猪葡萄球菌临床分离株感染仔猪渗出性皮炎模型的建立

doi:10.16431/j.cnki.1671-7236.2019.07.028

《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (7): 2110-2118.doi: 10.16431/j.cnki.1671-7236.2019.07.028

猪葡萄球菌临床分离株感染仔猪渗出性皮炎模型的建立

马炫炫^1,2, 任卫科³, 滕达^1,2, 池晶晶³, 王秀敏^1,2, 董志民³, 毛若雨^1,2, 郝娅^1,2, 李秀丽³, 王建华^1,2

1. 中国农业科学院饲料研究所基因工程研究室, 北京 100081;
2. 农业农村部饲料生物技术重点实验室, 北京 100081;
3. 天津市畜牧兽医研究所, 天津 300381

收稿日期:2019-01-31 出版日期:2019-07-20 发布日期:2019-07-22
通讯作者: 李秀丽, 王建华 E-mail:lxl5084@sina.com;wangjianhua@caas.cn
作者简介:马炫炫(1993-),女,河南三门峡人,硕士生,研究方向:抗菌肽及抗生素替代品,E-mail:1550010528@qq.com
基金资助:
国家自然科学基金"畜禽体表外用抗菌肽渗透机理及强化改造探索"（31872393）；中国农业科学院重大产出科研选题"饲用抗生素替代关键产品创制与产业化"（CAAS-ZDXT2018008）；国家农业科技创新工程"抗菌肽及抗生素替代品方向"（CAAS-ASTIP-2013-FRI-02）

Establishment of Model of Exudative Dermatitis of Piglets Infected with Clinical Isolated Staphylococcus hyicus

MA Xuanxuan^1,2, REN Weike³, TENG Da^1,2, CHI Jingjing³, WANG Xiumin^1,2, DONG Zhimin³, MAO Ruoyu^1,2, HAO Ya^1,2, LI Xiuli³, WANG Jianhua^1,2

1. Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China;
2. Key Laboratory of Feed Biotechnology, Ministry of Agriculture and Rural Areas, Beijing 100081, China;
3. Tianjin Animal Science and Veterinary Research Institute, Tianjin 300381, China

Received:2019-01-31 Online:2019-07-20 Published:2019-07-22

摘要/Abstract

摘要：

为建立研究仔猪渗出性皮炎发病机制及治疗制剂的动物模型，本研究采用PCR及BLAST序列比对方法对猪葡萄球菌437-2株毒素基因型进行鉴定，采用纸片扩散法检测其耐药性，通过不同浓度菌液人工感染仔猪试验分析该菌株的临床致病力、最小致病浓度、发病周期及组织病理学变化。结果显示，该分离菌株为ExhD毒素基因型，其毒素基因与德国分离株（GenBank登录号：AM94662.1）和俄罗斯分离株（GenBank登录号：AM950188.1）的ExhD基因同源性达99%；药敏试验结果显示，该菌株为多重耐药菌株，对青霉素、磺胺异噁唑、链霉素、阿奇霉素、红霉素、四环素、杆菌肽及诺氟沙星耐药；仔猪致病性试验结果显示，低剂量组在整个试验周期内仅注射部位表现出皮炎症状，而中、高剂量组在耳后皮下注射2 d后均出现显著的渗出性皮炎症状，病猪表现为耳后出现油皮，全身被毛粗糙，皮肤呈深褐色厚皮痂，有大片蜕皮和黄色液体渗出；感染仔猪的脏器和皮肤病理切片结果显示，低剂量组仔猪脏器无异常，耳部皮肤角质层角化过度，炎性细胞浸润，无棘皮层细胞分离，而中、高剂量组感染仔猪脏器具有一定程度的损伤，皮肤表皮棘细胞层出现明显的细胞分离。上述试验结果说明，利用猪葡萄球菌437-2株可成功建立仔猪渗出性皮炎模型，所需的最小攻菌浓度为1×10⁹ CFU/mL。

关键词: 仔猪渗出性皮炎; 猪葡萄球菌; 表皮脱落毒素; 仔猪渗出性皮炎模型

Abstract:

In order to provide a target animal model of exudative dermatitis of piglet for exploring the pathogenesis and therapeutic reagents,in this study,the toxin type of isolated Staphylococcus hyicus 437-2 was identified by PCR and BLAST sequence alignment,its drug resistance was detected by disk diffusion method,and the clinical pathogenicity,minimum pathogenic concentration,pathogenesis and histopathological changes caused by the strain were analyzed by artificially infecting piglets with different concentrations of bacterial liquid.The results showed that the strain carried the ExhD virulence gene,which showed 99% gene sequence homology with the isolated Germany strain (GenBank accession No.:AM94662.1) and Russian isolate (GenBank accession No.:AM950188.1).The drug resistant assay displayed S.hyicus 437-2 was a multi-drug resistant strain,which showed resistance to penicillin,sulfamethoxazole,streptomycin,azithromycin,erythromycin,tetracycline,bacitracin and norfloxacin.And the pathogenicity test of S.hyicus 437-2 on piglets showed that the low dose group displayed dermatitis symptoms only at the injection site throughout the experimental period,while significant exudative dermatitis symptoms appeared in the middle and high dose groups after subcutaneous injection for 2 d,which showed detailedly as oily skin behind the ear,rough fur,a large piece of suede and yellow liquid exuded,and thick dark brown skin.The results of pathological sections showed that organs in the low dose group were not damaged,while the skin tissue displayed excessive keratinization on the ear skin,inflammatory cell infiltration,and no cell separation.Meanwhile,the piglets in the middle and high dose groups all showed a certain degree of damage,the cell epidermal layer of the skin appeared obvious cell separation.This results indicated that this strain had strong pathogenicity and could successfully replicate exudative dermatitis on piglets.In addition,this result further indicated that the minimum bacterium concentration required for the establishment of exudative dermatitis model was 1×10⁹ CFU/mL.

Key words: piglet exudative dermatitis; Staphylococcus hyicus; exfoliative toxin; piglet exudative dermatitis model

中图分类号:

S858.28

马炫炫, 任卫科, 滕达, 池晶晶, 王秀敏, 董志民, 毛若雨, 郝娅, 李秀丽, 王建华. 猪葡萄球菌临床分离株感染仔猪渗出性皮炎模型的建立[J]. 《中国畜牧兽医》, 2019, 46(7): 2110-2118.

MA Xuanxuan, REN Weike, TENG Da, CHI Jingjing, WANG Xiumin, DONG Zhimin, MAO Ruoyu, HAO Ya, LI Xiuli, WANG Jianhua. Establishment of Model of Exudative Dermatitis of Piglets Infected with Clinical Isolated Staphylococcus hyicus[J]. , 2019, 46(7): 2110-2118.

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猪葡萄球菌临床分离株感染仔猪渗出性皮炎模型的建立

Establishment of Model of Exudative Dermatitis of Piglets Infected with Clinical Isolated Staphylococcus hyicus

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