DR.

REBECCA F SLYKERMAN (Orcid ID : 0000-0003-4942-8887)

Accepted Article
Article type : Regular Article

Effect of early probiotic supplementation on childhood cognition, behaviour and

mood a randomised, placebo-controlled trial

Short title: Probiotic supplementation

RF Slykerman1, J Kang2, N Van Zyl3, C Barthow2, K Wickens2, T Stanley4, C Coomarasamy3,

G Purdie 5, R Murphy6, J Crane2, & EA Mitchell3

1. Department of Psychological Medicine, University of Auckland, Private Bag 92019,

Auckland 1142

2. Department of Medicine, University of Otago, P O Box 7343, Wellington

3. Department of Paediatrics: Child and Youth Health, University of Auckland, Private

Bag 92019, Auckland 1142

4. Department of Paediatrics, University of Otago, P O Box 7343, Wellington

5. Dean’s Office, University of Otago, P O Box 7343, Wellington

6. Department of Medicine, University of Auckland, Private Bag 92019 Auckland 1142

Corresponding Author:

RF Slykerman, Department of Psychological Medicine, University of Auckland, Private Bag

92019, Auckland 1142, New Zealand

Email: r.slykerman@auckland.ac.nz

Phone: +64 9 923 1132

Fax: +64 9 373 7013

This article has been accepted for publication and undergone full peer review but has not
been through the copyediting, typesetting, pagination and proofreading process, which may
lead to differences between this version and the Version of Record. Please cite this article as
doi: 10.1111/apa.14590
This article is protected by copyright. All rights reserved.
 ABSTRACT

Aim: To determine whether probiotic supplementation in early life improves neurocognitive

Accepted Article
outcomes assessed at 11 years of age.

Methods: 474 children who were born March 2004-Aug 2005 participated in a two-centre

randomized placebo-controlled trial of infants at risk of developing allergic disease.

Pregnant women were randomized to take Lactobacillus rhamnosus strain HN001,

Bifidobacterium animalis subsp. lactis strain HN019 or placebo daily from 35 weeks

gestation until 6 months if breastfeeding, and their infants the same treatment from birth to 2

years. Intelligence, executive function, attention, depression and anxiety were assessed

when the children were 11 years of age.

Results: 342 (72.2%) children were assessed (HN001 n=109, HN019 n=118 and placebo

n=115). Overall, there were no significant differences in the neurocognitive outcomes

between the treatment groups.

Conclusion: HN001 and HN019 given in early life were not associated with neurocognitive

outcomes at 11 years of age in this study. However, we cannot exclude that other probiotics

may have a beneficial effect. Further clinical trials are indicated.

Key notes

 We conducted a randomised controlled study of early supplementation with

probiotics on later mood behaviour and cognition.

 At age 11 years, no significant difference was found in cognition, behaviour or mood

between children who had earlier received probiotics or placebo.

 Beneficial effects of other probiotic strains cannot be excluded.

This article is protected by copyright. All rights reserved.

 Funding: Fonterra Co-operative Group Ltd and Cure Kids
Accepted Article
List of Abbreviations

ABC Study: Auckland Birthweight Collaborative Study

AD: Asperger’s Disorder

ADHD: Attention Deficit Hyperactivity Disorder

AGA: Appropriate for gestational age

ASD: Autism Spectrum Disorder

AST: Attention Switching Test

BRIEF: Behavior Rating Inventory of Executive Function

CANTAB: Cambridge Neuropsychological Test Automated Battery

CES-DC: Center for Epidemiological Studies Depression Scale for Children

CFU (cfu lower case in text): Colony forming units

CPT: Continuous Performance Test

MASC:2 Multidimensional Anxiety Scale for Children – Second Edition

OST: One Touch Stockings of Cambridge Test

SDQ: Strengths and Difficulties Questionnaire

SGA: Small for gestation age

SWM: Spatial Working Memory

WSIC-IV: Wechsler Intelligence Scale for Children – Fourth Edition

INTRODUCTION

The gut-brain–axis refers to bidirectional links between the microbiota of the gastrointestinal

tract and the development and function of the central nervous system (1-3). The causal role

of gut bacteria in brain development and function has been demonstrated in germ free mice

who show exaggerated physiological reactions to stress, reversible through probiotic-

induced gut bacterial colonisation (4). Furthermore, the germ free mice also had decreased

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 amounts of brain-derived neurotrophic factor (BDNF) in their hippocampi, indicating that the

gut microbiota may also influence cognitive function, in particular memory. A recent review
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reported on 11 animal studies in which probiotics improved spatial memory function (5).

