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

摘要/Abstract

摘要：

以35株生猪屠宰场及养殖场来源的金黄色葡萄球菌分离菌株为试验材料，分析金黄色葡萄球菌对苯扎溴铵敏感性与生物被膜形成能力的相关性。采用微量肉汤稀释法检测金黄色葡萄球菌对苯扎溴铵的敏感性，结晶紫染色法检测生物被膜形成能力，基于全基因组测序结果分析屠宰环节的26株金黄色葡萄球菌多位点序列分型(multi-locus sequence typing，MLST)情况，以及消毒剂抗性与生物被膜形成相关基因的携带情况。结果表明，35株菌中，苯扎溴铵的最小杀菌浓度(minimum bactericidal concentration，MBC)范围为1~16 mg·L^-1，强成膜能力菌株共16株，占总数的45.71%(16/35)，其中93.75%(15/16)来自于屠宰链的下游。ST7是强成膜能力菌株的优势序列型(93.75%，15/16)。金黄色葡萄球菌生物被膜形成能力和苯扎溴铵MBC值呈正相关(相关系数为0.690)，而消毒剂抗性与相关基因携带情况无关。综上，金黄色葡萄球菌可通过形成生物被膜提高对消毒剂的耐受性，减弱了苯扎溴铵的消毒效果，增加了养殖和屠宰环节中细菌污染和传播的风险。

关键词: 金黄色葡萄球菌, 生物被膜, 苯扎溴铵, 相关性

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

中图分类号:

S852.61

崔恒洁, 覃金珑, 朱志豪, 鲍雪, 栗绍文, 孟宪荣. 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[J]. 畜牧兽医学报, 2024, 55(8): 3669-3677.

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.

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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	++

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