Dr. Guus Roeselers Nutricia Research, Gut Microbiology research group

THE MICROBIAL MATERNAL

HERITAGE:

THE JOURNEY OF BACTERIA IN EARLY LIFE

AND INFLUENCE ON MASTITIS AND ALLERGY

THE HUMAN MICROBIOME

An average adult has ~1.5 kilograms of gut bacteria

1013 cells per gram of colonic content

Microorganisms are an integrated part of our physiology

Ottman et al.(2012) Front Cell Infect Microbiol. 2:104

HUMAN GUT MICROBIOTA DIVERSITY

BACTERIA AND HOST INTERACTIONS

AT THE GUT EPITHELIAL BARRIER

The gut epithelium covers over 250 square

meters. Enough to cover a tennis court!

The gut epithelium separates us from the

microbiota but also interacts with it and thus

affects host immune status and physiology

‒ Barier function

‒ Adsorption

‒ Innate defense

‒ Mucus production

‒ Hormone secretion

CROSS TALK BETWEEN MICROBIOTA

AND IMMUNE SYSTEM

The lamina propria is connective tissue

constituted by stromal cells, blood vessels,

nerves, and immune cells.

Macrophages and dendritic cells are

strategically located next to the epithelial

layer

Sampling luminal antigens and orchestrating

the innate and adaptive immune response

Tolerance mechanisms prevent

reactions against the microbiota and

food antigens

A FRONTIER FOR PHARMACEUTICAL & NUTRITIONAL

DISCOVERY

Roeselers et al., 2012 Pharmacological Research

WHAT IS CAUSE & WHAT IS EFFECT?

How is the gut microbiota assembled?

Is microbiota assembly a deterministic or stochastic process?

What are the effects of host genotype, physiology, and environment?

How can we tell what aspects of this tremendous biodiversity matters?

How much is our microbiota/microbiome evolving as a reflection of our changing

lifestyles/biosphere?

What is therapeutic potential of the microbiota/microbiome?

How and when can/should we intentionally manipulate this system?

SOME ECOLOGICAL QUESTIONS

WHERE DOES THE MICROBIOTA COME FROM?

We are born sterile!

Colonization of the mammalian gut with microbes begins

immediately at birth.

Upon passage through the birth canal, infants are exposed to a

complex microbial population (Redondo-Lopez et al., 1990 Rev Infect Dis

12: 856–872)

Infants delivered through c- section have different microbiota

compared with vaginally delivered infants (Huurre et al., 2008

Neonatology 93: 236–240).

Acquisition of the microbiome in early life by vertical transmission,

and factors modifying mother-to-child microbial transmission.

1000 DAYS

Gonzalez et al 2011 EMBO reports

CESAREAN SECTION

05101520253035404550

Chad

Ethiopia

SierraLeone

Guinea

CentralAfricanRepublic

Tajikistan

Nepal

Hai

UnitedRepublicofTanzania

Liberia

Turkmenistan

Zim

babwe

Lesotho

Gabon

Algeria

Coted′Lvoire

Ghana

Azerbaijan

Tunisia

Philippines

UnitedArabEmirates

Kazakhstan

Guatemala

Honduras

Netherlands

Ukraine

Iceland

Qatar

Poland

Serbia

Sweden

Thailand

Panama

Jordan

France

Estonia

NewZealand

Lithuania

CostaRica

Denmark

Latvia

Romania

Peruc

Venezuela

China

Ireland

Colombia

Austria

Germ

any

Switzerland

Australia

Chile

Malta

Argen

na

Mexico

DominicanRepublic

Brazil

Cesareansec ondeliveryrate(%)

Cesareansec ondeliveryrate(%)

Dominguez-Bello et al. (2010) PNAS 11971–11975

Roduit C et al. Thorax 2009;64:107-113

*

CESAREAN SECTION

od

ds r

atio

(O

R)

for

asth

ma

at

8 y

ea

rs

Dicksved et al., ISME Journal 2008

N ~40 P < 10-5

Zoetendal et al., Microbial Ecology in Health & Disease 2001

Monozygotic twins have similar microbiota

HUMAN GENETICS

Breast-fed baby

0

10

20

30

40

50

60

70

80

90

100

0 5 10 15 20 25

Days

%

0

10

20

30

40

50

60

70

80

90

100

0 5 10 15 20 25

%

Days

Formula-fed baby

Breast-fed babies develop a

microbiota often dominated

by Bifidobacteria.

Formula-fed babies develop a

more “adult-like” microbiota.

Harmsen et al, 2000.JPGN 30:61-67

Human milk

EARLY LIFE NUTRITION

DIRECTS MICROBIOTA DEVELOPMENT

Formula (cow milk)

Bifidobacteria

Bacteroides

E. coli

DEVELOPMENT OF THE INTESTINAL MICROBIOTA

Birth 4 days 4-6 months 20 days Prepartal

Mode of delivery

Inoculation with maternal/

environmental microbiota

Genetics

Sterile?

