金黄色葡萄球菌对苯扎溴铵敏感性与生物被膜形成能力相关性分析

doi:10.11843/j.issn.0366-6964.2024.08.038

Abstract

Abstract:

Overall 35 strains of Staphylococcus aureus (S. aureus) isolated from one pig slaughterhouse and different pig farms were used as experimental materials to analyze the correlation between the susceptibility of benzalkonium bromide and the biofilm formation ability. Sensitivity to benzalkonium bromide and ability to form biofilms of S. aureus strains were determined using the micro-dilution broth method and the crystal violet staining test, respectively. The multi-locus sequence typing (MLST) and the carrying of genes related to disinfectant resistance and biofilm formation of 26 strains of S. aureus from the slaughterhouse were analyzed based on whole-genome sequences. The results showed that among the 35 strains, the minimum bactericidal concentration (MBC) of benzalkonium bromide ranged from 1 to 16 mg·L^-1, and there were 16 strains with strong biofilm-forming ability, accounting for 45.71% (16/35) of the total, of which 93.75% (15/16) were samples collected from downstream in the slaughter chain. ST7 was the dominant type of S. aureus with strong biofilm-forming ability (93.75%, 15/16). The benzalkonium bromide MBC value of S. aureus was positively correlated with biofilm formation ability (Correlation coefficient=0.690), the disinfectance has nothing to do with the gene. In conclusion, biofilm formation may increase S. aureus resistance to disinfectants, weaken the disinfection effect of benzalkonium bromide, thus enhance the risk of bacterial contamination and transmission in breeding and slaughtering.

Key words: Staphylococcus aureus, biofilm, benzalkonium bromide, correlation

CLC Number:

S852.61

Hengjie CUI, Jinlong QIN, Zhihao ZHU, Xue BAO, Shaowen LI, Xianrong MENG. Correlation Analysis of Benzalkonium Bromide Sensitivity and Biofilm Formation Ability in Staphylococcus aureus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3669-3677.

