牙周炎患犬口腔菌群差异性及卟啉单胞菌耐药性分析

doi:10.11843/j.issn.0366-6964.2025.02.041

摘要/Abstract

摘要：

牙周炎是犬常见的口腔疾病之一，会导致不可逆转的牙周组织丢失、牙周附着力降低，严重影响动物福利。本研究旨在了解牙周炎患犬和牙周健康犬口腔菌群丰度及药物敏感性差异，探究犬牙周炎特征性致病菌及其药物敏感性、耐药基因携带情况。通过采集牙周炎患犬与牙周健康犬牙菌斑，采用16S rDNA测序方法进行口腔菌群差异性分析、确定菌群药物敏感性、明确特征性致病菌。并对致病菌进行分离鉴定及药物敏感性试验。结果表明，与牙周健康犬相比，牙周炎患犬口腔菌群中龈下菌斑数目显著高于龈上菌斑，拟杆菌门和螺旋体门相对丰度有所增加，属水平上卟啉单胞菌属相对丰度明显增加，提示其可能为牙周炎潜在致病菌。菌群药敏结果显示，龈下需氧菌对阿莫西林、恩诺沙星等抗生素敏感性最高，甲硝唑敏感性最低；龈下厌氧菌对恩诺沙星敏感性中等，甲硝唑敏感性最低。分离到的可疑致病菌与猴卟啉单胞菌(P. macacae-DSM20710)相似性达98.89%，卟啉单胞菌对恩诺沙星敏感，含有大环内酯类(ermF)和四环素类(tetQ)两种耐药基因。犬牙周炎的临床治疗可首选恩诺沙星。

关键词: 犬牙周炎, 菌群分离鉴定, 药物敏感性试验, 耐药基因

Abstract:

Periodontitis is one of the common oral diseases in dogs, which can cause irreversible periodontal tissue loss and reduced periodontal adhesion, and seriously affect animal welfare. This study was designed to understand the drug resistance status oral flora in canine periodontitis; and to explore the drug sensitivity and drug resistance genes of characteristic pathogenic bacteria of canine periodontitis. This trial will be randomly selected periodontitis-infected dogs and healthy dogs to collect dental plaque and conduct drug sensitivity tests. In addition, 16S rDNA sequencing was performed on oral samples. The differences in oral flora between periodontitis-infected dogs and healthy dogs were explored, and the possible characteristic pathogenic bacteria causing canine periodontitis were identified. The drug susceptibility of the oral flora was determined. Then, the isolation, identification and drug sensitivity test were performed. The results showed that the relative abundance of Bacteroidetes and Spirochaeta increased at the level of oral flora, and the relative abundance of Porphyromonas at the genus level was significantly increased. It was speculated that the potential pathogenic bacteria of canine periodontitis was Porphyromonas. The subgingival aerobic bacteria had the highest sensitivity to amoxicillin and enrofloxacin, and had the lowest sensitivity to metronidazole. The sensitivity of subgingival anaerobes to enrofloxacin was medium, and the sensitivity to metronidazole was lowest. P. clinical is sensitive to enrofloxacin and contains two resistance genes ermF and tetQ. Enrofloxacin is preferred in the clinical treatment of canine periodontitis.

Key words: canine periodontitis, isolation and identification of flora, drug sensitivity test, drug resistance gene

中图分类号:

S852.6

杜清洁, 吴礼平, 张帆, 戴鹏秀, 冯献程, 张欣珂. 牙周炎患犬口腔菌群差异性及卟啉单胞菌耐药性分析[J]. 畜牧兽医学报, 2025, 56(2): 934-942.

DU Qingjie, WU Liping, ZHANG Fan, DAI Pengxiu, FENG Xiancheng, ZHANG Xinke. Difference Analysis of Oral Flora in Dogs with Periodontitis and Drug Resistance of Oral Porphyromonas[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 934-942.

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引物名称 Primer name		引物序列(5′→3′) Primer sequences	片段大小/bp Fragment size
通用引物 Universal primer	27F	AGAGTTTGATCCTGGCTCAG	1 450
通用引物 Universal primer	1492R	GGTTACCTTGTTACGACTT	1 450
特异性引物 Specific primer	C1-F	CGGTCCTATGGATTGTAAACT	367
	C1-R	CTGTTTGATACCCACGCTTT	367
	C2-F	ACGACGGTTCTATGGATTG	369
	C2-R	GTTTGATCCCCACGCTTT	369

耐药基因 Drug resistance gene	引物序列(5′→3′) Primer sequences	片段大小/bp Fragment size
ermF F	CGGGTCAGCACTTTACTATTG	466
ermF R	GGACCTACCTCATAGAGAAG
tetQ F	TTATACTTCCTCCGGCATCG	904
tetQ R	ATCGGTTCGAGAATGTCC
nim F	ATGTTCAGAGAAATGGGGCGTAAG	458
nim R	GCTTCCTTGCCTGTCATGTGCTC
cfxA F	CGTAGTTTTGAGTATAGCTTT	966
cfxA R	GATGTTGCCTATATATGTC

牙菌斑 Dental plaque	培养条件 Culture condition	药敏试纸 Drug sensitivity test paper	抑菌圈直径(d)/mm Inhibition zone diameter (d)	敏感性 Sensitivity
龈上菌斑 Supragingival plaque	需氧	恩诺沙星	26.90±1.52	极敏
		头孢氨苄	24.31±0.43	极敏
		阿奇霉素	20.34±0.51	极敏
		氨苄西林	20.34±1.12	极敏
		阿莫西林	19.89±0.69	极敏
		多西环素	14.69±0.07	中敏
		甲硝唑	14.25±0.25	中敏
	厌氧	恩诺沙星	24.74±0.04	极敏
		甲硝唑	23.34±9.96	极敏
		阿奇霉素	22.30±0.01	极敏
		阿莫西林	21.27±0.95	极敏
		头孢氨苄	21.02±0.28	极敏
		氨苄西林	17.64±0.68	高敏
		多西环素	15.40±0.26	高敏
龈下菌斑 Subgingival plaque	需氧	恩诺沙星	27.60±7.29	极敏
		头孢氨苄	25.42±0.58	极敏
		阿奇霉素	25.10±0.71	极敏
		阿莫西林	24.40±0.88	极敏
		氨苄西林	22.34±0.89	极敏
		多西环素	18.71±0.31	高敏
		甲硝唑	10.08±0.66	低敏
	厌氧	阿莫西林	24.16±0.77	极敏
		头孢氨苄	23.32±0.45	极敏
		氨苄西林	22.79±0.41	极敏
		阿奇霉素	20.70±0.69	极敏
		恩诺沙星	19.31±0.66	高敏
		多西环素	18.69±0.19	高敏
		甲硝唑	17.25±0.89	高敏

分类 Classification	药物名称 Medicines vocabularies	MIC/(μg·mL^-1)	结果 Result
氟喹诺酮类Fluoroquinolones	恩诺沙星	0125~0.25	S
β-内酰胺类Bata-lactam	阿莫西林	4~8	I
	头孢曲松钠	32~64	I
	头孢氨苄	32~64	I
四环素Tetracycline	盐酸多西环素	＞128	R
硝基咪唑Nitroimidazole	甲硝唑	＞128	R
氨基糖苷类Aminoglycosides	硫酸庆大霉素	32	R
大环内酯类Macrolides	阿奇霉素	＜8	R

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