Study on antibacterial and quorum quenching activity of a Pseudoalteromonas sp. DL3

doi:10.3969/j.issn.1004-1524.2023.01.06

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (1): 50-57.DOI: 10.3969/j.issn.1004-1524.2023.01.06

• Animal Science • Previous Articles Next Articles

Study on antibacterial and quorum quenching activity of a Pseudoalteromonas sp. DL3

ZHAO Saisai¹(), ZHANG Haojie¹, CAI Xiulei¹, WANG Lirong², SHEN Xiaoran², CAO Zhi¹, SHAN Hu¹()

1. College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, Shandong, China
2. Shandong Bilan Biotechnology Co., Ltd., Tai’an 271411, Shandong, China

Received:2022-05-24 Online:2023-01-25 Published:2023-02-21

Abstract

Abstract:

In this study, a strain DL3 was isolated from the microbial mucous membrane of antifouling attached steel sheet in aquaculture ponds. In order to clarify the activity of this strain and its potential application as beneficial bacteria, it was preliminarily identified by bacterial isolation culture, 16S rRNA sequencing and phylogenetic analysis. Oxford cup method and plate method were used to determine the bacteriostatic activity and quorum quenching activity. Based on phylogenetic analysis of the 16S rRNA gene sequence of strain DL3, this strain had the highest similarity with Pseudoalteromonas peptidolytica NBRC 101021T, with the closest affinity of 99.64%. The bacteriostasis test showed that the strain could significantly inhibit the growth of V. anguillarum and Pseudomonas aeruginosa, and showed that could inhibit the growth of Staphylococcus aureus, Escherichi coli, Salmonella, Streptococcus, Shigella, V. fluvialis, V. orientalis, V. harveyi and Bacillus pumilus to different degrees, and the antibacterial activity was strongest when cultured till 72 h. From the results of plate test, the strain DL3 had quorum quenching activity and could quench signal molecules such as 3-OXO-C6-HSL, C10-HSL, C12-HSL and 3-OXO-C14-HSL. In this study, the strain DL3 was successfully isolated and its antagonistic activity against pathogenic bacteria was clarified, which laid a foundation for further studies on the antibacterial mechanism of strain DL3 and its active products and the application of marine microorganisms in the prevention and control of animal bacterial diseases.

Key words: Pseudoalteromonas, antibacterial activity, quorum quenching

CLC Number:

S182

ZHAO Saisai, ZHANG Haojie, CAI Xiulei, WANG Lirong, SHEN Xiaoran, CAO Zhi, SHAN Hu. Study on antibacterial and quorum quenching activity of a Pseudoalteromonas sp. DL3[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 50-57.

Figures/Tables 5

Table 1 Names and numbers of 23 clinical pathogen strains

菌种名称 Strain Name	菌株号 Strain No.	数量 Number
Escherichia coli	E1	1
E. coli	E2	1
E. coli	E3	1
E. coli	E4	1
Staphylococcus aureus	PMJ4-1	1
S. aureus	PMJ10-1	1
S. aureus	PMJ30-2	1
S. aureus	TTB1-1	1
S. aureus	TTB2-1	1
S. aureus	QTB1-1	1
Vibrio anguillarum	VIB72	1
V. fluvialis	VIB292	1
V. harveyi	VIB295	1
V. orientalis	VIB303	1
Bacillus pumilus	FYB01	1
Shigella	T3	1
Pseudomonas aeruginosa	PA2	1
Pseudomonas aeruginosa	PA1	1
Salmonella	QS1	1
Streptococcus	SS-1	1
Streptococcus	SS-2	1
Streptococcus	SS-7	1
Streptococcus	SS-9	1
总计Total		23

Fig.1 Antibacterial activity of strain DL3

Fig.2 Phylogenetic tree of strain DL3 based on 16S rRNA gene sequences by neighbour-joining method “●” indicated that the corresponding nodes are also recovered in tree generated with the ML and MP algorithms; “○” indicated that the corresponding nodes were also recovered in tree generated with the ML algorithm.

Fig.3 Effects of different culture time on biomass, protein concentration and antibacterial activity of strain DL3 A, Effect of culture time on the biomass of strain DL3; B, Effect of culture time on the protein concentration of strain DL3; C. Effect of culture time on the antibacterial activity of strain DL3. “**” meant significant difference (P<0.01), “ns” meant no significant difference (P>0.05).

