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Volume 14
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10.3390/microorganisms14081606
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Article

Prophylactic Administration of Engineered Bacillus subtilis Expressing Mucosal Repair Factors Alleviates Pullorum Disease in Chicks

by
Fei Teng
1,†,
Yingying Ma
1,†,
Xinrui Li
1,
Rongyan Li
1,
Yang Yang
1,
Yue Yan
1,
Hongzhe Zhao
1,
Guiwei Li
2,
Yanping Jiang
1,
Jiaxuan Li
1,
Wen Cui
1 and
Xinyuan Qiao
1,*
1
Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development, Department of Preventive Veterinary, College of Veterinary, Northeast Agricultural University, 600 Changjiang Road, Harbin 150030, China
2
Institute of Rural Revitalization Science and Technology, Heilongjiang Academy of Agricultural Sciences, Harbin 150030, China
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Microorganisms 2026, 14(8), 1606; https://doi.org/10.3390/microorganisms14081606
Submission received: 5 June 2026 / Revised: 17 July 2026 / Accepted: 21 July 2026 / Published: 23 July 2026
(This article belongs to the Special Issue Microbial Interventions in Veterinary Medicine)
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Abstract

Salmonella pullorum (S. pullorum) remains a major enteric pathogen in young chicks, causing high mortality and severe economic losses in poultry production. This study evaluated a prophylactic strategy using engineered Bacillus subtilis (B. subtilis) expressing gallus trefoil factor 2 (gTFF2) and epidermal growth factor (gEGF) to protect chicks against S. pullorum infection. In vitro, gEGF significantly promoted UMNSAH/DF-1 (DF-1) cell proliferation, whereas both gTFF2 and gEGF enhanced epithelial cell migration. In vivo, oral administration of recombinant B. subtilis significantly reduced mortality from 75.0% and 66.7% in the PBS and pHT43 groups to 50%, 33.3%, and 25.0% in the gTFF2, gEGF, and gTFF2+gEGF groups, respectively. Cecal colonization of S. pullorum was reduced by 2–4 log10 CFU/g, accompanied by improved average daily gain and immune organ indices. Treatment also significantly decreased serum IL-6 levels and increased TGF-β levels (p < 0.05), indicating attenuation of the inflammatory response. Histological analysis showed significantly increased villus height (0.94–1.58-fold) and villus height-to-crypt depth ratio (1.47–3.25-fold) compared with the infected controls, together with markedly alleviated hepatic and intestinal lesions. The combined administration of gTFF2- and gEGF-expressing strains consistently exhibited the greatest protective efficacy. Collectively, these findings demonstrate that engineered probiotic-mediated delivery of mucosal repair factors represents a promising antibiotic-alternative strategy for preventing pullorum disease and improving intestinal health in poultry production.
Keywords: engineered probiotics; Bacillus subtilis; Salmonella pullorum; poultry health; mucosal protection; antibiotic alternatives engineered probiotics; Bacillus subtilis; Salmonella pullorum; poultry health; mucosal protection; antibiotic alternatives

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MDPI and ACS Style

Teng, F.; Ma, Y.; Li, X.; Li, R.; Yang, Y.; Yan, Y.; Zhao, H.; Li, G.; Jiang, Y.; Li, J.; et al. Prophylactic Administration of Engineered Bacillus subtilis Expressing Mucosal Repair Factors Alleviates Pullorum Disease in Chicks. Microorganisms 2026, 14, 1606. https://doi.org/10.3390/microorganisms14081606

AMA Style

Teng F, Ma Y, Li X, Li R, Yang Y, Yan Y, Zhao H, Li G, Jiang Y, Li J, et al. Prophylactic Administration of Engineered Bacillus subtilis Expressing Mucosal Repair Factors Alleviates Pullorum Disease in Chicks. Microorganisms. 2026; 14(8):1606. https://doi.org/10.3390/microorganisms14081606

Chicago/Turabian Style

Teng, Fei, Yingying Ma, Xinrui Li, Rongyan Li, Yang Yang, Yue Yan, Hongzhe Zhao, Guiwei Li, Yanping Jiang, Jiaxuan Li, and et al. 2026. "Prophylactic Administration of Engineered Bacillus subtilis Expressing Mucosal Repair Factors Alleviates Pullorum Disease in Chicks" Microorganisms 14, no. 8: 1606. https://doi.org/10.3390/microorganisms14081606

APA Style

Teng, F., Ma, Y., Li, X., Li, R., Yang, Y., Yan, Y., Zhao, H., Li, G., Jiang, Y., Li, J., Cui, W., & Qiao, X. (2026). Prophylactic Administration of Engineered Bacillus subtilis Expressing Mucosal Repair Factors Alleviates Pullorum Disease in Chicks. Microorganisms, 14(8), 1606. https://doi.org/10.3390/microorganisms14081606

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