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. 2023 Nov 7;11(11):1698.
doi: 10.3390/vaccines11111698.

The Safety and Protective Efficacy Evaluation of an Attenuated M. bovis-BoHV-1 Bivalent Vaccine in Rabbits

Sen Zhang  1   2   3 , Yisheng Zhang  1   2 , Guoxing Liu  1   2 , Chen Wang  1   2   3 , Yan Ji  4   5 , Jianguo Chen  1   2 , Changmin Hu  1   2 , Xi Chen  1   2 , Aizhen Guo  1   2   3 , Yingyu Chen  1   2   3
Affiliations

The Safety and Protective Efficacy Evaluation of an Attenuated M. bovis-BoHV-1 Bivalent Vaccine in Rabbits

Sen Zhang et al. Vaccines (Basel). .

Abstract

Bovine respiratory disease (BRD) is a global prevalent multifactorial infection primarily caused by viral and bacterial coinfections. In China, Mycoplasma bovis (M. bovis) and bovine herpesvirus type 1 (BoHV-1) are the predominant pathogens associated with BRD. Our previous study involved the development of attenuated M. bovis HB150 and BoHV-1 gG-/tk- vaccine strains, which were thoroughly assessed for their safety profiles and protective efficacy in cattle. In this study, we applied a combination of vaccines in varying ratios and used a rabbit model to determine the safety and protective efficacy. We used PCR/RT-PCR to detect the postimmunization and challenge shedding of M. bovis and BoHV-1. Additionally, we measured antibody titers and the expression of IFN-β and TNF-α to evaluate the humoral and cellular immune responses, respectively. Furthermore, we performed a histopathological analysis to assess lung damage. Our study provides evidence of the safety and effectiveness of the bivalent M. bovis-BoHV-1 vaccine in rabbits, particularly when applying a combination of 1.0 × 108 CFU of M. bovis HB150 and 1.0 × 106 TCID50 of the BoHV-1 gG-/tk- strain. The bivalent vaccine significantly enhanced both the long-term antibody immune response and cellular protection against the M. bovis and BoHV-1 challenge. These findings provide a valuable model for the potential application in cattle.

Keywords: BRD; BoHV-1; M. bovis; attenuated vaccine; protective efficacy.

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Conflict of interest statement

Y.J. is employed by the Spirit JinYu Biological Pharmaceutical Co., Ltd, Hohhot, China. The re-maining authors declare no potential conflicts of interest.

Figures

Figure 1
Figure 1
Rectal temperature of experimental rabbits pre- (A) and postchallenge by M. bovis HB0801 (B) and BoHV-1 HB06 (C).
Figure 2
Figure 2
Changes in the mean body weight of the rabbits in the M. bovis groups (A) and BoHV-1 (B) groups during the entire experimental period.
Figure 3
Figure 3
Detection of M. bovis HB0801 (A) and BoHV-1 HB06 (B) shedding in nasal swabs after the challenge. Nasal swabs were collected every day.
Figure 4
Figure 4
Humoral immune responses induced by the M. bovis–BoHV-1 bivalent vaccine. Serum samples were collected weekly to determine the (A) M. bovis serum antibody and (B) BoHV-1 serum neutralizing antibody. Variation is expressed as standard deviation.
Figure 5
Figure 5
Production of IFN-β (A) and TNF-α (B) after immunization. The cytokines were detected with commercial ELISA kits. Results are shown as means ± SEM.
Figure 6
Figure 6
Histopathological images of lung tissues from the experimental rabbits stained by H&E. (A–C) represent experimental groups with different antigenic ratios of 1:1, 1:2, and 2:1, respectively. (D–F) represent experimental groups of the M. bovis HB150 immunization vaccine, the unvaccinated group, and the blank control group as the negative control.
Figure 7
Figure 7
Histopathological images of lung tissues from the experimental rabbits stained by H&E. (A–C) represent experimental groups with different antigenic ratios of 1:1, 1:2, and 2:1, respectively. (D,E) represent experimental groups of the BoHV-1 gG-/tk- immunization vaccine and unvaccinated groups.
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