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DOI: 10.1542/peds.2007-1192 2008;122;8 Pediatrics Korpela, Tuija Poussa, Tuula Tuure and Mikael Kuitunen Kaarina Kukkonen, Erkki Savilahti, Tari Haahtela, Kaisu Juntunen-Backman, Riitta

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DOI: 10.1542/peds.2007-1192

2008;122;8

Pediatrics

Korpela, Tuija Poussa, Tuula Tuure and Mikael Kuitunen Kaarina Kukkonen, Erkki Savilahti, Tari Haahtela, Kaisu Juntunen-Backman, Riitta

Trial

Placebo-Controlled Prebiotic (Synbiotic) Treatment: Randomized, Double-Blind,

Long-Term Safety and Impact on Infection Rates of Postnatal Probiotic and

http://pediatrics.aappublications.org/content/122/1/8.full.html

located on the World Wide Web at:

The online version of this article, along with updated information and services, is

of Pediatrics All rights reserved Print ISSN: 0031-4005 Online ISSN: 1098-4275

Boulevard, Elk Grove Village, Illinois, 60007 Copyright © 2008 by the American Academy published, and trademarked by the American Academy of Pediatrics, 141 Northwest Point

publication, it has been published continuously since 1948 PEDIATRICS is owned,

PEDIATRICS is the official journal of the American Academy of Pediatrics A monthly

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Long-Term Safety and Impact on Infection Rates of

Postnatal Probiotic and Prebiotic (Synbiotic)

Treatment: Randomized, Double-Blind,

Placebo-Controlled Trial

Kaarina Kukkonen, MD a , Erkki Savilahti, MD, PhD b , Tari Haahtela, MD, PhD a , Kaisu Juntunen-Backman, MD, PhD a , Riitta Korpela, PhD c , Tuija Poussa, MSc d , Tuula Tuure, PhD e , Mikael Kuitunen, MD, PhD a

a Department of Pediatrics, Skin and Allergy Hospital, b Department of Pediatrics, Hospital for Children and Adolescents, and c Department of Pharmacology, Institute of Biomedicine, University of Helsinki, Helsinki, Finland; d STAT Consulting, Tampere, Finland; e Valio Research and Development, Helsinki, Finland

Financial Disclosure: Salaries Dr Kukkonen received and grants Dr Kuitunen received from the Clinical Research Institute of Helsinki University Central Hospital were funded by Valio Drs Korpela and Tuure were employed by Valio Research Centre Ms Poussa received consulting fees from Valio.

What’s Known on This Subject

Probiotics and prebiotics are known to modulate immune responses The accumulating

evidence of their health-promoting effects has led to increased consumption in infancy.

However, long-term follow-up and safety data for administration to newborn infants are

lacking.

What This Study Adds

This study documents safety and provides long-term follow-up data on probiotics and prebiotics administered to newborn infants The study suggests that feeding probiotics and prebiotics to allergy-prone infants may increase their resistance to respiratory infections.

ABSTRACT

OBJECTIVE.Live probiotic bacteria and dietary prebiotic oligosaccharides (together

termed synbiotics) increasingly are being used in infancy, but evidence of long-term

safety is lacking In a randomized, placebo-controlled, double-blind trial, we studied

the safety and long-term effects of feeding synbiotics to newborn infants

METHODS.Between November 2000 and March 2003, pregnant mothers carrying

in-fants at high risk for allergy were randomly assigned to receive a mixture of 4

probiotic species (Lactobacillus rhamnosus GG and LC705, Bifidobacterium breve Bb99,

and Propionibacterium freudenreichii ssp shermanii) or a placebo for 4 weeks before

delivery Their infants received the same probiotics with 0.8 g of

galactooligosaccha-rides, or a placebo, daily for 6 months after birth Safety data were obtained from

clinical examinations and interviews at follow-up visits at ages 3, 6, and 24 months

and from questionnaires at ages 3, 6, 12, and 24 months Growth data were collected

at each time point

RESULTS.Of the 1018 eligible infants, 925 completed the 2-year follow-up assessment

Infants in both groups grew normally We observed no difference in neonatal

morbidity, feeding-related behaviors (such as infantile colic), or serious adverse

events between the study groups During the 6-month intervention, antibiotics were

prescribed less often in the synbiotic group than in the placebo group (23% vs 28%)

Throughout the follow-up period, respiratory infections occurred less frequently in

the synbiotic group (geometric mean: 3.7 vs 4.2 infections)