Fig.4 Determination results of AHL degradation characteristics of strain DL3

References 31

[1]	黄智森. 浅析水产养殖病害及其药物控制[J]. 江西水产科技, 2019(3): 40-41.
	HUANG Z S. Analysis of aquaculture diseases and drug control[J]. Jiangxi Fishery Science and Technology, 2019(3): 40-41. (in Chinese)
[2]	HOSSAIN A, HABIBULLAH-AL-MAMUN M, NAGANO I, et al. Antibiotics, antibiotic-resistant bacteria, and resistance genes in aquaculture: risks, current concern, and future thinking[J]. Environmental Science and Pollution Research International, 2022, 29(8): 11054-11075. DOI PMID
[3]	马有录. 试论我国畜牧业生产中限制抗生素的使用问题[J]. 畜禽业, 2019, 30(8): 28.
	MA Y L. Restriction of antibiotic use in animal husbandry production in China[J]. Livestock and Poultry Industry, 2019, 30(8): 28. (in Chinese)
[4]	KALIA V C, et al. Quorum sensing inhibitors as antipathogens: biotechnological applications[J]. Biotechnology Advances, 2019, 37(1): 68-90. DOI PMID
[5]	MIDHUN S J, NEETHU S, VYSAKH A, et al. Antibacterial activity and probiotic characterization of autochthonous Paenibacillus polymyxa isolated from Anabas testudineus(Bloch, 1792)[J]. Microbial Pathogenesis, 2017, 113: 403-411. DOI URL
[6]	杨行, 章翔, 龙昊, 等. 假交替单胞菌对哈维氏弧菌拮抗作用初探[J]. 水产科学, 2019, 38(6): 833-838.
	YANG H, ZHANG X, LONG H, et al. Antagonistic mechanism of Pseudoalteromonas sp against Vibrio harveyi.[J]. Fisheries Science, 2019, 38(6): 833-838. (in Chinese with English abstract)
[7]	RICHARDS G P, WATSON M A, NEEDLEMAN D S, et al. Mechanisms for Pseudoalteromonas piscicida-induced killing of vibrios and other bacterial pathogens[J]. Applied and Environmental Microbiology, 2017, 83(11): AEM.00175-17.
[8]	SKOVHUS T L, RAMSING N B, HOLMSTRÖM C, et al. Real-time quantitative PCR for assessment of abundance of Pseudoalteromonas species in marine samples[J]. Applied and Environmental Microbiology, 2004, 70(4): 2373-2382. DOI URL
[9]	张天震, 刘伶普, 李文超, 等. 群体感应系统介导细菌生物膜形成的研究进展[J]. 生物加工过程, 2020, 18(2): 177-183.
	ZHANG T Z, LIU L P, LI W C, et al. Advances in quorum sensing regulating formation of biofilm[J]. Chinese Journal of Bioprocess Engineering, 2020, 18(2): 177-183. (in Chinese with English abstract)
[10]	CAO J G, et al. Purification and structural identification of an autoinducer for the luminescence system of Vibrio harveyi[J]. Journal of Biological Chemistry, 1989, 264(36): 21670-21676. DOI URL
[11]	NEALSON K H, PLATT T, HASTINGS J W. Cellular control of the synthesis and activity of the bacterial luminescent system[J]. Journal of Bacteriology, 1970, 104(1): 313-322. DOI PMID
[12]	LI S J, WU S, REN Y, et al. Characterization of differentiated autoregulation of LuxI/LuxR-type quorum sensing system in Pseudoalteromonas[J]. Biochemical and Biophysical Research Communications, 2022, 590: 177-183. DOI URL
[13]	崔天琦, 吕欣然, 孙梦桐, 等. 降解嗜水气单胞菌群体感应AHLs信号分子的乳酸菌筛选及淬灭作用[J]. 中国食品学报, 2020, 20(9): 19-29.
	CUI T Q, LYU X R, SUN M T, et al. Screening and quorum quenching of lactic acid bacteria with degrading Aeromonas hydrophila quorum sensing AHLs activity[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(9): 19-29. (in Chinese with English abstract)
[14]	GRANDCLÉMENT C, TANNIÈRES M, MORÉRA S, et al. Quorum quenching: role in nature and applied developments[J]. FEMS Microbiology Reviews, 2015, 40(1): 86-116. DOI URL
[15]	SAURAV K, COSTANTINO V, VENTURI V, et al. Quorum sensing inhibitors from the sea discovered using bacterial N-acyl-homoserine lactone-based biosensors[J]. Marine Drugs, 2017, 15(3): 53. DOI URL
[16]	袁宗胜. 毛竹内生拮抗细菌筛选及其16S rDNA鉴定[J]. 安徽农学通报, 2021, 27(17): 34-37.
	YUAN Z S. Screening of endophytic antagonistic bacteria in Phyllostachys edulis and identification of 16S rDNA[J]. Anhui Agricultural Science Bulletin, 2021, 27(17): 34-37. (in Chinese with English abstract)
[17]	王艺颖, 丛丽娜, 王伟, 等. 产生物活性物质海洋放线菌的筛选和鉴定[J]. 工业微生物, 2017, 47(3): 30-35.
