Whole Genome Sequencing and Genome Evolution Analysis of Capsular Serotype A and D Pasteurella multocida of Bovine

doi:10.13560/j.cnki.biotech.bull.1985.2024-0371

Abstract

Abstract:

【Objective】The capsular serotype of the Pasteurella multocida isolated from bovine in the Wulagai management area was identified. Whole genome sequencing and genome evolution analysis were performed for the strains with different serotypes. 【Method】Taking the lung tissue samples from bovine for pathogen isolation and purification, the 16S rRNA identification, capsular serotype typing, drug sensitivity experiments, and mouse pathogenicity experiments were performed. Whole-genome sequencing and genomic evolution analysis were performed for the strains with different serotypes. 【Result】Two strains of bovine Pasteurella multocida were successfully isolated, designated as Pm-YQ and Pm-SM. The Pm-YQ strain was capsular serotype A and the Pm-SM strain was capsular serotype D. Both strains presented relatively high virulence but low resistance to antibiotics. The whole genome lengths of the isolates were 2 274 102 bp and 2 244 957 bp, respectively, encoding 2 070 and 2 007 genes. The Pm-YQ strain belonged to sequence type ST 179, while Pm-SM strain belonged to ST 1. Through the construction of whole-genome phylogenetic tree, Pm-YQ strain was found to be closely related to three strains registered in GenBank from Germany, Denmark, and the United States, Pm-SM strain was found in the same evolutionary branch with a capsular serotype F of P. multocida strain isolated from Chongqing, China. 【Conclusion】The isolation, identification and whole genome sequencing of two strains with different capsular serotypes of bovine P. multocida were completed. The Pm-YQ strain was closely related to the German isolate, and the Pm-SM strain was relatively far related to the currently registered bovine P. multocida strains.

Key words: bovine, Pasteurella multocida, capsular serotype, drug-resistant gene, virulence gene, whole genome sequencing

WANG Zi, SHI Jin-chuan, WANG Yong-qiang, SUN Miao, MENG Ling-hao, GENG Chao, LIU Kai. Whole Genome Sequencing and Genome Evolution Analysis of Capsular Serotype A and D Pasteurella multocida of Bovine[J]. Biotechnology Bulletin, 2024, 40(12): 282-290.

Figures/Tables 11

References 37

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目的基因 Target gene	引物序列 Primer sequence（5'-3'）	扩增长度Amplification length/bp
16S rRNA	F: AGAGTTTGATCCTGGCTCAG R: GGTTACCTTGTTACGACTT	1 369
KMT1	F:ATCCGCTATTTACCCAGTGG R:GCTGTAAACGAACTCGCCAC	460
hyaD-hyaC	F:TGCCAAAATCGCAGTCAG R: TTGCCATCATTGTCAGTG	1 044
bcbD	F: CATTTATCCAAGCTCCACC R: GCCCGAGAGTTTCAATCC	760
dcbF	F:TTACAAAAGAAAGACTAGGAGCCC R: CATCTACCCACTCAACCATATCAG	657
ecbJ	F: TCCGCAGAAAATTATTGACTC R: GCTTGCTGCTTGATTTTGTC	511
fcbD	F:AATCGGAGAACGCAGAAATCAG R: TTCCGCCGTCAATTACTCTG	851

目的基因 Target gene	引物序列 Primer sequence（5'-3'）	扩增长度Amplification length/bp
16S rRNA	F: AGAGTTTGATCCTGGCTCAG R: GGTTACCTTGTTACGACTT	1 369
KMT1	F:ATCCGCTATTTACCCAGTGG R:GCTGTAAACGAACTCGCCAC	460
hyaD-hyaC	F:TGCCAAAATCGCAGTCAG R: TTGCCATCATTGTCAGTG	1 044
bcbD	F: CATTTATCCAAGCTCCACC R: GCCCGAGAGTTTCAATCC	760
dcbF	F:TTACAAAAGAAAGACTAGGAGCCC R: CATCTACCCACTCAACCATATCAG	657
ecbJ	F: TCCGCAGAAAATTATTGACTC R: GCTTGCTGCTTGATTTTGTC	511
fcbD	F:AATCGGAGAACGCAGAAATCAG R: TTCCGCCGTCAATTACTCTG	851

抗生素 Antibiotic	菌株编号Strain No.
抗生素 Antibiotic	Pm-YQ	Pm-SM
强力霉素	S（17）	I（10）
诺氟沙星	R（9）	S（21）
头孢噻肟	S（15）	S（20）
氟苯尼考	S（26）	S（21）
阿米卡星	I（10）	S（17）
庆大霉素	R（9）	S（16）
恩诺沙星	R（7）	S（25）
多粘菌素B	S（18）	S（13）
环丙沙星	S（22）	S（23）
青霉素	S（34）	S（34）
氧氟沙星	I（14）	S（21）
卡那霉素	R（0）	R（0）

抗生素 Antibiotic	菌株编号Strain No.
抗生素 Antibiotic	Pm-YQ	Pm-SM
强力霉素	S（17）	I（10）
诺氟沙星	R（9）	S（21）
头孢噻肟	S（15）	S（20）
氟苯尼考	S（26）	S（21）
阿米卡星	I（10）	S（17）
庆大霉素	R（9）	S（16）
恩诺沙星	R（7）	S（25）
多粘菌素B	S（18）	S（13）
环丙沙星	S（22）	S（23）
青霉素	S（34）	S（34）
氧氟沙星	I（14）	S（21）
卡那霉素	R（0）	R（0）

菌株名称Name of strain	基因岛Genomic island	位置Location	基因位点Position	长度Length/bp
Pm-YQ	Genomic-island-1	染色体Chromosome	845109-852871	7763
	Genomic-island-2	染色体Chromosome	1105564-1116467	10904
	Genomic-island-3	染色体Chromosome	1602281-1618085	15805
	Genomic-island-4	染色体Chromosome	1876215-1889636	13422
Pm-SM	Genomic-island-1	染色体Chromosome	160470-193937	33468
	Genomic-island-2	染色体Chromosome	953950-964232	10283
	Genomic-island-3	染色体Chromosome	1143374-1159357	15984