Figures/Tables 6

Table 1

Table 2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

References 30

1	史昕平, 陆利霞, 熊晓辉. 南京部分地区食品中金黄色葡萄球菌污染状况调查[J]. 食品安全质量检测学报, 2018, 9 (4): 723- 728.
	SHI X P , LU L X , XIONG X H . Investigation of Staphylococcus aureus pollution condition in food in several districts of Nanjing[J]. Journal of Food Safety & Quality, 2018, 9 (4): 723- 728.
2	IVBULE M , MIKLAŠEVIČS E , ČUPĀNE L , et al. Presence of Methicillin-resistant Staphylococcus aureus in slaughterhouse environment, pigs, carcasses, and workers[J]. J Vet Res, 2017, 61 (3): 267- 277. doi: 10.1515/jvetres-2017-0037
3	陈星雨. 湖北省生猪养殖场和屠宰场金黄色葡萄球菌流行情况及耐药性研究[D]. 武汉: 华中农业大学, 2020.
	CHEN X Y. Prevalence and drug resistance of Staphylococcus aureus in pig farms and slaughterhouses in Hubei province[D]. Wuhan: Huazhong Agricultural University, 2020. (in Chinese)
4	张卓娜, 孙琦, 杨艳伟. 国内市场季铵盐类消毒剂使用情况调查[J]. 中国消毒学杂志, 2018, 35 (1): 23- 25.
	ZHANG Z N , SUN Q , YANG Y W . Investigation on the using situation of quaternary ammonium disinfectant from domestic market[J]. Chinese Journal of Disinfection, 2018, 35 (1): 23- 25.
5	杨磊, 王祎, 杨郇垚, 等. 非洲猪瘟疫情影响下生猪定点屠宰企业消毒剂的选择[J]. 农业技术与装备, 2020, (10): 136- 137.
	YANG L , WANG Y , YANG X Y , et al. Selection of disinfectants for pig slaughtering enterprises under the influence of African swine plague[J]. Agricultural Technology & Equipment, 2020, (10): 136- 137.
6	陈姿颐. 武汉某生猪屠宰场金黄色葡萄球菌消毒剂抗性研究[D]. 武汉: 华中农业大学, 2020.
	CHEN Z Y. Study on disinfectant resistance of Staphylococcus aureus strains isolated from a pig slaughterhouse in Wuhan[D]. Wuhan: Huazhong Agricultural University, 2020. (in Chinese)
7	SCHILCHER K , HORSWILL A R . Staphylococcal biofilm development: structure, regulation, and treatment strategies[J]. Microbiol Mol Biol Rev, 2020, 84 (3): e00026- 19.
8	BRIDIER A , BRIANDET R , THOMAS V , et al. Resistance of bacterial biofilms to disinfectants: a review[J]. Biofouling, 2011, 27 (9): 1017- 1032. doi: 10.1080/08927014.2011.626899
9	POMARANSKI E K , SOTO E . The formation, persistence, and resistance to disinfectant of the Erysipelothrix piscisicarius biofilm[J]. J Aquat Anim Health, 2020, 32 (1): 44- 49. doi: 10.1002/aah.10097
10	STEPANOVIĆ S , VUKOVIĆ D , DAKIĆ I , et al. A modified microtiter-plate test for quantification of staphylococcal biofilm formation[J]. J Microbiol Methods, 2000, 40 (2): 175- 179. doi: 10.1016/S0167-7012(00)00122-6
11	BANKEVICH A , NURK S , ANTIPOV D , et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing[J]. J Comput Biol, 2012, 19 (5): 455- 477. doi: 10.1089/cmb.2012.0021
12	GUREVICH A , SAVELIEV V , VYAHHI N , et al. QUAST: quality assessment tool for genome assemblies[J]. Bioinformatics, 2013, 29 (8): 1072- 1075. doi: 10.1093/bioinformatics/btt086
13	SEEMANN T . Prokka: rapid prokaryotic genome annotation[J]. Bioinformatics, 2014, 30 (14): 2068- 2069. doi: 10.1093/bioinformatics/btu153
14	FITCH W M . Homology: a personal view on some of the problems[J]. Trends Genet, 2000, 16 (5): 227- 231. doi: 10.1016/S0168-9525(00)02005-9
15	JIANG X B , YU T , LIANG Y , et al. Efflux pump-mediated benzalkonium chloride resistance in Listeria monocytogenes isolated from retail food[J]. Int J Food Microbiol, 2016, 217, 141- 145. doi: 10.1016/j.ijfoodmicro.2015.10.022
16	RUSSELL A D . Bacterial resistance to disinfectants: present knowledge and future problems[J]. J Hosp Infect, 1999, 43 Suppl 1, S57- S68.
17	中华人民共和国农业农村部. NY/T 3384-2021畜禽屠宰企业消毒规范[S]. 北京: 中国农业出版社, 2021.