	WANG Y Y, CONG L N, WANG W, et al. Screening and identification of active substances from marine actinomycetes[J]. Industrial Microbiology, 2017, 47(3): 30-35. (in Chinese with English abstract)
[18]	王丽, 雷娟, 向文良, 等. 一株产紫色素放线菌的鉴定及其色素特性研究[J]. 应用与环境生物学报, 2009, 15(1): 139-142.
	WANG L, LEI J, XIANG W L, et al. Identification and characterization of purple pigment-producing actinomycete strain[J]. Chinese Journal of Applied & Environmental Biology, 2009, 15(1): 139-142. (in Chinese with English abstract)
[19]	蔡秀磊, 单虎, 于敏, 等. 褐杆菌L2的抑菌活性与抑菌蛋白研究[J]. 中国海洋大学学报(自然科学版), 2018, 48(10): 26-31.
	CAI X L, SHAN H, YU M, et al. The research of the antibacterial activity and protein of Phaeobacter L2[J]. Periodical of Ocean University of China, 2018, 48(10): 26-31. (in Chinese with English abstract)
[20]	汤开浩. 利用密度感应抑制技术控制水产养殖细菌性病害的基础研究[D]. 青岛: 中国海洋大学, 2014.
	TANG K H. The basic studies on quorum quenching as a novel tool to fight against bacterial infections in aquaculture[D]. Qingdao: Ocean University of China, 2014. (in Chinese with English abstract)
[21]	孙武, 马世科, 王书祥. 替抗在畜禽养殖业中的研究进展[J]. 饲料研究, 2021, 44(16): 141-144.
	SUN W, MA S K, WANG S X. Recent advances on antibiotic substitutes in livestock and poultry industry[J]. Feed Research, 2021, 44(16): 141-144. (in Chinese with English abstract)
[22]	粟胜兰, 饶正华. 长期用力综合施策推动禁抗减抗[J]. 中国畜牧业, 2020(22): 24-26.
	SU S L, RAO Z H. Promoting prohibition and reduction of antibiotics usage by long term comprehensive measures[J]. China Animal Industry, 2020(22): 24-26. (in Chinese)
[23]	GAUTHIER G, GAUTHIER M, CHRISTEN R. Phylogenetic analysis of the genera Alteromonas, Shewanella, and Moritella using genes coding for small-subunit rRNA sequences and division of the genus Alteromonas into two genera, Alteromonas(emended) and Pseudoalteromonas gen. nov., and proposal of twelve new species combinations[J]. International Journal of Systematic Bacteriology, 1995, 45(4): 755-761. DOI URL
[24]	于敏. 金丽假交替单胞菌JG1抑菌机理研究[D]. 青岛: 中国海洋大学, 2013.
	YU M. The antibacterial mechanism of Pseudoalteromonas flavipulchra JG1[D]. Qingdao: Ocean University of China, 2013. (in Chinese with English abstract)
[25]	HAYASHIDA-SOIZA G, UCHIDA A, MORI N, et al. Purification and characterization of antibacterial substances produced by a marine bacterium Pseudoalteromonas haloplanktis strain[J]. Journal of Applied Microbiology, 2008, 105(5): 1672-1677. DOI URL
[26]	WANG H L. Pseudoalteromonas probiotics as potential biocontrol agents improve the survival of Penaeus vannamei challenged with acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus[J]. Aquaculture, 2018, 494: 30-36. DOI URL
[27]	王枫林, 王秀华, 张宇哲, 等. 杀鱼假交替单胞菌2515的抗弧菌效果及在对虾养殖中的应用[J]. 中国水产科学, 2021, 28(7): 903-913.
	WANG F L, WANG X H, ZHANG Y Z, et al. Anti-Vibrio effect of Pseudoalteromonas piscicida 2515 and its application in shrimp culture[J]. Journal of Fishery Sciences of China, 2021, 28(7): 903-913. (in Chinese with English abstract)
[28]	HETTIARACHCHI S, LEE S J, LEE Y, et al. A rapid and efficient screening method for antibacterial compound-producing bacteria[J]. Journal of Microbiology and Biotechnology, 2017, 27(8): 1441-1448. DOI PMID
[29]	BUSETTI A, SHAW G, MEGAW J, et al. Marine-derived quorum-sensing inhibitory activities enhance the antibacterial efficacy of tobramycin against Pseudomonas aeruginosa[J]. Marine Drugs, 2014, 13(1): 1-28. DOI URL
[30]	唐杨. 假交替单胞菌胞外产物的特性分析及其抑菌蛋白的克隆与表达[D]. 上海: 上海海洋大学, 2017.
	TANG Y. Analysis of extracellular product characteristics of Pseudoalteromonas and cloning and expression of its antibacterial protein[D]. Shanghai: Shanghai Ocean University, 2017. (in Chinese with English abstract)
[31]	杨登辉, 孔里程, 孙建和, 等. 密度感应系统: 对细菌致病力的自行调控[J]. 微生物学通报, 2017, 44(12): 3007-3014.
	YANG D H, KONG L C, SUN J H, et al. Quorum sensing: an auto regulator for bacterial pathogenicity[J]. Microbiology China, 2017, 44(12): 3007-3014. (in Chinese with English abstract)

