冠突散囊菌Ec-12上清抑制小鼠的伤寒沙门菌机制的初步分析

doi:10.11843/j.issn.0366-6964.2024.02.031

Abstract

Abstract: To investigated the probiotic function of Eurotium cristatum(EC) in black tea and reveal the preliminary mechanism of inhibit Salmonella Typhimurium,we used dilution coating method to isolate EC, and analyzed its taxonomic status by morphology and ITS rRNA gene sequence. And then the agar diffusion inhibition test, thermal acid-base and UV stability test, electron microscopy and mouse animal test were used to evaluate the preliminary mechanism of inhibition of S. Typhimurium bacteria by this Ec-12 supernatant. A strain of Ec-12 was identified as E. cristatum based on morphology, growth characteristics and ITS rRNA gene sequence analysis, and named strain Ec-12. The supernatant of the crude isolate Ec-12 produced good inhibition at pH near neutral conditions with good stability. The supernatant of tEc-12 showed good bacterial inhibition at pH near neutral and good stability of inhibition. The inhibition of S. Typhimurium was observed by electron microscopy, which showed that the supernatant of the crude isolate Ec-12 caused deformation of the body and roughness of the surface layer of the bacterium. It also significantly reduced the rate of diarrhea and mortality caused by S. Typhimurium infection in mice, improved the integrity of mouse villi and increased the secretion of mucin (P<0.05), reduced the apoptosis of colonic cells (P<0.05), and inhibited the harm of S. Typhimurium to mice. The supernatant of Ec-12 strain acted on S. Typhimurium in the murine gastrointestinal tract, disrupting the structural integrity of S. Typhimurium, while acting on the murine intestine, improving the integrity of intestinal villi, promoting the secretion of colonic mucin, reducing the apoptosis of colonic cells, and thus reducing the diarrhea and death caused by S. Typhimurium. It has a probiotic potential.

Key words: Eurotium cristatum, supernatant, Salmonella Typhimurium, inhibition

CLC Number:

S852.6

LI Zhaolong, KONG Xiangrui, LIN Fengqiang, WANG Xiuping, ZHAO Ran, PENG Xiaoli, CHEN Changsong. Preliminary Study of the Mechanism of Inhibition of Salmonella Typhimurium by Ec-12 Supernatant[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 739-750.

