TY - JOUR
T1 - Evaluation of inhibitor activity of bacterial sialidase from Clostridium perfringens against Newcastle disease virus in the cell culture model using chicken embryo fibroblast
AU - Kurnia, Ryan Septa
AU - Setiawaty, Rahajeng
AU - Natih, Ketut Karuni Nyanakumari
AU - Nugroho, Christian Marco Hadi
AU - Silaen, Otto Sahat Martua
AU - Widyaningtyas, Silvia Tri
AU - Tarigan, Simson
AU - Ibrahim, Fera
AU - Sudarmono, Pratiwi Pudjilestari
N1 - Funding Information:
The authors express their appreciation to Dr. Sudarisman, Drh. Lily Natalia, and Drh. Adin Priadi from PT. Medika Satwa Laboratories, Bogor, Indonesia, for their support in microbiology laboratory facilities
Publisher Copyright:
© The authors. This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License (http://creativecommons.org/licenses/by/4.0)
PY - 2022/6
Y1 - 2022/6
N2 - Objective: The Newcastle disease virus (NDV) is an infectious disease that causes very high economic losses due to decreased livestock production and poultry deaths. The vaccine’s ineffectiveness due to mutation of the genetic structure of the virus impacts obstacles in controlling the disease, especially in some endemic areas. This study aimed to provide an alternative treatment for NDV infection by observing the viral replication inhibitor activity of Clostridium perfringens sialidase in primary chicken embryo fibroblast (CEF) cells. Materials and Methods: The virus was adapted in CEF monolayer cells, then collected thrice using the freeze–thaw method and stored at −20°C for the next step in the challenge procedure. C. perfringens crude sialidase was obtained, but it was further purified via stepwise elution in ion exchange using Q Sepharose® Fast Flow and affinity chromatography with oxamic acid agarose. The purified sialidase was tested for its toxicity, ability to breakdown sialic acid, stopping viral replication, and how treated cells expressed their genes. Results: According to this study, purified C. perfringens sialidase at dosages of 187.5, 93.75, and 46.87 mU effectively hydrolyzes CEF cells’ sialic acid and significantly inhibits viral replication on the treated cells. However, sialidase dosages of 375 and 750 mU affected the viability of monolayer CEF cells. Interestingly, downregulation of toll-like receptor (TLR)3 and TLR7 (p < 0.05) in the sialidase-treated group indicates viral endocytosis failure. Conclusions: By stopping endocytosis and viral replication in host cells, sialidase from C. perfringens can be used as an alternative preventive treatment for NDV infection.
AB - Objective: The Newcastle disease virus (NDV) is an infectious disease that causes very high economic losses due to decreased livestock production and poultry deaths. The vaccine’s ineffectiveness due to mutation of the genetic structure of the virus impacts obstacles in controlling the disease, especially in some endemic areas. This study aimed to provide an alternative treatment for NDV infection by observing the viral replication inhibitor activity of Clostridium perfringens sialidase in primary chicken embryo fibroblast (CEF) cells. Materials and Methods: The virus was adapted in CEF monolayer cells, then collected thrice using the freeze–thaw method and stored at −20°C for the next step in the challenge procedure. C. perfringens crude sialidase was obtained, but it was further purified via stepwise elution in ion exchange using Q Sepharose® Fast Flow and affinity chromatography with oxamic acid agarose. The purified sialidase was tested for its toxicity, ability to breakdown sialic acid, stopping viral replication, and how treated cells expressed their genes. Results: According to this study, purified C. perfringens sialidase at dosages of 187.5, 93.75, and 46.87 mU effectively hydrolyzes CEF cells’ sialic acid and significantly inhibits viral replication on the treated cells. However, sialidase dosages of 375 and 750 mU affected the viability of monolayer CEF cells. Interestingly, downregulation of toll-like receptor (TLR)3 and TLR7 (p < 0.05) in the sialidase-treated group indicates viral endocytosis failure. Conclusions: By stopping endocytosis and viral replication in host cells, sialidase from C. perfringens can be used as an alternative preventive treatment for NDV infection.
KW - Cell culture
KW - Clostridium perfringens
KW - In vitro
KW - Inhibit
KW - Sialidase
KW - Virus
UR - http://www.scopus.com/inward/record.url?scp=85133538969&partnerID=8YFLogxK
U2 - 10.5455/javar.2022.i600
DO - 10.5455/javar.2022.i600
M3 - Article
AN - SCOPUS:85133538969
SN - 2311-7710
VL - 9
SP - 335
EP - 345
JO - Journal of Advanced Veterinary and Animal Research
JF - Journal of Advanced Veterinary and Animal Research
IS - 2
ER -