CONCLUSION.Feeding synbiotics to newborn infants was safe and seemed to increase resistance to respiratory infections

during the first 2 years of life Pediatrics 2008;122:8–12

PROBIOTICS ARE LIVEmicrobes that, when ingested, may modulate systemic immune responses.1Their biological effects are strain specific, and prerequisites to their effects are viability and the ability to colonize.2Prebiotics are indigestible nutrients, such as galactooligosaccharides (GOSs) in human breast milk, that stimulate the growth and metabolic activity of beneficial bacteria in the gut flora but also may produce a direct immunologic effect.3,4 Long-term use of these immunomodulatory agents among infants has been beneficial in autoimmune and allergic disorders, such as inflammatory bowel diseases5 and atopic eczema.6 Their use is also associated with increased resistance to acute enteric and respiratory infections.7,8With the accumulating evidence of the benefits they produce, probiotics and prebiotics are added to dairy products, which results in long-term consumption9 among pregnant mothers and young infants

www.pediatrics.org/cgi/doi/10.1542/ peds.2007-1192

doi:10.1542/peds.2007-1192

This trial has been registered at www clinicaltrials.gov (identifier NCT00298337).

Key Words

probiotic, prebiotic, synbiotic, safety, growth, respiratory infections, antibiotics

Abbreviations

GOS— galactooligosaccharide OR— odds ratio

CI— confidence interval

Accepted for publication Nov 6, 2007 Address correspondence to Kaarina Kukkonen,

MD, Helsinki University Central Hospital, Skin and Allergy Hospital, Meilahdentie 2, PO Box

160, 00029 HUCH Helsinki, Finland E-mail: kaarina.kukkonen@hus.fi

PEDIATRICS (ISSN Numbers: Print, 0031-4005; Online, 1098-4275) Copyright © 2008 by the American Academy of Pediatrics

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Gut microbiota affect nutrient uptake and thereby

host energy metabolism.10 In view of that, probiotics

may have an impact on weight gain and growth

Probiotics and prebiotics are regulated mostly within the

context of food, not drugs Although they are generally

considered safe,11reporting on the long-term safety of

bac-terial strains proven to colonize and to induce a clinical

effect is warranted In this placebo-controlled study, we

document safety and provide long-term follow-up data

The synbiotics consisted of 4 probiotic strains and prebiotic

GOSs They were given in a double-blinded manner to

pregnant mothers and to their allergy-prone infants from

birth to the age of 6 months.12

METHODS

A detailed description of the study design appears

else-where.12 In brief, we enrolled 1223 pregnant mothers

carrying infants at high risk for allergy in an

allergy-prevention trial in Helsinki, Finland, between November

2000 and March 2003 Mothers took capsules containing

a mixture of Lactobacillus rhamnosus GG and LC705,

Bi-fidobacterium breve Bb99, and Propionibacterium

freuden-reichii ssp shermanii JS (8 –9⫻ 109colony-forming units

in each capsule) or a placebo twice daily for 4 weeks

before delivery For 6 months after birth, the infants

received daily 1 opened capsule of the same probiotics

and 0.8 g of GOSs (of bovine origin) in liquid form or

placebo (microcrystalline cellulose plus sugar syrup)

Parents received illustrated instructions to mix the

pro-biotic powder with liquid (water, breast milk, or

for-mula) in a teaspoon and to feed it to the infants with the

spoon Exclusion criteria included birth at⬍37 weeks of

gestation, being a B twin, and having a major

malfor-mation Mothers provided their written informed

con-sent, and the ethics committee at the local hospital

ap-proved the study protocol

The study pediatrician, blinded to group allocation,

examined the infants and interviewed the parents at

ages 3, 6, and 24 months At 3, 6, 12, and 24 months, the

parents completed questionnaires covering 0 to 3, 3 to 6,

6 to 12, and 12 to 24 months, respectively We inquired

about neonatal morbidity, feeding-related behaviors,

nutrition, the environment, and numbers of infections,

antibiotics, and other diseases The questionnaires were

delivered by mail except for the 3- to 6-month

question-naire, which was given in person at the 3-month visit

The questionnaires were returned during the study visits

(at 3, 6, and 24 months) or by mail (at 12 months) The

parents were advised to contact the study pediatrician in

the event of adverse reactions The infants’

anthropo-metric measures were obtained from primary health

care charts Growth measurements were converted to

SD scores with Pediator software (Tilator Ltd, S ¨akyl ¨a,

Finland), by using data for Finnish children as reference

data.13

All analyses used an intention-to-treat approach The

sample size calculations are presented elsewhere.12

An-thropometric measures were analyzed by using the t test

for independent samples The␹2test was used to

com-pare categorized or dichotomized conditions between

the groups The results are given as odds ratios (ORs)

with 95% confidence intervals (CIs) The numbers of infections and antibiotic courses were skewed to the

right and were logarithmically transformed The t test for

independent samples was then used for group compar-isons, and the results are presented as synbiotic/placebo ratios with 95% CIs The data were analyzed with SPSS 14.0 (SPSS, Chicago, IL)