However, a study of human participants found that those given probiotics had poorer

memory function on the Wechsler Memory Scale than those given placebo (6). The

colonisation and stabilisation of gut microbiota in human infants is influenced by multiple

factors including mode of delivery, breastfeeding and antibiotic use (7, 8). The development

of the gut, and the connections it has with the central nervous system, occurs at a time when

the brain is also rapidly developing. Further studies examining different cognitive outcomes

across childhood are needed to determine whether giving probiotics early in life could

improve intellectual outcomes.

Probiotic supplementation to introduce beneficial bacteria into the gut microbiome, thereby

influencing gut neurotransmitter production and inflammatory responses, has been one of

the ensuing areas of interest in the pursuit of possible novel therapies to improve health.

Animal studies manipulating the gut microbiota have shown effects on behaviour. An

increasing number of randomised controlled trials (RCT) of the effect of probiotics on various

outcomes in children and adults have shown mixed results. This has been the subject of a

recent review which concluded that more scientific evidence to support the positive narrative

reviews of the topic is required (9).

A study that followed a sample of 75 children who had received either probiotic

supplementation or placebo for the first 6 months of life reported that 6 of 35 children in the

placebo group had a diagnosis of Attention Deficit hyperactivity Disorder (ADHD) or

Asperger’s Disorder (AD) while none of the children in the probiotic group did. This

difference was statistically significant (10). Furthermore, there is some evidence that

supplementation with probiotics in children with Autism Spectrum Disorders (ASD) reduces

autism-related behaviours (11, 12); however a comprehensive review on the relationship

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 between the gut microbiota and ASD concluded that more RCTs were needed as direct

clinical evidence was limited (13).

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This study was stimulated by our observation that children given antibiotics in the first year of

life had more behavioural difficulties and more symptoms of depression in early childhood

(14). In addition, we have found that pregnant women given probiotics from 16 weeks

gestation to delivery had lower depression and anxiety scores 1-2 months postpartum (15).

The aim of the study was to examine the association between probiotics given in early life

and cognitive (intelligence and executive function), behavioural and mood outcomes at age

11 years. We hypothesised that children who received early life supplementation with

probiotics would have favourable outcomes at follow up. This was a secondary aim as the

original study was designed to investigate the relationship between probiotic

supplementation and later risk of developing allergic disease in childhood.

METHOD

Study design

The study has been described in detail previously (16). In brief, it was a two-centre

(Wellington and Auckland), double-blind, randomized, placebo-controlled trial of the effects

of probiotic supplementation on the development of eczema and atopic sensitization (the

primary outcome), and neurocognitive outcomes at 11 years of age (a post-randomisation

secondary outcome).

Participants

Pregnant women were invited to take part in the study if they or the infant’s father had a

history of treated asthma, eczema or hay fever. Women were ineligible for the study if they

planned to move from the study centre in the next two years, were already taking probiotic

supplements long-term, or intended to use these in the child, or intended to formula-feed

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 their infant. They were excluded if they delivered prior to 37 weeks gestation, had not taken

the study probiotics for ≥2 weeks before birth, their infant’s weight was less than the 3rd
Accepted Article
percentile for sex and gestation, their infant was placed in the neonatal unit for more than 48

hours or had serious congenital abnormalities at birth. If there were twins, only the heavier

was included in the study.

