志贺氏菌噬菌体ΦDS8对患病肉鸡的治疗及其肠道菌群的影响

doi:10.3969/j.issn.1004-1524.2022.07.05

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (7): 1386-1395.DOI: 10.3969/j.issn.1004-1524.2022.07.05

志贺氏菌噬菌体ΦDS8对患病肉鸡的治疗及其肠道菌群的影响

陈诗雨¹^,²(), 徐美余¹^,², 邓征宇¹^,², 王峰¹^,², 张麒麟¹^,², 邓先余¹^,², 林连兵¹^,²^,^*()

1.昆明理工大学生命科学与技术学院,云南昆明 650500
2.云南省高校饲用抗生素替代技术工程研究中心,云南昆明 650500

收稿日期:2021-04-22 出版日期:2022-07-25 发布日期:2022-07-26
通讯作者: 林连兵
作者简介:* 林连兵,E-mail: linlb@kmust.edu.cn
陈诗雨(1997—),女,安徽铜陵人,硕士研究生,研究方向为生物制药。E-mail: 1789611985@qq.com
基金资助:
云南省高校饲用抗生素替代技术工程研究中心

Treatment of diseased broilers with Shigella bacteriophage ΦDS8 and the effect on their intestinal flora

CHEN Shiyu¹^,²(), XU Meiyu¹^,², DENG Zhengyu¹^,², WANG Feng¹^,², ZHANG Qilin¹^,², DENG Xianyu¹^,², LIN Lianbing¹^,²^,^*()

1. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China
2. Engineering Research Center for Replacement Technology of Feed Antibiotics of Yunnan College, Kunming 650500, China

Received:2021-04-22 Online:2022-07-25 Published:2022-07-26
Contact: LIN Lianbing

摘要/Abstract

摘要：

志贺氏菌是一种常见的肠道致病菌,其引起的细菌病是危害公共卫生的人畜共患病之一。随着抗生素的滥用,志贺氏菌耐药性的问题难以控制,急需新的方法来解决,采用噬菌体杀灭耐药性细菌已日益受到重视。本研究建立肉鸡感染志贺氏菌模型,分别采用感染前2 h灌喂噬菌体液预防和感染2 h后灌喂噬菌体液两种方法对肉鸡进行治疗。灌喂志贺氏菌2 h后的肉鸡均出现腹泻状况,粪便中志贺氏菌含量最高达到1×10⁷ CFU·g^-1,优势菌群含量大幅下降,有害菌群占比上升。噬菌体治疗后,肉鸡粪便中噬菌体含量上升,志贺氏菌含量下降,粪便性状及肠道菌群恢复正常,且肉鸡存活率100%。没有噬菌体治疗的肉鸡存活率60%,解剖观察发现器官病变明显。噬菌体ΦDS8在治疗志贺氏菌株感染方面有良好的效果,其作为抗生素替代产品有较好的应用价值和前景。

关键词: 志贺氏菌, 噬菌体, 肉鸡, 肠道菌群

Abstract:

Shigella is a common enteric pathogenic bacterium, and the bacterial disease it causes is one of the zoonotic diseases that endanger public health safety. With the misuse of antibiotics, the problem of Shigella resistance is difficult to control and new approaches are urgently needed to solve it. The use of bacteriophages to kill drug-resistant bacteria has received increasing attention. In this study, the Shigella infection model of broiler chickens was established. The broiler chickens were treated with phage fluid 2 h before and after infection. The broilers fed with Shigella for 2 h all had diarrhea, with the highest Shigella content in feces of 1×10⁷ CFU·g^-1, a large decrease in the content of dominant flora and a rise in the proportion of harmful flora. After phage therapy, the fecal phage content of broilers increased, Shigella content decreased, the fecal traits and intestinal flora returned to the normal, and the broiler survival rate was 100%. The survival rate of broiler chickens without phage treatment was 60%, and anatomical observation revealed obvious organ lesions. Bacteriophage ΦDS8 has shown promising efficacy in the treatment of Shigella strain infection, and it has good application value and prospect as an alternative product to antibiotics.

Key words: Shigella, bacteriophage, broiler, becal microflora

中图分类号:

S816.7

陈诗雨, 徐美余, 邓征宇, 王峰, 张麒麟, 邓先余, 林连兵. 志贺氏菌噬菌体ΦDS8对患病肉鸡的治疗及其肠道菌群的影响[J]. 浙江农业学报, 2022, 34(7): 1386-1395.

