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Biomedical subjects

A C Ouwehand

Publications and source records attributed to A C Ouwehand.

At least 19 recordsLinked to original sources

Oral adhesion and survival of probiotic and other lactobacilli and bifidobacteria in vitro.

INTRODUCTION: Most probiotic products are consumed orally and hence it is feasible that the bacteria in these products may also attach to oral surfaces; however, the effects of these bacteria on the oral ecosystem are mostly unknown. Our aim was to evaluate the oral colonization potential of different probiotic, dairy, and fecal Lactobacillus and Bifidobacterium strains in vitro. METHODS: The binding of 17 Lactobacillus and seven Bifidobacterium strains to hydroxyapatite and microtitre wells coated with human saliva was tested. Binding of selected strains to human buccal epithelial cells and co-adherence with Fusobacterium nucleatum were also investigated. In addition, the survival in sterilized human whole saliva was examined. RESULTS: There was a large variation in binding to saliva-coated surfaces and buccal epithelial cells but all strains survived in saliva. The binding pattern of the probiotics did not differ from the binding of the fecal strains. F. nucleatum altered the binding of both the low-binding bifidobacteria and the high-binding lactobacilli. CONCLUSION: The differences in binding in vitro may indicate that there are also differences in the persistence of the different probiotic strains in the oral cavity in vivo.

Adult↗

Lactobacilli and enterococci--potential probiotics for dogs.

Forty strains of enterococci and forty strains of lactobacilli isolated from feces of 10 healthy dogs were tested for the antimicrobial activity, tolerance to bile and adhesion activity. The total count of fecal enterococci reached 5.5 log CFU/g and of lactobacilli 7.6 log CFU/g. Screening for production of bacteriocin-like substances showed an to partly inhibit the growth of Enterobacter sp. (hazy zones of inhibition). Ten strains of Enterococcus sp. and nine strains of Lactobacillus sp. were found without any inhibitory activity against all indicators used. Seven enterococcal strains and six lactobacilli with the broadest antimicrobial spectrum were selected for further probiotic assays. In the presence of 1% bile, the survival rate of selected enterococci (71.7-97.5%) was higher than that of lactobacilli (66.7-75.4%). The adhesion of strains to human intestinal mucus (5.1-8.2% by enterococci, 2.7-8.3% by lactobacilli) was found to be similar as adhesion to canine intestinal mucus (3.7-10.6% by enterococci, 2.1-6.0% by lactobacilli). Strain AD1, one lactobacillus isolate, reduced the higher level of serum cholesterol and alanine aminotransferase after oral administration to dogs suffering from diseases of the gastrointestinal tract.

Alanine Transaminase↗

Binding of aflatoxin B1 to cell wall components of Lactobacillus rhamnosus strain GG.

The surface of Lactobacillus rhamnosus strain GG (LGG) has previously been shown to bind aflatoxin B(1) (AFB(1)) effectively, it being a food-borne carcinogen produced by certain species of Aspergillus fungi. To establish which components of the cell envelope are involved in the AFB(1) binding process, exopolysaccharides and a cell wall isolate containing peptidoglycan were extracted from LGG and its AFB(1) binding properties were tested. LGG was also subjected to various enzymatic and chemical treatments and their effects on the binding of AFB(1) by LGG were examined. No evidence was found for exopolysaccharides, cell wall proteins, Ca(2+) or Mg(2+) being involved in AFB(1) binding. The AFB(1) binding activity of the cell wall isolate indicates that AFB(1) binds to the cell wall peptidoglycan of LGG or compounds tightly associated with the peptidoglycan.

Aflatoxin B1↗

Probiotics for the skin: a new area of potential application?

AIMS: The current study aimed at assessing, in vitro, the potential use of probiotics for the skin. METHODS AND RESULTS: Propionibacteria were chosen as potential probiotics as they are members of the normal cutaneous microbiota. Dairy strains were chosen because of their documented safe use. Production of anti-microbial substances was assessed, against selected skin pathogens. Only production of organic acids was detected. Two of the tested strains were found to exhibit high adhesion to human keratin, in vitro. Despite this high adhesion, no inhibition of skin pathogen adhesion to human keratin was observed. CONCLUSIONS: The current strains assessed may not be optimal for use as skin probiotics. However, the results of the study show that the methodology works for investigating this kind of application. SIGNIFICANCE AND IMPACT OF THE STUDY: Methods for selecting probiotics for potential application on the skin are presented.

