<i>In vivo</i> study on the persistence of transformed β-glucanase-producing <i>Lactobacillus</i> strains in the gastrointestinal tract of chickens

Abstract

The persistence and ability of transformed Lactobacillus strains to function and produce β-glucanase in the gastrointestinal tract of chickens was investigated. Chickens were fed diets supplemented with 10 9 cells/kg of transformed Lactobacillus strains which harboured plasmid pSA3 carrying a gene encoding β-glucanase. The transformed β-glucanase-producing Lactobacillus strains were L. brevis C10CpSA3b6, L.crispatus I12pSA3b6, L. brevis I23pSA3b6, L. fermentum I25pSA3b6, L. brevis I211pSA3b6 and L. brevis I218pSA3b6. At days 14 to 18, digesta samples from the gizzard, duodenum, jejunum, ileum and caecum were collected daily and the presence of the respective β-glucanase-producing Lactobacillus strains was determined by a PCR-based method. Results of the study showed that the β-glucanase-producing Lactobacillus strains could persist in the gastrointestinal tract of the chickens for about two days. The strains were found to be alive and active in vivo, in which β-glucanase activity was significantly (P<0.05) higher in chickens fed diets supplemented with β-glucanase-producing Lactobacillus strains. Higher enzyme activities were located in the jejunum and ileum, especially in birds fed β-glucanase-producing L. brevis C10CpSA3b6 and L. brevis I211pSA3b6. The results indicated that the Lactobacillus strains could be a potential vehicle to deliver enzymes into the gastrointestinal tract of chickens.

Lactobacillus Strain Ecology and Persistence within Broiler Chickens Fed Different Diets: Identification of Persistent Strains

“…While several articles have described Lactobacillus strains which are able to survive and persist within the GIT of poultry (8,16,31,42,46), the persistence ranged from 1 day through 4 weeks after cessation of administration. Ehrmann et al (16) described the persistence of their two selected strains within ducks for up to 28 days using only a single dose.…”

“…Lactobacilli are also excellent candidates as mucosal delivery vectors designed to express bioactive peptides in situ to reduce colonization by C. perfringens. The use of lactobacilli and other lactic acid bacteria (LAB) as live delivery vectors for therapeutic proteins has recently been reviewed (6, 53), but few studies have been conducted in chickens (46,55,56). An attenuated Salmonella enterica serovar Typhimurium delivery vector targeting C. perfringens has provided partial protection against experimental NE challenge (35, 57).…”

Coexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri

“…Lactobacillus probiotics have been used to deliver proteins (such as green fluorescent protein (GFP), therapeutic peptides, bacteriocins and enzymes) to the chicken gastrointestinal tract to further the understanding of the mechanisms of colonization and adhesion [ 22 , 23 ], treating disease and cancer [ 24 ], substituting antibiotics [ 25 ] or improving production performance [ 26 , 27 ]. As a delivery vector, Lactobacilli must be able to express and secrete proteins of interest abundantly and efficiently; thus, it is very important to select appropriate and effective transcriptional promoters and secretion signal peptides [ 28 ].…”

Transformation of, and Heterologous Protein Expression in, Lactobacillus agilis and Lactobacillus vaginalis Isolates from the Chicken Gastrointestinal Tract

“…Our results are consistent with other studies that have successfully utilized ldhL promoters to drive heterologous expression of the gfp gene within lactobacilli (20,26,56). There are exciting applications in using these strains to deliver proteins to the chicken GIT: either GFP, which could facilitate studies into the mechanism(s) of colonization (34,56), or therapeutic peptides, such as antigens to be used as mucosal vaccines (54), bacteriocins (38) as an alternative to antibiotics (22), or enzymes to increase production efficiency (25,42).…”