Nicotine is a potent extracellular polysaccharide inducer in Fusobacterium nucleatum biofilms

The purpose of this in vitro study was to analyze the influence of nicotine on the extracellular polysaccharides in Fusobacterium nucleatum biofilm. Methods: F. nucleatum (ATCC 10953) biofilms supplemented with different concentrations of nicotine (0, 0.5, 1, 2, 4, and 8 mg/mL) were grown in two different BHI broth conditions [no sucrose and 1% sucrose]. Extracellular polysaccharides assay, pH measurements, and a spectrophotometric assay were performed. Data were submitted for ANOVA and Tukey honestly significant difference analyses (HSD) tests (α =.05). Results: Extracellular polysaccharides synthesis was influenced by an interaction between nicotine concentrations and growth medium solution containing sucrose (P<.05). The pH values declined in the sucrose-exposed biofilm were greater than in the group exposed only to nicotine (P<.05). The biofilm exposed to sucrose and nicotine had a higher total biofilm growth (P<.05) than the nicotine-treated biofilm without sucrose. Conclusions: Regardless of sucrose exposure, biofilms exposed to different nicotine concentrations influenced the amount of extracellular polysaccharides

Exopolysaccharides from Klebsiella oxytoca: anti-inflammatory activity

Abstract Exopolysaccharides (EPS) produced by Klebsiella oxytoca are of environmental, pharmaceutical, and medicinal interest. However, studies about the anti-inflammatory activity of EPS produced by this microorganism still remain limited. The aim of this study was to produce, characterize, and evaluate the anti-inflammatory activity of EPS from K. oxytoca in a pleurisy model. Colorimetric analysis revealed that precipitated crude exopolysaccharides (KEPSC) and deproteinated exopolysaccharides (KEPS) present high levels of total carbohydrates (65.57% and 62.82%, respectively). Analyses of uronic acid (7.90% in KEPSC and 6.21% in KEPS) and pyruvic acid (3.01% in KEPSC and 1.68% in KEPS) confirm that the EPS are acidic. Gas chromatography-mass spectrometry analyses demonstrated that the EPS consisted of rhamnose (29.83%), glucose (11.21%), galactose (52.45%), and mannose (6.50%). The treatment of an experimental pleurisy model in rats through subcutaneous administration of 50, 100, 200, and 400 mg/kg of KEPS decreased both the volume of inflammatory exudate and the number of leukocytes recruited to the pleural cavity. The present data showed that EPS production by K. oxytoca using the method described is easy to perform and results in a good yield. In addition, we show that KEPS exhibit anti-inflammatory activity when administered subcutaneously in rats.

Fructosan form Paenibacillus kribbensis PS04 enhance disease resistance against Rhizoctonia solani and tobacco mosaic virus

BACKGROUND: Rice sheath blight (caused by Rhizoctonia solani) and tobacco mosaic virus are very important plant diseases, causing a huge loss in global crop production. Paenibacillus kribbensis PS04 is a broad-spectrum biocontrol agent, used for controlling these diseases. Previously, extracellular polysaccharides (EPS) from P. kribbensis PS04 had been purified and their structure was inferred to be fructosan. This study aimed to evaluate the effects of exogenous EPS treatment on plant­pathogen interactions. RESULTS: Plant defense genes such as phenylalanine ammonia-lyase, catalase, chitinase, allene oxide synthase, and PR1a proteins were significantly induced by exogenous EPS treatment. Moreover, subsequent challenge of EPSpretreated plants with the pathogens (R. solani or tobacco mosaic virus) resulted in higher expression of defenseassociated genes. Increased activities of defense-associated enzymes, total phenols, and flavonoids were also observed in EPS pretreated plants. The contents of malondialdehyde in plants, which act as indicator of lipid peroxidation, were reduced by EPS treatment. CONCLUSIONS: This study comprehensively showed that EPS produced from P. kribbensis PS04 enhances disease resistance in plants by the activation of defense-associated genes as well as through the enhancement of activities of defense-related enzymes.