Weaning

transition

to adult

Formula Human

milk

Dietary influence

Antibiotics

Stimulation of specific Bifidobacterium and Lactobacillus species;

Reduction in pathogenic bacteria.

Relief of gastrointestinal symptoms

Reduction of digestive problems

Nutricia developed an innovative concept of adding prebiotics

oligosaccharides to infant milks, in order to mimic the composition of

oligosaccharides in breast milk.

GALACTO- AND FRUCTO-OLIGOSACCHARIDES

AND GUT HEALTH

log

10

of

CF

U/g

we

t fa

ec

es

(m

ed

ian

, IQ

R)

3

0.8

(n=27)

0

(n=33)

0.4

(n=30)

10

0

1

2

4

5

6

7

8

9

Breast-fed infants

Lactobacillus

group difference:

p<0.05 *vs. 0

* *

Bifidobacterium

* *

group difference:

p<0.05

*vs. 0, # vs. 0.4

#

Supplementation levels of a unique mixture of Prebiotics

[g/100ml]

0.8

(n=27)

0

(n=33)

0.4

(n=30)

Breast-fed infants

5

12

2

3

4

6

7

8

9

10

11

log

10

of

CF

U/g

we

t fa

ec

es

(m

ed

ian

, IQ

R)

FORMULA WITH GOS/FOS:

MORE LIKE BREAST-FED INFANTS

Dose-dependent bifidogenic effect after feeding a scGOS/lcFOS

supplemented infant formula

Moro et al. (2002) J Pediatr Gastroenterol Nutr. 34:291-5.

SCGOS/LCFOS & MICROBIOTA METABOLISM

18

SCFA patterns similar to that of breast milk

sIgAP< 0.001

Control scGOS/lcFOS Breastfed0

250

500

750

22

24

31

Bakker-Zierikzee et al (2006) Pediatr Allergy Immunol. 17:134-40.

HIGHER FAECAL SC-IGA SECRETION IN

INFANTS FED ON PREBIOTIC INFANT

FORMULA

The origin of the bacteria present in breast milk has become a

controversial issue in the last years.

A possible route may include the establishment of the mothers' skin

microbiota and infants' oral microbiota into the mammary gland

Also Bacteria are postulated to translocate from the mothers'

intestine to the mammary gland via the mesenteric lymph nodes

BACTERIAL DIVERSITY OF HUMAN MILK

Mastitis is an inflammation (infectious or non-infectious) of the breast tissue

Staphylcoccus aureus is the most common etiological organism responsible,

but S. epidermidis and streptococcci are occasionally isolated as well.

Early stages of mastitis can present with local pain, redness, swelling, and

warmth, later stages also present with systemic symptoms like fever and flu-like

symptoms and in rare cases an abscess can develop.

WHAT IS MASTITIS?

Science supporting probiotics in Mastitis treatment

Significant reduction of bacterial taxa counts in breast milk after the probiotic treatment

A) daily 9 log10 L. fermentum CECT 5716 (n=127)

B) daily 9 log10 L. salivarius CECT 5713 (n=124)

C) Prescribed antibiotics (n=101)

TREATMENT OF INFECTIOUS MASTITIS DURING

LACTATION: ANTIBIOTICS VERSUS PROBIOTICS

Arroyo et al., (2010) Clin Infect Dis.50 12: 1551-1558.

352 women with mild infectious mastitis

….with a significant reduction in pain score

Breast pain score from 0 (most painful) to 10 (no pain)

Antibiotic group

At day 21, 27 women reported an intense pain (score 0–4),

45 women improved but still reported discomfort for

breastfeeding (5–7), and only 29 women recovered

completely (8–10)

TREATMENT OF INFECTIOUS MASTITIS DURING

LACTATION: ANTIBIOTICS VERSUS PROBIOTICS

A) L. fermentum CECT 5716 (n=127)

B) L. salivarius CECT 5713 (n=124)

C) Antibiotics (n=101)

Arroyo et al., (2010) Clin Infect Dis.50 12: 1551-1558.

IN SUMMARY

Microbial colonization in the first 1000 days of life may have a big influence on metabolic,

immune, and gut health

Following birth, the human intestine is rapidly colonized

Colonization is influenced by gestational age, mode of delivery (vaginal birth vs. c-section),

diet (breast milk vs. formula), sanitation, and antibiotics

Breastfed infants typically have a microbiota dominated by Bifidobacterium sp

Formula-fed infants may benefit from infant formulas containing prebiotic oligosacharides

Recent studies have shown that breast milk represents a continuous supply of bacteria to

the infant gut

Mammary dysbiosis may lead to mastitis, a condition that represents the first medical cause

for undesired weaning

The use of probiotic L. fermentum CECT5716 or L. salivarius CECT5713 appears an

efficient alternative to antibiotics for the treatment of infectious mastitis during lactation.