Intervention

The pregnant women were randomised to receive either Lactobacillus rhamnosus strain

HN001 (6X109 cfu/day), Bifidobacterium animalis subsp. lactis strain HN019 (9X109 cfu/day)

or placebo daily from 35 weeks gestation until 6 months if breastfeeding, and their infants

receive the same treatment from birth to 2 years (n=474). The capsules were

indistinguishable. The placebo capsule was identical in appearance and smell, and

contained dextran, salt and a yeast extract. The probiotic or placebo powder was in a

capsule, and the powder was given to the infant mixed with water, breast milk or formula,

and given via a teaspoon or syringe until solid food was introduced. It was then sprinkled on

food.

Randomisation

Randomisation was managed by a clinical trials pharmacist at Auckland City Hospital who

had no contact with the participants. Randomisation was stratified by study centre and

performed in blocks of 15 according to a computer-generated randomization list. Research

staff screened and enrolled eligible participants and provided enrolled participants with the

next available sequentially-numbered capsule container.

Data collection

Demographic information was collected at baseline. The children were seen at 3, 6, 12, 18

months and 2, 4 and 6 years (for the primary outcomes). The participants were blind to

treatment groups for assessments until all 2 year outcomes were assessed. For this study,

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 when children were aged 11 years, they were assessed using a wide range of individually

administered measures and, self-report questionnaires, and parents were asked to complete
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questionnaires regarding their child. Parents/guardians were aware whether their child had

received one of the probiotics or the placebo in the first two years of life. The staff

administering the assessments were blind to group allocation.

Outcome Measures

The Wechsler Intelligence Scale for Children – Fourth Edition (WISC-IV)

The WISC-IV was administered by psychologists. This generates a FullScale IQ (FSIQ)

which represents overall cognitive ability (17).

Behavior Rating Inventory of Executive Function (BRIEF)

Executive function was assessed using the BRIEF, which is designed for children aged 5-18

years (18). The parent form of the questionnaire was completed by a parent or caregiver.

Executive function is an umbrella term for the management (regulation, control) of cognitive

processes, including working memory, reasoning, task flexibility, and problem solving as well

as planning and execution.

Conners 3rd Edition™ (Conners 3™)

Conners 3 is based on its predecessor, the Conners’ Rating Scales–Revised (CRS–R™). It

offers a thorough assessment of ADHD (19). A parent or caregiver completed this

questionnaire.

The Strengths and Difficulties Questionnaire (SDQ)

The SDQ was completed by the parents. It is a 25 item questionnaire which is divided into

five subscales, emotional symptoms, conduct problems, hyperactivity/inattention, peer

relationship problems and prosocial behaviour (20).

Multidimensional Anxiety Scale for Children 2nd Ed (MASC 2)

The MASC 2 is a multi-rater assessment with 50-item self (MASC 2–SR) and parent (MASC

2–P) rating forms. The MASC 2 assesses the presence of symptoms related to anxiety

disorders in youth aged 8 to 19 years (21). The measure distinguishes between important

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 anxiety symptoms and dimensions that broadband measures do not capture. The MASC 2

aids in the early identification of anxiety-prone youth.

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The Centre for Epidemiological Studies Depression Scale for Children (CES-DC)

This 20 item self-report inventory yields scores ranging from 0 to 60 with higher scores being

indicative of increasing depression symptomology(22). Although a cut-off of =>15 is

recommended for screening, a cut off of =>24 is suggestive of moderate to severe

depression, which is more clinically relevant.

The Conners Continuous Performance Test 3rd Edition™ (Conners CPT 3™)

Conners CPT 3 is a task-oriented computerized assessment of attention-related problems in

individuals aged 8 years and older (23). The Conners CPT 3 provides objective information

about an individual’s performance in attention tasks, complementing information obtained

from rating scales such as the Conners 3. By indexing the respondent’s performance in

areas of inattentiveness, impulsivity, sustained attention, and vigilance, the Conners CPT 3

can be useful to the process of diagnosing Attention-Deficit/Hyperactive Disorder (ADHD).

