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Identification of Bifidobacterium animalis, Bifidobacterium adolescentis and Bifidobacterium bifidum from Stool of Children and Detection of Their Antibacterial Properties  [PDF]
Mansour Amin, Ahmad Farajzadeh Sheikh, Hamed Goodarzi, Mehdi Sormeh
Advances in Infectious Diseases (AID) , 2013, DOI: 10.4236/aid.2013.33029
Abstract:

Introduction and Objective: The genus Bifidobacterium can generally be found in quantity in the habitats such as human and animal gastrointestinal tract, dental caries, vagina and oral cavity. The aim of this study was to isolate Bifidobacterium from stool and determine their inhibitory effect against some pathogens. Materials and Methods: 130 samples were collected by wet swabs and kept in sterile tubes containing MRS broth media. And Bifidobacterium isolated from stool was enriched in Man-Rogosa-Sharpe medium (MRS) broth and isolated by growing on MRS agar medium and characterized by phenotypic characteristics and PCR technique at genus and species levels. The antimicrobial substance was extracted from ethyl acetate solvent and the antimicrobial activity against some pathogenic bacteria, such as Salmonella typhi and Shigella sonnei, were investigated. Results: Eleven Bifidobacterium bifidum and four Bifidobacterium adolescentis, which were isolated from fresh stool, were identified by PCR. Antimicrobial substance from MRS broth medium was extracted. This antimicrobial compound showed a potent inhibitory activity against four tested bacteria. These bacteria produced acetic acid and lactic acid as inhibitory substances that were different from bacteriocins. Conclusion: Fresh stool may be used as a source of antimicrobial lactic acids bacteria, Bifidobacterium bifidum and adolescentis as two probiotics can establish themselves in gut and urogenital tract to prevent the human body from adverse effects of pathogens.

Screening and identification of probiotic bifidobacterium from Mongolian children
蒙古族儿童源益生特性双歧杆菌的筛选及鉴定

Pengfei Gao,Zhihong Sun,Shiwei M,Qiushi Wang,Jie Gao,Cengyuan Deng,Heping Zhang,
高鹏飞
,孙志宏,麻士卫,王秋实,高杰,邓承远,张和平

微生物学报 , 2009,
Abstract: Objective] Bifidobacterium has important physiological activity in gastrointestinal tract of human and animals, and the tolerability of acid, intestinal juice and oxgall bile salt are the key factors that influence the function of living bacteria preparation. In this study, feces of 12 healthy Mongolian children were selected to isolate Bifidobacterium with probiotic properties. Methods] Bifidobacterium was isolated from feces by selective medium and identified by physiological biochemical test. The tolerability of acid, intestinal juice and oxgall bile salt of 11 Bifidobacteria were studied and B. animalis V9 was identified further by molecular biological approach 16S rDNA. Results] Eleven Bifidobacteria isolated from feces of 12 healthy Mongolian children were identified as B. adolescentis (A1, H3, G4, A8 and V10), B. longum (C6, C7 and D11), B. pseudocatenlatum (B2), B. bifidum (G5), B. animalis (V9). B. animalis V9 had the best acid tolerance with the survival rate 92.4% in artificial gastric juice at pH2.0 for 3h, whereas others had lower than 31.25%. B. animalis V9 also had good survival rate (99.7%) in artificial intestinal juice at pH8.0 for 8h after anaerobic cultured 3h in artificial gastric juice at pH2.0, and tolerated oxgall bile salt at concentration of 0.3%. B. animalis V9 was identified further by molecular biological approach 16S rDNA and result showed the homologies of B. animalis V9 was 99% with Bifidobacterium animalis subsp. lactic BB12. Conclusion] B. animalis V9 had good probiotic properties to be potentiallyused in dairy products and health products.
Production of Functional Kariesh Cheese by Microencapsulation of Bifidobacterium adolescentis ATCC 15704
A.M. Abd-Elhamid
Advance Journal of Food Science and Technology , 2012,
Abstract: Bifidobacterium adolescentis ATCC 15704 was microencapsulated with rennet gelation of milk proteins. Encapsulated and free Bifidobacterium adolescentis ATCC 15704 under simulated gastrointestinal conditions and bile tolerance were estimated. The survivability of encapsulated Bifidobacterium adolescentis ATCC 15704 and the changes of organic acids in Kariesh cheese during 2 weeks of cold storage were determined. The study demonstrated that microencapsulation using rennet gelation of milk proteins increased the survival of Bifidobacterium adolescentis ATCC 15704 in simulated gastric conditions. Furthermore, the results showed that, the survival of encapsulated Bifidobacterium adolescentis ATCC 15704 in bile solution was significantly enhanced (p<0.05) compared to the free Bifidobacterium adolescentis ATCC 15704. Microencapsulation protected Bifidobacterium adolescentis ATCC 15704 in Kariesh cheese during cold storage as compared to free Bifidobacterium adolescentis ATCC 15704. In addition, the capsules made from rennet gelation of milk proteins had a positive effect in organic acids during cold storage.