	Ministry of Agriculture and Rural Affairs of the People's Republic of China. NY/T 3384-2021 Specification for disinfection of livestock and poultry slaughtering enterprises[S]. Beijing: China Agriculture Press, 2021. (in Chinese)
18	WORTHING K A , MARCUS A , ABRAHAM S , et al. Qac genes and biocide tolerance in clinical veterinary methicillin-resistant and methicillin-susceptible Staphylococcus aureus and Staphylococcus pseudintermedius[J]. Vet Microbiol, 2018, 216, 153- 158. doi: 10.1016/j.vetmic.2018.02.004
19	DE QUADROS C L , MANTO L , MISTURA E , et al. Antimicrobial and disinfectant susceptibility of Salmonella serotypes isolated from swine slaughterhouses[J]. Curr Microbiol, 2020, 77 (6): 1035- 1042. doi: 10.1007/s00284-020-01904-9
20	FLEMMING H C , WUERTZ S . Bacteria and archaea on Earth and their abundance in biofilms[J]. Nat Rev Microbiol, 2019, 17 (4): 247- 260. doi: 10.1038/s41579-019-0158-9
21	MARINO M , FRIGO F , BARTOLOMEOLI I , et al. Safety-related properties of Staphylococci isolated from food and food environments[J]. J Appl Microbiol, 2011, 110 (2): 550- 561. doi: 10.1111/j.1365-2672.2010.04909.x
22	LEBEAUX D , GHIGO J M , BELOIN C . Biofilm-related infections: bridging the gap between clinical management and fundamental aspects of recalcitrance toward antibiotics[J]. Microbiol Mol Biol Rev, 2014, 78 (3): 510- 543. doi: 10.1128/MMBR.00013-14
23	张阳. 猪源金黄色葡萄球菌的毒力基因分布, 生物被膜形成和凝血致病性的研究[D]. 广州: 华南理工大学, 2018.
	ZHANG Y. The distribution of virulence genes, biofilm formation and coagulation pathogenicity of porcine Staphylococcus aureus[D]. Guangzhou: South China University of Technology, 2018. (in Chinese)
24	JUZWA W , MYSZKA K , BIAŁAS W , et al. Investigation of the effectiveness of disinfectants against planktonic and biofilm forms of P. aeruginosa and E. faecalis cells using a compilation of cultivation, microscopic and flow cytometric techniques[J]. Biofouling, 2015, 31 (7): 587- 597. doi: 10.1080/08927014.2015.1075126
25	唐子云, 胡健欣, 陈进, 等. 动物源金黄色葡萄球菌生物被膜形成能力与分子分型关系研究[J]. 中国农业科学, 2022, 55 (3): 602- 612.
	TANG Z Y , HU J X , CHEN J , et al. Relationship between biofilm formation and molecular typing of Staphylococcus aureus from animal origin[J]. Scientia Agricultura Sinica, 2022, 55 (3): 602- 612.
26	KOCOT A M , OLSZEWSKA M A . Interaction of Pseudomonas aeruginosa and Staphylococcus aureus with Listeria innocua in dual species biofilms and inactivation following disinfectant treatments[J]. LWT, 2020, 118, 108736. doi: 10.1016/j.lwt.2019.108736
27	RESCH A , FEHRENBACHER B , EISELE K , et al. Phage release from biofilm and planktonic Staphylococcus aureus cells[J]. FEMS Microbiol Lett, 2005, 252 (1): 89- 96. doi: 10.1016/j.femsle.2005.08.048
28	WANG R , SCHMIDT J W , HARHAY D M , et al. Biofilm formation, antimicrobial resistance, and sanitizer tolerance of Salmonella enterica strains isolated from beef trim[J]. Foodborne Pathog Dis, 2017, 14 (12): 687- 695. doi: 10.1089/fpd.2017.2319
29	WESSELS S , INGMER H . Modes of action of three disinfectant active substances: a review[J]. Regul Toxicol Pharmacol, 2013, 67 (3): 456- 467. doi: 10.1016/j.yrtph.2013.09.006
30	GANESHNARAYAN K , SHAH S M , LIBERA M R , et al. Poly-N-acetylglucosamine matrix polysaccharide impedes fluid convection and transport of the cationic surfactant cetylpyridinium chloride through bacterial biofilms[J]. Appl Environ Microbiol, 2009, 75 (5): 1308- 1314. doi: 10.1128/AEM.01900-08

