International Journal Probiotics and Prebiotics, Volume 1 Number 2 May 2007

COMMENTARY
83 Improvement of Intestinal Sugar Uptake Through Probiotic Fermentation And Subsequent Induced Hepatic Gluconeogenesis
GER BONGAERTS, RENÉ SEVERIJNEN, THEO WAGENER

REVIEW ARTICLES
87 Probiotics: Are They Incredible Panaceas? On The Science Behind Beneficial Non-Pathogenic Microbial Agents
GER BONGAERTS AND RENÉ SEVERIJNEN

97 Probiotics and Prebiotics In Inflamatory Bowel Disease: Existing Evidence and Potential Benefits
PAOLO GIONCHETTI, FERNANDO RIZZELLO, CLAUDIA MORSELLI, ROSY TAMBASCO, CARLO CALABRESE AND MASSIMO CAMPIERI

105 Pathogenicity Of Fusobacterium Nucleatum; General Aspects Of Its Virulence
MARIO J. AVILA-CAMPOS AND VIVIANE NAKANO

RESEARCH ARTICLES
113 Adaptation To Lactose Intolerance May Not Be Achieved By Long Term Ingestion Of A Multi- Species Containing Probiotic: An Extended Preliminary Study
PAULA MALOLEPSZY, IAN SHRIER AND ANDREW SZILAGYI

121 Sorbitol, Maltodextrin, Inulin And Bifidobacterium Infantis Modify Serum Lipid Profiles, Intestinal Microbial Population And Organic Acids Concentration In Rats
MIN-TZE LIONG AND NAGENDRA P. SHAH

131 Probiotic Traits Of Lactobacillus Plantarum Strains Isolated From Shmen, A Traditional Butter Made From Camel Milk
KACEM MOURAD

137 The Effects Of Selenium Enriched Enterococcus Faecium M-74 On Methotrexate Treatment Of Rats With Adjuvant Arthritis
JOZEF ROVENSKÂ, KAROL ŠVÍKA, MÁRIA STANÈÍKOVÁA, RICHARD IŠTOKA, LIBOR EBRINGERB, MIROSLAV FERENÈÍKC

CLINICAL CASE REPORT
145 Lactobacillus Fermentum Bacteremia In A Seriously Ill Premature Short Small Bowel Patient During Probiotic Lactobacillus Casei Therapy
GER P.A. BONGAERTS, SABINE L.A.G. VRANCKEN, JOS M.TH. DRAAISMA, CORRY M.R. WEEMAES, AND RENÉ S.V.M. SEVERIJNEN







Commentary
83-86 Improvement Of Intestinal Sugar Uptake Through Probiotic Fermentation And Subsequent Induced Hepatic Gluconeogenesis
GER BONGAERTS, RENÉ SEVERIJNEN, THEO WAGENER


After consumption of digestible carbohydrates by people with insufficient intestinal sugar uptake plenty of sugar is available for intestinal bacteria. Normally this will cause increased fermentation by common intestinal bacteria. Consequently these people may feel miserable due to both an enormous gas production and adverse (non-gaseous) fermentation products. Our studies on the short small bowel syndrome as well as on cancer cachexia did us postulate that in the presence of an adequately functioning hepatic gluconeogenesis probiotic therapy with high numbers of lactic acid bacteria improves the uptake of sugars. In the proximal part of the small bowel, especially the duodenum and the first part of the jejunum, the available digestible sugars are fermented at a high rate by the consumed probiotics. Next, instead of the sugars the bowel wall takes up the produced lactic acid. As soon as the lactic acid concentration in blood passes a threshold, the hepatic gluconeogenesis is activated. This means that the liver (i) removes lactic acid from blood, (ii) indirectly increases blood pH and intracellular pH, and (iii) increases the supply of energy-rich glucose into host cells, tissues and organs. Thus, insufficient intestinal sugar uptake can be bypassed by probiotic therapy.



















