Lactobacillus plantarum IFPL935 improves colonic metabolism
in a simulator of the human intestinal microbial ecosystem
during feeding with red wine phenolic compounds
E. Barroso1*, T. van de Wiele2, A. Jiménez-Girón1, I. Muñoz-González1, M.V. Moreno-Arribas1, B. Bartolomé1,
1
1
1
C. Peláez , M.C Martínez-Cuesta , T. Requena
1Instituto
de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), Madrid, Spain.
2LabMET, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
*elvira.barroso@csic.es
In the present work we have evaluated the adaptation of the colonic microbiota developed in two parallel five-stage in vitro gastrointestinal
simulators (TWIN-SHIME) to long-term feeding with a polyphenol-rich red wine extract and the combination of this extract with a potential
probiotic strain Lactobacillus plantarum IFPL935.
Experimental period
Control: pH, Tª, feed
TWIN-SHIME
Day 1: TWIN-SHIME inoculation with the same faecal sample.
Days 1- 22: Stabilitation period.
Days 1t- 11t: Treatment period. Supplementation of Provinols ® and Provinols ® + L. plantarum IFPL935
Day 12: End of supplementation.
Days 12-20: Washout period.
SAMPLING: Stabilitation, Treatment and Washout period
colon vesssels ascendens (AC), transversum (TC) and descendens (DC)
Chemical analysis
Microbial ecology
GC: Short Chain Fatty Acids (SCFA: Acetate, Propionate, Butyrate) Fermentation activity.
Real Time PCR (qPCR).
UPLC-DAD-ESI-TQ: Metabolism of wine polyphenolics. Metabolic evolution.
BUTYRATE PRODUCTION
Butyrate (mg/mL)
800
Provinols®
800
AC
700
600
Provinols® + L. plantarum IFPL935
800
TC
700
MICROBIAL COUNTS by qPCR
(counts are expressed in average log copy number/mL)
500
500
400
Provinols + L. plantarum IFPL 935
400
400
300
AC Provinols
300
300
200
200
200
100
100
100
0
0
0
1t
2t
3t
4t
5t
9t
11t 12
13
16
19
Ti me peri od
Tota l Ba cteri a
AC
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
Wa s hout
Sta b.
Trea t. da ys
Trea t. da ys
600
500
22
Compa rtment
700
600
20
DC
Ba cteri a l group
20
22
1t
2t
Days
3t
4t
5t
9t
11t 12
13
16
19
TC
TC Provinols + L. plantarum IFPL
935
DC Provinols + L. plantarum IFPL
935
TC Provinols
DC Provinols
Lactobacillus s pp.
DC
AC
TC
20
22
1t
2t
3t
4t
Days
5t
9t
11t 12
13
16
19
Days
DC
• The polyphenolic extract addition (day 1t) caused a sharp reduction of the fermentation activity (SCFAs) in the three colon vessels.
• Significant increase of butyrate production during the polyphenols treatment and the washout period in vessels TC and DC treated with
L. plantarum IFPL935 compared with the vessels without the strain.
Bifidobacterium s pp.
AC
TC
MICROBIAL CATABOLISM OF WINE POLYPHENOLS
DC
L. plantarum IFPL935
(Tabasco et al., 2011)
Clostridium coccoidesEubacterium rectale group
1-(3’,4’-dihydroxyphenyl)-3-(2’’,4’’,6’’-trihydroxyphenyl)propan-2-ol
(+) – catechin
4-hydroxy-5-(3’,4’-dihydroxyphenyl)valeric acid
AC
TC
4-Hydroxy-5-(phenyl)-valeric acid
DC
0,6
AC
0,4
0,2
0
TC
(+)-Catechin (µg/mL)
0
2
4
6
8 10 12 14 16 18 20
Days
0,8
0,6
0,4
0,2
0
0,8
0
2
4
6
8
Days
0,6
DC
10 12 14 16 18 20
0,4
0,2
0
0
2
4
6
8
10 12 14 16 18 20
Treatment
0,1
0,08
0,06
0,04
0,02
0
Washout
Treatment
60
Washout
Clostridium leptum
AC
40
0
2
4
6
8 10 12 14 16 18 20
Days
0,1
0,08
0,06
0,04
0,02
0
0,1
0
2
4
6
8 10 12 14 16 18 20
Days
0,08
0,06
4-Hydroxy-5-(phenyl)-valeric acid (µg/mL)
Washout
4-Hydroxy-5-(3’,4’-dihydroxyphenyl)-valeric acid (µg/mL)
Treatment
0,8
80
20
0
0
0
Days
Days
Provinols®
2
4
6
8 10 12 14 16 18 20
Days
60
0
8 10 12 14 16 18 20
8 10 12 14 16 18 20
DC
AC
TC
80
20
6
6
Ruminococccus s pp.
