Microbial composition and fecal fermentation end products from colicky infants – a probiotic supplementation pilot

Figures & data

Table I.  Demographic characteristics of the infants at enrolment.

Parameter	Colicky	Control
Gender
Males	3	3
Females	6	6
Age (weeks)
Mean	3	4
Range	2–5	1–6
Mode of delivery
Vaginal	8	6
Cesarean	1	3
Birth weight (kg)
Mean	4.2	4.2
Range	3.3–5.0	3.7–4.6
Type of feeding
Breast-fed	9	8
Formula-fed	0	1*
Colicky cry (h/week)
Mean	16	−†
Range	9–28	−†

*Fed with both breast milk and formula.

†Sporadic, infrequent.

Table II.  Microbial findings (log cfu/g wet weight) in nine colicky and nine control infants at baseline.

	Control			Colicky
Parameter	n	Mean	Range	n	Mean	Range
Total anaerobic growth	9	10.2	8.5–10.7	9	10.3	8.8–10.7
Total lactobacilli	6	9.0	5.0–9.5	7	9.5	3.4–10.3
Non-LGG lactobacilli	6	8.9	5.0–9.5	7	9.5	3.4–10.3
L. rhamnosus GG	1	8.2	0.0–9.2	0
Bifidobacteria	8	10.0	8.3–10.7	8	9.9	8.6–10.5
Clostridium spp.	2	7.8	8.5	4	7.5	6.5–8.3
Gram-positive rods†	4	8.8	6.0–9.6	2	8.8	8.0–9.8
Bacteroides fragilis group	6	9.3	5.3–10.0	7	9.6	5.0–10.3
Gram-negative rods‡	1	3.0	0.0–4.0	2	7.3	3.0–8.3
Gram-positive cocci	3	8.6	8.3–9.5	1	7.0	0.0.–8.0
Gram-negative cocci	1	8.1	0.0–9.0	3	9.0	8.0–10.0
Total aerobic growth	9	9.1	8.0–9.4	9	9.5	7.0–10.1
Coliform, indole-positive	3	9.0	8.0–9.4	8*	9.3	4.0–10.0
Coliform, indole-negative	2	7.9	2.0–8.9	1	7.5	0.0–6.6
Pseudomonas spp.	3	2.6	2.7–3.5	4	5.6	2.2–6.5
Staphylococcus aureus	5	6.5	2.6–7.3	4	5.9	1.3–6.8
Coagulase-negative staphylococci	7	8.0	6.7–8.5	9	8.2	5.3–8.7
Enterococci	5	7.5	7.0–8.0	2	8.1	8.7–8.8
Streptococci	2	7.2	7.8–8.0	0
Micrococcus spp.	1	5.0	0.0–6.0	0
Corynebacteria	3	8.0	4.0–8.9	0
Bacillus spp.	3	8.5	4.0–9.3	0
Yeasts	2	2.3	2.0–3.2	0

*p < 0.05.

†Includes Actinomyces spp., Eubacterium-like organisms, and Propionibacterium spp.

‡Includes Bilophila spp., Fusobacterium spp., and Prevotella spp.

Figure 1.  Proportions of bacterial cellular fatty acids (CFAs) of total fatty acids (mean area percentage and SD) and number of infants carrying them among nine colicky and nine control infants at baseline. Only CFAs comprising at least 0.5% of the total CFAs are presented. *Difference between groups was significant (p < 0.05). Dma, dimethyl acetal.

Figure 2.  Fecal volatile and nonvolatile short chain fatty acids (SCFAs) (mean mmol/l and SD) and hydrogen and carbon dioxide gases (percentage of total gas volume, SD) produced at baseline in 48 h fermentation in peptone-yeast extract broth (PY) and peptone-yeast extract-glucose broth (PYG) by nine colicky and nine control infants. Also minor amounts (1–5 mmol/l) of isovaleric, isobutyric, isocapronic, and phenylacetic acids were produced from PY and pronionic acid from PYG (not shown). No capronic or valeric acids were detected. *Difference between the groups is in the limit of being significant, p = 0.051.

Table III.  Microbial key findings (log cfu/g wet weight) in colicky infants before and after supplementation with the probiotic (five infants) or placebo (four infants).