Review Articles 87-96 Probiotics: Are They Incredible Panaceas? On The Science Behind Beneficial Non-Pathogenic Microbial Agents
GER BONGAERTS AND RENÉ SEVERIJNEN

This review focuses on the science behind the beneficial effects of probiotics on both the intestinal flora and on the host's condition. These probiotics concern non-pathogenic microorganisms, especially lactic acid bacteria, such as lactococci, lactobacilli and bifidobacteria, as well as yeasts, such as Saccharomyces cerevisiae and Saccharomyces boulardii. For a good understanding we have characterized the site of primary action, its specific flora and the specific properties of the various probiotics. The common property of all living probiotics implies fermentative energy generation from sugars with only a low energy yield and consequently a high fermentation rate. The nature of the probiotics determines the fermentation products and thus the specific probiotic fermentation may involve (i) only 2 moles lactic acid from 1 mole glucose (lactococci and obligate homolactic lactobacilli), (ii) nearly 2 moles lactic acid with a small amount of CO2 from 1 mole glucose (facultative heterolactic lactobacilli), (iii) only 1 mole lactic acid, 1 mole ethanol and 1 mole CO2 from 1 mole glucose (obligate heterolactic lactobacilli), (iv) 2 moles lactic acid and 3 moles acetic acid from 2 moles glucose (bifidobacteria), (v) 2 moles ethanol and 2 moles CO2 from 1 mole glucose (yeasts). A relevant specific fermentationlinked aspect is the stereo-isomeric conformation of lactic acid, i.e., D- or L-lactic acid, especially since in neonates and young children because of too low D-2-hydroxy-acid dehydrogenase activity D-lactic acid may behave as a toxic compound that is comparable with acetaldehyde in case of ethanol intoxication. Another lactic acid-linked factor that may contribute to human health concerns the human hepatic gluconeogenesis since it lowers the intestinal lactic acid load. Other factors that may contribute to human health concern bacterial production of anti-microbial compounds such as nisin and bacteriocines, mechanisms that inactivate microbial toxins, inactivating binding of toxins and viruses, microbial deconjugation of bile acids, and induction of antibodies that cross-react with cell wall components of pathogens and thus contribute to phagocytosis-mediated eradication. Consequences for therapeutic manipulation are discussed as it regards moment of intake, composition of preparation, therapeutic dose, rational choice, clinical limitations and risks.



















97-104 Probiotics And Prebiotics In Inflammatory Bowel Disease: Existing Evidence And Potential Benefits
PAOLO GIONCHETTI, FERNANDO RIZZELLO, CLAUDIA MORSELLI, ROSY TAMBASCO, CARLO CALABRESE AND MASSIMO CAMPIERI

The rationale for using probiotics and prebiotics in Inflammatory Bowel Disease is based on convincing evidence that implicates intestinal bacteria in the pathogenesis of these diseases. Probiotics are "living organisms, which upon ingestion in certain numbers, exert health benefits beyond inherent basic nutrition. Several mechanisms have been proposed to account for the action of probiotics. VSL#3, a highly concentrated cocktail of probiotics has been shown to be effective in the prevention of pouchitis onset and relapses. Results on the use of probiotics in ulcerative colitis are promising, both in terms of the prevention of relapses and the treatment of mild-to-moderate attacks. Results in Crohn's disease are not yet clear because of conflicting data and the limited number of well-performed studies. Prebiotics are dietary substances, usually nondigestible carbohydrates, which beneficially affect the host by selectively stimulating the growth and activity of protective commensal enteric bacteria prebiotics are functionally equivalent to probiotic bacteria. Prebiotics are an exciting potential treatment for Inflammatory Bowel Disease patients. They offer a safe and cost-effective approach and may be considered for long-term treatment; however, experimental evidence supporting the use of these nutriceuticals is still limited.



















105-112 Pathogenicity Of Fusobacterium Nucleatum: General Aspects Of Its Virulence
MARIO J. AVILA-CAMPOS AND VIVIANE NAKANO

F. nucleatum constitutes an important part of the subgingival microbiota of gingivitis and periodontitis. It is present in larger amounts in adults than in children. This organism exhibits several biological activities and participates in a broad range of bacterial coaggregations in oral cavity cavity. Moreover, F. nucleatum play an important role in serious infections in other parts of the body, thus an accurate identification of these species will be a great importance not only for taxonomic reasons but also for appropriate treatment of infections, since the antimicrobial susceptibility vary widely. It must be emphasized that the etiology of periodontal disease is complex and multifactorial, but although much has been learned of the involvement of F. nucleatum very little is known of the exact reactions in pathogenesis. Certainly, further studies of functions and structure relations of F. nucleatum OMPs may contribute significantly to progress.



















