基于16S rDNA高通量测序技术分析受水灾影响猪场水样的菌群特征

Investigation of Bacterial Communities in Water Samples from Flood-affected Pig Farms Using 16S rDNA High-throughput Sequencing Technology

2021年7月,河南省部分地区突发特大水灾,致使受灾区域内的猪场损失惨重,本研究旨在探究水灾发生后猪场水样中存在的病原菌,为猪场防控可能出现的疾病提供技术支撑。通过实地采样和委托采样的方式,获得了26个受灾猪场中的48份水样,包括19份猪饮用水、22份场内积雨水和7份废弃污水,采用菌落计数和高通量测序技术,对样品的细菌总数和菌群丰度进行分析研究,同时对采集的水样开展消毒剂杀菌试验。结果发现:饮用水、积雨水和污水中的平均含菌量分别为1.55×10^(5)、1.88×10^(8)和3.70×10^(9)CFU·mL^(-1),其中污水中的细菌总数最多,其次为积雨水,饮用水中的细菌总数最少,但远超国家饮用水的安全标准。本研究在不同种类的水样中均检测到了链球菌属、埃希氏杆菌属、弓形杆菌属、不动杆菌属和假单胞菌属等潜在的致病菌属,以及嗜低温弓形杆菌、猪链球菌、大肠杆菌、鲍曼不动杆菌和豚鼠气单胞菌等致病菌。杀菌试验显示,氯制剂终浓度达到2500mg·L^(-1),作用30 min即可杀灭积雨水和污水中所有的细菌。本研究发现,水灾后养猪场内的水样中含有多种致病菌,规模化猪场可选用含氯消毒剂对场内污染的水体进行消杀,以确保人和猪群的健康安全。

In July 2021,a severe flood occurred in some area of Henan Province,pig farms in these areas suffered heavy losses.The purpose of the current research is to explore the bacteria in the flood-affected water samples of pig farms,so as to provide technical support for the prevention and control of possible diseases.Different types of water samples including 19 drinking water,22 rainwater,and 7 waste sewage samples were collected by field sampling and airmail from 26 farms in the affected areas.Using colony counting and high-throughput sequencing technology,the total number and the abundance of bacteria in the collected samples were analyzed,and disinfection tests were carried out for the collected water samples.The results showed that the total bacterial counts in drinking water,rainwater and sewage were 1.55×10^(5),1.88×10^(8)and 3.70×10^(9)CFU·mL^(-1),respectively.The total number of bacteria in the sewage was the highest,followed by the rainwater,and that of the drinking water was the lowest,but still higher than the national standard of drinking water.By mean of high-throughput detection technology,potential pathogenic genus such as Streptococcus,Escherichia,Arcobacter,Actinobacter and Pseudomonas,as well as waterborne pathogenic bacteria such as Arcobacter cryaerophilus,Streptococcus suis,Escherichia coli,Acinetobacter baumannii and Aeromonas caviae,were detected in water samples.Bactericidal tests showed that when the final concentration of chlorine reaches 2500 mg·L^(-1),all bacteria in rainwater and sewage were killed within 30 minutes.It was demonstrated that water samples in pig farms after floods contained a variety of pathogenic bacteria and pig farms can choose chlorine-containing disinfectants to kill the bacteria in polluted water in the farm to ensure the health of humans and pigs.