Adhesion of Lactobacillus plantarum 423 and Lactobacillus salivarius 241 to the intestinal tract of piglets, as recorded with fluorescent in situ hybridization (FISH), and production of plantaricin 423 by cells colonized to the ileum

Maré, L.; Wolfaardt, Gideon; Dicks, Leon M. T.

doi:10.1111/j.1365-2672.2006.02835.x

Cited by 39 publications

(24 citation statements)

References 66 publications

(105 reference statements)

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“…Previous studies found L. plantarum to predominantly persist in the cecum and colon (5,57). In another study, L. plantarum 423 was reported to adhere to mucus isolated from the ileum and cecum of piglets (58). The CFU counts confirmed that the cecum and colon were the major sites of L. plantarum 423 and E. mundtii ST4SA colonization (Fig.…”

Section: Discussionsupporting

confidence: 65%

Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice

Zyl

Deane

Dicks

2015

Appl Environ Microbiol

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Lactic acid bacteria (LAB) are natural inhabitants of the gastrointestinal tract (GIT) of humans and animals, and some LAB species receive considerable attention due to their health benefits. Although many papers have been published on probiotic LAB, only a few reports have been published on the migration and colonization of the cells in the GIT. This is due mostly to the lack of efficient reporter systems. In this study, we report on the application of the fluorescent mCherry protein in the in vivo tagging of the probiotic strains Enterococcus mundtii ST4SA and Lactobacillus plantarum 423. The mCherry gene, encoding a red fluorescent protein (RFP), was integrated into a nonfunctional region on the genome of L. plantarum 423 by homologous recombination. In the case of E. mundtii ST4SA, the mCherry gene was cloned into the pGKV223D LAB/Escherichia coli expression vector. Expression of the mCherry gene did not alter the growth rate of the two strains and had no effect on bacteriocin production. Both strains colonized the cecum and colon of mice. Lactic acid bacteria (LAB) are common inhabitants of a healthy human gastrointestinal tract (GIT). Lactobacillus spp. are used as starter cultures in many fermented foods and are well known for their probiotic properties and the exclusion of pathogens from the GIT (1-6). Some strains have been closely associated with the treatment of gastrointestinal disorders, lactose intolerance, and stimulation of the immune system (7,8).Despite the increasing consumer interest in probiotic LAB, the mechanisms whereby these bacteria exert their beneficial effects in the GIT are not well understood (9). Although simulated gastrointestinal models (10-14) produced valuable data on the colonization of probiotic LAB and the exclusion of pathogens, the method remains an in vitro approach and is not a true reflection of in situ conditions. Labeling of probiotic bacteria with genes expressing fluorophores allows studying colonization and competition between gut microorganisms in vivo (15)(16)(17)(18)(19).Since the discovery of the green fluorescent protein (GFP) by Shimomura et al. (20), a number of genetic variants that emit light at longer wavelengths, and that are more suited for in vivo animal studies, have been described (21-23). The mCherry red fluorescence protein (RFP), a variant of the Discosoma red (DsRed) protein (24,25), is excited at wavelengths longer than 600 nm and is photostable (26-28). Studies conducted with Bacillus subtilis (29) and Escherichia coli (30) showed that expression of the mCherry protein, even at high levels, had no effect on the cell's physiology. Tauer and coworkers (31) used the mCherry gene to study recombinant protein expression in Lactobacillus plantarum. Mohedano and coworkers (32) adapted the broad-host-range vector pNZ8048 to study gene expression in Lactobacillus and showed that the mCherry construct could be used to study complex promoter induction mechanisms, such as bacteriocin production by Lactobacillus acidophilus. The role that lactococcin 972 (...

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Section: Discussionsupporting

confidence: 65%

Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice

Zyl

Deane

Dicks

2015

Appl Environ Microbiol

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“…Esto les permite pasar a través del tracto gastrointestinal sin sufrir alteraciones en su comportamiento fisiológico. Así, para que una cepa probiótica ejerza una óptima actividad es imprescindible que sea capaz de manifestar una actividad metabólica que le permita colonizar el intestino delgado; por lo que la tolerancia a bilis, es un criterio esencial de selección de bacterias lácticas (27).…”

Section: Discussionunclassified

Caracterización de bacterias probióticas aisladas del intestino grueso de cerdos como alternativa al uso de antibióticos