Evaluation of culture media for counts of Bifidobacterium animalis in the presence of yoghurt bacteria
Moriya, Juliana;Fachin, Luciano;Gandara, Ana Lourdes Neves;Viotto, Walkiria Hanada;
Brazilian Journal of Microbiology , 2006, DOI: 10.1590/S1517-83822006000400021
Abstract: much attention has been paid to the use of bifidobacterium sp. in yoghurts due to their excellent therapeutic and nutritional properties. however, in order to present health properties, it is recommended that each commercial product should indicate the minimum daily amount required for it to confer health benefit(s) based on in vitro and human studies. besides the inherent low growth of bifidobacterium sp in milk, there is a need for a reliable method for counting bifidobacterium sp in the presence of yoghurt bacteria. this study evaluated the use of the media m-mrs, mrs-nnlp and rcpb ph5 aimed at counting the number of bifidobacterium animalis subesp. lactis in the presence of streptococcus thermophilus and lactobacillus delbrueckii subsp bulgaricus after yoghurt fermentation. the m-mrs medium was not selective, allowing for the growth of l. delbrueckii subsp bulgaricus. mrs-nnlp medium presented a good selectivity for b. animalis bb12 with a slight reduction in the cell count of this microrganism when compared it to the standard mrs medium in pure culture. mrs-nnlp medium was considered a good option to enumerate b. animalis bb12 although the reduction found in pure culture due to the low difference between the counts. the medium rcpb ph5 presented differentiated growth of b. animalis bb12 in relation to the yoghurt bacteria and a cell recovery equal to that of the standard mrs, being considered the best option to enumerate bifidobacterium sp in the presence of yoghurt bacteria.
Randomized controlled trial of live lactobacillus acidophilus plus bifidobacterium bifidum in prophylaxis of diarrhea during radiotherapy in cervical cancer patients
Imjai Chitapanarux, Taned Chitapanarux, Patrinee Traisathit, Sudkaneung Kudumpee, Ekkasit Tharavichitkul, Vicharn Lorvidhaya
Radiation Oncology , 2010, DOI: 10.1186/1748-717x-5-31
Abstract: Patients who were undergoing pelvic radiotherapy concurrent with weekly cisplatin were randomly assigned to a study drug or placebo, in a double-blind study. Diarrhea was graded weekly according the Common Toxicity Criteria (CTC) system. Stool consistency and white and red blood cell count in stool were also assessed. The primary endpoint was to reduce the incidence of diarrhea, defined by a CTC grade 2 or more, and the need for anti-diarrheal medication.A total of 63 patients were enrolled. Grade 2 -3 diarrhea was observed in 45% of the placebo group (n = 31) and 9% of the study drug group (n = 32) (p = 0.002). Anti-diarrheal medication use was significantly reduced in the placebo group (p = 0.03). The patients in the study drug group had a significantly improved stool consistency (p < 0.001).Live lactobacillus acidophilus plus bifidobacterium bifidum reduced the incidence of radiation-induced diarrhea and the need for anti-diarrheal medication and had a significant benefits on stool consistency.Radical radiation therapy to pelvic malignancy carries a risk of complications to normal tissues around the tumor. Acute complications affecting the gastrointestinal tract occur in approximately 80% of patients, but they are usually mild and only rarely affect the treatment planning [1]. One of the most common acute complications of pelvic radiotherapy is acute inflammatory change in the small intestine leading to gastrointestinal symptoms during treatment because healthy bowel tissue is encompassed in the radiation field. Acute symptoms include diarrhea, abdominal pain, tenesmus and nausea, usually starting during the second or third week of radiotherapy [1,2]. Because of this the incidence of malnutrition in patients who receive pelvic radiotherapy is 11-33% and up to 83% of patients lose weight during treatment [3]. The development of late gastrointestinal symptoms following pelvic radiotherapy is not entirely dose related, but depends on a complex interaction between ph