RESULTS Study Groups

Of the 1223 randomly assigned mothers, 156 refused to participate, and 49 of their infants (plus 14 B twins) were ineligible Of these, 8 infants in the synbiotic group and 7 in the placebo group were born prematurely to mothers who had started the intervention The baseline characteristics of the 1018 intention-to-treat infants were comparable between the study groups (Table 1) A

total of 939 infants (synbiotic, n ⫽ 468; placebo, n ⫽

471) completed the 6-month follow-up evaluation, and

925 (synbiotic, n ⫽ 461; placebo, n ⫽ 464) completed

the 2-year follow-up evaluation

Neonatal Morbidity

We observed no significant differences in parent-re-ported neonatal morbidity of any cause for infants in the synbiotic group, compared with those in the placebo group (Table 2)

Infantile Colic and Defecation

Infantile colic, defined as cryingⱖ4 hours per day for ⱖ3 days per week,14 occurred in 4% and similar but less-frequent crying (once or twice per week) occurred in 10% of each group Defecatingⱖ3 times per day was less

frequent in the synbiotic group (18% vs 29%; P⬍ 001)

Tolerance and Adverse Events

Feeding-related behaviors (vomiting, constipation, ex-cessive crying, and abdominal discomfort) occurred

sim-TABLE 1 Baseline Demographic and Clinical Characteristics of the

Infants in the Intention-to-Treat Synbiotic and Placebo Groups

Synbiotic

(n⫽ 506)

Placebo

(n⫽ 512)

Birth weight, mean ⫾ SD, g 3595 ⫾ 483 3593 ⫾ 484 Birth length, mean ⫾ SD, cm 50.5 ⫾ 2.0 50.6 ⫾ 1.9

Daily exposure to tobacco smoke, % 32 29 Partially breastfed for ⱖ6 mo, % 71 68 Total duration of breastfeeding, mean ⫾ SD, mo 8.6 ⫾ 5.4 8.2 ⫾ 5.0 Attending day care before 2 y of age, % 50 51 Firstborn child in the family, % a 58 52

aP⬍ 05 between the intention-to-treat groups, but no difference occurred between the study groups included in this safety analysis (at follow-up times of ⱖ3 months) No significant differ-ences in other baseline variables were observed.

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ilarly in the study groups (data shown elsewhere).12

Symptoms that caused discontinuation of the

interven-tion are presented in Table 2 One of the 6 infants with

difficulties swallowing the powder experienced a

chok-ing event associated with chok-ingestion of the powder but

recovered completely Any other reason for

hospitaliza-tion after discharge from the maternity hospital to 2

years of age (Table 2) was likely unrelated to the

inter-vention

Growth

The anthropometric measures at the ages of 6 months

and 2 years, showing similar normal growth in the 2

groups, are presented in Table 3

Infections and Antibiotics

During the intervention (0 – 6 months), we observed no

significant difference between the synbiotic and placebo

groups in the occurrence (at least once) of respiratory

infections (66% vs 68%), middle ear infections (15% vs

19%), or gastroenteritis (13% vs 14%) However, fewer

infants received antibiotics in the synbiotic group than in

the placebo group (23% vs 28%; OR: 0.74; 95% CI:

0.55–1.00; P⫽ 049)

After the intervention, during the follow-up period

(6 –24 months), respiratory infections occurred less

fre-quently in the synbiotic group (93%) than in the

.023) The total number of respiratory infections was

significantly lower in the synbiotic group (geometric

mean: 3.7 vs 4.2 infections; ratio: 0.87; 95% CI: 0.79 –

0.97; P⫽ 009) In these respective groups, middle ear

infections occurred in 72% vs 76% (ratio: 0.83; 95% CI:

0.62–1.11; P ⫽ 204) The total number of middle ear

infections tended to be lower in the synbiotic group

(geometric mean: 1.7 vs 1.9 infections; ratio: 0.89; 95%

CI: 0.78 –1.01; P ⫽ 068) Gastroenteritis was equally common in the synbiotic and placebo groups (74% vs 71%; geometric mean: 1.3 vs 1.2 episodes; ratio: 1.02;

period (6 –24 months), most infants received antibiotics, with no significant difference between the synbiotic group (80%) and the placebo group (83%); the geomet-ric mean number of antibiotic courses was 2.2 vs 2.4

(ratio: 0.92; 95% CI: 0.81–1.05; P⫽ 206)

DISCUSSION

We showed that treatment of mothers with probiotics during late pregnancy and treatment of their healthy, allergy-prone infants with synbiotics for 6 months after birth were safe Infants in both treatment groups grew normally, and no difference in morbidity related to syn-biotics occurred In fact, the synsyn-biotics seemed to im-prove the infants’ resistance to respiratory infections; during their first 6 months of life, they were prescribed antibiotics less frequently than were infants receiving placebo and, thereafter to the age of 2 years, they expe-rienced fewer respiratory infections

This is the largest randomized, clinical trial on probi-otics and prebiprobi-otics given to pregnant mothers and their newborn infants Probiotics and prebiotics have gener-ally been well tolerated, but we documented the safety

of their prenatal use with respect to neonatal morbidity Our trial with baseline-comparable treatment groups, good adherence to the treatment, and successful probi-otic bacterial colonization was initiated when the use of probiotics and prebiotics in infant foods was still uncom-mon.12

Vaginal flora is crucial to the initial colonization of the newborn gut after normal delivery,15and treating preg-nant mothers with probiotics promotes newborn coloni-zation with the same bacteria.16Perinatal exposure may

be vital to the probiotic effect, because postnatal

admin-istration of Lactobacillus acidophilus showed no preventive

effect on atopy.17In addition, the diversity of gut

micro-TABLE 3 Anthropometric Measurements at 6 and 24 Months of Age

for Infants Who Received Synbiotics or Placebo During the First 6 Months of Life

Visit Synbiotic Group

(n⫽ 446) a

Placebo Group

(n⫽ 456) b Age, mean ⫾ SD, d 6 mo 183 ⫾ 10 184 ⫾ 11

24 mo 736 ⫾ 17 735 ⫾ 20 Length, mean ⫾ SD, cm 6 mo 68.4 ⫾ 2.4 68.4 ⫾ 2.4

24 mo 88.4 ⫾ 3.2 88.6 ⫾ 3.1 Length SD scores, mean ⫾ SD 6 mo 0.00 ⫾ 0.97 ⫺0.04 ⫾ 0.98

24 mo 0.28 ⫾ 1.01 0.34 ⫾ 0.96 Weight, mean ⫾ SD, kg 6 mo 8.16 ⫾ 0.98 8.09 ⫾ 0.95

24 mo 12.8 ⫾ 1.5 12.8 ⫾ 1.4 Head circumference, mean ⫾ 6 mo 43.9 ⫾ 1.3 43.9 ⫾ 1.3

SD, cm 24 mo 49.4 ⫾ 1.5 49.5 ⫾ 1.7 Growth measurements were converted to SD scores by using data on Finnish children as reference data 13

a Growth data were available for 442 infants at the age of 24 months.

b Growth data were available for 449 infants at the age of 24 months.

TABLE 2 Neonatal Morbidity and Reasons for Discontinuation of

the 6-Month Intervention in the Study Groups

No of Infants Synbiotic

(n⫽ 506)

Placebo

(n⫽ 512) Neonatal morbidity

Other (meconium plug obstruction, patent

ductus arteriosus, or neonatal hepatitis)

Reasons for discontinuing the intervention

Difficulty in swallowing the product 2 2

Reasons for hospitalization at 0–2 y

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biota is important for maturation of the immune