G¹,

F²,

T³

2009

Rev MVZ Córdoba

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RESUMENObjetivo. Caracterizar bacterias ácido lácticas probióticas aisladas del intestino grueso de cerdos adultos como posible alternativa al uso de antibióticos. Materiales y métodos. Se incluyeron muestras de intestino grueso de 20 cerdos adultos, para el aislamiento de bacterias lácticas (BAL). La caracterización de los parámetros bioquímicos-fisiológicos de las BAL contemplo la evaluación del pH del fermento, determinación de ácidos orgánicos por HPLC y detección de isómeros de ácido láctico. Los procedimientos microbiológicos evaluaron la capacidad de inhibición del crecimiento de patógenos y la actividad antagónica del probiótico vs antibiótico con su interacción competitiva. Se utilizaron pruebas API CHL 50 para la identificación de las BAL que se complementaron con la producción de gas, actividad de catalasa y viabilidad a diferentes concentraciones de sales biliares y temperaturas. Resultados. El plan de análisis permitió seleccionar dos aislados que se identificaron como: Lactobacillus plantarum 1 H1 y Lb. plantarum 1 H2. Las pruebas microbiológicas evidenciaron que las BAL seleccionadas inhibieron principalmente los enteropatógenos: Escherichia coli, Salmonella typhimurium y Clostridium perfringens. Las pruebas bioquímicas fueron compatibles con el género Lactobacillus, mostrando el siguiente perfil: Catalasa negativas; no productoras de gas; productoras de isómeros DL; resistentes a sales biliares y pH óptimo de crecimiento 3.5 a 38 o C. Conclusiones. Los dos aislados de Lactobacillus plantarum 1 reunieron las características para el uso potencial como probióticos. Se recomienda su uso como complemento de terapias con antibióticos y en la elaboración de inóculos para aplicación en raciones alimenticias de lechones destetos, sin embargo, estudios complementarios se requieren para evaluar el efecto directo del probiótico como sustituto del uso de antibióticos in vivo.Palabras clave: Lactobacillus plantarum, probióticos, antibióticos.

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“…Some investigators have investigated survival or activity of bacteriocin-producing strains but not bacteriocin stability in intestinal simulation studies (2,21). While Maré et al (15) confirmed plantaricin production in the ileal section of a gastrointestinal model, the solution used appeared to lack digestive enzymes. In the present study, RLSM was added (1:1) to simulated gastric juice or porcine gastric digesta (pH 2) and to simulated ileum juice or ileum digesta, and each of the mixtures was incubated at 37°C for 2 and 5 h, respectively.…”

mentioning

confidence: 99%

Fate of the Two-Component Lantibiotic Lacticin 3147 in the Gastrointestinal Tract

Gardiner

Rea

O'Riordan

et al. 2007

Appl Environ Microbiol

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The component peptides of lacticin 3147 were degraded by ␣-chymotrypsin in vitro with a resultant loss of antimicrobial activity. Activity was also lost in ileum digesta. Following oral ingestion, neither of the lacticin 3147 peptides was detected in the gastric, jejunum, or ileum digesta of pigs, and no lacticin 3147 activity was found in the feces. These observations suggest that lacticin 3147 ingestion is unlikely to have adverse effects, since it is probably inactivated during intestinal transit.Lacticin 3147 is a lantibiotic produced by Lactococcus lactis (16,23). Its antimicrobial activity is mediated by the combined action of its component peptides, LtnA1 and LtnA2 (29). Like other bacteriocins, lacticin 3147 has a number of potential applications for both food and biomedicine (8). In particular, it offers advantages for use in foods, and previous studies have demonstrated its efficacy, both as a food biopreservative and in improving food quality (9, 27). While its broad spectrum offers certain advantages, it may also be a cause for concern, given that it has inhibited all gram-positive bacteria tested to date, including prominent members of the resident gut microbiota such as Lactobacillus, Enterococcus, and Streptococcus spp. (23). Indeed, the direct addition of lacticin 3147 to a human fecal fermentation gut model causes a dramatic reduction in gram-positive populations after only 30 min (22). Consequently, the ingestion of lacticin 3147 or other broad-spectrum bacteriocins in foods has the potential to perturb the commensal gut microbiota. Therefore, as part of a risk assessment for bacteriocins with potential applications in foods, it is important to determine their fate in the gastrointestinal tract (GIT) in vivo. This type of information is also important when considering potential therapeutic applications of orally delivered bacteriocins. This study evaluates the fate of lacticin 3147 in the digestive tract, initially by means of in vitro/ex vivo simulation studies and subsequently in vivo using the pig as a model. A food-grade lacticin 3147 ingredient developed in our laboratory to facilitate food applications was employed (17,18). This spray-dried lacticin 3147 skim milk powder was manufactured as described previously (22) and reconstituted (10%, wt/vol) in sterile distilled water to prepare the reconstituted lacticin 3147 skim milk (RLSM) used in experiments.To test lacticin 3147's sensitivity to a range of proteolytic enzymes representative of those found in the mammalian GIT, RLSM was incubated at 37°C for 3 h with 25 mg/ml trypsin,

show abstract

Adhesion of Lactobacillus plantarum 423 and Lactobacillus salivarius 241 to the intestinal tract of piglets, as recorded with fluorescent in situ hybridization (FISH), and production of plantaricin 423 by cells colonized to the ileum

Cited by 39 publications

References 66 publications

Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice

Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice

Caracterización de bacterias probióticas aisladas del intestino grueso de cerdos como alternativa al uso de antibióticos

Fate of the Two-Component Lantibiotic Lacticin 3147 in the Gastrointestinal Tract

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