Evaluation of anti-biofouling potential of Viburnum opulus extracts / Evaluación del potencial anti bioincrustante de extractos de Viburnum opulus

In this study the in vitro investigation of the inhibitory effect of ethanol extract of Viburnum opulus L. bark sample on Streptococcus mutans planctonic cells and biofilm has been intended. A Scanning electron microscopy analysis has been performed in order to investigate the inhibitory effect of the extract on Streptococcus mutans biofilms. Furthermore, the Exopolysaccharide and dextran production of this bacteria have been identified in the presence of the extract. It has been found out that the bark extract with the concentration of 2,5 mg/mL is able to inhibit more than 50% of the cells in the different times development phases. According to this, the exopolymeric matrix on the biofilm surface disperses and the Exopolysaccharide and dextran production get lowered in the presence of bark extract compared to the control group. It is considered that this extract can be used as an alternative approach for the new chemotherapeutic strategies against tooth decay.

En este estudio se investigó el efecto inhibitorio in vitro del extracto de etanólico de una muestra de corteza de Viburnum opulus L. en biopelículas de células planctónicas de Streptococcus mutans. Se realizó un análisis de microscopía electrónica de barrido para investigar el efecto inhibitorio del extracto sobre las biopelículas de Streptococcus mutans. Además, se identificó la producción de exopolisacárido y dextrano de esta bacteria en presencia del extracto. Se descubrió que el extracto de corteza con una concentración de 2,5 mg/ml inhibió más del 50% de las células en las diferentes fases de desarrollo. Consecuentemente, la matriz exopolimérica en la superficie de la biopelícula se dispersa y la producción de exopolisacárido y dextrano se reduce en presencia de extracto de corteza en comparación con el grupo de control. Se sugiere que este extracto puede ser usado como un enfoque alternativo para las nuevas estrategias quimioterapéuticas contra la carie dental.

Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

Abstract DNA vaccines have been evaluated as an option to prevent several diseases. In this study, the capacity of the xanthan biopolymer to improve the DNA vaccines immune response, administered intramuscularly, was evaluated. The experimental vaccines consisted of genes encoding fragments of the proteins LigA and LigB of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Copenhageni strain Fiocruz L1-130. The humoral immune response was evaluated by indirect ELISA. Cytokine expression levels were determined by RT-qPCR. Compared to the control group, the IgG antibody levels of animals immunized with pTARGET/ligAni and pTARGET/ligBrep plasmids associated with xanthan biopolymer were significantly higher than the control group. Additionally, there was a significant increase in IL-17 expression in animals vaccinated with pTARGET/ligBrep and xanthan.

Application of Films Based on Chitosan and Xanthan Gum in Refrigerated Fish Conservation

Abstract This research aims to determine the efficiency of chitosan and xanthan gum films in conservation of croaker fillets kept in refrigeration for 9 days. Proximal composition, loss of mass, color, pH, TVB-N (Total Volatile Bases) and microbiological profile were assessed. The films were prepared with chitosan and xanthan gum in varying mass proportions 100:0, m:m (C100XG0); 60:40, m:m (C60XG40); 50:50, m:m (C50XG50). They presented the respective values for moisture content, water solubility, thickness and water vapor permeability: 24.59%, 19.50%, 0.086 mm and 11.45gm-1.s-1.Pa-1for C100XG0; 24.58%; 20.27%, 0.091 mm and 10.41 gm-1.s-1.Pa-1for C60XG40; 22.11%, 22.06%, 0.089 mm and 10.68 gm-1.s-1.Pa-1 forC50XG50.The films were made in small bags format capable to hold about 20 g of fish fillets. A control sample was prepared in parallel, using polyethylene bags under the same storage conditions. The results showed that the chitosan films combined with xanthan gum had excellent antimicrobial properties, capable of preserving the quality of chilled fish fillets during the studied period, since it inhibited the growth of Staphylococcus coagulase-positive, Salmonella spp and coliforms at 45 ° C. Mass loss of the croaker fillets was not significantly affected by xanthan gum addition to the films. On the other hand, xanthan gum addition affected pH and color parameters of the corvina fillets. It was also verified that the combination of these two polymers promoted the reduction of N-BVT, being the C50XG50 film that presented the best response.