CHEN Shiyu, XU Meiyu, DENG Zhengyu, WANG Feng, ZHANG Qilin, DENG Xianyu, LIN Lianbing. Treatment of diseased broilers with Shigella bacteriophage ΦDS8 and the effect on their intestinal flora[J]. Acta Agriculturae Zhejiangensis, 2022, 34(7): 1386-1395.

图/表 10

参考文献 34

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分组 Group	动物数量 Animal number	每只灌喂细菌量 Amount of bacteria per single animal/CFU	每只灌喂噬菌体量 Bacteriophage volume per single animal/PFU	灌喂噬菌体治疗时间点 Time point of phage therapy/h
阴性对照组Negative control	15	—	—	—
阳性对照组Positive control group	15	1×10⁸	—	—
噬菌体治疗组Therapeutic group	15	1×10⁸	1×10⁸	4
噬菌体预防组Prophylactic group	15	1×10⁸	1×10⁸	0
噬菌体治疗对照组Therapeutic control	15	—	1×10⁸	4
噬菌体预防对照组Prophylactic control	15	—	1×10⁸	0

分组 Group	动物数量 Animal number	每只灌喂细菌量 Amount of bacteria per single animal/CFU	每只灌喂噬菌体量 Bacteriophage volume per single animal/PFU	灌喂噬菌体治疗时间点 Time point of phage therapy/h
阴性对照组Negative control	15	—	—	—
阳性对照组Positive control group	15	1×10⁸	—	—
噬菌体治疗组Therapeutic group	15	1×10⁸	1×10⁸	4
噬菌体预防组Prophylactic group	15	1×10⁸	1×10⁸	0
噬菌体治疗对照组Therapeutic control	15	—	1×10⁸	4
噬菌体预防对照组Prophylactic control	15	—	1×10⁸	0

分组Group	0 h	2 h	4 h	6 h	12 h	24 h	48 h
阴性对照组Negative control	-	-	-	-	-	-	-
阳性对照组Positive control	-	+	+	++	+++	+++	++
噬菌体治疗组Therapeutic group	-	+	++	++	+	-	-
噬菌体预防组Prophylactic group	-	+	++	++	+	-	-
噬菌体治疗对照组Therapeutic control	-	-	-	-	-	-	-
噬菌体预防对照组Prophylactic control	-	-	-	-	-	-	-

分组Group	0 h	2 h	4 h	6 h	12 h	24 h	48 h
阴性对照组Negative control	-	-	-	-	-	-	-
阳性对照组Positive control	-	+	+	++	+++	+++	++
噬菌体治疗组Therapeutic group	-	+	++	++	+	-	-
噬菌体预防组Prophylactic group	-	+	++	++	+	-	-
噬菌体治疗对照组Therapeutic control	-	-	-	-	-	-	-
噬菌体预防对照组Prophylactic control	-	-	-	-	-	-	-

分组 Group	死亡数量 Number of deaths	存活率 Survival rate/%
阴性对照组Negative control	0	100
阳性对照组Positive control	6	60
噬菌体治疗组Therapeutic group	0	100
噬菌体预防组Prophylactic group	1	93
噬菌体治疗对照组Therapeutic control	0	100
噬菌体预防对照组Prophylactic control	0	100

志贺氏菌噬菌体ΦDS8对患病肉鸡的治疗及其肠道菌群的影响

Treatment of diseased broilers with Shigella bacteriophage ΦDS8 and the effect on their intestinal flora

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参考文献 34

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样品 Sample	Ace指数 Ace	Chao1指数 Chao1	辛普森指数 Simpson	香农指数 Shannon	覆盖率 Coverage
0-A	268.734	272.7	0.313 2	2.627 4	0.983 6
0-B	312.564	286.2	0.091 8	3.568 5	0.966 8
2-A	229.834	236.4	0.113 0	3.157 7	0.984 8
2-B	282.828	269.3	0.084 4	3.404 6	0.982 3
12-A	222.394	169.1	0.08 23	3.524 2	0.990 7
12-B	207.277	217.3	0.353 1	2.121 1	0.976 8
48-A	224.704	235.7	0.536 9	1.779 6	0.998 2
48-B	207.677	299.1	0.111 0	3.074 0	0.991 7

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