云南地区血清7型猪胸膜肺炎放线杆菌的分离鉴定及生物学特性分析

doi:10.16431/j.cnki.1671-7236.2026.01.045

Abstract

Abstract:

Objective This experiment aimed to study the biological characteristics of Actinobacillus pleuropneumoniae from pigs in Yunnan. Method The bacteria was isolated from the lung of pigs suffering from respiratory diseases in a pig farm located at Honghe， Yunnan. The bacterial isolate was identified through biochemical tests， 16S rDNA sequence analysis and PCR amplification. The biological characteristics of the isolate， including serotypes， pathogenicity， drug sensitivity and resistance genes， were analyzed. Result A Gram-negative coccobacillus was isolated from the lung samples of the diseased pig and was numbered YN240724. The biochemical identification results showed that the isolate decomposed glucose， fructose， sucrose， xylose and mannitol to produce acid， and the oxidase， urease and nitrate reduction tests were positive. These biochemical characteristics were the same as those of the reference strain of Actinobacillus pleuropneumoniae. The results of the 16S rDNA gene sequence analysis showed that the 16S rDNA sequence similarity between the isolate and the type strain ATCC 27088^T of Actinobacillus pleuropneumoniae and other reference strains of Actinobacilluspleuropneumoniae was 100% and 99.6%-100%， respectively. The isolate formed the same evolutionary branch with all the reference strains of Actinobacillus pleuropneumoniae. Based on these characteristics， the isolate YN24074 was identified as Actinobacillus pleuropneumoniae. The serotype identification results showed that the isolate was Actinobacillus pleuropneumoniae serotype 7. The results of the drug resistance gene detection showed that the isolate carried the β-lactam resistance genes bla_CIT and bla_TEM， as well as the tetracycline resistance gene tetM. The results of the antimicrobial susceptibility test showed that the isolate was sensitive to aminoglycosides （gentamicin and amikacin）， quinolones （ofloxacin）， chloramphenicols （florfenicol）， tetracyclines （tetracycline）， sulfonamides （sulfamethoxazole）， and cephalosporins （cefotaxime and cefuroxime）， but resistant to penicillins （penicillin， ampicillin and piperacillin） and glycopeptides （vancomycin）. The pathogenicity test results showed that the median lethal dose （LD₅₀） of the isolate for mice was 7.5×10⁴ CFU， indicating a relatively strong pathogenicity. Conclusion In this study， a strain of porcine Actinobacillus pleuropneumoniae serotype 7 from Yunnan was isolated. This strain carried the resistance genes bla_CIT， bla_TEM and tetM， and showed resistance to various antibacterial agents， with strong pathogenicity. This test results could provide important references for the treatment and immunization prevention and control of porcine contagious pleuropneumonia in pig farms of Yunnan.

Key words: pigs; Actinobacillus pleuropneumoniae; isolation and identification; serotype; pathogenicity

CLC Number:

S855.1

LI Fuxiang, SONG Jianling, LI Zhanhong. Isolation， Identification and Biological Characterization of Actinobacillus pleuropneumoniae Serotype 7 from Pigs in Yunnan[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(1): 499-508.

Figures/Tables 9

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References 29

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血清型 Serotypes	引物 Primers	引物序列 Primer sequences （5′→3′）	产物大小 Product size/bp	目的基因 Target genes
1	AP1F	CTGGAGTAATTACGGCGACTATTCC	959	cps1B
1	AP1R	AGGAGAAGCTAGTAGTACTTGCATTTTC	959	cps1B
5	AP5F	AGCCACAAGACCCGAATGGTATAATG	825	cps5B
5	AP5R	CCATCAAATGCAGCTTCAAGGAGC	825	cps5B
7	AP7F	TTGGAATGGATTCATGATTGGGC	601	cps7E
7	AP7R	CGGAAATGGCCTATTGAAAAACG	601	cps7E
15	AP15F	GCAACTTGGAGAACATGGTTAAATCAAG	1 595	cps15B/cps15C
15	AP15R	CAACCCTCCAATGTAAGCGAAGG	1 595	cps15B/cps15C

项目 Items	结果 Results	项目 Items	结果 Results
葡萄糖 Glucose	＋	水杨素 Sulicin	―
果糖 Fructose	＋	甘露醇 Mannitol	＋
蔗糖 Saccharose	＋	山梨醇 Sorbitol	―
木糖 Xylose	＋	硝酸盐还原 Nitrate reduction	＋
蕈糖 Mushroom sugar	―	尿素酶 Urease	＋
棉子糖 Raffinose	―	氧化酶 Oxidase	＋
菊糖 Inulin	―	过氧化氢酶 Catalase	―

抗菌药物类别 Class of antibacterial agents	抗菌药物 Antibacterial agents	抑菌圈直径 Inhibition zone diameter/mm	结果 Results
青霉素类 Penicillins	青霉素 Penicillin（10 units）	6	R
	氨苄西林 Ampicillin （10 μg）	6	R
	哌拉西林 Piperacillin （100 μg）	6	R
氨基糖苷类 Aminoglycosides	阿米卡星 Amikacin （30 μg）	17	S
	卡那霉素 Kanamycin （30 μg）	14	I
	庆大霉素 Gentamicin （30 μg）	15	S
	链霉素 Streptomycin （10 μg）	12	I
喹诺酮类 Quinolones	氧氟沙星 Ofloxacin （5 μg）	34	S
大环内酯类 Macrolides	红霉素 Erythromycin （15 μg）	21	I
酰胺醇类 Chloramphenicols	氟苯尼考 Florfenicol （30 μg）	35	S
四环素类 Tetracyclines	四环素 Tetracycline （30 μg）	18	S
磺胺类 Sulfonamides	磺胺甲噁唑 Sulfamethoxazole （30 μg）	25	S
糖肽类 Glycopeptides	万古霉素 Vancomycin （30 μg）	6	R
头孢菌素类 Cephalosporins	头孢呋辛 Cefuroxime （30 μg）	32	S
	头孢噻肟 Cefotaxime （30 μg）	34	S

剂量 Dose/CFU	死亡率 Mortality/%	半数致死量 LD₅₀/CFU
3.0×10⁶	100（10/10）	7.5×10⁴
3.0×10⁵	90（9/10）
3.0×10⁴	20（2/10）
3.0×10³	0（0/10）

Isolation， Identification and Biological Characterization of Actinobacillus pleuropneumoniae Serotype 7 from Pigs in Yunnan

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