Role of extracellular DNA in Enterococcus faecalis biofilm formation and its susceptibility to sodium hypochlorite

Abstract Objective This study investigated the role of extracellular deoxyribonucleic acid (eDNA) on Enterococcus faecalis ( E. faecalis ) biofilm and the susceptibility of E. faecalis to sodium hypochlorite (NaOCl). Methodology E. faecalis biofilm was formed in bovine tooth specimens and the biofilm was cultured with or without deoxyribonuclease (DNase), an inhibitor of eDNA. Then, the role of eDNA in E. faecalis growth and biofilm formation was investigated using colony forming unit (CFUs) counting, eDNA level assay, crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. The susceptibility of E. faecalis biofilm to low (0.5%) or high (5%) NaOCl concentrations was also analyzed by CFU counting. Results CFUs and biofilm formation decreased significantly with DNase treatment (p<0.05). The microstructure of DNase-treated biofilms exhibited less structured features when compared to the control. The volume of exopolysaccharides in the DNase-treated biofilm was significantly lower than that of control (p<0.05). Moreover, the CFUs, eDNA level, biofilm formation, and exopolysaccharides volume were lower when the biofilm was treated with DNase de novo when compared to when DNase was applied to matured biofilm (p<0.05). E. faecalis in the biofilm was more susceptible to NaOCl when it was cultured with DNase (p<0.05). Furthermore, 0.5% NaOCl combined with DNase treatment was as efficient as 5% NaOCl alone regarding susceptibility (p>0.05). Conclusions Inhibition of eDNA leads to decrease of E. faecalis biofilm formation and increase of susceptibility of E. faecalis to NaOCl even at low concentrations. Therefore, our results suggest that inhibition of eDNA would be beneficial in facilitating the efficacy of NaOCl and reducing its concentration.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization

Abstract Objectives: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. Methodology: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). Results: M. urundeuva All. at 100, 10 and 0.1 μg/mL and Q. grandiflora Mart. at 100 and 0.1 μg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 μg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 μg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. Conclusions: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Exopolysaccharide production from Bacillus velezensis KY471306 using statistical experimental design

Abstract Exopolysaccharide (EPS) biopolymers produced by microorganisms play a crucial role in the environment such as health and bio-nanotechnology sectors, gelling agents in food and cosmetic industries in addition to bio-flocculants in the environmental sector as they are degradable, nontoxic. This study focuses on the improvement of EPS production through manipulation of different culture and environmental conditions using response surface methodology (RSM). Plackett-Burman design indicated that; molasses, yeast extract and incubation temperature are the most effective parameters. Box-Behnken RSM indicated that; the optimum concentration for each parameter was 12% (w/v) for molasses, 6 g/L yeast extract and 30 °C for incubation temperature. The most potent bacterial isolate was identified as Bacillus velezensis KY498625. After production, EPS was extracted, purified using DEAE-cellulose, identified using Fourier transform infrared (FTIR), gel permeation chromatography (GPC) and gas chromatography-mass spectroscopy (GC-MS). The result indicated that; it has molecular weight 1.14 × 105 D consisting of glucose, mannose and galactose.

Bacterial cellulose to reinforce urethrovesical anastomosis. A translational study

Abstract Purpose: To evaluate the efficacy of the cellulosic exopolysaccharide membrane (CEM) as a urethral reinforcement for urethrovesical anastomosis. Methods: Twenty eight rabbits were submitted to urethrovesical anastomosis with or without CEM reinforcement. The animals were divided into 4 groups: C7, CEM7, C14 and CEM14: (C= only anastomosis or CEM = anastomosis + CEM), evaluated after 7 weeks, and 14 weeks. The biointegration and biocompatibility of CEM were evaluated according to stenosis, fistula, urethral wall thickness, urethral epithelium, rate of inflammation and vascularization. Results: Between the two experimental groups, the difference in the number of stenosis or urinary fistula was not statistically significant. The morphometric analysis revealed preservation of urethral lumen, well adhered CEM without extrusion, a controlled inflammatory process and implant vascularization. The urothelium height remained constant over time after CEM reinforcement and the membrane wall was thicker, statistically, after 14 weeks. Conclusion: The absence of extrusion, stenosis or urinary fistula after 14 weeks of urethrovesical anastomosis demonstrates cellulosic exopolysaccharide membrane biocompatibility and biointegration with tendency to a thicker wall.

