Muhammet Ekin AZBAZDAR, Görkem AKINCI, Erkan GÜLER, Zeynep Ahsen KOÇER

The effect of sediment composition and polyethylene glycol precipitation on the detection limit of H6N2 influenza virus in sediment samples

Influenza A viruses (IAVs) are naturally carried by wild aquatic birds and generally cause asymptomatic gastroenteric disease in their natural reservoir hosts. Because the viruses follow oral-fecal route in the avian host, they could be shed into water bodies through feces. Furthermore, IAVs that are secreted to abiotic sources might be preserved in the environment for a period, facilitating the transmission of viruses between individuals or species. Viral stability could be affected by several factors such as pH, salinity, and temperature of water. Therefore, this study aims to investigate the lowest amount of infectious IAVs that could be detected in sediment samples via molecular and virus isolation methods, and to compare the sediment composition with the efficiency of detection/isolation of IAVs and viral persistence. For this purpose, an H6N2 virus (A/Aquatic bird/Gediz Delta/1/2018) of avian origin was used for artificially seeding the sediment samples that were collected from Gediz Delta, Izmir, Turkey. Molecular methods showed that lower amount of H6N2 virus could be detected in sediment sample collected from freshwater area (FS) in comparison with the sediment samples that were collected from salty water area (SS). Furthermore, virus precipitation method using polyethylene glycol increased the efficiency of virus isolation by 10-fold in FS, but not in SS. On the other hand, although the detection limit for IAVs was higher in SS than in FS, viral fitness was better maintained in SS. Moreover, high number of cations in the composition of SS along with larger surface area facilitated virus adsorption on SS complicating the virus to detach from sediment particles. Thus, the result of this study remarks that the environmental origin of abiotic sources could affect the viral stability and fitness; therefore, it could affect the transmission dynamics of the virus in different environments.

Keywords:

influenza A virus sediment, environmental sources, environmental factors, H6N2 virus,