来源 Source	菌株分离数/株 No. of strains	菌株编号(2019-) Strain No.(2019-)
养殖场生猪鼻腔拭子 Nasal swabs from farms	9	01100、01101、01103、01109、01115、0262、0277、0284、0289
电麻环节生猪鼻腔拭子 Nasal swabs after stunning	3	01159、01161、01211
脱毛环节生猪体表拭子 Carcass swabs after dehairing	7	01376、01396、02316、02321、02330、02331、02349
烫毛池水样 Scald water sample	1	02S3
卸头环节生猪鼻腔拭子 Nasal swabs after head-removal	3	01229、01262、01264
劈半环节生猪肉样 Pork samples after splitting	2	0145、0159
劈半环节生猪体表拭子 Carcass swabs after splitting	4	01417、01422、01455、02401
冷库环节生猪肉样 Pork samples from the cold storage	2	0162、0166
冷库环节生猪体表拭子 Carcass swabs from the cold storage	4	01505、01545、02488、02550

菌株编号(2019-) Strain No.(2019-)	来源 Source	苯扎溴铵敏感性 Sensitivity to benzalkonium bromide		生物被膜检测 Detection of biofilms
菌株编号(2019-) Strain No.(2019-)	来源 Source	MBC/(mg·L^-1)	MBC/MBC_{ATCC 29213}	D_{630 nm}	成膜能力 Biofilm formation ability
0145	劈半环节肉样	16	8	1.108	+++
0159	劈半环节肉样	16	8	0.708	+++
0162	冷库环节肉样	16	8	0.178	++
0166	冷库环节肉样	16	8	0.993	+++
01159	电麻环节鼻腔拭子	4	2	0.198	++
01161	电麻环节鼻腔拭子	4	2	0.125	+
01211	电麻环节鼻腔拭子	1	0.5	0.074	+
01229	卸头环节鼻腔拭子	1	0.5	0.161	++
01262	卸头环节鼻腔拭子	4	2	0.073	+
01264	卸头环节鼻腔拭子	16	8	1.626	+++
01376	脱毛环节体表拭子	4	2	0.079	+
01396	脱毛环节体表拭子	2	1	0.175	++
01417	劈半环节体表拭子	16	8	1.352	+++
01422	劈半环节体表拭子	16	8	1.190	+++
01455	劈半环节体表拭子	16	8	0.662	+++
01505	冷库环节体表拭子	16	8	1.076	+++
01545	冷库环节体表拭子	16	8	1.631	+++
02316	脱毛环节体表拭子	8	4	1.299	+++
02321	脱毛环节体表拭子	1	0.5	0.204	++
02330	脱毛环节体表拭子	1	0.5	0.195	++
02331	脱毛环节体表拭子	8	4	2.020	+++
02349	脱毛环节体表拭子	8	4	1.757	+++
02401	劈半环节体表拭子	8	4	2.055	+++
02488	冷库环节体表拭子	8	4	0.468	+++
02550	冷库环节体表拭子	2	1	0.143	+
02S3	烫毛池水样	8	4	1.345	+++
01100	养殖场鼻腔拭子	4	2	0.114	+
01101	养殖场鼻腔拭子	2	1	0.145	++
01103	养殖场鼻腔拭子	2	1	0.145	++
01109	养殖场鼻腔拭子	2	1	0.159	++
01115	养殖场鼻腔拭子	4	2	0.151	++
0262	养殖场鼻腔拭子	4	2	0.170	++
0277	养殖场鼻腔拭子	2	1	0.348	+++
0284	养殖场鼻腔拭子	2	1	0.119	+
0289	养殖场鼻腔拭子	2	1	0.169	++