Characterization of recombinant β-fructofuranosidase from Bifidobacterium adolescentis G1
Toshima Omori, Keiji Ueno, Kei Muramatsu, Masanori Kikuchi, Shuichi Onodera, Norio Shiomi
Chemistry Central Journal , 2010, DOI: 10.1186/1752-153x-4-9
Abstract: Using the information of cscA gene from Bifidobacterium adolescentis ATCC 15703, cscA gene from B. adolescentis G1 was cloned and sequenced. The N-terminal amino acid sequence of purified β-FFase from B. adolescentis G1 was identical to the deduced amino acid sequences of cscA gene from B. adolescentis G1. To confirm the translated product of the cscA gene, the recombinant protein was expressed in Escherichia coli. Molecular mass of the purified recombinant enzyme was estimated to be about 66,000 by SDS-PAGE and 60,300 by MALDI TOF-MS. The optimum pH of the enzyme was 5.7 and the enzyme was stable at pH 5.0-8.6. The thermostability of the enzyme was up to 50°C. The Km (mM), Vmax (μmol/mg of protein/min), k0 (sec-1) and k0/Km(mM-1 sec-1) for 1-kestose, neokestose, nystose, fructosylnystose, sucrose and inulin were 1.7, 107, 107.5, 63.2, and 1.7, 142, 142.7, 83.9, and 3.9, 152, 152.8, 39.2, and 2.2, 75, 75.4, 34.3, and 38, 79, 79.4, 2.1, and 25.9, 77, 77.4, 3.0, respectively. The hydrolytic activity was strongly inhibited by AgNO3, SDS, and HgCl2.The recombinant enzyme had similar specificity to the native enzyme, high affinity for 1-kestose, and low affinity for sucrose and inulin, although properties of the recombinant enzyme showed slight difference from those of the native one previously described.Bifidobacteria are saccharolytic anaerobes generally present in human intestine. Growth of bifidobacteria is selectively promoted by prebiotics [1].Fructo-oligosaccharides, such as 1-kestose, nystose and fructosylnystose, consist of β-2,1-linked fructose to sucrose, and they are naturally contained in artichoke tubers [2], chicory roots [3] and burdock roots [4,5]. These saccharides have been produced and commercially manufactured from sucrose with bacterial fructosyltransferase [6] and β-fructofuransidases (β-FFases) [7-9], and have been on the market as prebiotics. Fructo-oligosaccharides are not hydrolyzed by digestive enzymes of mammalian origin, so they are able to
Perfil de Susceptibilidade Antimicrobiana de Bifidobacterium bifidum Bb12 e Bifidobacterium longum Bb46
Flávio Henrique Ferreira Barbosa,Andréia Mar?al da Silva,Rinaldo Duarte,Jacques R. Nicoli (ori.)
Revista de Biologia e Ciências da Terra , 2001,
Abstract: Bifidobacterium constitutes the most recent group of bacteria used as dietetic adjunct presenting probiotic properties such as prevention and treatment of infectious intestinal disorder. An expected characteristic for the selection of a probiotic is its antibiotic sensibility discarding the possibility of resistance transmission in the digestive ecosystem and allowing the elimination of the biotherapeutic, if necessary. In this study, the susceptibility to antibiotics of two Bifidobacterium species was tested. Bifidobacterium bifidum Bb12 and Bifidobacterium longum Bb46 have been isolated from a commercial lyophilized preparation (DVS – Christian Hansen Lab., Horsholm, Denmark). Bacteroides fragilis ATCC 25285 was used as control. The following antibiotic classes were tested: b -lactam, lincosomide, macrolide, phenolic, glycopeptide, tetracycline and aminoglycoside. The susceptibility assays of the antibiotics were performed using the standard agar dilution method (NCCLS-2000), with some modification. After inoculation and incubation during 48 hours at 37oC, in an anaerobic chamber, the absence or presence of growth on the plates was observed. All the tests were realized in duplicate. The results showed that the both B. bifidum Bb12 and B. longum Bb46 were sensible to all of the antibiotics tested, and for all of the concentrations used, except to gentamicin. Concluding, the possibility of a transmission of antibiotic resistance from these probiotics to pathogenic bacteria can be considered to be low during the use of these biotherapeutics.