CANTAB

Designed by Cambridge Cognition, the CANTAB is a battery of cognitive and executive

function tests in tablet form (24). This study used three tasks from the CANTAB battery.

Attention Switching Test (AST): This is a measure of a participant’s ability to switch their

attention efficiently and ability to filter out distractions.

One Touch Stockings of Cambridge (OTS): This is a measure of executive function ability

particularly spatial planning and working memory. The variable “responses solved in first

move” is an accuracy measure where higher scores represent better performance.

Spatial Working Memory (SWM): This test requires the retention and manipulation of

visuospatial information. It yields a strategy score.

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 Statistical Analysis

Data were analysed as intent-to-treat. Statistical analysis was conducted in SAS 9.4 using
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ANOVA and two-sample t-test or Kruskal-Wallis test/Wilcoxon score test. There were 9

different tests and 28 subscales examined.

Sample size

Sample size was limited by the size of the original study and the participation rate. There

was on average 114 participants in each group. This would allow us to detect a 0.37

standard deviation difference between two groups with 80% power at the 5% level of

significance.

Ethics

The study was approved by the Central Health and Disability Ethics Committee

(15/CEN/75/AM02). The parent or guardian and the child gave written consent.

Trial registration: Australian New Zealand Clinical Trials Registry: ACTRN12607000518460

RESULTS

There were 474 eligible participants at birth. Participant retention at 2, 4 and 6 years of age

was 446 (94.1%), 425 (89.7%) and 412 (86.9%) respectively. At 11 years of age 342

children (72.2% of those eligible at birth) were assessed on cognitive outcomes (HN001

n=109 (69.4%), HN019 n=118 (74.7%) and placebo n=115 (72.3%)). There was no

difference between those who responded at 11 years of age and those who did not

according to treatment and placebo group assignment (p=0.58). Table 1 shows the mode of

delivery, income and breastfeeding status of respondents by treatment group assignment,

there was no difference between groups on any of these variables. Figure 1 shows the

number of women enrolled and assessed at each stage of the study.

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 Table 2 shows the outcome measure results according to study group assignment: a p-value

of ≤0.05 was considered a statistically significant result. Overall, there were no significant
Accepted Article
differences between groups for the WISC-IV, Conners 3™, MASC 2, SDQ, Conners CPT

3™ and the tasks from CANTAB battery (Attention Switching Test, One Touch Stockings of

Cambridge and Spatial Working Memory). For the BRIEF Behavior Regulation index and the

CES-DC scores there was a significant difference between groups in the opposite direction

to that expected: those in the probiotic group had higher scores (indicating more problems)

than those in the placebo group.

DISCUSSION

We conducted a randomised, placebo-controlled trial of the potential effect of early life

supplementation with two probiotics on cognitive, behavioural and mood outcomes later in

childhood. Results showed no significant benefit on any of the outcome measures in either

of the probiotic supplemented groups. This study had a large sample size relative to other

RCT of probiotic use, and employed a range of self-reported, observer-reported and

objective outcome measures.

The lack of a significant effect of probiotics on these outcomes is consistent with a recent

systematic review of the evidence for probiotic supplementation, which concluded there were

only a few clinical trials with small samples and the results were mixed (9). Response rates

were similar in each group and ranged from 69.4% to 74.7% of those enrolled at birth. This

suggests that response bias is unlikely to account for these negative findings. There are

potential explanations for the lack of a positive association between probiotics in early life

and later neurocognitive outcomes. It is possible that probiotic supplementation that ceases

after the first two years of life may be insufficient to influence longer term neurocognitive

outcome in healthy children. A recent paper which measured growth and development at

age 5 following early feeding with a hydrolysed protein formula fortified with Lactobacillus

rhamnosus GG also found no difference between placebo and probiotic- treated groups (25).

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 A more recent trial in adult males also found no significant benefit of the probiotic

Lactobacillus rhamnosus (JB-1) on cognitive measures or reported stress (26). Other studies
Accepted Article
in adults have shown positive effects of probiotic use on mood with shorter latencies

between supplementation and outcome measurement (Rao & Messoundi).