PDF

___

Abbaszadegan, M., Huber, M. S., Gerba, C. P. & Pepper, I. L. (1993). Detection of enteroviruses in groundwater with the polymerase chain reaction. Applied and Environmental Microbiology, 59(5): 1318-1324. https://doi.org/10.1128/aem.59.5.1318-1324.1993
Bitton, G. (1975). Adsorption of viruses onto surfaces in soil and water. Water Research, 9(5-6): 473-484. https://doi.org/10.1016/0043-1354(75)90071-8
Brown, J. D., Goekjian, G., Poulson, R., Valeika, S. & Stallknecht, D. E. (2009). Avian influenza virus in water: Infectivity is dependent on pH, salinity and temperature. Veterinary Microbiology, 136(1-2): 20-26. https://doi.org/10.1016/j.vetmic.2008.10.027
Debode, F., Marien, A., Janssen, É., Bragard, C. & Berben, G. (2017). The influence of amplicon length on real-time PCR results. Base, 21(1): 3-11. https://doi.org/10.25518/1780-4507.13461
Dublineau, A., Batéjat, C., Pinon, A., Burguière, A. M., Leclercq, I. & Manuguerra, J. C. (2011). Persistence of the 2009 pandemic influenza a (H1N1) virus in water and on non-porous surface. PLoS One, 6(11): e28043. https://doi.org/10.1371/journal.pone.0028043
Eisfeld, A. J., Neumann, G. & Kawaoka, Y. (2014). Influenza A virus isolation, culture and identification. Nature Protocols, 9(11): 2663-2681. https://doi.org/10.1038/nprot.2014.180
Gillim-Ross, L., Santos, C., Chen, Z., Aspelund, A., Yang, C.F., Ye, D., Jin, H., Kemble, G. & Subbarao, K. (2008). Avian influenza H6 viruses productively infect and cause illness in mice and ferrets. Journal of Virology, 82(21): 10854-10863. https://doi.org/10.1128/JVI.01206-08
Gonchar, K. A., Agafilushkina, S. N., Moiseev, D. V., Bozhev, I. V., Manykin, A. A., Kropotkina, E. A., Gambaryan, A. S. & Osminkina, L. A. (2020). H1N1 influenza virus interaction with a porous layer of silicon nanowires. Materials Research Express, 7(3): 035002. https://doi.org/10.1088/2053-1591/ab7719
Guan, J., Chan, M., Ma, B., Grenier, C., Wilkie, D. C., Pasick, J., Brooks, B. W. & Spencer, J. L. (2008). Development of methods for detection and quantification of avian influenza and Newcastle disease viruses in compost by real-time reverse transcription polymerase chain reaction and virus isolation. Poultry Science, 87(5): 838-843. https://doi.org/10.3382/ps.2007-00195
Guven, D. E. & Akinci, G. (2013). Effect of sediment size on bioleaching of heavy metals from contaminated sediments of Izmir Inner Bay. Journal of Environmental Sciences, 25(9): 1784-1794. https://doi.org/10.1016/S1001-0742(12)60198-3
Ito, T., Okazaki, K., Kawaoka, Y., Takada, A., Webster, R. G. & Kida, H. (1995). Perpetuation of influenza A viruses in Alaskan waterfowl reservoirs. Archives of Virology, 140(7): 1163-1172. https://doi.org/10.1007/BF01322743
Lang, A. S., Kelly, A. & Runstadler, J. A. (2008). Prevalence and diversity of avian influenza viruses in environmental reservoirs. Journal of General Virology, 89(2): 509-519. https://doi.org/10.1099/vir.0.83369-0
Lickfett, T. M., Clark, E., Gehring, T. M. & Alm, E. W. (2018). Detection of Influenza A viruses at migratory bird stopover sites in Michigan, USA. Infection Ecology & Epidemiology, 8(1): 1474709. https://doi.org/10.1080/20008686.2018.1474709
Matrosovich, M. & Klenk, H. D. (2003). Natural and synthetic sialic acid-containing inhibitors of influenza virus receptor binding. Reviews in Medical Virology, 13(2): 85-97. https://doi.org/10.1002/rmv.372
Mercan, Y., Atim, G., Kayed, A. E., Azbazdar, M. E., Kandeil, A., Ali, M. A., Rubrum, A., McKenzie, P., Webby, R. J., Erima, B., Wabwire-Mangen, F., Ukuli, Q. A., Tugume, T., Byarugaba, D. K., Kayali, G., Ducatez, M. F. & Koçer, Z. A. (2021). Molecular characterization of closely related H6N2 Avian Influenza Viruses isolated from Turkey, Egypt, and Uganda. Viruses, 13(4), 607. https://doi.org/10.3390/v13040607
Miller, H. K. & Schlesinger, R. W. (1955). Differentiation and purification of influenza viruses by adsorption on aluminum phosphate. The Journal of Immunology, 75(2): 155-160.
Reed, L. J. & Muench, H. (1938). A simple method of estimating fifty per cent endpoints. American Journal of Epidemiology, 27(3): 493-97. https://doi.org/10.1093/oxfordjournals.aje.a118408
Schrader, C., Schielke, A., Ellerbroek, L. & Johne, R. (2012). PCR inhibitors–occurrence, properties and removal. Journal of Applied Microbiology, 113(5): 1014-1026. https://doi.org/10.1111/j.1365-2672.2012.05384.x
Stallknecht, D. E., Shane, S. M., Kearney, M. T. & Zwank, P. J. (1990a). Persistence of avian influenza viruses in water. Avian Diseases, 34(2): 406-411. https://doi.org/10.2307/1591428
Stallknecht, D. E., Kearney, M. T., Shane, S. M. & Zwank, P. J. (1990b). Effects of pH, temperature, and salinity on persistence of avian influenza viruses in water. Avian Diseases, 34(2): 412-418. https://doi.org/10.2307/1591429
Wang, G., Deng, G., Shi, J., Luo, W., Zhang, G., Zhang, Q., Liu, L., Jiang, Y., Li, C., Sriwilaijaroen, N. & Hiramatsu, H. (2014). H6 influenza viruses pose a potential threat to human health. Journal of Virology, 88(8): 3953-3964. https://doi.org/10.1128/JVI.03292-13
Warren, J., Neal, A. & Rennels, D. (1966). Adsorption of myxoviruses on magnetic iron oxides. Proceedings of the Society for Experimental Biology and Medicine, 121(4): 1250-1253. https://doi.org/10.3181/00379727-121-31020
Webster, R.G., Yakhno, M., Hinshaw, V.S., Bean, W.J. & Murti, K.C. (1978). Intestinal influenza: Replication and characterization of influenza viruses in ducks. Virology, 84(2): 268-278. https://doi.org/10.1016/0042-6822(78)90247-7
Webster, R. G., Bean, W. J., Gorman, O. T., Chambers, T. M. & Kawaoka, Y. (1992). Evolution and ecology of influenza A viruses. Microbiology Reviews, 56(1): 152-179. https://doi.org/10.1128/mr.56.1.152-179.1992
WHO. (2002). World Health Organization manual on animal influenza diagnosis and surveillance (No. WHO/CDS/CSR/NCS/2002.5). World Health Organization.
WHO. (2021). Information for the molecular detection of influenza viruses. Retrieved on March 18, 2022 from https://www.who.int/teams/global-influenza-programme/laboratory-network/quality-assurance/eqa-project/information-for-molecular-diagnosis-of-influenza-virus
Williamson, K. E., Wommack, K. E. & Radosevich, M. (2003). Sampling natural viral communities from soil for culture-independent analyses. Applied and Environmental Microbiology, 69(11): 6628-6633. https://doi.org/10.1128/AEM.69.11.6628-6633.2003
Zowalaty, M. E. E., Chander, Y., Redig, P. T., El Latif, H. K. A., Sayed, M. A. E. & Goyal, S. M. (2011). Selective isolation of avian influenza virus (AIV) from cloacal samples containing AIV and Newcastle disease virus. Journal of Veterinary Diagnostic Investigation, 23(2), 330-332. https://doi.org/10.1177/104063871102300222