Evaluation of thymol activity against pseudorabies virus in vitro and its action mode

doi:10.3969/j.issn.1004-1524.2023.01.05

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (1): 41-49.DOI: 10.3969/j.issn.1004-1524.2023.01.05

• Animal Science • Previous Articles Next Articles

Evaluation of thymol activity against pseudorabies virus in vitro and its action mode

SONG Tianhao¹(), PANG Lianfeng¹, CHEN Lingshuang², DENG Huidan¹, XU Zhiwen¹, ZHU Ling¹, REN Zhihua¹, DENG Junliang¹^,^*()

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2. Animal Disease Prevention and Control Center, Agricultural and Rural Affairs Bureau of Ganluo County, Liangshan Yi Autonomous Prefecture, Sichuan Province, Ganluo 616850, Sichuan, China

Received:2022-01-05 Online:2023-01-25 Published:2023-02-21

Abstract

Abstract:

In this experiment, BHK-21 cells were used as the research model to explore the anti pseudorabies virus (PRV) activity and action mode of thymol at different mass concentrations in vitro. The maximum no-cytotoxic concentration (MNTC) of thymol on BHK-21 cells was detected by CCK-8 method, and the half maximal inhibitory concentration (IC₅₀) was calculated to be (212.789±1.652) μg·mL^-1. The half effective concentration (EC₅₀) of thymol against PRV was determined as (30.710±0.303) μg·mL^-1 and the therapeutic index (TI) of thymol was 6.929, which indicated that thymol was a high-efficiency and low-toxicity anti PRV drug. The effects of thymol on virus titer and one-step growth curve of PRV were detected by CPE observation method. The results showed that thymol could significantly reduce the virus titer of PRV and the virulence of PRV after proliferation in BHK-21 cells in a dose-dependent manner. The quantitative PCR was used to detect the gene copies of PRV gE in BHK-21 cells after different treatments to study the action mode of anti PRV effect of thymol in vitro. It was shown that the thymol exerted its antiviral activity mainly by inhibiting the proliferation of PRV in BHK-21 cells, as well as it could directly kill some PRV virus particles.

Key words: thymol, pseudorabies virus, antiviral activity in vitro, action mode

CLC Number:

S859

SONG Tianhao, PANG Lianfeng, CHEN Lingshuang, DENG Huidan, XU Zhiwen, ZHU Ling, REN Zhihua, DENG Junliang. Evaluation of thymol activity against pseudorabies virus in vitro and its action mode[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 41-49.

Figures/Tables 9

References 20

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无水乙醇体积分数 Volume fraction of ethanol/%	D₄₅₀
10	0.089±0.008 e
8	0.456±0.017 d
6	0.957±0.003 c
4	1.033±0.005 b
2	1.074±0.011 a
0	1.068±0.005 a

无水乙醇体积分数 Volume fraction of ethanol/%	D₄₅₀
10	0.089±0.008 e
8	0.456±0.017 d
6	0.957±0.003 c
4	1.033±0.005 b
2	1.074±0.011 a
0	1.068±0.005 a

无水乙醇体积分数 Volume fraction of ethanol/%	D₄₅₀
4	0.717±0.007 b
3	0.523±0.011 c
2	0.514±0.005 c
1	0.517±0.013 c
0(CK-1E)	1.205±0.007 a
0(CK-2E)	0.524±0.006 c

无水乙醇体积分数 Volume fraction of ethanol/%	D₄₅₀
4	0.717±0.007 b
3	0.523±0.011 c
2	0.514±0.005 c
1	0.517±0.013 c
0(CK-1E)	1.205±0.007 a
0(CK-2E)	0.524±0.006 c

百里香酚质量浓度 Mass concentration of thymol/(μg·mL^-1)	相对存活率 Relative survival rate/%
256	41.934±0.507^*
128	72.165±0.548^*
64	92.200±0.683^*
32	94.447±0.584^*
16	96.667±0.639
8	98.499±0.219
0	100

Evaluation of thymol activity against pseudorabies virus in vitro and its action mode

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稀释倍数的常用对数值 Common logarithm of dilution multiple	重复数 Repetition number	CPE数 CPE number	累计CPE数 Accumulative CPE number	无CPE数 No-CPE number	累计无CPE数 Accumulative no-CPE number	累计总数 Total number	比例 Proportion/%
-1	6	6	43	0	0	43	100
-2	6	6	37	0	0	37	100
-3	6	6	31	0	0	31	100
-4	6	6	25	0	0	25	100
-5	6	6	19	0	0	19	100
-6	6	6	13	0	0	13	100
-7	6	5	7	1	1	8	87.5
-8	6	2	2	4	5	7	28.6
-9	6	0	0	6	11	11	0
-10	6	0	0	6	17	17	0

百里香酚质量浓度 Mass concentration of thymol/(μg·mL^-1)	相对抑制率 Relative inhibition rate/%
64	78.881±1.162^*
32	51.986±0.567^*
16	18.497±0.964^*
8	8.482±1.133^*
4	5.094±0.644
2	2.365±0.300
0(CK-2)	100^*
0(CK-1)	0

百里香酚质量浓度 Mass concentration of thymol /(μg·mL^-1)	-lg(TCID₅₀)/mL^-1
64	4.775±0.121 e
32	5.326±0.066 d
16	5.744±0.105 c
8	6.225±0.120 b
4	6.500±0.136 a
2	6.409±0.079 a
0(CK-1)	6.545±0.079 a

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