Because our sample was comprised of children who did not have mood, behavioural or

cognitive disorders of clinical magnitude, it is possible that the probiotic effect demonstrated

in clinical populations was not evident here. However, we have previously found that

probiotics given to healthy pregnant women from 16 weeks to delivery had lower depression

and anxiety scores at 1-2 months postpartum (15).

Cognition includes skills such as information processing that contributes to decision making,

problem solving and executive function. Due to the multifactorial nature of cognitive

development it is possible that probiotic supplementation does not have a large enough

effect to be demonstrated in measures of cognitive ability. There are few studies linking the

gut microbiota to cognition. There are a small number of probiotics studies in mice which

have been shown to improve impaired spatial memory and social cognition (1). Results from

a previously published study showing that antibiotics in early life is associated with

depression, behavioural problems and reduced scores of intelligence and reading in later

childhood is suggestive, but causation could not be established, as antibiotic exposure may

be indicative of ill-health and subsequent neurocognitive development (14). In this study,

antibiotic use was high (>80%) but did not differ between groups (16).

A potential limitation of this study was that parents were not blind to the treatment their child

had received, as they had been informed of their child’s group assignation 7–8 years

previously. This potentially may have altered their responses on parent-completed

questionnaires. However, our study also included a range of individually administered

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 assessments, self-report measures in addition to parent ratings and results were consistent

in demonstrating no benefit of having received either probiotic over placebo.

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Finally, there are a wide range of probiotics used in clinical trials and it is possible that

HN001 and HN019 did not show an effect where other species or strains might. Previous

animal studies indicate that infection and gut inflammation increase anxiety-like and

depressive-like behaviours. Probiotics, especially L. rhamnosus or Bifidobacterium longum,

can be used to reverse this. Depression has been strongly associated with increased

immune and inflammatory activation (27, 28). L. rhamnosus has immunological and anti-

inflammatory activity, as we have previously shown by reducing the prevalence of eczema

(16, 29, 30) and more recently gestational diabetes mellitus (31). We found that HN001

halved the risk of developing eczema, but this was not seen with HN019. The 10 clinical

trials reviewed by Romijn & Rucklidge (9) used various species of Lactobacillus.

CONCLUSIONS

We performed a randomised controlled trial of early probiotic supplementation on later

neurocognitive outcomes in childhood using a large sample size and a range of outcome

measures. We found no benefit of either probiotic over placebo. Further research into the

short and long term effects of a variety of probiotics is needed.

Acknowledgements

We are grateful to the Wellington and Auckland families who generously gave their time to

participate in the study.

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 Funding

Funding was provided by Fonterra Co-operative Group, New Zealand and Cure Kids New
Accepted Article
Zealand. EA Mitchell is supported by Cure Kids New Zealand. The funders had no role in the

study design, data collection and analysis, the decision to publish or preparation of the

manuscript.

Conflict interest

No authors have a conflict of interest.

REFERENCES

1. Borre YE, O'Keeffe GW, Clarke G, Stanton C, Dinan TG, Cryan JF. Microbiota and

neurodevelopmental windows: implications for brain disorders. Trends Mol Med 2014; 20:

509-18.

2. Clarke G, O'Mahony SM, Dinan TG, Cryan JF. Priming for health: gut microbiota acquired

in early life regulates physiology, brain and behaviour. Acta Paediatr 2014; 103: 812-9.

3. Foster JA, McVey Neufeld KA. Gut-brain axis: how the microbiome influences anxiety and

depression. Trends Neurosci 2013; 36: 305-12.

4. Sudo N, Chida Y, Aiba Y, Sonoda J, Oyama N, Yu XN, et al. Postnatal microbial

colonization programs the hypothalamic-adrenal-pituitary system for stress response in

mice. J Physiol 2004; 558: 263-75.

5. Wang H, Lee I, Braun C, Enck P. Effect of Probiotics on Central Nervous System

Functions in Animals and Humans: A Systematic Review. J Neurogastroenterol Motil 2016;

22: 589-605.