Comparative genomics reveals diverse capsular polysaccharide synthesis gene clusters in emerging Raoultella planticola

Raoultella planticola is an emerging zoonotic pathogen that is associated with rare but life-threatening cases of bacteremia, biliary tract infections, and urinary tract infections. Moreover, increasing antimicrobial resistance in the organism poses a potential threat to public health. In spite of its importance as a human pathogen, the genome of R. planticola remains largely unexplored and little is known about its virulence factors. Although lipopolysaccharides has been detected in R. planticola and implicated in the virulence in earlier studies, the genetic background is unknown. Here, we report the complete genome and comparative analysis of the multidrug-resistant clinical isolate R. planticola GODA. The complete genome sequence of R. planticola GODA was sequenced using single-molecule real-time DNA sequencing. Comparative genomic analysis reveals distinct capsular polysaccharide synthesis gene clusters in R. planticola GODA. In addition, we found bla TEM-57 and multiple transporters related to multidrug resistance. The availability of genomic data in open databases of this emerging zoonotic pathogen, in tandem with our comparative study, provides better understanding of R. planticola and the basis for future work.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm

Abstract Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. Objective The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual-species biofilm and to evaluate its potential ability on affecting dental health. Material and methods The effect of H. pylori supernatant on single-species and dual-species biofilm was measured by colony forming units counting and fluorescence in situ hybridization (FISH) assay, respectively. The effect of H. pylori supernatant on S. mutans and S. sanguinis extracellular polysaccharides (EPS) production was measured by both confocal laser scanning microscopy observation and anthrone-sulfuric acid method. The effect of H. pylori supernatant on S. mutans gene expression was measured by quantitative real-time PCR (qRT-PCR) assays. Results H. pylori supernatant could inhibit both S. mutans and S. sanguinis biofilm formation and EPS production. S. sanguinis inhibition rate was significantly higher than that of S. mutans. Finally, S. mutans bacteriocin and acidogenicity related genes expression were affected by H. pylori culture supernatant. Conclusion Our results showed that H. pylori could destroy the balance between S. mutans and S. sanguinis in oral biofilm, creating an advantageous environment for S. mutans, which became the dominant bacteria, promoting the formation and development of dental caries.

Yield and rheological properties of exopolysaccharide from a local isolate: Xanthomonas axonopodis pv. vesicatoria

Background: The aim of the present study was to evaluate gum productivity of a local strain, Xanthomonas axonopodis pv. vesicatoria, isolated from pepper plant, and its rheological behavior for the first time compared to the standard strain, Xanthomonas campestris DSM 19000 (NRRL B-1459). The influence of operational conditions (agitation rate and inoculum volume) on gum production and rheological properties of gums from the Xanthomonas strains were investigated. Results: The isolated strain of Xanthomonas showed similar xanthan yield compared to the standard strain. Furthermore, this study clearly confirmed that gum yield depended on bacterial strain, agitation rate, and inoculum size. The most suitable conditions for the gum production in an orbital shaker in terms of agitation rate and inoculum size were 180 rpm and 5%, respectively, resulting in an average production of 10.96 and 11.19 g/L for X. axonopodis pv.vesicatoria and X. campestris DSM 19000, respectively. Regarding the rheological properties, Ostwald-de-Waele and power law models were used to describe flow and oscillatory behavior of the gum solutions, respectively. Consistency of the novel gum solution remarkably was much higher than the commercial xanthan gum solution. Flow and oscillatory behavior and their temperature ramps showed that weak gel-like structure could be obtained with less gum concentrations when the novel gum was used. Conclusion: Therefore, yield and technological properties of the aqueous solutions of the exopolysaccharide synthesized by X. axonopodis pv. vesicatoria were observed to be more suitable for industrial production.

Alginate overproduction and biofilm formation by psychrotolerant Pseudomonas mandelii depend on temperature in Antarctic marine sediments

Background: In recent years, Antarctica has become a key source of biotechnological resources. Native microorganisms have developed a wide range of survival strategies to adapt to the harsh Antarctic environment, including the formation of biofilms. Alginate is the principal component of the exopolysaccharide matrix in biofilms produced by Pseudomonas, and this component is highly demanded for the production of a wide variety of commercial products. There is a constant search for efficient alginate-producing organisms. Results: In this study, a novel strain of Pseudomonas mandelii isolated from Antarctica was characterized and found to overproduce alginate compared with other good alginate producers such as Pseudomonas aeruginosa and Pseudomonas fluorescens. Alginate production and expression levels of the alginate operon were highest at 4°C. It is probable that this alginate-overproducing phenotype was the result of downregulated MucA, an anti-sigma factor of AlgU. Conclusion: Because biofilm formation is an efficient bacterial strategy to overcome stressful conditions, alginate overproduction might represent the best solution for the successful adaptation of P. mandelii to the extreme temperatures of the Antarctic. Through additional research, it is possible that this novel P. mandelii strain could become an additional source for biotechnological alginate production.

