Modification of the phosphoketolase assay for rapid identification of bifidobacteria

Orban, J. I.; Patterson, John A.

doi:10.1016/s0167-7012(00)00133-0

Cited by 117 publications

(72 citation statements)

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“…F6PPK is the key enzyme in this pathway and is considered a taxonomic marker for identification of species of Bifidobacterium and related genera (Biavati & Mattarelli, 2012). F6PPK activity was determined according to the method described by Scardovi (1986) and modified by Orban & Patterson (2000). All the isolates possessed F6PPK activity.…”

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confidence: 99%

Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus)

Modesto

Michelini

Stefanini

et al. 2014

International Journal of Systematic and Evolutionary Microbiology

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Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus) Six Gram-positive-staining, microaerophilic, non-spore-forming, fructose-6-phosphate phosphoketolase-positive bacterial strains with a peculiar morphology were isolated from faecal samples of baby common marmosets (Callithrix jacchus). Cells of these strains showed a morphology not reported previously for a bifidobacterial species, which resembled a coiled snake, always coiled or ring shaped or forming a 'Y' shape. Strains MRM 3/1 T and MRM 4/2 were chosen as representative strains and characterized further. The bacteria utilized a wide range of carbohydrates and produced urease. Glucose was fermented to acetate and lactate. Strain MRM 3/1 T showed a peptidoglycan type unique among members of the genus Bifidobacterium. The DNA base composition was 64.7 mol% G+C. Almost-complete 16S rRNA, hsp60, clpC and rpoB gene sequences were obtained and phylogenetic relationships were determined. Comparative analysis of 16S rRNA gene sequences showed that strains MRM 3/1 T and MRM 4/2 had the highest similarities to Bifidobacterium scardovii DSM 13734 T (94.6 %) and Bifidobacterium stellenboschense DSM 23968 T (94.5 %). Analysis of hsp60 showed that both strains were closely related to B. stellenboschense DSM 23968 T (97.5 % similarity); however, despite this high degree of similarity, our isolates could be distinguished from B. stellenboschense DSM 23968 T by low levels of DNA-DNA relatedness (30.4 % with MRM 3/1 T ). Strains MRM 3/1 T and MRM 4/2 were located in an actinobacterial cluster and were more closely related to the genus Bifidobacterium than to other genera in the family Bifidobacteriaceae. On the basis of these results, strains MRM 3/1 T and MRM 4/2 represent a novel species within the genus Bifidobacterium, for which the name Bifidobacterium aesculapii sp. nov. is proposed; the type strain is MRM 3/1 T (5DSM 26737 T 5JCM 18761 T ).

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Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus)

Modesto

Michelini

Stefanini

et al. 2014

International Journal of Systematic and Evolutionary Microbiology

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“…The isolates from Vaginalis agar were identified as G. vaginalis according to Piot et al (1982), using the results from Gram stain, catalase and oxidase tests, starch and hippurate hydrolysis, and a-and b-galactosidase activity. Activity of fructose-6-phosphate phosphoketolase was also evaluated (Orban & Patterson, 2000).…”

Section: Methodsmentioning

confidence: 99%

Characteristics of Lactobacillus and Gardnerella vaginalis from women with or without bacterial vaginosis and their relationships in gnotobiotic mice

