CD163基因敲除猪对胸膜肺炎放线杆菌、猪链球菌和副猪嗜血杆菌的易感性研究

doi:10.11843/j.issn.0366-6964.2024.12.014

摘要/Abstract

摘要：

旨在探究在CD163基因的SRCR5区域插入1个碱基的CD163基因敲除(CD163 gene knockout, CD163-KO)型猪对3种关键细菌的易感性相较于野生型(wild-type, WT)猪是否会有所变化，评估CD163-KO型猪的生物安全性。本研究选用均为50日龄、体重与健康状况等相似的17头CD163-KO型大白猪和17头WT型大白猪，其中2头CD163-KO型大白猪和2头WT型大白猪作为空白对照，不做任何感染处理。试验猪只均无胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae, APP)、猪链球菌(Streptococcus suis，SS)及副猪嗜血杆菌(Haemophlius parasuis, HPS)感染史与接种史。试验猪只随机分为3组，每组均有5头CD163-KO型猪和WT型猪，A组接种APP1型菌株、B组接种SS2型菌株、C组接种HPS13型菌株。连续观察8 d，记录猪只临床症状表现与死亡情况；试验猪只死亡后及时解剖，观察心脏、胸腔、肺部等重要部位病变情况；采集肺样并提取肺部中的细菌DNA，用于定量PCR检测并统计分析与对比CD163-KO型与WT型死亡大白猪肺中3种细菌分别的相对DNA拷贝量。依据试验猪只临床症状表现可得，各组试验猪只均成功感染相应的细菌，且各组内的CD163-KO型猪与WT型猪的存活率均无显著差异(P>0.05)；依据死亡猪只肺部病变情况及细菌定量PCR检测结果可得，CD163-KO型死亡猪肺与WT型死亡猪肺中APP1型、SS2型及HPS13型菌株的相对DNA拷贝量均无显著差异(P>0.05)。本研究的结果是CD163-KO型大白猪生物安全性的有力证据，证明CD163基因敲除后没有提高猪只对APP、SS和HPS的易感性，不会显著影响猪只抵抗细菌的能力，为CD163-KO型猪的生物安全性评估提供强有力的支持。

关键词: CD163, 基因敲除, 大白猪, 猪繁殖与呼吸综合征病毒, 胸膜肺炎放线杆菌, 猪链球菌, 副猪嗜血杆菌, 生物安全评估

Abstract:

This study aimed to investigate whether pigs with a CD163 gene knockout (CD163-KO) resulting from a single base insertion in the SRCR5 domain of the CD163 gene exhibit altered susceptibility to 3 crucial bacterial pathogens compared to their wild-type (WT) counterparts and to evaluate the biosafety profile of CD163-KO pigs. This study selected a total of 34 Large White pigs, 17 CD163-KO pigs and 17 wild-type pigs, all at the age of 50 days with similar body weights and health statuses. Among them, 2 CD163-KO Large White pigs and 2 WT Large White pigs were used as blank controls without any infection treatment. None of the pigs had a history of infection or vaccination against Actinobacillus pleuropneumoniae (APP), Streptococcus suis (SS), and Haemophilus parasuis (HPS). The pigs were randomly divided into 3 groups of 5 CD163-KO and 5 WT pigs each: group A received APP serotype 1, group B received SS serotype 2, and group C received HPS serotype 13. The pigs were observed continuously for 8 days, the clinical symptoms and mortality were recorded. Upon death, the pigs were promptly necropsied to observe lesions in critical organs such as the heart, thorax, and lungs. Lung samples were collected, and bacterial DNA was extracted from the lungs for quantitative PCR analysis. The relative DNA copy numbers of the 3 bacterial species in the lungs of deceased CD163-KO and WT pigs were statistically analyzed and compared. The primary aim of this study was to evaluate whether CD163-KO pigs exhibit altered susceptibility to these 3 key bacterial pathogens compared to their WT counterparts, ultimately assessing the biosafety profile of CD163-KO pigs. Based on the clinical manifestations observed in the experimental pigs, all groups successfully contracted the respective bacterial infections, and there were no significant differences in survival rates between CD163-KO and WT pigs within each group (P>0.05). Furthermore, according to the pathological findings in the lungs of deceased pigs and the results of quantitative PCR analysis for bacterial DNA, no significant differences were detected in the relative DNA copy numbers of APP serotype 1, SS serotype 2, and HPS serotype 13 strains in the lungs between deceased CD163-KO and WT pigs (P>0.05). The findings of this study serve as robust evidence for the biological safety of CD163-KO Large White pigs, demonstrating that the knockout of the CD163 gene does not enhance the susceptibility of pigs to APP, SS, and HPS, nor does it significantly compromise their ability to resist these bacterial infections, which provides strong support for the biological safety assessment of CD163-KO pigs.

Key words: CD163, gene knockout, Large White pig, porcine reproductive and respiratory syndrome virus, Actinobacillus pleuropneumoniae, Streptococcus suis, Haemophilus parasuis, biosafety assessment

中图分类号:

S828.2

梁国濠, 胡丹丹, 钟海文, 张健, 杨德鸿, 吴珍芳, 杨化强, 张献伟. CD163基因敲除猪对胸膜肺炎放线杆菌、猪链球菌和副猪嗜血杆菌的易感性研究[J]. 畜牧兽医学报, 2024, 55(12): 5478-5488.

LIANG Guohao, HU Dandan, ZHONG Haiwen, ZHANG Jian, YANG Dehong, WU Zhenfang, YANG Huaqiang, ZHANG Xianwei. Study on the Susceptibility of CD163 Gene Knockout Pigs to Actinobacillus pleuropneumoniae, Streptococcus suis, and Haemophilus parasuis[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5478-5488.

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组别 Group		数量 Number	菌株 Strain	注射量/(mL·头^-1) Injection volume	菌液浓度/(cfu·mL^-1) Concentration of bacterium	攻毒方式 Attack mode
A	A1	5头CD163-KO型	APP101	2	2.2×10⁸^[24]	气管注射
	A2	5头WT型
B	B1	5头CD163-KO型	SS201	2	8.0×10⁷^[25]	腹腔注射
	B2	5头WT型
C	C1	5头CD163-KO型	HPS1301	2	3.6×10⁹^[26]	腹腔注射
	C2	5头WT型

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