抗菌肽NZ2114对来源于奶牛乳房炎的无乳链球菌及其生物膜的消杀作用

doi:10.16431/j.cnki.1671-7236.2020.07.035

Abstract

Abstract: In order to explore the antimicrobial activity and antimicrobial mechanism of antimicrobial peptide NZ2114 against Streptococcus agalactiae (S.agalactiae),S.agalactiae ATCC 13813 was used.The antimicrobial properties and mechanism of antimicrobial peptide NZ2114 were evaluated by means of minimum inhibitory concentration (MIC),time-killing curves,post antibiotic effect (PAE) and electron microscopy.The inhibition and elimination effects of S.agalactiae on biofilms and persister bacteria were further analyzed.The results showed that the MIC value of NZ2114 against S.agalactiae ATCC 13813 was 0.23 μmol/L,while the values of NZ2114 against CAU-FRI 1,2 and 3 strains,which were clinical isolated from mastitis cow,were all 0.11 μmol/L.The MIC values of vancomycin against the above-mentioned four strains were all 0.67 μmol/L,so the antimicrobial activity of NZ2114 were 2.91 and 6.09 times than that of vancomycin,respectively.Meanwhile,2×MIC and 4×MIC of NZ2114 could kill 99.9% S.agalactiae in 0.5 h and the pharmacodynamic time was up to 270 min.The result of scanning electron microscope (SEM) showed that the surface of the bacteria appeared to shrink or even break after the treatment of NZ2114,and the biofilms were eliminated.NZ2114 effectively inhibited early biofilms by 99.9% in 2×MIC,and eradicated mature biofilms by 99.9% in 32×MIC.At the same time,the bacteria (approximately 99.9%) in biofilm were efficiently killed by NZ2114 in 16×MIC,and that of the persisters which were resistant to vancomycin were eliminated by 99% in 0.5×MIC of NZ2114.Confocal laser scanning microscopy (CLSM) also demonstrated the potent antimicrobial and anti-biofilm activity of NZ2114 (biofilm from 28.48 μm reduced to 10.32 μm).The above results showed that NZ2114 had high bactericidal activity against S.agalactiae ATCC 13813 with fast speed,strong ability to inhibit/eradicate the biofilm,and high bactericidal activity against persisters in early and mature biofilms,and its effects were all significantly higher than those of vancomycin.NZ2114 had the potential to develop as a novel antibacterial agent for mastitis treatment caused by S.agalactiae.

Key words: antimicrobial peptide NZ2114; cow mastitis; Streptococcus agalactiae; antimicrobial activity; biofilm

CLC Number:

S859.79⁺7

SHAN Yuxue, YANG Na, TENG Da, WANG Xiumin, MAO Ruoyu, HAO Ya, MA Xuanxuan, FAN Huan, WANG Jianhua. The Disinfection Effect of Antimicrobial Peptide NZ2114 on Streptococcus agalactiae Isolated from Bovine Mastitis and Its Biofilm[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2284-2294.

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The Disinfection Effect of Antimicrobial Peptide NZ2114 on Streptococcus agalactiae Isolated from Bovine Mastitis and Its Biofilm

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