梅花鹿源A型产气荚膜梭菌的分离鉴定及对小鼠的致病性分析

doi:10.11843/j.issn.0366-6964.2025.08.035

Abstract

Abstract:

Clostridium perfringens is an important pathogen that endangers the deer breeding industry. In this study, a strain of Clostridium perfringens was isolated from clinical samples obtained from deer using selective identification media. The strain was identified through 16S rRNA sequencing, toxin-type polymerase chain reaction (PCR), and biochemical assays. An analysis of antimicrobial resistance was also performed. To establish a murine infection model, mice were subjected to intraperitoneal injection with the isolated strain, and subsequent observations were made regarding incidence rates, mortality, and histopathological alterations. The findings indicated that the isolated strain was identified as Clostridium perfringens type A, aligning with the biochemical characteristics of Clostridium perfringens. Furthermore, this strain exhibited resistance to the antibiotics amikacin, polymyxin B, streptomycin, kanamycin, clindamycin, and cotrimoxazole. The mice of the experimental group exhibited poor vitality, characterized by unkempt fur and reduced mobility. Examination revealed hepatosplenomegaly, thinning of the intestinal wall, and the presence of yellow material within the small intestine. The median lethal dose was determined to be 7.99×10⁷ CFU·mL^-1. Histopathological analysis indicated hyperemia and hemorrhage across various tissues and organs, infiltration of inflammatory cells, partial villous atrophy and loss in the small intestine, and structural disintegration.

Key words: Clostridium perfringens, deer, isolation, mouse model

CLC Number:

S852.616.3

ZENG Shengxin, SONG Chengqi, SHEN Kaiyuan, HAO Guoxin, WANG Yakun, WANG Xin, WANG Xiaoxu, LIU Zhijie, LIU Yongbo, LIU Yongsheng, YANG Shunli, FU Zhixin. Isolation, Identification of a Deer Clostridium perfringens and Pathogenicity Analysis in Mouse[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3976-3984.

Figures/Tables 7

Table 1

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基因 Genes	引物序列(5′→3′) Primer sequences	片段长度/bp Product length
16S rRNA	F-AGAGTTTGATCCTGGCTCAG	1 465^[7]
16S rRNA	R- GGTTACCTTGTTACGACTT	1 465^[7]
cpa	F-GCTAATGTTACTGCCGTTGA	324
cpa	R-CCTCTGATACATCGTGTAAG	324
cpb	F-GCGAATATGCTGAATCATCTA	196
cpb	R-GCAGGAACATTAGTATATCTTC	196
etx	F-GCGGTGATATCCATCTATTC	655
etx	R-CCACTTACTTGTCCTACTAAC	655
itx	F-ACTACTCTCAGACAAGACAG	446
itx	R-CTTTCCTTCTATTACTATACG	446
NetB	F-GCTGGTGCTGGAATAAATGC	384
NetB	R-TCGCCATTGAGTAGTTTCCC	384
cpe	F-GGAGATGGTTGGATATTAGG	233
cpe	R-GGACCAGCAGTTGTAGATA	233

药物种类 Drug type	药物名称 Drugs	缩写 Abbreviations	药物含量/(μg·片^-1) Drug content	结果 Results
氨基糖苷类 Aminoglycoside	阿米卡星 Amikacin	AMK	30	R
	链霉素 Streptomycin	S	10	R
	卡那霉素 Kanamycin	KAN	30	R
	庆大霉素 Gentamicin	GM	120	S
β-内酰胺类 β-lactams	氨苄西林 Ampicillin	AMP	10	S
	头孢氨苄 Cefalexin	CN	30	S
	苯唑西林 Oxacillin	OX	1	R
	头孢唑林 Cefazolin	CZ	30	S
	头孢他啶 Ceftazidime	CAZ	30	S
	青霉素 Penicillin	PEN	10	S
四环素类 Tetracyclines	强力霉素 Doxycycline	DO	30	S
	米诺环素 Minocycline	MI	30	S
	四环素 Tetracycline	TET	30	S
多肽类 Polypeptide	万古霉素 Vancomycin	VAN	30	R
多肽类 Polypeptide	多黏菌素B Polymyxin B	PB	300 IU	S
氯霉素类 Chloram phenicols	氟苯尼考 Florfenicol	FFC	30	S
喹诺酮类 Quinolones	环丙沙星 Ciprofloxacin	CIP	5	I
林可酰胺类 Lincosamides	克林霉素 Clindamycin	CC	2	R
大环内酯类 Macrolides	红霉素 Erythromycin	E	15	R
碳青霉烯类 Carbapenem	亚胺培南 Imipenem	IPM	10	S
磺胺类 Sulfonamides	复方新诺明 Compound sulfamethoxazole	SXT	25	R

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Isolation, Identification of a Deer Clostridium perfringens and Pathogenicity Analysis in Mouse

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