This article is protected by copyright. All rights reserved.

 6. Benton D, Williams C, Brown A. Impact of consuming a milk drink containing a probiotic

on mood and cognition. Eur J Clin Nutr 2007; 61: 355-61.

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7. Yassour M, Vatanen T, Siljander H, Hämäläinen A, Härkönen T, Ryhänen SJ, et al.

Natural history of the infant gut microbiome and impact of antibiotic treatment on bacterial

strain diversity and stability. Sci Trans Med 2016; 8: 343ra81.

8. O'Toole PW, Claesson MJ. Gut microbiota: Changes throughout the lifespan from infancy

to elderly. Int Dairy J 2010; 20: 281-91.

9. Romijn AR, Rucklidge JJ. Systematic review of evidence to support the theory of

psychobiotics. Nutr Rev 2015; 73: 675-93.

10. Partty A, Kalliomaki M, Wacklin P, Salminen S, Isolauri E. A possible link between early

probiotic intervention and the risk of neuropsychiatric disorders later in childhood: a

randomized trial. Pediatr Res 2015; 77: 823-8.

11. Hsiao EY, McBride SW, Hsien S, Sharon G, Hyde ER, McCue T, et al. Microbiota

modulate behavioral and physiological abnormalities associated with neurodevelopmental

disorders. Cell 2013; 155: 1451-63.

12. Parracho HM, Gibson GR, Knott F, Bosscher D, Kleerebezem M, McCartney AL. A

double-blind, placebo-controlled, crossover-designed probiotic feeding study in

childrendiagnosed with Autustic Spectrum Disorders. Int J Probiotics Prebiotics 2010; 5: 69.

13. Li Q, Zhou JM. The microbiota-gut-brain axis and its potential therapeutic role in autism

spectrum disorder. Neuroscience 2016; 324: 131-9.

14. Slykerman RF, Thompson J, Waldie KE, Murphy R, Wall C, Mitchell EA. Antibiotics in

the first year of life and subsequent neurocognitive outcomes. Acta Paediatr 2017; 106: 87-

94.

This article is protected by copyright. All rights reserved.

 15. Slykerman RF, Hood F, Wickens K, Thompson JMD, Barthow C, Murphy R, et al. Effect

of Lactobacillus rhamnosus HN001 in pregnancy on postpartum symptoms of depression

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and anxiety: A randomised double-blind placebo-controlled trial. EBioMedicine 2017; 24:

159-65.

16. Wickens K, Black PN, Stanley TV, Mitchell E, Fitzharris P, Tannock GW, et al. A

differential effect of 2 probiotics in the prevention of eczema and atopy: a double-blind,

randomized, placebo-controlled trial. J Allergy Clin Immunol 2008; 122: 788-94.

17. Wechsler D. Wechsler Intelligence Scale for Children Fourth Edition. San Antonio, TX:

Psychological Corporation. 2003.

18. Gioia GA, Isquith PK, Guy SC, Kenworthy L. Behavior rating inventory of executive

function. Lutz, FL: PsychologicalAssessment Resources. 2000.

19. Conners CK. Conner 3™.

20. Goodman R. The Strengths and Difficulties Questionnaire: a research note. J Chil

Psychol Psychiatry Allied Disc 1997; 38: 581-6.

21. March JS. Multidimensional Anxiety Scale for Children Second Edition. 2012.

22. Faulstich ME, Carey MP, Ruggiero L, Enyart P, Gresham F. Assessment of depression

in childhood and adolescence: an evaluation of the Center for Epidemiological Studies

Depression Scale for Children (CES-DC). Am J Psychiatry 1986; 143: 1024-7.

23. Conners CK. Conners Continuous Performance Test 3rd Edition.

24. Cambridge Neuropsychological Test Automated Battery (CANTAB).

This article is protected by copyright. All rights reserved.

 25. Scalabrin D, Harris C, Johnston WH, Berseth CL. Long-term safety assessment in

children who received hydrolyzed protein formulas with Lactobacillus rhamnosus GG: a 5-
Accepted Article
year follow-up. Eur J Pediatr 2017; 176: 217-24.

