Two Lactobacillus Species Inhibit the Growth and α-Toxin Production of Clostridium perfringens and Induced Proinflammatory Factors in Chicken Intestinal Epithelial Cells in Vitro

Abstract

Clostridium perfringens is the causative pathogen of avian necrotic enteritis. Lactobacillus spp. are well-characterized probiotics with anti-microbial and immune-modulatory activities. In the present study, we investigated the effects of L. acidophilus and L. fermentum on the growth, α-toxin production and inflammatory responses of C. perfringens. In in vitro culture experiments, both lactobacilli inhibited the growth of C. perfringens (P < 0.01), accompanied with a decrease in pH (P < 0.01). Supernatants from lactobacilli cultures also suppressed the growth of C. perfringens during 24 h of incubation (P < 0.01), but this inhibitory effect disappeared after 48 h. Both lactobacilli decreased the α-toxin production of C. perfringens (P < 0.01) without influencing its biomass, and even degraded the established α-toxin (P < 0.01). Lower environmental pH reduced the α-toxin production as well (P < 0.01). Preincubation with L. acidophilus decreased the attachment of C. perfringens to cells (P < 0.01) with the cell cytotoxicity being unaffected. Both lactobacilli pretreatment reduced the up-regulation of proinflammatory factors, peptidoglycan (PGN) receptors and nuclear factor kappa B (NF-κB) p65 in C. perfringens-challenged chicken intestinal epithelial cells (P < 0.05). In conclusion, L. acidophilus and L. fermentum inhibited the pathological effects of C. perfringens in vitro conditions.

Effect of probiotic treatment on the clinical course, intestinal microbiome, and toxigenic Clostridium perfringens in dogs with acute hemorrhagic diarrhea

“…acidophilus and L . fermentum [ 45 , 46 ]. The probiotic mixture used in our study may be associated with an increase of beneficial bacteria and decrease of possible pathogenic bacteria due to similar mechanisms.…”

Effects of Lactobacillus plantarum on intestinal integrity and immune responses of egg-laying chickens infected with Clostridium perfringens under the free-range or the specific pathogen free environment

“…Environment improvement (intensive feeding patterns) and the extensive use of antibiotics have played an important role in preventing CP infection and the incidence of NE in the past decades [33]. Antibiotics are considered to be effective measures to reduce the incidence of NE, but due to ban on feeding growth-promoting antibiotics in Europe, there has been an increase in the incidences of NE [14]. Owing to the ban on the use of antibiotics in feed industry, researchers have searching for alternatives to help growth-promoting and prevention of the incidence of NE.…”

Research Note: Effect of synbiotic supplementation on caecal Clostridium perfringens load in broiler chickens with different necrotic enteritis challenge models

“…Probiotic species outcompete pathogenic bacteria by secreting bacteriocins and reducing the pH of the intestine ( Dobson et al., 2012 ). C. perfringens is highly sensitive to environmental pH where CP proliferation is greatly reduced at pH below 5.1 ( Guo et al., 2017 ). Lactic acid bacteria have been shown to have bactericidal and bacteriostatic action through production of bacteriocins, and the optimal pH for lactic acid bacteria is 5.5 of which they produce lactic acid to further decrease the pH of the media ( Hutkins and Nannen, 1993 ).…”

“…Supernatant from L. reuteri reduced the in vitro proliferation of CP in this study. An earlier report identified 2 different lactobacillus strains, Lactobacillus acidophilus and L. fermentum , reduced the in vitro proliferation of CP ( Guo et al., 2017 ). This inhibitory reaction was confirmed by treating the P. acidilactici supernatants with proteolytic enzymes, which reversed the inhibitory effects of P. acidilactici ( Jager and Harlander, 1992 ).…”

“…Lactobacillus probiotics not only decrease the production of alpha toxin by CP but also degraded the alpha toxin ( Guo et al., 2017 ). Furthermore, in vitro preincubation of chicken intestinal cells with L. acidophilus decreases the attachment of CP to the intestinal cells.…”