References

[1] 陈云兰, 于汉寿, 吕毅, 等. 康砖和青砖茶中散囊菌的分离、鉴定及其生物学特性研究[J]. 茶叶科学, 2006, 26(3):232-236.
CHEN Y L, YU H S, LV Y, et al. Investigation on the isolation, identification and the biological characteristic of Eurotium fungi in the Kangzhuan and Qingzhuan brick tea[J]. Journal of Tea Science, 2006, 26(3):232-236.(in Chinese)
[2] 刘天囡. 冠突散囊菌分离培养及接种发酵夏秋茶初步探究[D]. 南京:南京农业大学, 2016.
LIU T N. Study on isolated cultrue and artificial inoculation of E. cristatum to ferment summer and autumn tea[D]. Nanjing:Nanjing Agricultural University, 2016.(in Chinese)
[3] 关美玲, 刘仲华, 刘素纯, 等. 黑茶发花过程中冠突散囊菌对茶叶中游离氟含量的影响[J]. 茶叶科学, 2022, 31(5):386-390.
GUAN M L, LIU Z H, LIU S C, et al. Effect of Eurotium Cristatum strains on content of free-state fluorine in tea duringthe fungus growing pr ocess of brick tea[J]. Journal of Tea Science, 2022, 31(5):386-390.(in Chinese)
[4] 吕嘉枥, 郑欣欣, 马亚宁. 冠突散囊菌液体发酵产降脂类物质的初步研究[J]. 陕西科技大学学报, 2014, 32(6):115-118.
LV J L, ZHENG X X, MA Y N. Study on lipid-lowering substances from Eurotium cristutum by liquid fermentation[J]. Journal of Shaanxi University of Science & Technology, 2014, 32(6):115-118.(in Chinese)
[5] 李兆龙, 陈常颂, 孔祥瑞, 等. 冠突散囊菌茶对高糖、高脂大鼠的血糖和血脂相关因素的影响[J]. 中国兽医学报, 2022, 42(9):1889-1894.
LI Z L, CHEN C S, KONG X R, et al. Effects of Canticata tea on blood glucose and related factors in high glucose and fat rats[J]. Chinese Journal of Veterinary Science, 2022, 42(9):1889-1894.(in Chinese)
[6] 刘子音, 许爱清, 李宗军, 等. 茯砖茶中冠突散囊菌及其代谢产物研究进展[J]. 茶叶通讯, 2010, 37(1):23-26.
LIU Z Y, XU A Q, LI Z J, et al. Research progress on Eurotium cristataum and its metabolites in Fuzhuan tea[J]. Tea Communication, 2010, 37(1):23-26.(in Chinese)
[7] 陈桂梅. 冠突散囊菌研究进展[J]. 西北农林科技大学学报:自然科学版, 2012, 40(3):179-184.
CHEN G M. Research progress on Eurotium cristatum[J]. Journal of Northwest A & F University:Natural Science Edition, 2012, 40(3):179-184.(in Chinese)
[8] 杨娟娟. 冠突散囊菌固体发酵及其次级代谢产物影响因素的研究[D]. 阿拉尔:塔里木大学, 2016.
YANG J J. The study on the influence factors of solid fermentation and the secondary metabolites of Eurotium cristatum[D]. Alaer:Tarim University, 2016.(in Chinese)
[9] 邓放明, 龚淑莉, 杨伟丽. 冠突散囊菌胞外多糖生物活性高通量筛选试验[J]. 食品与机械, 2007, 23(6):48-51, 113.
DENG F G, GONG S L, YANG W L. Study on bioactivity of extracellular polysaccharide of Eurotium cristatum by high throughput screening[J]. Food & Machinery, 2007, 23(6):48-51, 113.(in Chinese)
[10] 彭晓赟, 梁法亮, 李冬利, 等. 茯砖茶中冠突散囊菌的次级代谢产物及其生物活性研究[J]. 中草药, 2013, 44(14):1881-1886.
PENG X Y, LIANG F L, LI D L, et al. Secondary metabolites of Eurotium cristatum from Fu Brick Tea and their biological activities[J]. Chinese Traditional and Herbal Drugs, 2013, 44(14):1881-1886.(in Chinese)
[11] 李响. 冠突散囊菌发酵工艺的优化及其免疫活性的初步研究[D]. 贵阳:贵州大学, 2019.
LI X. Preliminary study on the optimization of fermentation technology and immune activity of Eurotium cristatum[D]. Guiyang:Guizhou University, 2019.(in Chinese)
[12] 王亚丽, 秦俊哲, 黄亚亚, 等. 冠突散囊菌对茯砖茶品质形成的影响[J]. 食品与发酵工业, 2018, 44(4):194-197.
WANG Y L, QIN J Z, HUANG Y Y, et al. Effect of the quality of Fuzhuan brick tea fermented in Eurotium cristatum[J]. Food and Fermentation Industries, 2018, 44(4):194-197.(in Chinese)
[13] 尹旭敏. 四川"金花"菌生物学特性及其发酵液态茶工艺的初步研究[D]. 成都:四川农业大学, 2006.
YIN X M. Primary study on biological characteristic from Sichuan and fermentation technique of liquid tea of "Gloden flower" fungus[D]. Chengdu:Sichuan Agricultural University, 2006.(in Chinese)
[14] 蔡正安, 刘素纯, 刘杏益, 等. 冠突散囊菌在不同茶类及几种植物材料上"发花"的研究[J]. 茶叶科学, 2010, 30(4):263-268.
CAI Z A, LIU S C, LIU X Y, et al. Study of the growth of Eurotium cristatum fungi on some kinds of tea and plant materials[J]. Journal of Tea Science, 2010, 30(4):263-268.(in Chinese)
[15] 严蒸蒸, 余婷, 黄悠然, 等. 温度和水活度对冠突散囊菌菌丝生长和产孢的影响[J]. 菌物研究, 2018, 16(4):219-227.
YAN Z Z, YU T, HUANG Y R, et al. Influence of temperature and water activity on mycelial growth and conidia formation of Eurotium cristatum isolated from Chinese brick tea[J]. Journal of Fungal Research, 2018, 16(4):219-227.(in Chinese)
[16] 周杨艳, 李雨枫, 白雪, 等. 茯砖茶中优势微生物在不同培养基的差异性比较[J]. 中国酿造, 2014, 33(9):75-80.