Production of xanthanases by paenibacillus spp.: complete xanthan degradation and possible applications

Background: A number of microorganisms and their enzymes have been reported as xanthan depolymerizers. Paenibacillus species are well-known polysaccharide hydrolyzing bacteria. However, Paenibacillus alginolyticus and Paenibacillus sp. XD are the only species in the genus which are now known to degrade xanthan

Objectives: Complete biodegradation of the xanthan exopolysaccharide is a rarely found capability among microorganisms. The aim of this study is to survey xanthanase producing bacteria with an appropriate bioactivity for the biopolymer degradation under different environmental conditions

Materials and Methods: The bacteria were isolated based on viscosity reduction of the xanthan solution. Bacterial isolates were identified using rep-PCR [repetitive element-based genomic fingerprinting] and 16S rDNA sequencing. Xanthanases were characterized by measuring their activity at different temperatures, pH values, and NaCl concentrations. Degradation of other polysaccharides and xanthan degradation products were investigated based on the screening plate method and TLC [thin-layer chromatography], respectively

Results: Six isolates from different Paenibacillus species with a complete xanthan degrading capability were isolated from Urmia Lake. Phylogenetic analysis placed these strains within the genus Paenibacillus with the closest relatives that were found to be P. nanensis, P. phyllosphaerae, P. agaridevorans, P. agarexedens, and P. taohuashanense. These isolates displayed different levels of the xanthan biodegradation activity in temperatures ranging from 15 to 55 [degree]C and pH values from 4 to 11. Xanthanolytic activity was generally prevented in presence of NaCl [> 0.1 mol.L-1]. Furthermore, the isolated Paenibacillus spp. could degrade several other polysaccharides including xylan, CMC [carboxymethyl cellulose], starch, alginate, and pectin

Conclusion: Novel strains of the six different Paenibacillus species that were introduced in the present study are able to produce xanthanases with interesting characteristics. In light of the results from this study, special applications, particularly in healthcare, medicine, and the environment is hereby proposed for these enzymes

Effect of carbon source on production, characterization and bioactivity of exopolysaccharide produced by Phellinus vaninii Ljup

ABSTRACT The effect on different three carbon source (i.e. glucose, fructose and sucrose) on production, chemical characterization and antioxidant activity of exopolysaccharide (EPS) produced by Phellinus vaninii Ljup was investigated in this study. Amongst carbon sources examined, glucose and sucrose were favorable for the mycelia growth, while the maximum EPS yield was achieved when sucrose was employed. The predominant carbohydrate compositions in EPSs identified were gluconic acid, glucose, mannose and galactose acid. Then, FT-IR spectral analysis revealed prominent characteristic groups in EPSs. EPSs molecule exist as nearly globular shape form in aqueous solution. The variation also affects antioxidant activities by investigated by using hydroxyl and DPPH radical scavenging assay. Sucrose was best carbon source from the viewpoint of antioxidant activity due to the relatively high contents of galactose in the EPS with moderate molecular weight and polydispersity.

Exopolysaccharide dispelled by calcium hydroxide with volatile vehicles related to bactericidal effect for root canal medication

ABSTRACT Objective: Enterococcus faecalis is the dominant microbial species responsible for persistent apical periodontitis with ability to deeply penetrate into the dentin. Exopolysaccharides (EPS) contribute to the pathogenicity and antibiotic resistance of E. faecalis. Our aim was to investigate the antimicrobial activity of calcium hydroxide (CH), camphorated parachlorophenol (CMCP), and chlorhexidine (CHX) against E. faecalis in dentinal tubules. Material and Methods: Decoronated single-canal human teeth and semicylindrical dentin blocks were incubated with E. faecalis for 3 weeks. Samples were randomly assigned to six medication groups for 1 week (n=10 per group): CH + 40% glycerin-water solution (1:1, wt/vol); CMCP; 2% CHX; CH + CMCP (1:1, wt/vol); CH + CMCP (2:3, wt/vol); and saline. Bacterial samples were collected and assayed for colony-forming units. After dentin blocks were split longitudinally, confocal laser scanning microscopy was used to assess the proportion of viable bacteria and EPS production in dentin. Results: CMCP exhibited the best antimicrobial activity, while CH was the least sensitive against E. faecalis (p<0.05). CHX showed similar antimicrobial properties to CH + CMCP (1:1, wt/vol) (p>0.05). CH combined with CMCP inhibited EPS synthesis by E. faecalis, which sensitized biofilms to antibacterial substances. Moreover, increasing concentrations of CMCP decreased EPS matrix formation, which effectively sensitized biofilms to disinfection agents. Conclusion: The EPS matrix dispelled by CH paste with CMCP may be related to its bactericidal effect; the visualization and analysis of EPS formation and microbial colonization in dentin may be a useful approach to verify medicaments for antimicrobial therapy.