Teixeira

Carvalho

Arantes

et al. 2012

Journal of Medical Microbiology

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The objectives of the present study were to evaluate in vitro the production of antagonistic compounds against Gardnerella vaginalis by Lactobacillus strains isolated from women with or without bacterial vaginosis (BV), and to select one of the better Lactobacillus producers of such a substance to be tested in vivo using a gnotobiotic animal model challenged with one of the more sensitive G. vaginalis isolates. A total of 24 isolates from women with and without BV were identified as G. vaginalis. A higher frequency (P,0.05) of this bacterium was observed in women with BV (56.7 %) when compared to healthy women (17.6 %). A total of 86 strains of Lactobacillus were obtained from healthy women and women with BV. Lactobacillus strains were more frequently present (P,0.05) in healthy women (97.5 %) than in women with BV (76.7 %). Lactobacillus crispatus was the predominating strain in both healthy women and women with BV. Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus gasseri and Lactobacillus vaginalis were isolated with an intermediate frequency in the two groups. In vitro antagonism assays were performed using as indicators 17 reference strains and the G. vaginalis strains isolated from women with BV and from healthy women. Lactobacillus isolated from healthy women showed the higher antagonistic activity against all the indicator strains when compared with isolates from women with BV. Concerning the indicator strains, G. vaginalis found in women with BV was more resistant to the antagonism, particularly when Lactobacillus isolates from women with BV were used as producer strains. A high vaginal population level of G. vaginalis was obtained by intravaginal inoculation of germ-free mice, and this colonization was accompanied by vaginal histopathological lesions. A tenfold decrease in vaginal population level of G. vaginalis and a reduction of histological lesions were observed when the pathogenic challenge was performed in mice previously monoassociated with an L. johnsonii strain. Concluding, results of the present study suggest that progression of G. vaginalis-associated BV depends in part on a simultaneous presence of Lactobacillus populations with a low antagonistic capacity and of a G. vaginalis strain with a high resistance to this antagonism. The results could also explain why G. vaginalis is frequently found in the vaginal ecosystem of healthy women.

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“…The spread-plating of 100 µL from each dilution was prepared and the plates were incubated at 37°C for 72 hours under anaerobic conditions (14) using a commercial anaerobic atmosphere generation system (Anaerobac from Probac ® ), followed by counting of bifidobacteria colonies in the plates containing between 30 and 300 colony-forming units (CFU). For the confirmation of the Bifidobacterium genus, Gram staining was made, as well as catalase proof and fructose-6-phosphate phosphoketolase (F6PPK) reaction, as stated by Orban & Patterson (2000), for all isolated colony types (18) .…”

Section: Determination Of Faecal Bifidobacteria Content and Phmentioning

confidence: 99%

Effects of Probiotic Intake on Intestinal Bifidobacteria of Celiac Patients

Martinello

Roman

Souza

2017

Arq. Gastroenterol.

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-Background -Healthy individuals exhibit a significantly higher concentration of faecal bifidobacteria in comparison to celiac patients.Even though there are potential benefits in probiotic usage, they have been little explored as an adjunctive therapy in celiac disease. Objective -This study aimed at the comparison of faecal bifidobacteria concentration and pH among celiac patients and healthy subjects before and after the daily intake of 100 g of yogurt containing probiotic for a thirty-day period. Methods -Feces from 17 healthy subjects and 14 celiac patients were analyzed, in which stool culture was performed for the isolation and quantification of faecal bifidobacteria. Furthermore, Gram's method was employed for the microscopic analysis of the colonies, while the identification of the Bifidobacterium genus was made through determination of the fructose-6-phosphate phosphoketolase enzyme. Faecal pH was measured using a calibrated pHmeter. Results -Faecal bifidobacteria concentration before probiotic consumption was significantly higher in healthy individuals (2.3x10 8 ±6.3x10 7 CFU/g) when compared to celiac patients (1.0x10 7 ±1.7x10 7 CFU/g). Faecal pH values did not show a significant difference. After the daily consumption of probiotic-containing yogurt both groups showed a significant increase in the concentration of faecal bifidobacteria, but healthy subjects presented significantly higher bifidobacteria concentrations (14.7x10 8 ±0.2x10 8 CFU/g) than the celiac group (0.76x10 8 ±0.1x10 8 CFU/g). The obtained pH values from both groups were not significantly different, being 7.28±0.518 for the celiac patients and 7.07±0.570 for healthy individuals after the probiotic intake. Conclusion -The probiotic supplementation significantly increased the number of bifidobacteria in the feces of celiac patients, although it was not sufficient to reach the concentration found in healthy individuals prior to its consumption.

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Modification of the phosphoketolase assay for rapid identification of bifidobacteria

Cited by 117 publications

References 2 publications

Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus)

Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus)

Characteristics of Lactobacillus and Gardnerella vaginalis from women with or without bacterial vaginosis and their relationships in gnotobiotic mice

Effects of Probiotic Intake on Intestinal Bifidobacteria of Celiac Patients

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