The phylogenetic analysis of RNA-dependent RNA polymerase gene of chronic bee paralysis virus in Turkey

honey bee chronic bee paralysis virus RT-PCR phylogenic analysis Turkey

Chronic bee paralysis virus (CBPV) is a major contributor to global bee colony losses. However, among the RNA viruses infecting honeybees, CBPV is still not classified. In this study, samples of asymptomatic adult honey bees were randomly collected from 45 apiaries in three provinces (Burdur, Antalya, and Isparta) in Turkey. Five of the 45 samples were determined to be positive by reverse transcription polymerase chain reaction (RT-PCR). In addition, three positive samples from Burdur, Isparta, and Antalya were confirmed by sequencing. We constructed a phylogenetic tree that was divided into five main groups based on the RNA-dependent RNA polymerase gene region. The Turkish strains TUR/BUR/CBPV1, TUR/ISPT/CBPV2, and TUR/ANT/CBPV3 and the Turkish strains from obtained previous studies formed a different cluster. The sequence homology results of a phylogenetic analysis showed that the Turkish strains shared 97-98.9% of their nucleotide identity and had 96.9-99% similarity rates with each other. The strains obtained in this study and the Turkish strains detected in previous studies were also 81-84.6% similar to European, Chinese, and Uruguay strains. This research underlines the presence of CBPV in apparently healthy Turkish bee colonies and the remarkable differences in Turkish CBPV strains. Further investigation is needed to identify the molecular characterization, complete genome sequence, and pathogenesis of Turkish CBPV strains.

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Antunez K, Alessandro BD, Corbella E, Zunino, P (2005) Detection of chronic bee paralysis virus and acute bee paralysis virus in Uruguayan honeybees. J Invertebr Pathol. 90: 69-72.
Bailey L, Gibbs AJ, Woods RD (1968) The purification and properties of chronic bee-paralysis virus. J Gen Virol 2: 251-260.
Bailey L, Ball BV, Perry JN (1983) Honeybee paralysis: its natural spread and its diminished incidence in England and Wales. J Apic Res 22:191-195.
Bailey L, Gibbs AJ, Woods RD (1963) Two viruses from adult honey bees (Apis mellifera Linnaeus). Virology 21: 390-395.
Ball BV (1999) Paralysis In: Bee disease diagnosis. Options Mediteranneennes: pp 81-89.
Berenyi O, Bakonyi T, Derakhshifar I, Koeglberger H, Nowotny N (2006) Occurrence of six honeybee viruses in diseased austrian apiaries. Appl Environ Microbiol. 72: 2414-2420.
Blanchard P, Olivier V, Iscache AL, Celle O, Schurr F, Lallemand P, Ribière M (2007a) Improvement of RT-PCR detection of chronic bee paralysis virus (CBPV) required by the description of genomic variability in French CBPV isolates. J Invertebr Pathol 97: 182-185.
Blanchard P, Ribière M, Celle O, Lallemand P, Schurr F, Olivier V, Iscache AL, Faucon JP (2007b) Evaluation of a real-time two-step RTPCR assay for quantitation of Chronic bee paralysis virus (CBPV) genome in experimentally-infected bee tissues and in life stages of a symptomatic colony. J Virol Methods 141(1): 7-13. doi: 10.1016/j.jviromet.2006.11.021.
Blanchard P, Schurr F, Olivier V, Celle O, Antùnez K, Bakonyi T, Berthoud H, Haubruge E, Higes M, Kasprzak S, Koeglberger H, Kryger P, Thiéry R, Ribière M (2009) Phylogenetic analysis of the RNA-dependent RNA polymerase (RdRp) and a predicted structural protein (pSP) of the Chronic bee paralysis virus (CBPV) isolated from various geographic regions. Virus Res 144(1-2): 334-8. doi: 10.1016/j.virusres.2009.04.025.
Brutscher LM, McMenamin AJ, Flenniken ML (2016) The buzz about honey bee viruses. PLoS Pathog 12: e1005757 4.