sys-tem.18Among these infants, total counts of bifidobacteria

and lactobacilli were significantly higher in the active

group, which indicates the effectiveness of our

interven-tion.12

Some parents encountered problems in administering

the powder to their newborn infants Although parents

had received illustrated instructions to mix the powder

with liquid, the powder caused a choking event in one

infant Therefore, if the preparation is administered as a

powder, parents should receive personal instruction to

mix the powder with adequate quantities of breast milk

or formula The GOS syrup alone was easily

adminis-tered and had no such disadvantage

We chose structured questionnaires and interviews

rather than diaries, to enhance compliance during the

long follow-up period This limited the detailed

informa-tion available on the durainforma-tion and severity of infectious

diseases but allowed us to compare incidences of

infec-tions

We observed no benefit of the synbiotics in

feeding-related behaviors During our intervention, the majority

of infants (70%) were breastfed Breast milk contains

large quantities of GOSs (0.8 g/100 mL), which is

note-worthy when our results are compared with the benefits

of GOSs for bottle-fed infants.19,20 More than

simethi-cone, Lactobacillus reuteri has ameliorated infantile colic

in breastfed infants.21We observed no such effect,

how-ever The overall incidence of infantile colic (4%) was

lower than that in the aforementioned trials20,21and less

than the 9% incidence of infantile colic in a

community-based trial.22 Parents of colicky infants in our study

re-ceived counseling from our trained nurses and thus

might have felt more confident in handling such

symp-toms

Gut microbiota contribute to the host’s energy

me-tabolism.10Probiotic bacteria may enhance the uptake of

nutrients and thereby increase nutritional status (ie,

im-prove growth and iron status).23 In Estonia, bottle-fed

infants who received L rhamnosus GG-enriched formula

for 6 months grew better than did those who received

regular formula.24In the United States, growth was

sim-ilar in 3- to 24-month-old infants who received

Bi-fidobacterium lactis and Streptococcus thermophilus or a

pla-cebo.11 Consistently, the normal growth observed in

both our study groups did not support improved growth

with probiotics in otherwise well-nourished infants

The occurrence of fewer respiratory infections with

our synbiotics is in line with the results of a large

ran-domized trial in which L rhamnosus GG improved

resis-tance to respiratory infections in infants attending day

care.8In an Israeli multicenter trial, L reuteri and B lactis

provided no protection against respiratory infections

among children in day care, but the use of L reuteri was

associated with fewer prescribed antibiotics.25 In

con-trast, newborn Australian infants who received L

aci-dophilus postnatally for 6 months received no protection

from atopy or respiratory infections but were more likely

to be given antibiotics.17The lower frequency of

antibi-otic use among infants in day care who received formula

containing S thermophilus plus B lactis11 agrees with the

results of our study, indicating fewer antibiotic courses throughout the intervention We infer that feeding syn-biotics promotes maturation of the immune system, which results in 13% risk reduction for respiratory in-fections from 6 to 24 months of age

Our synbiotics failed to prevent episodes of diarrhea, which were rare (14%) during the intervention In the

Finnish study among children in day care, L rhamnosus

GG in milk failed to reduce the already low incidence of gastroenteritis.8

Our cohort consisted of infants who were genetically

at risk for atopy Some researchers have proposed that immune maturation in atopic infants is delayed,26 be-cause they exhibit weaker antibody responses to vac-cines27 and their resistance to respiratory infections is compromised.28Our results support the idea that probi-otics and prebiprobi-otics may enhance immune maturation and protect infants against respiratory pathogens.1

CONCLUSIONS

This study indicates that the use of live probiotic bacteria and prebiotic nutrients, even when administered to newborn infants over the long term, carries no risks Feeding probiotic bacteria to urban westernized infants genetically prone to atopy may increase their resistance

to infection Additional in vivo studies are warranted to identify the immunologic mechanisms that produce these benefits

ACKNOWLEDGMENTS

This study was supported by the Helsinki University Central Hospital Research Funds and Valio (Helsinki, Finland)

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STUDY: MOM’S MARKET VALUE AT $117,000

“Boston—If a stay-at-home mom could be compensated in dollars rather than personal satisfaction and unconditional love, she’d rake in a nifty sum of nearly $117,000 a year That’s according to a pre-Mother’s Day study released Thursday by Salary.com, a Waltham, Mass.-based firm that studies workplace compensation The eighth annual survey calculated a mom’s market value by studying pay levels for 10 job titles with duties that a typical mom performs, ranging from housekeeper and day care center teacher to van driver, psy-chologist and chief executive officer One stay-at-home mom said the six-figure salary sounds a little low.”

Burlington Free Press May 9, 2008

Noted by JFL, MD

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DOI: 10.1542/peds.2007-1192

2008;122;8

Pediatrics

Korpela, Tuija Poussa, Tuula Tuure and Mikael Kuitunen Kaarina Kukkonen, Erkki Savilahti, Tari Haahtela, Kaisu Juntunen-Backman, Riitta

Trial

Placebo-Controlled Prebiotic (Synbiotic) Treatment: Randomized, Double-Blind,

Long-Term Safety and Impact on Infection Rates of Postnatal Probiotic and

Services

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