title: Response of porcine intestinal in vitro organ culture tissues following exposure to Lactobacillus plantarum JC1 and Salmonella enterica serovar Typhimurium SL1344.
abstract: The development of novel intervention strategies for the control of zoonoses caused by bacteria such as Salmonella spp. in livestock requires appropriate experimental models to assess their suitability. Here, a novel porcine intestinal in vitro organ culture (IVOC) model utilizing cell crown (CC) technology (CCIVOC) (Scaffdex) was developed. The CCIVOC model was employed to investigate the characteristics of association of S. enterica serovar Typhimurium strain SL1344 with porcine intestinal tissue following exposure to a Lactobacillus plantarum strain. The association of bacteria to host cells was examined by light microscopy and electron microscopy (EM) after appropriate treatments and staining, while changes in the proteome of porcine jejunal tissues were investigated using quantitative label-free proteomics. Exposure of porcine intestinal mucosal tissues to L. plantarum JC1 did not reduce the numbers of S. Typhimurium bacteria associating to the tissues but was associated with significant (P < 0.005) reductions in the percentages of areas of intestinal IVOC tissues giving positive staining results for acidic mucins. Conversely, the quantity of neutrally charged mucins present within the goblet cells of the IVOC tissues increased significantly (P < 0.05). In addition, tubulin-α was expressed at high levels following inoculation of jejunal IVOC tissues with L. plantarum. Although L. plantarum JC1 did not reduce the association of S. Typhimurium strain SL1344 to the jejunal IVOC tissues, detection of increased acidic mucin secretion, host cytoskeletal rearrangements, and proteins involved in the porcine immune response demonstrated that this strain of L. plantarum may contribute to protecting the pig from infections by S. Typhimurium or other pathogens.
authors.affiliation: Department for Food and Environmental Safety, Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom. j.collins@imperial.ac.uk
authors.forename: J W
authors.initials: JW
authors.lastname: Collins
authors.forename: N G
authors.initials: NG
authors.lastname: Coldham
authors.forename: F J
authors.initials: FJ
authors.lastname: Salguero
authors.forename: W A
authors.initials: WA
authors.lastname: Cooley
authors.forename: W R
authors.initials: WR
authors.lastname: Newell
authors.forename: R A
authors.initials: RA
authors.lastname: Rastall
authors.forename: G R
authors.initials: GR
authors.lastname: Gibson
authors.forename: M J
authors.initials: MJ
authors.lastname: Woodward
authors.forename: R M
authors.initials: RM
authors.lastname: La Ragione
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D058491:Bacterial Load; D054884:Host-Pathogen Interactions; D007422:Intestines / Q000737:chemistry / Q000382:microbiology* / Q000473:pathology*; D048191:Lactiplantibacillus plantarum / Q000254:growth & development* / Q000276:immunology*; D008853:Microscopy; D009077:Mucins / Q000032:analysis; D009924:Organ Culture Techniques; D020543:Proteome / Q000032:analysis; D012486:Salmonella typhimurium / Q000254:growth & development* / Q000472:pathogenicity*; D013552:Swine; D014404:Tubulin / Q000032:analysis
chemical_list: D009077:Mucins; D020543:Proteome; D014404:Tubulin