婴儿源益生性双歧杆菌的筛选及肠道定殖性研究  [PDF]
韩俊燕
- , 2016, DOI: 10.13982/j.mfst.1673-9078.2016.10.003
Abstract: 本研究旨在从婴儿粪便中筛选出具有潜在益生特性的双歧杆菌,并探究其肠道定殖情况,为双歧杆菌的产品开发提供优良的菌株。采用MRS培养基对样品进行分离纯化,菌株经F6PPK检测及16S rDNA测序鉴定,之后进行模拟胃肠液、胆盐耐受性、对食源性致病菌(大肠杆菌、沙门氏菌、单增李斯特菌等)的抑制及对HT-29细胞的粘附能力测定,将筛选出的菌株进行动物实验,测定其肠道定殖能力。分离到的27株双歧杆菌,经分子生物学鉴定为7个不同的种:Bifidobacterium longum、Bifidobacterium breve、Bifidobacterium bifidum、Bifidobacterium pseudocatenulatum、Bifidobacterium infantis、Bifidobacterium animalis和Bifidobacterium adolescentis。体外实验表明,B. longum A9、B. breve A4、B. bifidum B6、B. longum C6、B. adolescentis F8和B. infantis H6等具有较强的潜在益生特性;动物实验表明,B. infantis H6和B. longum C6具有较强的肠道定殖能力。B. longum C6和B. infantis H6有望作为优良的益生性菌株,应用于双歧杆菌的产品开发。
Bifidobacterium strains with potential probiotic properties were screened from infant feces and their colonization ability in the intestine was explored, to provide potential probiotic strains for the development of Bifidobacterium products. De Man, Rogosa, and Sharpe (MRS) medium was used to isolate and purify the samples, and detection of fructose-6-phosphate phosphoketolase (F6PPK) and analysis of 16S rDNA sequences were performed on the strains. The probiotic properties of the strains, including tolerance to simulated gastrointestinal fluids and bile salts, inhibition of foodborne pathogenic bacteria (Escherichia coli, Salmonella spp., Listeria monocytogenes, etc.,) and adhesion capacity to HT-29 cells were determined. The screened Bifidobacterium strains with potential probiotic properties were used in animal experiments to measure their intestinal colonization ability. After isolation, 27 Bifidobacterium strains were identified from seven Bifidobacterium species using molecular biology: Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium pseudocatenulatum, Bifidobacterium infantis, Bifidobacterium animalis, and Bifidobacterium adolescentis. In vitro experiments showed that B. longum A9, B. breve A4, B. bifidum B6, B. longum C6, B. adolescentis F8, and B. infantis H6 had stronger potential probiotic properties. The animal experiments showed that B. infantis H6 and B. longum C6 had stronger gut colonization ability. The results demonstrate that B. infantis H6 and B. longum C6 have potential as good probiotic Bifidobacterium strains that can be applied to the development of Bifidobacterium products.
Growth characteristics modeling of Bifidobacterium bifidum using RSM and ANN
Meena, Ganga S.;Gupta, Suneel;Majumdar, Gautam C.;Banerjee, Rintu;
Brazilian Archives of Biology and Technology , 2011, DOI: 10.1590/S1516-89132011000600023
Abstract: the aim of this work was to optimize the biomass production by bifidobacterium bifidum 255 using the response surface methodology (rsm) and artificial neural network (ann) both coupled with ga. to develop the empirical model for the yield of probiotic bacteria, additional carbon and nitrogen content, inoculum size, age, temperature and ph were selected as the parameters. models were developed using ? fractional factorial design (ffd) of the experiments with the selected parameters. the normalized percentage mean squared error obtained from the ann and rsm models were 0.05 and 0.1%, respectively. regression coefficient (r2) of the ann model showed higher prediction accuracy compared to that of the rsm model. the empirical yield model (for both ann and rsm) obtained were utilized as the objective functions to be maximized with the help of genetic algorithm. the optimal conditions for the maximal biomass yield were 37.4 °c, ph 7.09, inoculum volume 1.97 ml, inoculum age 58.58 h, carbon content 41.74% (w/v), and nitrogen content 46.23% (w/v). the work reported is a novel concept of combining the statistical modeling and evolutionary optimization for an improved yield of cell mass of b. bifidum 255.
Comparision of growth of Lactobacillus acidophilus and Bifidobacterium Bifidum species in media suplemented with selected saccharides including prebiotics
Kamila Goderska,Jacek Nowak,Zbigniew Czarnecki
Acta Scientiarum Polonorum : Technologia Alimentaria , 2008,
Abstract: The performed investigations evaluated the growth dynamics of Lactobacillus acidophilus DSM 20079, Lactobacillus acidophilus DSM 20242, Bifidobacterium bifidum DSM 20082, Bifidobacterium bifidum DSM 20215, Bifidobacterium bifidum DSM 20239, Bifidobacterium bifidum DSM 20456 in media supplemented with various saccharides, including prebiotic preparations. The addition of saccharides to the medium exerted a significant influence not only on the number of bacterial cells but also on their acid-creating capability. Glucose, lactose and saccharose turned out to be the easiest available saccharides for all the bacterial strains tested. In the media supplemented with these sugars the highest numbers of bacterial cells were determined. At the shortage of mono- and disaccharides, all strains of the bacteria tested were capable of utilising the prebiotic preparations as sources of carbon and energy in the media. The amount and isomeric forms of lactic acid produced by Lactobacillus acidophilus DSM 20079 and Bifidobacterium bifidum DSM 20239 were determined. Both strains meet the requirements adopted by the WHO and produce more than 70% lactic acid L(+) in the media with the addition of various saccharides. Lactobacillus acidophilus DSM 20079 was found to produce significantly higher amount of lactic acid in different media.
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