Study on antibacterial and quorum quenching activity of a Pseudoalteromonas sp. DL3

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 5

References 31

Related Articles 10

Recommended Articles

Metrics

Comments

[1]	TANG Xiao, MA Ming. Antibacterial activity and stability of Extracts of four common rice dumplings leaves [J]. Acta Agriculturae Zhejiangensis, 2022, 34(6): 1287-1397.
[2]	WANG Lifang, YE Liang, XIE Zhongwen, DANG Xiangli. Antibacterial activity of tea antimicrobial peptide extraction and its effect on preservation of chilled meat [J]. Acta Agriculturae Zhejiangensis, 2022, 34(10): 2268-2276.
[3]	JIANG Yuhang, XIN Weigang, ZHANG Qili, DENG Xianyu, WANG Feng, LIN Lianbing. Isolation and identification of fungi from mildewed feed corn and study on anti-mildew and antifungal effects of lactobacillin [J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1283-1291.
[4]	LI Yu, LIU Cuijun, AI Lunqiang, FANG Xinyue, LIU Yu, YUAN Mingyuan, RAN Yalan, HE Meijun. Study on volatile components and antibacterial activity of ethyl acetate component of Fritillaria hupehensis Hsiao et K. C. Hsia [J]. Acta Agriculturae Zhejiangensis, 2021, 33(6): 1088-1094.
[5]	YUE Denggao, CHENG Liang, GUO Qingyun. Antibacterial activities and optimization of fermentation conditions of lipopeptides produced by Aureobasidium pullulans PA-2 [J]. Acta Agriculturae Zhejiangensis, 2021, 33(3): 479-489.
[6]	WEN Huiping, XIAO Jianzhong, LEI Weimin, JI Jiana. Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.H by HPLC combined with response surface methodology and its antibacterial activity [J]. , 2018, 30(2): 298-306.
[7]	JIANG Han, LIU Jiao, LI Ping, GU Qing. Heterologous expression and purification of plantaricin JK and analysis of its antibacterial activity [J]. , 2017, 29(2): 332-337.
[8]	QIAN Zhuoquan;YIN Haozhen;HUANG Xiaolin;LIN Shaozhen;NAN Haihan;*. Antibacterial and antioxidant bioassay of three kinds of algae [J]. , 2014, 26(2): 0-384387.
[9]	WU Jing;LIANG Yong\\|li;SHI Yyu\\|ying;LI Yyu\\|feng;MA Xiu\\|li;JIANG Yi\\|fei;SONG Min\\|xun;* . Fusion expression of Silkie Cathelicidins genes in Escherichia coli and detection of their antibacterial activity [J]. , 2013, 25(6): 0-1214.
[10]	LIU Hui-hui;LI Shu-ping;ZHAO Qian;ZHAO Shu-jiang;*. Identification of marine fungal strain NJ0104 with the anti-microbial activities of their metabolites to bacterial pathogen of Pseudosciaena [J]. , 2012, 24(5): 0-807.