摘要
目的 通过构建开放性创伤性脑损伤合并海水浸泡后颅内感染大鼠模型,检测经天然海水浸泡后大鼠颅内感染的发生率及病原菌种类.方法 从福建平潭观音奥港口取海水17000 ml,进行海水细菌培养、宏基因组测序和动物模型制作.66只雄性SD大鼠,随机分为3组:TBI+无菌人工海水浸泡(ASI)组(n=6)、对照(sham)+天然海水浸泡(NSI)组(n=30)和TBI+NSI组(n=30).造模后经枕大池抽取脑脊液进行细菌培养,根据培养结果统计感染率及病原菌种类.结果 海水宏基因组测序提示,海水菌群结构为弧菌属、假交替单胞菌属、远洋杆菌属、嗜冷菌属等;海水细菌培养提示,样本海水中存在副溶血弧菌、溶藻弧菌、创伤弧菌、大肠杆菌、金黄色葡萄球菌和肺炎克雷伯杆菌等.TBI+NSI组动物中,感染副溶血弧菌感染和大肠杆菌各6只,溶藻弧菌5只,铜绿假单胞菌1只,其中部分大鼠出现混合感染(3只).TBI+NSI组共感染21只,感染率70％(21/30),与sham+NSI组相比(P＜0.01);sham+NSI组感染副溶血弧菌2只,大肠杆菌1只,溶藻弧菌1只,肺炎克雷伯杆菌1只,感染率17％(5/30).TBI+ASI组大鼠未出现感染.结论 本研究成功建立了开放性创伤性脑损伤合并海水浸泡后颅内感染大鼠模型,初步明确了开放性创伤性脑损伤合并海水浸泡后颅内感染的致病菌多为革兰氏阴性菌,以海洋弧菌和肠道菌群为主.
Abstract
Objective By constructing a rat model of open traumatic brain injury combined with intracranial infection after immersion of seawater,we aimed to detect the incidence rate and bacterial species of intracranial infection in rats following immersion of natural seawater.Methods A total of 17 000ml seawater was collected from Guanyinao port,Pingtan,Fujian Province,for bacterial culture,metagenome sequencing and animal model making.Sixty-six male SD rats were randomly divided into 3 groups:TBI+sterile group with artificial seawater immersion(ASI)(n=6)、control(sham)+group with natural seawater immersion(NSI)(n=30)and TBI+NSI group(n=30).Subsequently,cerebrospinal fluid was collected from the cisterna magna for bacterial culture,and the infection rate and pathogenic bacteria species were counted according to outcomes of the culture.Results Outcomes of seawater metagenome sequencing indicated that structures at genus level of bacterial flora were Vibrio,Pseudoxanthomonas,Pelagibacter,Psychrophilic bacteria,etc.There were Vibrio parahaemolyticus,Vibrio alginolyticus,Vibrio vulnificus,Escherichia coli,Staphylococcus aureus,Klebsiella pneumoniae and so on in the sample seawater according to above-mentioned bacterial cultivation of seawater.In the TBI+NSI group,quantities of subjects infected with Vibrio parahaemolyticus,Escherichia coli,Vibrio alginolyticus,and Pseudomonas aeruginosa were 6,6,5 and 1 respectively,Some rats showed mixed infections(3).A total of 21 mice were infected in the TBI+NSI group,with an infection rate of 70％(21/30),compared to the sham+NSI group(P＜0.01).In the sham+TBI group,numbers of subjects infected with Vibrio parahaemolyticus,Escherichia coli,Vibrio alginolyticus,and Klebsiella pneumoniae were 2,1,1 and 1 respectively,with the infection rate of 17％(5/30).By contrast,SD rats in the TBI+ASI group were not infected.Conclusion In this study,a rat model of open craniocerebral injury combined with intracranial infection after seawater immersion was successfully established,and the fact that the majority of pathogenic bacteria in open traumatic brain injury combined with intracranial infection after seawater immersion were Gram-negative bacteria was preliminarily defined.Additionally,involved bacteria were primarily Marine Vibrio and intestinal flora.
基金项目
联勤医学重点专科(神经外科)项目(2022ZL01)
联勤保障部队第九○○医院指令性项目战创伤救治专项(2022ZL03)
NETL
NSTL
万方数据