Upadhyay Abhinav, Venkitanarayanan Kumar
Department of Animal Science, University of Connecticut, Horsebarn Hill Rd Ext., Unit 4040, Storrs, CT, 06269, USA.
J Nat Med. 2016 Jul;70(3):667-72. doi: 10.1007/s11418-016-0990-4. Epub 2016 Apr 19.
Listeria monocytogenes is a major foodborne pathogen that causes life-threatening illnesses in humans. With emergence of antibiotic resistance in L. monocytogenes, there is considerable interest in testing the efficacy of alternative therapies for controlling listeriosis in humans. This study investigated the efficacy of three phytochemicals, namely trans-cinnamaldehyde (TC), carvacrol (CR), and thymol (TY) in reducing L. monocytogenes virulence in the recently established invertebrate model, Galleria mellonella. In addition, the effect of phytochemicals on the transcription of antimicrobial peptide genes in G. mellonella (responsible for host defense) was investigated using real-time quantitative polymerase chain reaction. G. mellonella larvae were inoculated with L. monocytogenes (10(5) CFU/larvae) either with or without the subinhibitory concentration (chemical concentration not inhibiting bacterial growth) of phytochemicals. The larvae were incubated at 37 °C for 5 days, and their mortality was scored at 24-h intervals. The transcriptional response of the defense genes was studied in inoculated and uninoculated larvae at 6 h post challenge. The experiments were repeated at least six times with replicates. All phytochemicals enhanced the survival rates of G. mellonella infected with lethal doses of L. monocytogenes (P < 0.05). CR and TC at 0.01 % concentration were found to be the most effective treatments, and increased larval survival rates by 80 % and 50 %, respectively, on day 5 (P < 0.05). The phytochemicals also upregulated the expression of antimicrobial peptide genes in G. mellonella larvae challenged with L. monocytogenes (P < 0.05). Results suggest that TC, CR, and TY could potentially be used to control listeriosis. Further investigation in an appropriate mammalian model is warranted.
单核细胞增生李斯特菌是一种主要的食源性病原体,可导致人类患上危及生命的疾病。随着单核细胞增生李斯特菌出现抗生素耐药性,人们对测试控制人类李斯特菌病的替代疗法的疗效产生了浓厚兴趣。本研究调查了三种植物化学物质,即反式肉桂醛(TC)、香芹酚(CR)和百里香酚(TY)在最近建立的无脊椎动物模型——大蜡螟中降低单核细胞增生李斯特菌毒力的效果。此外,还使用实时定量聚合酶链反应研究了植物化学物质对大蜡螟中抗菌肽基因转录(负责宿主防御)的影响。将大蜡螟幼虫接种单核细胞增生李斯特菌(10⁵CFU/幼虫),同时添加或不添加植物化学物质的亚抑制浓度(不抑制细菌生长的化学浓度)。将幼虫在37°C下孵育5天,并每隔24小时记录其死亡率。在攻击后6小时,对接种和未接种的幼虫中防御基因的转录反应进行了研究。实验至少重复六次,并设有重复样本。所有植物化学物质均提高了感染致死剂量单核细胞增生李斯特菌的大蜡螟的存活率(P < 0.05)。发现浓度为0.01%的CR和TC是最有效的处理方法,在第5天分别使幼虫存活率提高了80%和50%(P < 0.05)。植物化学物质还上调了受单核细胞增生李斯特菌攻击的大蜡螟幼虫中抗菌肽基因的表达(P < 0.05)。结果表明,TC、CR和TY可能可用于控制李斯特菌病。有必要在合适的哺乳动物模型中进行进一步研究。