Research Articles
113-120 Adaptation To Lactose Intolerance May Not Be Achieved By Long Term Ingestion Of A Multi- Species Containing Probiotic: An Extended Preliminary Study
PAULA MALOLEPSZY, IAN SHRIER AND ANDREW SZILAGYI

Continued consumption of lactose in intolerant people can lead to symptomatic adaptation, putatively through colonic bacterial changes. Whether adaptation can be achieved in vivo by expanding lactic acid producing bacterial populations without metabolic adaptation is still unclear. We used the multispecies high concentration probiotic VSL3 in 43 participants to attempt to clarify this issue. Four groups of lactase non-persistent subjects underwent a single blinded 42 day trial with 50g lactose challenges at the beginning and end of intervention. The groups were as follows: Control (N=12), Consumption of 60ml milk BID (N=12), VSL3 900x109, BID (N=9) and VSL3+ 60ml milk BID (N=10). Breath hydrogen and symptoms were recorded for 4.5Hrs with each test. A 3-day recall diet survey targeting lactose was used to control for spontaneous alterations in lactose intake prior to each test and showed no substantial changes. Analysis by ANOVA of changes in hydrogen or symptoms showed no significant effect in any of the groups. In conclusion symptomatic adaptation to lactose in lactase non persistent subjects is not corrected by VSL3 and may require additional metabolic priming to bacterial population expansion. Further evaluation of the mechanism of colonic adaptation is warranted.



















121-130 Sorbitol, Maltodextrin, Inulin And Bifidobacterium Infantis Modify Serum Lipid Profiles, Intestinal Microbial Population And Organic Acids Concentration In Rats
MIN-TZE LIONG AND NAGENDRA P. SHAH

The aims of this study were to evaluate the effectiveness of synbiotics containing Bifidobacterium and the prebiotics sorbitol, maltodextrin and/or inulin on serum lipid profiles, intestinal microbiology, intestinal organic acids concentration and the translocation ability of Bifidobacterium using male Wistar rats (n = 30). Rats were given five high-cholesterol diets namely the control (without probiotic and prebiotics), and those containing Bifidobacterium infantis and sorbitol (BS), B. infantis and maltodextrin (BM), B. infantis and inulin (BI), and B. infantis and all three prebiotics (BSMI). Rats on the BM diet decreased total serum cholesterol, triglycerides and low-density lipoprotein (LDL) cholesterol level compared to the control, possibly due to increased production of propionic acid. However, symbiotic diets did not contribute to any change in HDL-cholesterol level compared to those on the control diet. Diet BS, BM and BI increased the total Bifidobacterium population in the cecum and colon, accompanied by increased concentration of acetic acid. These led to decreased counts of total aerobes, Escherichia coli and bacteroides in those intestinal segments. There was no translocation of Bifidobacterium to the spleen, liver and kidney suggesting that the synbiotics used were safe for consumption. Results from this study showed that the individual synbiotic diets (BS, BM and BI) beneficially altered serum cholesterol level and intestinal microbial population to varying degrees depending on the prebiotic, while the combination of Bifidobacterium and all prebiotics (BFMI) did not perform better than when the prebiotics were used independently.



















131-136 Priobiotic Traits Of Lactobacillus Plantarum Strains Isolated From Shmen, A Traditional Butter Made From Camel Milk
KACEM MOURAD