26. Kelly JR, Allen AP, Temko A, Hutch W, Kennedy PJ, Farid N, et al. Lost in translation?

The potential psychobiotic Lactobacillus rhamnosus (JB-1) fails to modulate stress or

cognitive performance in healthy male subjects. Brain Behav Immun 2017; 61: 50-9.

27. Berk M, Williams LJ, Jacka FN, O'Neil A, Pasco JA, Moylan S, et al. So depression is an

inflammatory disease, but where does the inflammation come from? BMC Med 2013; 11:

200.

28. Maes M. Depression is an inflammatory disease, but cell-mediated immune activation is

the key component of depression. Prog Neuropsychopharmacol Biol Psychiatry 2011; 35:

664-75.

29. Wickens K, Black P, Stanley TV, Mitchell E, Barthow C, Fitzharris P, et al. A protective

effect of Lactobacillus rhamnosus HN001 against eczema in the first 2 years of life persists

to age 4 years. Clin Exp Allergy 2012; 42: 1071-9.

30. Wickens K, Stanley TV, Mitchell EA, Barthow C, Fitzharris P, Purdie G, et al. Early

supplementation with Lactobacillus rhamnosus HN001 reduces eczema prevalence to 6

years: does it also reduce atopic sensitization? Clin Exp Allergy 2013; 43: 1048-57.

31. Kristin L Wickens, Christine A Barthow, Rinki Murphy, Peter R Abels, Robyn M Maude,

Peter R Stone, et al. Early pregnancy probiotic supplementation with Lactobacillus

rhamnosus HN001 may reduce the prevalence of gestational diabetes mellitus: a

randomised controlled trial. Brit J Nutr 2017; 117: 804.

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Accepted Article
Table 1. Mode of delivery, income and breastfeeding status by treatment and placebo group for the 342
participants who responded at 11 years of age.

Treatment Group

HN019 n(%) HN001 n(%) Placebo n(%)

Mode of delivery

Caesarean 57 (36.1%) 46 (29.3%) 50 (31.5%)

Non-Caesarean 101 (63.9%) 111 (70.7%) 109 (68.6%)

Breastfeeding

Yes 154 (97.5%) 153 (97.5%) 152 (95.6%)

No 4 (2.5%) 4 (2.6%) 7 (4.4%)

Income

<$30,000 5 (3.2%) 3 (1.9%) 5 (3.3%)

$30,000-$60,000 21 (13.6%) 25 (16.2%) 16 (10.5%)

>$60,000 129 (83.2%) 126 (81.8%) 132 (86.3%)

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ccepted Articl
Table 2: Cognitive, behavioural and mood outcomes at age 11 years by treatment group.

HN019 vs. HN001 vs.

HN019 HN001 Placebo placebo Placebo Overall†
WISC-IV
‡ ‡ ‡
Full Scale IQ N 104 97 97 0.17 0.17 0.31
110 (104, 107 (102,
Median (IQR) 109 (103, 116.5) 119) 116)

BRIEF Parent T-scores

Behavior Regulation Index N 118 109 115 0.008 0.051 0.023
Median (IQR) 52 (45, 61) 51 (43, 60) 48 (43, 54)
Metacognition Index N 118 109 115 0.2 0.66 0.44
Median (IQR) 52 (45, 58) 50 (44, 58) 49 (44, 57)
Global Executive Composite N 118 109 115 0.054 0.21 0.15
Median (IQR) 52 (46, 60) 51 (44, 58) 49 (44, 54)