Cagirgan AA, Yazici Z (2020) Development of a multiplex RT-PCR assay for the routine detection of sevenRNA viruses in Apis mellifera. JVirol Methods 281: 113858. doi:10.1016/j.jviromet.2020.113858
Cagirgan AA, Yildirim Y, Usta A (2020) The investigation of Israil acute bee paralysis virus, sacbrood virus, kashmir bee virus and chronic bee paralysis virus in honeybees (Apis mellifera). Eurasian J Vet Sci 36:2, 96-10. doi:10.15312/EurasianJVetSci.2020.265
Cagirgan, AA (2018) Ege bölgesinde virus nedenli arı hastalıklarının multipleks polimeraz zincir reaksiyonu ile araştırılması (Investigation of virus-induced bee diseases in Aegean Region with multiplex polymerase chain reaction).
tezSorguSonucYeni.jsp (accessed 9 September 2020).
Gumusova Okur S, Albayrak H, Kurt M, Yazici Z (2010) Prevalence of three honey bee viruses in Turkey. Vet Arhiv 80 (6): 779-785.
Celli G, Maccagnani B (2003) Honey bees as bioindicators of environmental pollution. Bull Insectology 56:137-139.
Chauzat MP, Cauquil L, Roy L, Franco S, Hendrikx P, Ribière-Chabert M (2013) Demographics of the European apicultural industry. PLoS One 13: 8(11): e79018. doi: 10.1371/journal.pone.0079018.
Chen YP, Siede R (2007) Honey Bee Viruses. Adv Virus Res 70: 33-80.
Chevin A, Coutard B, Blanchard P, Dabert-Gay AS, Ribière-Chabert M, Thiéry R (2015) Characterisation of structural proteins from chronic bee paralysis virus (CBPV) using mass spectrometry. Viruses 23:7(6): 3329-44. doi: 10.3390/v7062774.
Cornman RS, Tarpy DR, Chen Y, Jeffreys L, Lopez D, Pettis JS, vanEngelsdorp D, Evans JD (2012) Pathogen webs in collapsing honey bee colonies. PLoS One 2012: 7(8): e43562. doi: 10.1371/journal.pone.0043562.
Dittes J, Schäfer MO, Aupperle-Lellbach H, Mülling CKW, Emmerich IU (2020) Overt Infection with Chronic Bee Paralysis Virus (CBPV) in Two Honey Bee Colonies. Vet Sci22: 7(3):142.
Food and Agriculture Organization Corporate Statistical Database (FAOSTAT), 2018. Data of Turkey beehives in 2018. (accessed 03 August 2020).
Forgach P, Bakonyi T, Tapaszti Z, Nowotny N, Rusvai M (2008) Prevalence of pathogenic bee viruses in Hungarian apiaries: situation before joining the European Union. J Invertebr Pathol. 98: 235-238.
Gajger IT, Kolodziejek J, Bakonyi T, Nowotny N (2014) Prevalence and distribution patterns of seven different honeybee viruses in diseased colonies: a case study from Croatia. Apidologie 45: 701-706.
Ghorani M, Ghalyanchi Langeroudi A, Madadgar O, Rezapanah M, Nabian S, Khaltabadi Farahani R, Maghsoudloo H, Forsi M, Abdollahi H, Akbarein H (2017) Molecular identification and phylogenetic analysis of chronic bee paralysis virus in Iran. Vet Res Forum 8(4):287-292.
Gisder S, Genersch E (2016) Viruses of commercialized insect pollinators. J Invertebr Pathol doi:10.1016/j.jip.2016.07.010 5.
Kalayci G, Cagirgan AA, Kaplan M, Pekmez K, Beyazit A, Ozkan B, Yesiloz B, Arslan F (2020) The role of viral and parasitic pathogens affected by colony losses in Turkish apiaries. Kafkas Univ Vet Fak Derg 26(5): 671-677. doi: 10.9775/kvfd.2020.24154
Kalayci G, Cagirgan AA, Pekmez K, Ozkan B, Kaplan M (2019) Molecular detection and phylogenetic analysis of the honeybee (Apis mellifera) sacbroodvirus in Turkey. Turk J Vet Anim Sci 43: 551-554.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35: 1547-1549.
