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肉桂油和银纳米粒子对牛乳腺炎分离的无乳链球菌的抗菌和抗生物膜潜力:克服耐药性的新途径。

Antimicrobial and antibiofilm potentials of cinnamon oil and silver nanoparticles against Streptococcus agalactiae isolated from bovine mastitis: new avenues for countering resistance.

机构信息

Department of Microbiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Sharkia, 44511, Egypt.

Department of Animal Medicine, Infectious Diseases, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Sharkia, Egypt.

出版信息

BMC Vet Res. 2021 Mar 31;17(1):136. doi: 10.1186/s12917-021-02842-9.

Abstract

BACKGROUND

Streptococcus agalactiae (S. agalactiae) is a contagious pathogen of bovine mastitis. It has financial implications for the dairy cattle industry in certain areas of the world. Since antimicrobial resistance increases in dairy farms, natural antimicrobials from herbal origins and nanoparticles have been given more attention as an alternative therapy. Hence, this study reported the antimicrobial and antibiofilm potentials of cinnamon oil, silver nanoparticles (AgNPs), and their combination against multidrug-resistant (MDR) S. agalactiae recovered from clinical bovine mastitis in Egypt.

RESULTS

Our findings revealed that 73% (146/200) of the examined milk samples collected from dairy cows with clinical mastitis were infected with Streptococci species. Of these, 9.59% (14/146) were identified as S. agalactiae and categorized as MDR. S. agalactiae isolates expressed four virulence genes (Hyl, cylE, scpB, and lmb) and demonstrated an ability to produce biofilms. Cinnamon oil showed high antimicrobial (MICs ≤0.063 μg /mL) and antibiofilm (MBIC = 4 μg/mL) potentials against planktonic and biofilms of S. agalactiae isolates, respectively. However, AgNPs showed reasonable antimicrobial (MICs ≤16 μg/mL) and relatively low antibiofilm (MBIC = 64 μg/mL) activities against screened isolates. Synergistic antimicrobial or additive antibiofilm interactions of cinnamon oil combined with AgNPs were reported for the first time. Scanning electron microscope (SEM) analysis revealed that biofilms of S. agalactiae isolates treated with cinnamon oil were more seriously damaged than observed in AgNPs cinnamon oil combination. Moreover, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) showed that cinnamon oil exerted a remarkable down-regulation of pili biosynthesis genes (pilA and pilB) and their regulator (rogB) against S. agalactiae biofilms, meanwhile the AgNPs cinnamon oil combination demonstrated a lower efficacy.

CONCLUSIONS

This is an in vitro preliminary approach that documented the antibiofilm potential of cinnamon oil and the inhibitory activity of cinnamon oil and its combination with AgNPs against MDR S. agalactiae recovered from clinical mastitis. Further in vivo studies should be carried out in animal models to provide evidence of concept for implementing these alternative candidates in the treatment of dairy farms infected by streptococcal mastitis in the future.

摘要

背景

无乳链球菌(S.agalactiae)是牛乳腺炎的一种传染性病原体。在世界某些地区,它对奶牛养殖业有经济影响。由于奶牛场的抗药性增加,来自草药来源的天然抗菌剂和纳米颗粒作为替代疗法受到了更多关注。因此,本研究报道了肉桂油、银纳米颗粒(AgNPs)及其组合对从埃及临床奶牛乳腺炎中分离的多药耐药(MDR)无乳链球菌的抗菌和抗生物膜潜力。

结果

我们的研究结果显示,从患有临床乳腺炎的奶牛采集的 200 份牛奶样本中,有 73%(146/200)受到链球菌属的感染。其中,9.59%(14/146)被鉴定为无乳链球菌,且被归类为 MDR。无乳链球菌分离株表达了 4 种毒力基因(Hyl、cylE、scpB 和 lmb),并表现出产生生物膜的能力。肉桂油对浮游生物和生物膜的无乳链球菌分离株均表现出较高的抗菌(MICs≤0.063μg/ml)和抗生物膜(MBIC=4μg/ml)潜力。然而,AgNPs 对筛选出的分离株表现出合理的抗菌(MICs≤16μg/ml)和相对较低的抗生物膜(MBIC=64μg/ml)活性。肉桂油与 AgNPs 联合使用的协同抗菌或相加抗生物膜相互作用是首次报道。扫描电子显微镜(SEM)分析显示,与肉桂油- AgNPs 联合处理的无乳链球菌生物膜相比,肉桂油处理的生物膜受到更严重的破坏。此外,逆转录定量聚合酶链反应(RT-qPCR)显示,肉桂油对无乳链球菌生物膜中菌毛生物合成基因(pilA 和 pilB)及其调节因子(rogB)表现出显著的下调作用,而肉桂油- AgNPs 联合使用的效果较低。

结论

这是一项体外初步研究,记录了肉桂油的抗生物膜潜力,以及肉桂油及其与 AgNPs 的组合对从临床乳腺炎中分离的多药耐药无乳链球菌的抑制活性。应在动物模型中进行进一步的体内研究,为将来在感染链球菌乳腺炎的奶牛场中实施这些替代候选药物提供概念验证。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc96/8010958/f217ea8fa04b/12917_2021_2842_Fig1_HTML.jpg

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