Bacteria↗

Probiotics: on-going research on atopic individuals.

The challenge for the modern health care system is to fight against the increasing prevalence of atopic disease. The introduction of scientifically composed probiotic functional foods for prophylactic or therapeutic purposes could be one solution. Probiotics are live microbial food supplements or components of bacteria which have beneficial effects on human health. Specific strains have been demonstrated to exert powerful anti-pathogenic, anti-inflammatory and anti-allergic effects. The hygiene hypothesis suggests that atopic disease may arise from a lack of counterbalancing microbial exposure at an early age. The initial compositional development of the gut microflora is considered a key determinant of the development of both the immune responder phenotype and normal gut barrier functions. The regulatory role of probiotics in human allergic disease was first emphasised in the demonstration of a suppressive effect on lymphocyte proliferation and interleukin-4 generation in vitro. Subsequently, a significant improvement in the clinical course of atopic eczema was reported in infants given a probiotic-supplemented diet. The potential of probiotics to reduce the risk of atopic disease has recently been demonstrated in a double-blind, placebo-controlled study: probiotics administered pre- and postnatally for 6 months to at-risk subjects reduced the prevalence of atopic eczema to half of that observed in infants receiving placebo. Ongoing research is directed towards the development of novel techniques to characterise the gut microflora. Future research will clarify the mechanisms to control specific physiological processes in the evolution of atopic disease in at-risk populations or in the management of allergic diseases.

Diet↗

Assessment of adhesion properties of novel probiotic strains to human intestinal mucus.

Potential new probiotic strains Lactobacillus brevis PELI, L. reuteri ING1, L. rhamnosus VTT E-800 and L. rhamnosus LC-705 were assessed for their adhesion properties using the human intestinal mucus model. The effect on the adhesion of exposure to acid and pepsin and to milk were tested to simulate gastric and food processing conditions, and the effect of different growth media on adhesion was tested. The properties of the four strains were compared to the well-investigated probiotic L. rhamnosus strain GG. Three of the tested strains showed significant adhesion properties in the mucus model, while L. brevis PELI had intermediate adhesion and L. rhamnosus LC-705 adhered poorly. Pretreatment with different milks decreased the adhesion and low pH and pepsin treatment reduced the adhesion of all tested strains except L. rhamnosus LC-705. No competitive exclusion of pathogenic Salmonella typhimurium or Escherichia coli SfaII was observed. The results indicate that major differences exist between tested proposed probiotic strains. The growth media and the food matrix significantly affect the adhesive ability of the tested strains. This has previously not been taken into account when selecting novel probiotic strains.

Acids↗

Differences in composition and mucosal adhesion of bifidobacteria isolated from healthy adults and healthy seniors.

Fifty-one Bifidobacterium strains were isolated from the feces of healthy adults (30-40 years old) and seniors (older than 70 years of age). B. adolescentis, B. breve, B. infantis, and B. longum were isolated from the healthy adults and B. adolescentis and B. longum from elderly subjects. The tested bacteria bound, in vitro, to intestinal mucus in a strain dependent manner. The strains isolated from healthy adults, and especially B. adolescentis, bound better to intestinal mucus than those isolated from seniors. These results indicate that the mucosal adhesive properties of the human Bifidobacterium flora were reduced with the aging of the host. This shift to a Bifidobacterium flora with reduced adhesive abilities may explain the decrease in bifidobacteria levels in the intestinal microflora of aging people.

Adult↗

Comparison of mucosal adhesion and species identification of bifidobacteria isolated from healthy and allergic infants.

