Development and efficacy evaluation of a SP01-adjuvanted inactivated Escherichia coli mutant vaccine against bovine coliform mastitis

Genomics, Bioengineering
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Xinyu Li# Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.

Chongyu Tian# College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.

Hui Han# Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.

Bochang Shi Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.++The fifth medical center of PLA General Hospital, Beijing 100039, China.

Huabin Li College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.

Yuan Liu College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.++The fifth medical center of PLA General Hospital, Beijing 100039, China.

Wei He Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.

Chunhe Li Inner Mongolia Huaxi Biotechnology Co, Ltd., Inner Mongolia 010010, China++State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Huhehaote Inner Mongolia 010070, China.

Min Li Inner Mongolia Huaxi Biotechnology Co, Ltd., Inner Mongolia 010010, China++Ximu Bio-technology Co., LTD, Beijing 100010, China.

Yuanqiang Zheng Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Huhehaote Inner Mongolia 010058, China.

Bo Li The fifth medical center of PLA General Hospital, Beijing 100039, China.

Liyan Zhao People's Liberation Army Navy's No.971 Hospital, Qingdao Shandong 266071, China.

Naxin Zheng The fifth medical center of PLA General Hospital, Beijing 100039, China.

Fang Yan* College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu Shanxi 030800, China.

Zhongpeng Zhao* Department of Immunology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.

# contributed equally to this work


Abstract

Objective: Escherichia coli (E. coli) is one of the most common pathogens causing clinical mastitis in cattle, and there is no available vaccine to prevent this disease in China; therefore, there is an urgent need to develop an E. coli vaccine against bovine clinical mastitis. 

Methods: the candidate vaccine (Ch-O111-1) and challenge (LZ06) strains were screened from the milk samples of the cows with clinical mastitis. Furthermore, to extend the cross-protection of the Ch-O111-1 strain, the galE gene fragment of the Ch-O111-1 strain was deleted by homologous recombination between the Ch-O111-1 strain and pCVD442/△galE plasmid, which was identified by the conventional methods, including PCR, SDS-PAGE and sequencing. The Ch-O111-1/ΔgalE (Z9) strain was characterized by extensive cross-reactivity and attenuated virulence, and we prepared inactivated Z9 vaccines with different adjuvants. 

Results: Immunization of inactivated Z9 antigen induced adjuvant-, dosage-, inoculation times-dependent antibody titers in cows and mice. Furthermore, immunization with SP01-adjuvanted inactivated Z9 vaccine protected cows from LZ06 caused severe clinical mastitis and mice from LZ06 caused death. 

Conclusion: SP01-adjuvanted inactivated Z9 vaccine was successfully developed, which could protect the cows from Escherichia coli caused severe mastitis.

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