref] [PubMed]

15. Nei, M. and Kumar, S. (2000) Molecular Evolution and Phylogenetic. Oxford University Press, New York. p231.

16. Suprihati, E. (2013) Phylogenetic Analysis of the Cytochrom b Gene of Leucocytozoon sp. in Domestic Chicken in Endemic Areas of Indonesia. Doctoral Dissertation, Universitas Airlangga.

17. Ishtiaq, F., Gering, E., Rappole, J.H., Rahmani, A.R., Jhala, Y.V., Dove, C.J., Milensky, C., Olson, S.L., Peirce, M.A. and Fleischer, R.C. (2007) Prevalence and diversity of avian hematozoan parasites in Asia: A regional survey. J. Wildl Dis., (43): 382-398. [Crossref] [PubMed]

18. Mahajan, B., Noiva, R., Yadava, A., Zheng, H., Majam,V., Mohan, K.V.K., Haynes, J.D., Nakhasi, H. and Kumar, S. (2006) Protein disulfide isomerase assisted protein folding in malaria parasites. Int. J. Parasitol., (36): 1037-1048. [Crossref] [PubMed]