Fifty bifidobacteria strains were isolated from fecal samples of allergic and age matched healthy infants. Allergic infants were found to have an adult type Bifidobacterium flora with high levels of Bifidobacterium adolescentis. Healthy infants had a typical infant Bifidobacterium flora with high levels of Bifidobacterium bifidum. These isolates were tested for their adhesive properties to human intestinal mucus. The adhesion of the fecal bifidobacteria from healthy infants was significantly higher (P<0.0001) than for allergic infants. This suggests a correlation between allergic disease and the composition of the intestinal bifidobacteria flora which has reduced adhesive abilities to the intestinal mucus. Therefore, dietary supplementation of bifidobacteria typical for healthy infants, may be beneficial in the treatment of allergic disorders.

Bacterial Adhesion↗

Good adhesion properties of probiotics: a potential risk for bacteremia?

The ability to adhere to human intestinal mucus was tested for lactic acid bacteria of clinical blood culture, human fecal and dairy origin. The blood culture isolates were found to adhere better than the dairy strains. Of the Lactobacillus rhamnosus strains (nine clinical, 10 fecal and three dairy), blood culture isolates adhered better than the fecal strains. Although these results indicate a trend for blood culture isolates to bind to intestinal mucus in higher numbers than strains of dairy and human fecal origin, other factors are also likely to be involved in the etiology of lactobacillemia since some of the clinical Lactobacillus isolates exhibited a relatively low level of adhesion.

Adult↗

Characterization of the properties of human- and dairy-derived probiotics for prevention of infectious diseases in fish.

The present study aimed to investigate the potential probiotic properties of six lactic acid bacteria (LAB) intended for human use, Lactobacillus rhamnosus ATCC 53103, Lactobacillus casei Shirota, Lactobacillus bulgaricus, L. rhamnosus LC 705, Bifidobacterium lactis Bb12, and Lactobacillus johnsonii La1, and one for animal use, Enterococcus faecium Tehobak, for use as a fish probiotic. The strains for human use were specifically chosen since they are known to be safe for human use, which is of major importance because the fish are meant for human consumption. The selection was carried out by five different methods: mucosal adhesion, mucosal penetration, inhibition of pathogen growth and adhesion, and resistance to fish bile. The adhesion abilities of the seven LAB and three fish pathogens, Vibrio anguillarum, Aeromonas salmonicida, and Flavobacterium psychrophilum, were determined to mucus from five different sites on the surface or in the gut of rainbow trout. Five of the tested LAB strains showed considerable adhesion to different fish mucus types (14 to 26% of the added bacteria). Despite their adhesive character, the LAB strains were not able to inhibit the mucus binding of A. salmonicida. Coculture experiments showed significant inhibition of growth of A. salmonicida, which was mediated by competition for nutrients rather than secretion of inhibitory substances by the probiotic bacteria as measured in spent culture liquid. All LAB except L. casei Shirota showed tolerance against fish bile. L. rhamnosus ATCC 53103 and L. bulgaricus were found to penetrate fish mucus better than other probiotic bacteria. Based on bile resistance, mucus adhesion, mucus penetration, and suppression of fish pathogen growth, L. rhamnosus ATCC 53103 and L. bulgaricus can be considered for future in vivo challenge studies in fish as a novel and safe treatment in aquaculture.

Aeromonas↗

Adherence of probiotic bacteria to human intestinal mucus in healthy infants and during rotavirus infection.

The concentration of fecal mucin and the adhesion of specific probiotics and their combinations in the intestinal mucus of infants during and after rotavirus diarrhea and in healthy children were determined. Mucus was prepared from fecal samples from 20 infants during and after rotavirus diarrhea and from 10 healthy age-matched children. Mucin concentration was determined, and the adhesion of five probiotics-Lactobacillus rhamnosus GG, Lactobacillus casei Shirota, Lactobacillus paracasei F19, Lactobacillus acidophilus LA5, and Bifidobacterium lactis Bb12-and their combinations was tested in vitro. The mean concentrations of fecal mucin during and after rotavirus diarrhea, 15.2 and 14.1 mg/g, were comparable to that in healthy children, 14.9 mg/g. The adherence of probiotics ranged from 1 to 34% in healthy subjects as indicated for the following strains: L. rhamnosus GG, 34%; B. lactis Bb12, 31%; L. acidophilus LA5, 4%; L. paracasei F19, 3%; and L. casei Shirota, 1% (P = 0.0001). The distinctive pattern of probiotic adherence was not influenced by rotavirus diarrhea. The adhesion of Bb12 in the presence of GG increased from 31 to 39% in healthy infants (P = 0.018) and in episodes of diarrhea increased from 26 to 44% (P = 0.001). Rotavirus diarrhea does not decrease the production of fecal mucin or with respect to the adhesion of probiotic bacteria tested in vitro. Combination of specific probiotic strains may enhance adherence in a synergistic manner. Optimal clinical application of these interactions may offer novel therapeutic guidelines for the treatment and prevention of gastrointestinal infections.

