Isolation of bifidobacteria for blood group secretor status targeted personalised nutrition

Figures & data

Table 1. Growth media and conditions for bacteria cultivation

Agar medium	For which bacteria	Incubation temp. (°C)	Incubation aerobic/anaerobic	Incubation time (h)
Beerens	Bifidobacteria	37	Anaerobic	72
RB	Bifidobacteria	37	Anaerobic	72
Rogosa	Lactobacilli	37	48 h aerobic/24 h anaerobic	72
LAMVAB	Lactobacilli	37	Anaerobic	72
RCBA	Anaerobes	37	Anaerobic	72
BBE	Bacteroides	37	Anaerobic	72

RB (raffinose-bifìdobacterium agar), LAMVAB (Lactobacillus anaerobic MRS with vancomycin and bromocresol), RCBA (reinforced Clostridial agar with China blue and horse blood) and BBE (bacteroides bile esculin agar).

Table 2. Survival of intestinal bacteria (number of cfu's) in the TIM-1 samples in non-secretor pool and secretor pool

Non-secretor	Total number of cfu in the samples						Secretor	Total number of cfu in the samples
Sample	Beerens	RB	Rogosa	LAMVAB	RCBA	BBE	Sample	Beerens	RB	Rogosa	LAMVAB	RCBA	BBE
intake	4.1E + 08	3.2E + 09	3.0E + 09	4.1E + 07	2.9E + 10	2.2E + 09	intake	6.5E + 08	3.4E + 09	8.4E + 09	1.3E + 07	2.3E + 10	3.4E + 08
Ie-1	1.7E + 05	1.9E + 07	1.9E + 07	3.4E + 03	4.8E + 07	0.0E + 00	Ie-1	1.0E + 07	4.8E + 07	4.1E + 07	3.5E + 04	4.1E + 08	0.0E + 00
Ie-2	2.4E + 05	2.1E + 07	1.3E + 07	6.8E + 03	1.6E + 08	0.0E + 00	Ie-2	5.1E + 06	5.1E + 07	3.3E + 07	1.7E + 04	1.3E + 09	3.4E + 06
Ie-3	1.4E + 05	2.4E + 07	1.6E + 07	3.5E + 03	5.2E + 08	0.0E + 00	Ie-3	5.4E + 06	6.5E + 07	3.0E + 07	2.4E + 04	4.1E + 09	0.0E + 00
Total survival	5.5E + 05	6.5E + 07	4.8E + 07	1.4E + 04	7.3E + 08	0.0E + 00	Total survival	2.1E + 07	1.6E + 08	1.0E + 08	7.5E + 04	5.8E + 09	3.4E + 06
% Survival	0.1	2.0	1.6	0.0	2.5	0.0	% Survival	3.2	4.8	1.2	0.6	25.0	1.0

Table 3. Bifidobacteria and lactobacilli isolates collected from the TIM-1 model experimentation sample plates

	No. of RAPD types (no. of isolates)		Species
Group	Secretor	Non-secretor	Secretor	Non-secretor
Bifidobacteria	31 (276)	11 (196)	6	5
Lactobacilli	21 (126)	8 (65)	9	6

Table 4. Substrate utilization, enzyme activity, and bile tolerance of bifidobacteria isolated from secretor and non-secretor individuals

Pool	No. of positive isolates/total no. of isolates (%)
	Growth*	Rosco Diatabs
Species	Fucose	Lacto-n-biose	Glucose	Lactose	Sucrose	Mannose	α-Glucosidase	β-Glucosidase	α-Galactosidase	β-Galactosidase	α-Fucosidase**	Bile resistance
Secretor
Bifidobacterium sp.	0/1	0/1	1/1	1/1	1/1	1/1	1/1	1/1	1/1	1/1	–	0/1
B. adolescentis	4/8	2/8	6/8	8/8	7/8	3/8	8/8	8/8	8/8	8/8	0/3	2/8
B. catenulatum	2/4	2/4	4/4	4/4	4/4	3/4	4/4	3/4	4/4	4/4	0/1	2/4
B. longum	0/8	0/8	6/8	8/8	4/8	5/8	8/8	3/8	8/8	6/8	0/3	0/8
B. animalis	0/4	0/4	4/4	4/4	4/4	4/4	4/4	3/4	4/4	4/4	0/1	4/4
All	6/25 (24%)	4/25 (16%)	21/25 (84%)	25/25 (100%)	20/25 (80%)	16/25 (64%)	25/25 (100%)	18/25 (72%)	25/25 (100%)	23/25 (92%)	0/8 (0%)	8/25 (32%)
Non-secretor
Bifidobacterium sp.	0/1	0/1	0/1	1/1	0/1	1/1	1/1	1/1	1/1	1/1	–	1/1
B. adolescentis	2/3	2/3	2/3	3/3	3/3	3/3	3/3	3/3	3/3	3/3	0/3	1/3
B. catenulatum	0/2	0/2	2/2	2/2	2/2	1/2	2/2	2/2	2/2	2/2	–	2/2
B. longum	0/4	0/4	3/4	4/4	2/4	2/4	4/4	4/4	4/4	4/4	0/3	1/4
B. animalis	0/1	0/1	1/1	1/1	1/1	1/1	1/1	1/1	1/1	1/1	0/1	1/1
All	2/11 (18%)	2/11 (18%)	8/11 (73%)	11/11 (100%)	8/11 (73%)	8/11 (73%)	11/11 (100%)	11/11 (100%)	11/11 (100%)	11/11 (100%)	0/7 (0%)	6/11 (55%)

*Bioscreen growth marked positive if (OD after growth with 0.5% substrate–OD after growth with 0.5% glucose) > 0.

**Due product shortage, α-fucosidase production was measured only from total of 15 strains.

– States for not measurement.