Structure of xanthan gum and cell ultrastructure at different times of alkali stress

Abstract The effect of alkali stress on the yield, viscosity, gum structure, and cell ultrastructure of xanthan gum was evaluated at the end of fermentation process of xanthan production by Xanthomonas campestris pv. manihotis 280-95. Although greater xanthan production was observed after a 24 h-alkali stress process, a lower viscosity was observed when compared to the alkali stress-free gum, regardless of the alkali stress time. However, this outcome is not conclusive as further studies on gum purification are required to remove excess sodium, verify the efficiency loss and the consequent increase in the polymer viscosity. Alkali stress altered the structure of xanthan gum from a polygon-like shape to a star-like form. At the end of the fermentation, early structural changes in the bacterium were observed. After alkali stress, marked structural differences were observed in the cells. A more vacuolated cytoplasm and discontinuities in the membrane cells evidenced the cell lysis. Xanthan was observed in the form of concentric circles instead of agglomerates as observed prior to the alkali stress.

Selection of superior bifidobacteria in the presence of rotavirus

The main purpose of this study was to investigate bifidobacteria flora in fecal samples from children with rotavirus infection and determine the significance of their selected probiotic properties for improvement of health status. Enzyme-linked immunosorbent assay was used to identify rotavirus antigen in fecal samples from 94 patients with gastroenteritis and from 30 without gastroenteritis. Bifidobacteria were identified by selective media, gram reaction, colony morphology, fructose-6-phosphate phosphoketolase enzyme activity and classical identification tests. Exopolysaccharide (EPS) production was identified by phenol-sulphuric acid method. The modified method was then used to identify the quantity of taurocholic and glycocholic acid deconjugation and cholesterol elimination of the strains. Thirty-five of the 94 fecal samples were found positive for rotavirus antigen (37.23%). Bifidobacteria were identified in 59 of the samples. The EPS production ranges were 29.56-102.21 mg/L. The cholesterol elimination rates ranged between 8.36-39.22%. Furthermore, a positive and strong correlation was determined between EPS production and the presence of cholesterol (r=0.984, P<0.001). The deconjugation rates for the sodium glycocholate group was higher than the sodium taurocholate group. Rotavirus (+) bifidobacteria strains had higher EPS production, deconjugation rate and cholesterol elimination compared to bifidobacteria strains isolated from children in the rotavirus (-) sample and without gastroenteritis. Significant differences were observed among groups in all parameters (P<0.05). Given the increased number of rotavirus cases in Turkey and worldwide, it is very important to add superior bifidobacteria in the diets of infected children to improve the intestinal and vital functions.

Effect of pH in the survival of Lactobacillus salivarius strain UCO_979C wild type and the pH acid acclimated variant

Background Bacterial acclimation involves cellular changes permitting the survival of a microorganism to prolonged acid pH exposure. The general aim of this work is to support this idea by determining the effect of pH in the survival of the human gastric derived probiotic strain Lactobacillus salivarius UCO_979C-1 (wild type) and L. salivarius UCO_979C-2 (acclimation to pH 2.6), which possesses anti-Helicobacter pylori properties. Results To assess this aim, the exopolysaccharide production through the phenol-sulfuric acid method was evaluated. Moreover, morphological and structural changes by transmission and scanning electron microscopy were observed. The bacterial survival was measured by viable count. The results showed that the acclimated variant strain synthesized higher levels of exopolysaccharide (690 ± 0.03 mg/L) more than the wild type (450 ± 0.12 mg/L). In addition, the acclimated variant preserved the viable count at pH 2.6 for 48 h, whereas the wild type strain decreases after 6 h and was non-viable at 24 h. Conclusion The results suggest that the acid stress acclimation of the strain L. salivarius UCO_979C-1 modified some cellular properties making this strain potentially useful as a gastric probiotic.