Bacterial Adhesion↗

Chemical, physical and enzymatic pre-treatments of probiotic lactobacilli alter their adhesion to human intestinal mucus glycoproteins.

Intestinal mucus glycoproteins extracted from faeces of healthy adult subjects were used as a substratum for bacterial adhesion to investigate the effects of physical, chemical and enzymatic pre-treatments of the bacteria on their adhesion. The strains studied were Lactobacillus acidophilus 1 (LCI, Nestlé), L. rhamnosus strain GG (ATCC 53103), L. rhamnosus LC-705, and L. casei strain Shirota (Yakult, Yakult Ltd). Hereafter the strains are referred to as LA1, LGG, LC-705, and Shirota, respectively. Strains LA1 and LGG adhered greatly whereas the adhesion of strains LC-705 and Shirota to intestinal mucus glycoproteins was low. Adhesion of LA1 and LGG was reduced by boiling, autoclaving and by pepsin and trypsin treatments suggesting that the bacterial protein structures are essential for their adhesion. Treatment in ethanol and in propanol prior to adhesion significantly increased the adhesion of LA1 and LC-705, respectively. Adhesion of Shirota strain was not altered by any of the treatments.

Bacterial Adhesion↗

The mucus binding of Bifidobacterium lactis Bb12 is enhanced in the presence of Lactobacillus GG and Lact. delbrueckii subsp. bulgaricus.

The ability to adhere to mucosal surfaces is related to many probiotic health effects. In the presence of Lactobacillus GG or Lact. bulgaricus, the adhesion of Bifidobacterium lactis Bb12 to a mucus model was more than doubled. Other tested lactobacilli did not affect the adhesion, nor was the adhesion of the lactobacilli influenced by the bifidobacteria. Co-aggregation between Bif. lactis Bb12 and the tested lactobacilli was insignificant and does not explain the observed effect. The results suggest that combinations of probiotics strains may have synergistic adhesion effects. Such specific strain combinations should also be assessed in clinical studies.

Bacterial Adhesion↗

Wheat or rye supplemented diets do not affect faecal mucus concentration or the adhesion of probiotic micro-organisms to faecal mucus.

In a crossover study, volunteers consumed a diet supplemented with either wheat or rye bread. Levels of faecal mucus were determined and the adhesion of five selected probiotics to immobilized intestinal mucus, was assessed. No significant difference in the concentration of faecal mucus or adhesion of the probiotics to the mucus was observed during the two dietary periods. However, male subjects were observed to excrete significantly more mucus during the rye bread supplemented diet. From these observations it was concluded that these two diets, with a low or high fibre content, did not change the potential binding sites for the tested probiotics or affect the level of mucus excreted in the faeces, however, on a high fibre diet men excrete more faecal mucus.

Bacterial Adhesion↗

Adhesion of inactivated probiotic strains to intestinal mucus.

It has been suggested that probiotics should be viable in order to elicit beneficial health effects. Inactivation of probiotics has been suggested to interfere with the binding to the mucosa and thereby with the immune modulating activity of probiotics. The effect of different inactivation methods on the mucus adhesion of nine probiotic strains was studied. Inactivation by heat or gamma-irradiation generally decreased the adhesive abilities. However, heat treatment increased the adhesion of Propionibacterium freudenreichii and gamma-irradiation enhanced the adhesion of Lactobacillus casei Shirota. Inactivation by u.v. was not observed to modulate the adhesion of the tested strains and it was concluded to be the most appropriate method for studying non-viable probiotics and preparing control products.

Bacterial Adhesion↗