ZHOU Y Y, LI Y F, BAI X, et al. Difference comparison of dominate microorganisms from Fuzhuan brick tea in different media[J]. China Brewing, 2014, 33(9):75-80.(in Chinese)
[17] 刘婷, 陈然, 陈頔, 等. 用CGMCC No. 8730冠突散囊菌发酵的金花黑茶对两种血脂异常模型大鼠体重和血脂的作用[J]. 卫生研究, 2016, 45(4):637-642.
LIU T, CHEN R, CHEN D, et al. Effect of CGMCC No. 8730 Eurotium cristatum fermented dark tea extract on body weight and blood lipid in two dyslipidemia rat models[J]. Journal of Hygiene Research, 2016, 45(4):637-642.(in Chinese)
[18] CHEN G J, XIE M H, DAI Z Q, et al. Kudingcha and Fuzhuan brick tea prevent obesity and modulate gut microbiota in high-fat diet fed mice[J]. Mol Nutr Food Res, 2018, 62(6):1700485.
[19] DU H P, WANG Q, YANG X B. Fu brick tea alleviates chronic kidney disease of rats with high fat diet consumption through attenuating insulin resistance in skeletal muscle[J]. J Agric Food Chem, 2019, 67(10):2839-2847.
[20] KANG D D, SU M, DUAN Y W, et al. Eurotium cristatum, a potential probiotic fungus from Fuzhuan brick tea, alleviated obesity in mice by modulating gut microbiota[J]. Food Funct, 2019, 10(8):5032-5045.
[21] XU L J, MENG W, CAO C, et al. Antibacterial and antifungal compounds from marine fungi[J]. Mar Drugs, 2015, 13(6):3479-3513.
[22] DU F Y, LI X, LI X M, et al. Indolediketopiperazine alkaloids from Eurotium cristatum EN-220, an endophytic fungus isolated from the marine alga Sargassum thunbergii[J]. Mar Drugs, 2017, 15(2):24.
[23] 霍娟, 李玲, 易杨华, 等. 海洋生物内生真菌中蒽醌类次生代谢产物的研究[J]. 中国海洋药物, 2010, 29(3):48-54.
HUO J, LI L, YI Y H, et al. Anthraquinone secondary metabolites from endophytic fungi[J]. Chinese Journal of Marine Drugs, 2010, 29(3):48-54.(in Chinese)
[24] 覃金球. 冠突散囊菌发酵液对金黄色葡萄球菌及其生物被膜的作用[D]. 南宁:广西医科大学, 2019.
QIN J Q. Effect of the fermentation broth of Eurotium cristatum on Staphylococcus aureus and its biofilm[D]. Nanning:Guangxi Medical University, 2019.(in Chinese)
[25] SHI J, LIU J X, KANG D D, et al. Isolation and characterization of benzaldehyde derivatives with anti-inflammatory activities from Eurotium cristatum, the dominant fungi species in fuzhuan brick tea[J]. ACS Omega, 2019, 4(4):6630-6636.
[LL] [26] DU F Y, LI X M, SONG J Y, et al. Anthraquinone derivatives and an orsellinic acid ester from the marine alga-derived endophytic fungus Eurotium cristatum EN-220[J]. Helv Chim Acta, 2014, 97(7):973-978.
[27] SHANENKO E F, EFREMENKOVA O V, MUKHAMEDZANOVA T G, et al. Using the Eurotium cristatum fungus for preparing fermented herbal teas[J]. J Pharm Nutr Sci, 2020, 10(6):341-361.
[28] 黄群, 陈林杰, 李彦坡, 等. 冠突散囊菌黑茶发酵液对消化酶活性影响的研究[J]. 微生物学通报, 2007, 34(5):917-920.
HUANG Q, CHEN L J, LI Y P, et al. Study on the effect of dark tea fermentation liquid with Eurotium cristatum on the activity of digestive enzyme[J]. Microbiology China, 2007, 34(5):917-920.(in Chinese)
[29] MA Y R, LI X Z, YANG X, et al. Comparative study on functional properties of mulberry black tea fermented by Phellinus igniarius and Eurotium cristatum[J]. Med Plants, 2022, 13(3):14-17.
[30] 胡婷婷, 刘刚, 邓钱江, 等. 基电子鼻、电子舌技术评价金花菌固态发酵对青刺尖茶感官品质的影响[J]. 现代食品科技, 2017, 33(6):285-292.
HU T T, LIU G, DENG Q J, et al. Effect of solid-state fermentation by Eurotium cristatum on the sensory quality of Prinsepia utilis tea, using E-nose/E-tongue techniques[J]. Modern Food Science Technology, 2017, 33(6):285-292.(in Chinese)
[31] STAAB J F, LEMME-DUMIT J M, LATANICH R, et al. Co-culture system of human enteroids/colonoids with innate immune cells[J]. Curr Protoc Immunol, 2020, 131(1):e113.
[32] AWASTHI V D, GOINS B, KLIPPER R, et al. Accumulation of PEG-liposomes in the inflamed colon of rats:potential for therapeutic and diagnostic targeting of inflammatory bowel diseases[J]. J Drug Target, 2002, 10(5):419-427.
[33] EASTWOOD M A. GASTROINTESTINAL T. Structure and function of the colon[J]. Encycl Human Nutr (Second Edition), 1998, 17:945-953.
[34] NAKAJIMA Y, BAUDRY N, DURANTEAU J, et al. Microcirculation in intestinal villi[J]. Am J Respir Crit Care Med, 2001, 164(8):1526-1530.

