金银花、连翘及其药对对马链球菌马亚种耐药基因和毒力基因的影响

doi:10.16431/j.cnki.1671-7236.2022.02.033

Abstract

Abstract: 【Objective】 This study was to investigate the effects of honeysuckle, forsythia and their drug pair (honeysuckle-forsythia 1∶1) on drug resistance genes and virulence genes of Streptococcus equi subsp. equi (SEE) carrying fneB virulence gene in Northern Xinjiang.【Method】 The water extract of 1 g/mL honeysuckle, forsythia and honeysuckle, forsythia drug pair 1∶1 (honeysuckle-forsythia 1∶1) were tested by traditional Chinese medicine ratio concentration gradient test.The concentration gradient test of traditional Chinese medicine was carried out, and the three strains were co-cultured with L1 strain carrying fneB virulence gene, D1 strain carrying lytA+fneB+ply virulence genes and Y1 strain carrying qnrA+bla_TEM+fneB genes.The effects of traditional Chinese medicine on drug resistance genes bla_TEM, qnrA and virulence genes ply, fneB of SEE strains were detected.192 SPF Kunming mice were divided into 16 groups: Control group (normal saline), honeysuckle group, forsythia group, honeysuckle-forsythia 1∶1 group, negative control group (Y1, L1 and D1 strain groups) and three strains were co-cultured with honeysuckle, forsythia and honeysuckle-forsythia 1∶1 respectively.The drugs or bactering fluid in each group were injected intraperitoneally with 0.5 mL per mouse for antibacterial test in vivo, and the effects of drugs on pathogenicity and fneB virulence gene in mice were detected.【Result】 The optimal concentration of Chinese herbal medicine was as follows∶Water extract of Chinese herbal medicine∶THB medium∶bacteria solution (D_{600 nm} value was 0.6) was 1 000 μL∶500 μL∶20 μL.The drug resistance gene and virulence gene were not detected in the three SEE strains co-cultured with the aqueous extract of traditional Chinese medicine, and their morphological structure did not change.The results of mouse pathogenicity test showed that the survival rates of negative control group were 16.7%, 8.3% and 0, respectively.The survival rate of mice in the co-culture of L1 strain+forsythia group and L1 strain+honeysuckle group were 83.3% and 75.0%, respectively.The survival rate of mice in the co-culture group of honeysuckle-forsythia 1∶1 and 3 SEE strains were 41.7%, 16.7% and 50.0%, respectively.The results of pathological anatomy of mice showed that the livers of other groups were normal except that the livers of mice inoculated with the original SEE strain was swollen and congestion, and the edge was blunt and round.The results of fneB virulence gene detection showed that Y1+honeysuckle, Y1+forsythia, Y1+honeysuckle-forsythia 1∶1, L1+honeysuckle, L1+honeysuckle-forsythia 1∶1, D1+honeysuckle, D1+forsythia and D1+honeysuckle-forsythia groups carried fneB virulence gene, but not in L1+forsythia group, which indicated that the SEE strains in mice had the ability to regain fneB virulence gene again, and the strain had anti-virus rejuvenation, and its mechanism needed to be further studied.【Conclusion】 honeysuckle, forsythia and their drug pair could weak the virulence gene and eliminate the drug resistance gene of SEE strain, which had guiding significance for the prevention and treatment of horse adenosis disease, and provided theoretical support for reducing and replacing resistance.

Key words: honeysuckle; forsythia; Streptococcus equi subsp. equi; drug resistance genes; virulence genes

CLC Number:

S859.3

MAI Zhanhai, LU Yabin, LI Jianlong, ZHAO Lu, SUN Danlan, TONG Panpan, KUANG Ling. Effects of Honeysuckle, Forsythia and Their Drug Pair on Resistance Genes and Virulence Genes of Streptococcus equi subsp. equi[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 709-717.

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Effects of Honeysuckle, Forsythia and Their Drug Pair on Resistance Genes and Virulence Genes of Streptococcus equi subsp. equi

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