[1]	Yue LI, Changchun ZHANG, Guangyu LIU, Mengyuan GAO, Chaojun FU, Jiabao XING, Sijia XU, Qiyuan KUANG, Jing LIU, Xiaopeng GAO, Heng WANG, Lang GONG, Guihong ZHANG, Yankuo SUN. Application and Analysis of Meta-transcriptomics Sequencing Technology in the Diagnosis of Viral Diarrhea Diseases in Piglets [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3579-3589.
[2]	XIAN Tingting, LIU Yan, CAO Xin, FENG Tao. Analysis of the Changes of Vaginal Microflora and Serum Pro-inflammatory Cytokines and Their Correlation in Sows with Endometritis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3688-3698.
[3]	Yinuo WANG, Dan XU, Jianhua YANG, Yang LIU, Yaofu TIAN, Xiaoling ZHAO. Research on a Breeding Method for Predicting Meat Yield Performance of Broilers Based on Ultrasound Measurement of Pectoral Muscle Thickness [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 2901-2912.
[4]	LONG Tanghui, ZHOU Jianghui, ZHAN Yanbo, ZHANG Jian, ZHAO Xianghui, LI Yanjiao, OUYANG Kehui, QIU Qinghua. Research Progress on LuxS/AI-2 Quorum Sensing of Rumen Microbe in Ruminants [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1893-1903.
[5]	HE Xiaolan, ZHAO Yankun, MENG Lu, LIU Huimin, GAO Jiaojiao, ZHENG Nan. Research Progress in Heteroresistance of Staphylococcus aureus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1432-1445.
[6]	XIA Shuwen, CHEN Kunlin, SHEN Yangyang, AN Zhenjiang, ZHAO Fang, DING Qiang, ZHONG Jifeng, LIN Zhiping, WANG Huili. The Estimation of Genetic Parameters for Longevity Traits of Holstein Cows in Jiangsu Region [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1030-1039.
[7]	WU Zihao, CAI Yilong, TUO Haixin, CHEN Wei. Pathogenicity Analysis of a PVL⁺ ST22 Staphylococcus aureus Isolated from Equine Raw Milk [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 718-726.
[8]	BI Yazhen, SHANG Mingyu, HU Wenping, ZHANG Li. Correlation and Regression Analysis among Growth Traits and Association Analysis of TRHDE Gene Polymorphism with Growth Traits in Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1415-1428.
[9]	GUO Mingpeng, MENG Yuan, WANG Honghao, WANG Yuan, CHE Leijie, SHEN Li, WANG Xi. Estimation of Genetic Parameters of Body Weight and Body Size Traits in Jinnan Cattle at Different Growth Stages [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1452-1464.
[10]	HAN Yunzhen, YANG Wenpan, HONG Yuan, LONG Yi, LIU Xiangjie, FAN Xiaoping, LI Wenjing, DENG Zheng, LIU Minghui, ZHENG Sumei, RUAN Guorong, DING Nengshui. Estimating the Inbreeding Coefficient of Huai Pigs by Different Methods Based on Chip Data [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 947-955.
[11]	YU Shiqiang, LI Liuxue, ZHAO Xiaobo, ZHAO Huiying, TU Yan, ZHAO Yuchao, JIANG Linshu. Differences and Correlations of Lactation Performance in Chinese Holstein Dairy Cows at Different Lactation Stages and Somatic Levels [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1003-1014.
[12]	HAO Ruochen, TANG Minjia, LIU Guangliang, ZHANG Yan, MUHAMMAD Shoaib, SHANG Ruofeng, CAO Zongxi, PU Wanxia. Distribution and Genotyping of Major Enterobacteriaceae Bacteria in Milk Sources and Dairy Farm Environment of Hainan Province [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5184-5197.
[13]	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.
[14]	WU Zhouhui, WANG Yu, DU Heng, WANG Zhiwen, XIAO Shuang, WU Jinliang, WANG Zhen. Analysis of the Antibacterial Sensitization Activity of Tirapazamine against Multi-Drug Resistant Salmonella [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4362-4371.
[15]	JIANG Nansong, JI Xing, WANG Yaxin, SUN Chengtao, WANG Yang, CHEN Hongmei, CHENG Longfei, HUANG Yu, WU Congming. Prevalence and Transduction of Prophages in Methicillin-resistant Staphylococcus aureus ST9 of Swine Origin [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 338-350.

Correlation Analysis of Benzalkonium Bromide Sensitivity and Biofilm Formation Ability in Staphylococcus aureus

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 30

Related Articles 15

Recommended Articles

Metrics

Comments