Oral delivery of Bacillus subtilis spores expressing Clonorchis sinensis paramyosin protects grass carp from cercaria infection.

Abstract

Clonorchis sinensis (C. sinensis), an important fishborne zoonotic parasite threatening public health, is of major socioeconomic importance in epidemic areas. Effective strategies are still urgently expected to prevent against C. sinensis infection. In the present study, paramyosin of C. sinensis (CsPmy) was stably and abundantly expressed on the surface of Bacillus subtilis spores. The recombinant spores (B.s-CotC-CsPmy) were incorporated in the basal pellets diet in three different dosages (1 × 105, 1 × 108, 1 × 1011 CFU/g pellets) and orally administrated to grass carp (Ctenopharyngodon idella). The immune responses and intestinal microbiota in the treated grass carp were investigated. Results showed that specific anti-CsPmy IgM levels in sera, skin mucus, bile, and intestinal mucus, as well as mRNA levels of IgM and IgZ in the spleen and head kidney, were significantly increased in B.s-CotC-CsPmy-1011 group. Besides, transcripts levels of IL-8 and TNF-αin the spleen and head kidney were also significantly elevated than the control groups. Moreover, mRNA levels of tight junction proteins in the intestines of B.s-CotC-CsPmy-1011 group increased. Potential pathogenetic bacteria with lower abundance and higher abundances of candidate probiotics and bacteria associated with digestion in 1 × 1011 CFU/g B.s-CotC-CsPmy spores administrated fishes could be detected compared with control group. The amount of metacercaria in per gram fish flesh was statistically decreased in 1 × 1011 CFU/g B.s-CotC-CsPmy spores orally immunized group. Our work demonstrated that B. subtilis spores presenting CsPmy on the surface could be a promising effective, safe, and needle-free candidate vaccine against C. sinensis infection for grass carp.

• Authors: Hengchang Sun, Mei Shang, Zeli Tang, Hongye Jiang, Huimin Dong, Xinyi Zhou, Zhipeng Lin, Cunbin Shi, Pengli Ren, Lu Zhao, Mengchen Shi, Lina Zhou, Houjun Pan, Ouqin Chang, Xuerong Li, Yan Huang, Xinbing Yu
• Journal: Applied microbiology and biotechnology (Appl Microbiol Biotechnol) Vol. 104 Issue 4 Pg. 1633-1646 (Feb 2020) ISSN: 1432-0614 [Electronic] Germany
• PMID: 31912200 (Publication Type: Journal Article)

Chemical References

• Antibodies, Helminth
• Immunoglobulin M
• Tropomyosin
• Vaccines

Topics

• Administration, Oral
• Animal Feed (microbiology)
• Animals
• Antibodies, Helminth (blood)
• Bacillus subtilis
• Carps (immunology, parasitology)
• Cercaria (immunology)
• Clonorchiasis (immunology, prevention & control, veterinary)
• Clonorchis sinensis
• Fish Diseases (parasitology, prevention & control)
• Immunoglobulin M (immunology)
• Intestines (immunology)
• Spores, Bacterial
• Tropomyosin (genetics, immunology)
• Vaccines (administration & dosage, immunology)