Conners3 Parent T-scores

Inattention N 116 107 114 0.11 0.51 0.26
Median (IQR) 51 (44, 59.5) 48 (43, 57) 48 (43, 57)
Impulsivity/Hyperactivity N 116 107 114 0.36 0.23 0.46
Median (IQR) 50 (43, 64) 51 (45, 61) 51 (43, 55)
Learning Problems N 116 107 114 0.66 0.43 0.74
Median (IQR) 47 (42, 56) 47 (42, 51) 49 (42, 55)
Executive Function N 116 107 114 0.11 0.54 0.26
Median (IQR) 51 (44, 58) 49 (42, 56) 47.5 (43, 55)
Defiance/Aggression N 116 107 114 0.76 0.36 0.70
Median (IQR) 49 (43, 59) 51 (43, 59) 49 (43, 55)
Peer Relationships N 116 107 114 0.82 0.91 0.97
Median (IQR) 43 (42, 58) 43 (43, 55) 43 (43, 55)

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ccepted Articl
SDQ Parent T-score
Total Difficulties Score N 118 109 114 0.025 0.21 0.078
Median (IQR) 6 (3, 10) 6 (3, 9) 5 (2, 8)

MASC2 Parent T-score

Total anxiety score N 117 108 114 0.056 0.47 0.18
Median (IQR) 50 (44, 55) 49 (42, 57) 47 (43, 53)

MASC2 Self Report T-score

‡ ‡ ‡
Total anxiety score N 114 106 113 0.11 0.57 0.29
Median (IQR) 51 (45, 58) 49 (42, 56) 48 (42, 56)

CES-DC
Total score N 116 107 113 0.006 0.095 0.023
Median (IQR) 11 (7, 18) 10 (6, 16) 8 (5, 13)

CPT3 T-scores
‡ ‡ ‡
Detectability N 104 96 97 0.46 0.47 0.70
Median (IQR) 56 (48, 61) 56 (49, 61) 57 (49, 63)
Omissions N 104 96 97 0.74 0.23 0.42
Median (IQR) 52 (45, 65) 49 (45, 58) 50 (46, 66)
‡ ‡ ‡
Commissions N 104 96 97 0.33 0.85 0.59
Median (IQR) 52 (47, 57) 53 (47, 59) 54 (48, 59)
Perseverations N 104 96 97 0.98 0.7 0.89
Median (IQR) 50 (44, 67) 50 (47, 62) 50 (46, 63)
HIT reaction time N 104 96 97 0.64 0.83 0.73
Median (IQR) 44 (38, 50) 42 (38, 47) 42 (37, 50)
HIR reaction times standard deviation N 104 96 97 0.38 0.3 0.54
Median (IQR) 55 (49, 64) 56 (50, 64) 53 (48, 61)

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ccepted Articl
Variability N
Median (IQR)
102
50 (45, 59)
94
53 (45, 63)
93
49 (45, 61)
0.93

‡
0.45

‡
0.66

‡
Hit reaction time block change N 104 95 96 0.88 0.81 0.97
Median (IQR) 52 (45, 59) 52 (45, 59) 50 (43, 60)
‡ ‡ ‡
Hit reaction time inter-stimulus-interval change N 104 96 97 0.22 0.095 0.26
Median (IQR) 54 (46, 59) 54 (46, 60) 50 (43, 58)

CANTAB
‡ ‡ ‡
AST contingency cost N 102 95 89 0.98 0.62 0.61
Median (IQR) 45 (10, 84) 47 (18, 68) 44 (17, 74)
‡ ‡ ‡
AST switching cost N 102 95 89 0.80 0.98 0.66
224 (115, 202 (131,
Median (IQR) 220 (139, 296) 339) 327)
AST response latency N 102 95 89 0.96 0.41 0.68
579 (518, 580 (478,
Median (IQR) 576 (512, 653) 691) 655)
OTS problems solved in first trial N 102 95 89 0.58 0.97 0.84
Median (IQR) 11 (9, 12) 11 (9, 12) 11 (9, 12)
SWM strategy score N 102 95 89 0.6 0.98 0.83
Median (IQR) 29 (26, 31) 29 (23, 31) 28 (25, 31)
†
ANOVA/two-sample t-test or Kruskal-Wallis test/Wilcoxon rank sum test used
‡
Parametric test

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Accepted Article

This article is protected by copyright. All rights reserved.