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四肢开放性白猪海水浸泡伤的致病菌特征研究

Pathogenic bacteria characteristics of limb open injuries by seawater immersion in white pigs

摘要目的:建立现场海水浸泡伤动物感染模型,研究致病菌种类,为海水浸泡伤的诊疗提供依据。方法:于舟山某码头设立9个海水取水点,用无菌海水采样器取水。采用宏基因组测序法检测实验海域海水中所有的细菌种类,并作为海水对照组(SW组)。选取健康白猪16只,均通过手术造成左胫骨开放性骨折,按照实验设计分为实验组( n=10)和对照组( n=6)。实验组白猪创面于海水中浸泡2 h,而对照组白猪旷置于无菌手术室。2 h后所有实验动物包扎伤口,定期换药。术后7 d取伤口深部软组织、骨组织行病理学检查并观察感染情况,采用微生物多样性分析法检测致病菌种类并进行差异性分析。 结果:实验组10只白猪皆感染,对照组6只白猪中2只感染;与SW组比较,实验组菌群数量明显减少;与对照组比较,实验组菌群数量明显增多( P<0.05或 P<0.01)。所有感染的白猪均检出坏死性梭杆菌、孔氏创伤球菌、猪链球菌、大肠埃希菌等机会致病菌,实验组除检出海水中专有的狭义梭状杆菌、脱硫弧菌、哈维氏弧菌外,还检出大芬戈尔德菌、化脓性拟杆菌等多种厌氧菌。 结论:开放性创伤经海水浸泡后具有较高的感染率,且为机会致病菌、海水细菌和厌氧菌的混合感染,故早期彻底的外科清创和使用覆盖厌氧菌的广谱抗生素对控制感染具有重要意义。

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abstractsObjective:To establish the animal model of infection due to seawater immersion injury and study the species of pathogenic bacteria,so as to provide a basis for the diagnosis and treatment of seawater immersion injuries.Methods:Sea water was collected from nine points at a wharf in Zhoushan city by sterile seawater sampler. Metagenomic sequencing was used to detect all bacterial species in the seawater,and the seawater was used as a control(SW group). Sixteen healthy white pigs were selected and underwent surgery for making open fracture of the left tibia,and then they were divided into experimental group( n=10)and control group( n=6),according to the experimental protocol. The wounds of the experimental group were immersed in seawater for 2 h,while the wounds of the control group were kept in a sterile operating room. Two hours later,all the wounds of the experimental animals were bandaged and the dressings were changed regularly. Seven days after surgery,the deep soft tissue and bone tissue were taken for pathological examination and the infection was observed. The species of pathogenic bacteria were detected by microbial diversity analysis and the differences were analyzed. Results:All ten pigs in the experimental group were infected,and two of six pigs in the control group were infected. Compared with the SW group,the number of bacterial communities was significantly decreased;while compared with the control group,the number of bacterial communities was significantly increased( P<0.05 or P<0.01). Opportunistic pathogens,such as Clostridium necrotizing, Traumatococcus coonii, Streptococcus suis,and Escherichia coli,were detected in all the infected white pigs. The experimental group was also found to have some seawater-specific bacteria,such as Clostridium difficile, Desulfovibrio vibrio,and Vibrio harveyi,as well as a variety of anaerobic bacteria,such as Macrofingordelia and Bacteroides pyogenes. Conclusion:The open wound after seawater immersion can lead to infections with high incidence,which are the mixture of opportunistic pathogens,seawater bacteria,and anaerobic bacteria. Therefore,early and thorough surgical debridement and the use of broad-spectrum antibiotics,covering anaerobic bacteria,are of great significance for infection control.

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