	Probiotic supplementation						Placebo supplementation
	Before			After			Before			After
Bacteria	n	Mean	Range	n	Mean	Range	n	Mean	Range	n	Mean	Range
Total anaerobes	5	10.3	8.8–10.6	5	10.7*	10.2–10.8	4	10.4	9.9–10.7	4	10.5	9.8–10.9
Bifidobacteria	5	9.9	8.6–10.5	5	10.4*	10.0–10.5	3	9.8	9.8–10.0	3	10.2	8.8–10.7
L. rhamnosus GG	0	0		5	8.6*	7.5–8.9	0	0		1	3.6	0.0–4.2
Non-LGG lactobacilli	4	9.0	3.4–9.7	3	9.6	2.0–10.3	4	9.7	8.6–10.3	3	8.4	3.1–9.0
Total lactobacilli	4	9.0	3.4–9.7	5	9.6	7.5–10.3	4	9.7	8.6–10.3	4	8.4	3.1–9.0
Enterococci	0	0		4	7.9*	5.0–8.5	2	8.5	8.7–8.8	3	8.2	7.9–8.5
Total aerobes	5	9.5	7.7–10.0	5	9.7	8.5–10.2	4	9.5	7.0–10.1	4	9.3	8.3–9.7

*p<0.05.

Table IV.  Fecal volatile and nonvolatile short chain fatty acids (SCFA mmol/l) and hydrogen and carbon dioxide gases (% of gas volume) produced in 48 h fermentation in peptone-yeast extract broth (PY) and peptone-yeast extract-glucose broth (PYG) before and after probiotic (five infants) or placebo (four infants) supplementation.

	Probiotic supplementation						Placebo supplementation
	Before			After			Before			After
Products	n	Mean	Range	n	Mean	Range	n	Mean	Range	n	Mean	Range
PY
Acetic	5	119	33–232	5	97	15–231	4	63	11–127	4	160	102–201
Propionic	5	35	22–66	5	30	8.0–74	4	18	3.6–47	4	31	19–52
Isobutyric	1	0.3	0–1.4	1	0.7	0–3.4	1	1.5	0–6.1	2	0.4	0–1.6
Butyric	4	13	0–33	5	13	0.8–32	3	7.7	0–21	4	24	0–42
Isovaleric	2	0.7	0–2.4	2	1.4	0–7.0	2	1.2	0–3.5	1	1.3	0–5.2
Valeric	0	0		0	0		1	<0.1	0–0.01	0	0
Isocapronic	0	0		0	0		1	1.5	0–5.9	1	1.5	0–5.9
Lactic	1	0.9	0–4.4	0	0		0	0		0	0
Succinic	2	3.3	0–8.9	0	0		2	3.3	0–6.9	2	4.5	0–7.9
Phenylacetic	0	0		1	0.8	0–3.9	1	0.5	0–1.8	1	0.5	0–1.8
H2	5	3.5	0.4–5.6	3	0.6	0–4.5	4	4.3	2.1–6.2	4	3.4	3.4–4.6
CO2	5	11	10–12	5	11	9.1–13	4	12	9.0–14	4	9.7	8.0–14
PYG
Acetic	5	148	6.6–462	5	436	53–962	4	74	17–206	4	80	19–181
Butyric	1	1.6	0–8.1	1	3.6	0–18	0	0		1	23	0–90
Lactic	5	159	89–247	5	196	185–211	4	159	73–278	4	157	99–233
Succinic	4	1.4	0–3.2	5	4.8	1.0–15	4	2.5	0.8–4.8	4	8.6	0.1–3.3
H2	5	1.5	0.2–5.0	5	1.8	0.2–4.2	4	5.6	2.8–9.3	4	4.6	1.2–7.5
CO2	5	5.6	2.5–8.6	5	5.3	1.1–11	4	10.9	6.8–15	4	9.5	6.3–13

Note that statistics were not performed.

Table V.  Mean crying times (SD) (h/week) preceding the baseline (week 0) and at the second week of the probiotic or placebo supplementation (week 2).

	Week 0		Week 2		Difference
Type of crying	Probiotic	Placebo	Probiotic	Placebo	Probiotic	Placebo
Moderate/severe	16.1 (2.8)	16.2 (7.3)	9.6 (6.5)	6.8 (3.7)	−6.5 (40%)	−9.4 (58%)
Mild	11.6 (6.6)	22.0 (10.9)	13.7 (5.3)	23.9 (16.3)	2.1 (19%)	1.9 (9%)
Total	27.7 (7.9)	38.3 (13.9)	23.4 (4.8)	30.7 (19.2)	−4.3 (16%)	−7.6 (20%)