Abstract
Weaning of piglets could increase the risk of infecting with Gram-negative pathogens, which can further bring about a wide array of virulence factors including the endotoxin lipopolysaccharide (LPS). It is in common practice that the use of antibiotics has been restricted in animal husbandry. Alkaline phosphatase (AKP) plays an important role in the detoxification and anti-inflammatory effects of LPS. This study investigated the protective effects of AKP on intestinal epithelial cells during inflammation. Site-directed mutagenesis was performed to modulate the AKP activity. The enzyme activity tests showed that the activity of the DelSigD153G-D330N mutants in B. subtilis was nearly 1,600 times higher than that of the wild-type AKP. In this study, an in vitro LPS-induced inflammation model using IPEC-J2 cells was established. The mRNA expression of interleukin-(IL-) 6, IL-8, and tumor necrosis factor-α (TNF-α) were extremely significantly downregulated, and that of ASC amino acid transporter 2 (ASCT-2), zonula occludens protein-1 (ZO-1), and occludin-3 (CLDN-3) were significantly upregulated by the DelSigD153G-D330N mutant compared with LPS treatment. This concludes the anti-inflammatory role of AKP on epithelial membrane, and we are hopeful that this research could achieve a sustainable development for the pig industry.
Highlights
Weaning is a critical stage of mammalian postpartum growth and intestinal development
Compared with the non-induced control, the Alkaline phosphatase (AKP) activity of the recombinants derived from E. coli was nearly increased by two times, while that from porcine and B. subtilis was not increased (Figure 1A)
Results revealed that the activity of the DelSigD153G-D330N mutants were the highest, which was 32 times higher than that of the DelSigphoA (Figure 1D)
Summary
Weaning is a critical stage of mammalian postpartum growth and intestinal development. The early stages of weaning are usually associated with decreased performance (Pié et al, 2004: 6417) and increased incidence of intestinal diseases and diarrhea in animals (Fairbrother et al, 2005: 17-39), which would subsequently result in increasing the risk of infecting with Gram-negative pathogens, such as E. coli, S. typhimurium, etc. It is necessary to prevent gut disorder and diseases by maintaining proper barrier function. Farmers were gravitated to add different antibiotics into the feeds to prevent or reduce pathogenic infections. Considering the mentioned issues, it is compelling to develop effective alternatives to replace antibiotics in animal feeds and to promote high-quality development of animal husbandry. Antibiotic alternatives include probiotics, organic acids, auxins, prebiotics, synbiotics, enzymes, antimicrobial peptides, hyperimmune eggs antibodies, clay, and metal (Gadde et al, 2017: 26-45)
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