犬猫皮肤病细菌性病原分离鉴定及天然产物抑菌效果分析

Isolation and Identification of Skin Pathogenic Bacteria from Dogs and Cats and Analysis of Antibacterial Effect of Natural Products

【目的】分离鉴定武汉市患皮肤病犬猫细菌性病原,并探索其对传统抗菌药物与天然活性产物藤黄酸(GA)和6-溴靛玉红-3’-肟(BIO)的敏感性。【方法】对患皮肤病犬猫采样并分离病原,通过生长特性观察、革兰氏染色镜检、PCR等方法鉴定并利用SPF小鼠验证致病性;通过药敏纸片验证其对传统药物的耐药性,并测定天然产物对其最小抑菌浓度(minimum inhibitory concentration,MIC)值。【结果】分离得到2株金黄色葡萄球菌、3株伪中间型葡萄球菌、2株猫葡萄球菌、1株犬链球菌及1株奇异变形杆菌。SPF小鼠皮肤创伤感染验证分离菌株均有致病性。犬链球菌及奇异变形杆菌对各自受试药物均敏感;葡萄球菌对复方新诺明、青霉素、红霉素、四环素、左氧氟沙星、苯唑西林、庆大霉素、克林霉素及氯霉素存在不同程度耐药。天然活性产物GA和BIO对上述9株菌均具有良好抑菌效果,且除分离菌株F5外GA对分离菌株的MIC值均小于BIO。【结论】本研究共分离得到5种、9株犬猫皮肤细菌。犬链球菌、奇异变形杆菌对传统抗菌药物均敏感,部分葡萄球菌存在耐药。GA和BIO对犬猫皮肤病原菌均有明显抑菌活性,显示其可作为防控犬猫细菌性皮肤病的候选药物。

【Objective】The purpose of the experiment was to isolate and identify the bacterial pathogens of dogs and cats suffering from skin diseases in Wuhan,and to explore their sensitivity to traditional antibiotics and natural active products gambogic acid(GA)and 6-bromoindirubin-3’-oxime(BIO).【Method】Skin pathogen bacteria from dogs and cats were collected and isolated,as well as growth characteristics,Gram staining microscope observation and PCR were determined.SPF mice were used to verify pathogenicity.The drug resistance to traditional drugs was verified by drug sensitive paper,and the minimum inhibitory concentration(MIC)of natural products was measured.【Result】2 strains of Staphylococcus aureus,3 strains of Staphylococcuspseudointermediate,2 strains of Staphylococcus felis,1 strain of Streptococcus canis,and 1 strain of Proteus mirabillis were isolated.The pathogenicity was verified by skin infection in SPF mice.Streptococcus canis and Proteus mirabillis were sensitive to the tested drugs.Staphylococcus aureus had different resistance to cotrimoxazole,penicillin,erythromycin,tetracycline,levofloxacin,oxacillin,gentamicin,clindamycin and chloramphenicol.The natural active products of GA and BIO had good antibacterial effects on the pathogenic bacteria,and except strain F5,the MIC of GA was universally lower than that of BIO.【Conclusion】In the study,a total of 9 pathogenic bacteria were isolated from dogs and cats.Streptococcus canis and Proteus mirabilis were sensitive to traditional antimicrobial agents,and some Staphylococcus strains were resistant to them.GA and BIO had obvious antibacterial effects against skin pathogenic bacteria.It showed that GA and BIO could be used as potential antibacterial drugs to prevent and control skin bacterial disease in dogs and cats.