Preliminary Study of the Mechanism of Inhibition of Salmonella Typhimurium by Ec-12 Supernatant

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 14

Recommended Articles

Metrics

Comments

[1]	ZHANG Shengying, LIU Zhongli, GUO Aijiang, WANG Shuai. The Effect of Seven Protease Inhibitors on Activity of DNA Damage Inducible 1 Protein in Echinococcus multilocularis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2273-2280.
[2]	WANG Dong, LIU Kexin, HE Yanjun, DENG Shouxiang, LIU Yun, MA Weiming. Effects of Dietary Sodium Humate Supplementation on Liver Tissue Inflammation and Antioxidant Capacity of Salmonella Typhimurium-Infected Broilers [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 629-639.
[3]	QIN Lei, WU Huimin, XU Qiqi, CHEN Wanzhao, WANG Dong, LI Hongbo, XIA Panpan, LIU Zepeng, XIA Lining. Effect of Exogenous Drug-Resistant Salmonella Typhimurium on Intestinal Flora in Healthy Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2158-2169.
[4]	JIANG Zenghai, TENG Lin, HE Anwen, LIU Yanyan, YUE Min, HE Qigai. Genomic Analysis of Salmonella Typhimurium Isolates and Salmonella Serotype 4, [5], 12: i:- Isolates from Pig-borne Food Chain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1199-1209.
[5]	LI Lili, CHEN Kaifeng, CHEN Bing, ZHOU Zhouping, WANG Nanwei, QU Xiaoyun, XU Chenggang, LIAO Ming, ZHANG Jianmin. Regulatory Role of STM1827 in the Biofilm Formation and Environmental Stress of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5207-5217.
[6]	CAO Qingguo, GUO Qin, PENG Kai, CAI Jiahui, REN Chengwan, LI Yunting. The Inhibition and Mechanism of the Combination of Berberine Hydrochloride and Levofloxacin on MRSA [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3208-3220.
[7]	CAO Li, CHENG Runan, WU Zhouhui, WANG Jiawei, WANG Yu, ZHANG Yonghong, WU Qingmin, WANG Zhen. Construction and Biological Characteristics of sapC Gene Deletion Strain of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2282-2289.
[8]	ZHANG Lin, LU Fang, FU Hengfeng, JIANG Xidi, WEI Qiling, QI Caili, GAO Haixia, LI Lin. Regulation Mechanism of BaeSR on Fluoroquinolones Resistance in Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 894-903.
[9]	WU Liting, LIU Banhong, TIAN Yuan, WANG Juan, BAO Hongduo, ZHOU Yan, PANG Maoda, WANG Ran, ZHANG Hui. Identification and Environmental Inhibition Effect of Clostridium perfringens Phage [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3550-3560.
[10]	HE Yirong, ZHANG Yijie, YANG Wei, CHEN Xiaocong, ZHANG Jiaxiang, CHEN Peifu. Isolation, Identification and Biological Properties of a Lytic Phage against Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 763-771.
[11]	QI Xing-cai, QIN Xiao-dong, GAN Xiao-li, ZHANG Shu-min, MA You-ji, LI Zhi-yong. Inhibition of Type O Foot-and-mouth Disease Virus ON Strain Replication by Lentivirus-mediated siRNA [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 360-367.
[12]	LI Wen-liang, MAO Li, DENG Jia-wu, HAO Fei, LI Ji-zong, YANG Lei-lei, ZHANG Wen-wen, JIANG Jie-yuan. Porcine Anti-viral Protein Viperin Inhibits the Replication of Classical Swine Fever Virus in PK-15 Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 139-146.
[13]	YIN Zhi-hong，WANG En-feng， BAI Rui，PANG Quan-hai. Effect of RNAi on MC1R Expression in Mouse Embryo Fibroblasts [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(2): 218-224.
[14]	NI Bo, BAI Fang-fang, WEI Yan-na, LIU Mao-jun, FENG Zhi-xin,XIONG Qi-yan, WEI Jian-zhong, SHAO Guo-qing. Comparative Analysis of the Inhibitory Effects of Lipid Associated Membrane Proteins from Four Different Strains of Mycoplasma hyopneumoniae on Porcine Alveolar Epithelial Cell Lines [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(8): 1283-1287.