0
0,02
4
4
80
60
40
20
0
40
2
2
Days
0,04
0
TC
DC
Provi nol s
Provi nol s + L. plantarum IFPL935
1-5
9-11
7,32
7,06
6,86
7,17
7,60
7,26
7,06
7,48
7,43
7,11
7,34
7,57
6,79
5,94
6,12
5,91
6,63
5,84
6,29
6,37
6,31
5,68
6,21
6,49
5,74
4,94
5,94
6,19
5,97
5,66
6,31
6,59
6,03
5,48
6,23
6,33
6,31
5,37
6,27
6,18
6,54
6,04
6,14
6,28
6,19
5,70
5,66
(0,21)
(0,40)
(0,20)
(0,21)
(0,09)
(0,25)
(0,36)
(0,01)
(0,09)
(0,38)
(0,35)
(0,16)
(0,29)
(0,31)
(0,47)
(0,45)
(0,08)
(0,18)
(0,18)
(0,10)
(0,01)
(0,23)
(0,29)
(0,14)
(0,28)
(0,42)
(0,15)
(0,09)
(0,12)
(0,20)
(0,41)
(0,21)
(0,32)
(0,13)
(0,30)
(0,02)
(0,14)
(0,42)
(0,10)
(0,13)
(0,07)
(0,06)
(0,45)
(0,03)
(0,26)
(0,20)
(0,60)
7,06
7,34
7,00
7,20
7,42
6,93
7,19
7,51
7,34
6,86
7,33
7,24
6,57
6,23
6,16
6,73
6,51
6,01
6,74
6,68
6,44
5,58
6,55
6,45
5,73
4,19
5,74
5,59
5,72
4,93
6,23
6,19
5,69
4,96
6,13
6,18
6,20
5,09
6,06
6,02
6,26
5,98
6,34
6,28
6,25
5,62
6,38
(0,14)
(0,26)
(0,22)
(0,12)
(0,21)
(0,48)
(0,08)
(0,27)
(0,18)
(0,59)
(0,04)
(0,31)
(0,44)
(0,35)
(0,40)
(0,10)
(0,09)
(0,51)
(0,02)
(0,07)
(0,08)
(0,50)
(0,02)
(0,35)
(0,42)
(0,51)
(0,00)
(0,16)
(0,10)
(0,53)
(0,18)
(0,20)
(0,21)
(0,49)
(0,32)
(0,45)
(0,05)
(0,38)
(0,04)
(0,15)
(0,14)
(0,17)
(0,19)
(0,20)
(0,10)
(0,57)
(0,21)
Wa s hout
Sta b.
Trea t. da ys 1-5
Trea t. da ys 9-11
Wa s hout
Sta b.
Trea t. da ys 1-5
6,32
3,66
2,91
3,70
3,76
5,33
4,35
(0,07)
(0,21)
(0,41)
(0,31)
(0,12)
(0,10)
(0,27)
6,10
2,78
1,90
1,39
1,80
5,18
4,33
(0,26)
(0,26)
(0,61)
(0,09)
(0,07)
(0,05)
(0,33)
Trea t. da ys 9-11
Wa s hout
4,60 (0,30)
5,15 (0,19)
5,18 (0,27)
5,16 (0,04)
Sta b.
Trea t. da ys 1-5
Trea t. da ys 9-11
5,40 (0,13)
4,62 (0,32)
4,83 (0,49)
5,40 (0,13)
4,55 (0,64)
5,43 (0,23)
Wa s hout
Sta b.
Trea t. da ys 1-5
Trea t. da ys 9-11
Wa s hout
Sta b.
Trea t. da ys 1-5
5,34
3,23
2,75
3,23
3,30
5,15
4,28
5,17
2,72
2,84
2,25
2,49
5,10
4,24
Trea t. da ys 9-11
Wa s hout
4,34 (0,19)
4,86 (0,54)
4,65 (0,38)
4,79 (0,16)
Sta b.
Trea t. da ys 1-5
Trea t. da ys 9-11
Wa s hout
6,00
5,27
5,45
5,45
5,95
5,05
5,47
5,62
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
1-5
9-11
(0,26)
(0,15)
(0,24)
(0,25)
(0,14)
(0,15)
(0,42)
(0,14)
(0,38)
(0,46)
(0,66)
(0,21)
(0,20)
(0,28)
(0,08)
(0,13)
(0,06)
(0,46)
(0,05)
(0,70)
(0,39)
(0,11)
• A decrease in the microbial community is observed the first days of treatment, related
with the reduction of butyrate and total SCFAs (data not shown).
0
2
4
6
8 10 12 14 16 18 20
Days
Provinols® + L. plantarum IFPL935
Catabolism of (+)-catechin.
• The evolution of the content of (+)-catechin during the treatment and washout periods shows disappearance of this
compound in TC and DC, in accordance to the progressive appearance of phenolic metabolites starting after 4-5 days of
treatment.
• L. plantarum tend to favor the catabolism of (+)-catechin, showing a more rapid disappearance of the precursor to the
formation of further-intermediate products.
• The increase of butyrate was associated with a faster recovery of the butyrate-producing
bacteria, Clostridium leptum, Clostridium coccoides-Eubacterium rectale group and
Ruminococcus on TC and DC vessels in the SHIME supplemented with L. plantarum IFPL935.
Conclusion
L. plantarum IFPL935 shows a protective effect on butyrate-producing bacteria and is able to
improve colonic metabolism by promoting the formation of biologically significant compounds
Reference:
Tabasco, R., Sánchez-Patán, F., Monagas, M., Bartolomé, B., Moreno-Arribas, M.V., Peláez, C., Requena, T. 2011 Effect of
grape polyphenols on lactic acid bacteria and bifidobacteria growth:resistance and metabolism. Food Microbiol. 28: 1345-1352.
This work was supported by Research Projects AGL2009-13361-C02-00, RM2011-00003-00-00 and Consolider
Ingenio 2010 FUN-C-FOOD-CSD2007-00063, and program ALIBIRD P2009/AGR-1469.