total of 40 strains of Lactobacillus plantarum previously isolated from the traditional butter made from camel milk (shmen) were characterized for the most typical probiotic criteria (resistance to bile, acidic pH values, freeze-drying, acidifying activity, proteolytic activity, antagonistic activity and bacteriocin production). Of these, 28 strains were tolerant to 2% oxgall with survival percentage ranged between 75 ± 1.8 and 69 ± 2.1%. Among these, 18 strains were resistant to pH 2.0 (71 to 54%) after 2h incubation period. Of the acid and bile tolerant strains (18 strains) examined for antagonistic activity, 4 (L. plantarum SH5, SH12, SH24 and SH32) were able to inhibit several strains of L. plantarum with inhibitions zones wider than 15 mm in diameter. Only crud extracts (CE) from L. plantarum SH12 (CE1) and L. plantarum SH24 (CE2) were inhibitory against the sensitive strains tested. Antibacterial activity of crud extracts showed a bactericidal effect on the sensitive strains and it was completely lost after treatment with proteolytic enzymes (a-chymotrypsin and protenase K) which reflecting the proteinaceous nature of inhibitory agents (bacteriocin like substances). The CE2 produced by L. plantarum SH24 showed inhibition of a wide range of lactic acid bacteria and non-lactic acid bacteria such as Pseudomonas, Erwinia and Propionilbacterium. However, the activity of CE1 (L. plantarum SH12) was restricted to related species. Also, the antibacterial producing L. plantarum SH12 and SH24 strains have showed fast acidification activity (0.921 ± 0.29 and 1.075 ± 0.23 mmol/l ± sd of lactic acid, respectively), a good proteolytic activity (5.45± 0.21 and 3.77± 0.25 mg l-1 tyrosine at 72 h, respectively) and high survival percentage after freeze-drying.



















137-144 The Effects of Selenium Enriched Enterococcus Faecium M-74 On Methotrexate Treatment Of Rats With Adjuvant Arthritis
JOZEF ROVENSKÂ, KAROL ŠVÍKA, MÁRIA STANÈÍKOVÁA, RICHARD IŠTOKA, LIBOR EBRINGERB, MIROSLAV FERENÈÍKC

The efficacy of combination therapy with methotrexate (MTX) and probiotic bacteria Enterococcus faecium M-74 enriched with organic selenium (EFSe) in rats with adjuvant arthritis was determined. Rats with adjuvant arthritis were administered the following drugs: MTX (0.3 mg/kg 2-times weekly, orally); lyophilised Enterococcus faecium M-74 enriched with selenium (15 mg/kg, 5 days a week, orally); and a combination of MTX plus EFSe for a period of 50 days from the immunisation. Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling, arthrogram score, bone erosions, whole body bone mineral density (BMD) and radiographic scores were measured in the rats as variables of inflammation and destructive arthritis-associated changes. Treatment with MTX and with the combination therapy with MTX + EFSe significantly inhibited markers of both inflammation and arthritis. Significant differences in favour of combination therapy with MTX + EFSe as compared to MTX alone were seen in serum albumin concentration, hind paw swelling and arthrogram score. Reductions in radiographic scores were also more pronounced in the combination therapy group. Combination therapy, but not MTX alone, inhibited the reduction of BMD in arthritic rats; treatment with EFSe alone had no significant effect on adjuvant arthritis in rats. The potent therapeutic effect of low dosage methotrexate therapy in combination with selenium-enriched Enterococcus faecium M-74 on adjuvant arthritis in rats was shown.



















Clinical Case Report
145-158 Lactobacillus Fermentum Bacteremia In A Seriously Ill Premature Short Small Bowel Patient During Probiotic Lactobacillus Casei Therapy
GER P.A. BONGAERTS, SABINE L.A.G. VRANCKEN, JOS M.TH. DRAAISMA, CORRY M.R. WEEMAES, AND RENÉ S.V.M. SEVERIJNEN

A pre- and dysmature born girl acquired a severe sepsis from a necrotizing enterocolitis. After surgical resection, 38 cm of small bowel was preserved. Multiple septic periods due to central venous catheters infections and increased bowel permeability as a result of the short bowel, caused a life-threatening condition. Antifungal medicine failed to eradicate Candida albicans. We restored her natural gastric acid-mediated intestinal resistance by stopping ranitidine and administered probiotics, first Lactobacillus casei Shirota, later a mixture of 60% bifidobacteria and 40% lactobacilli and lactococci. During probiotic therapy two positive blood cultures contained lactobacilli that were proven not to be of probiotic origin. The effect of the probiotic therapy is explained.



















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