Li BB, Deng S, Yang DH, Hou CS, Diao QY (2017) Complete sequences of the RNA 1 and RNA 2 segments of chronic bee paralysis virus strain CBPV-BJ detected in China. Arch Virol 8: 2451-2456.
Morse RA, Calderone NW (2000). The value of honey bees as pollinator of US crops in 2000. Bee Cult 128: 2-15.
Muhire BM, Varsani A, Martin DP (2014) SDT: a virus classification tool based on pairwise sequence alignment and identity calculation. PloS one 9: e108277. doi:10.1371/journal.pone.0108277.
Nielsen SL, Nicolaisen M, Kryger P (2008). Incidence of acute bee paralysis virus, black queen cell virus, chronic bee paralysis virus, deformed wing virus, Kashmir bee virus and sacbrood virus in honey bees (Apis mellifera) in Denmark. Apidologie 39: 310-314.
Olivier V, Blanchard P, Chaouch S, Lallemand P, Schurr F, Celle O, Dubois E, Tordo N, Thiéry R, Houlgatte R, Ribière M (2008) Molecular characterisation and phylogenetic analysis of Chronic bee paralysis virus, a honey bee virus. Virus Res 132(1-2): 59-68. doi: 10.1016/j.virusres.2007.10.014.
Ribière M, Lallemand P, Iscache AL, Schurr F, Celle O, Blanchard P, Olivier V, Faucon JP (2007) Spread of infectious chronic bee paralysis virus by honeybee (Apis mellifera L.) feces. Appl Environ Microbiol 73(23): 7711-6. doi: 10.1128/AEM.01053-07.
Ribiere M, Triboulot C, Mathieu L, Aurieres C, Faucon JP, Pepin M (2002) Molecular diagnosis of chronic bee paralysis virus infection. Apidologie 33: 339-351.
Schuster S, Zirkel F, Kurth A, van Cleef KW, Drosten C, van Rij RP, Junglen SA (2014) Unique nodavirus with novel features: mosinovirus expresses two subgenomic RNAs, a capsid gene of unknown origin, and a suppressor of the antiviral RNA interference pathway. J Virol 88(22): 13447-59. doi: 10.1128/JVI.02144-14.
Sguazza GH, Reynaldi FJ, Galosi CM, Pecoraro MR (2013) Simultaneous detection of bee viruses by multiplex PCR. J Virol Methods 194(1-2):102-6. doi: 10.1016/j.jviromet.2013.08.003.
Tehel A, Brown MJF, Paxton RJ (2016) Impact of managed honey bee viruses on wild bees. Curr Opin Virol 19: 16-22 6.
Tentcheva D, Gauthier L, Zappulla N, Dainat B, Cousserans F, Colin ME, Bergoin M (2004) Prevalence and seasonal variations of six bee viruses in Apis mellifera L. and Varroa destructor mite populations in France. Appl Environ Microbiol 70: 7185-7191.
vanEngelsdorp D, Hayes JJ, Underwood RM, Caron RM, Pettis J (2011) A survey of managed honey bee colony losses in the USA, fall 2009 to winter 2010. J Apic Res 50: 1-10.
Wu YY, Jia RH, Wang Q, Dai LP, Diao YQ, Xu FS, Wang X, Zhou T (2015). Multiple virus infections and the characteristics of chronic bee paralysis virus in diseased honey bees (Apis mellifera L.) in China. J Apic Sci 59: 95-106. doi:10.1515/JAS-2015-0026
Yildirim Y, Cagirgan AA, Usta A (2020) Phylogenetic analysis of sacbrood virus structural polyprotein and non-structural RNA dependent RNA polymerase gene: differences in Turkish strains. J Invertebr Pathol 176: 107459.
Youssef I, Schurr F, Goulet A, Cougoule N, Ribiere-Chabert M, Darbon H, Thiery R, Dubois E (2015) RNA 1 and RNA 2 genomic segments of chronic bee paralysis virus are infectious and induce chronic bee paralysis disease. J immunol Res. 423493. doi:10.1155/2015/423493