MY SLIDES ARE AVAILABLE THROUGH TODAYS ......My day job is starting companies for infectious disease...
Transcript of MY SLIDES ARE AVAILABLE THROUGH TODAYS ......My day job is starting companies for infectious disease...
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Herewith declared conflict of interest as a
non-exclusive and independent expert
advisor to numerous industrial, NGO,
private and public academic instutions and
(EU) consortia (over 160 clients).
MY SLIDES ARE AVAILABLE THROUGH TODAYS ORGANISERS
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BLOCKCHAIN-FACILITATED SHARING TO ADVANCE OUTBREAK R&D
van der Waal, M. B., Ribeiro, C. D. S., Ma, M., Haringhuizen, G. B., Claassen, E., & van de Burgwal, L. H. (2020). Science, 368(6492), 719-721.
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My day job is
starting
companies for
infectious
disease
diagnostics and
vaccines in a
One Health
setting mainly
for zoonoses
(animal to man)
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Ik werk sinds 1980 aan de immunologie van
slijmvliezen van de darm
en sinds 1987 aan melkzuur bacterieenGerritse, Posno, Schellekens, Boersma & Claassen E. (1990) Oral administration of TNP-lactobacillus conjugates in mice:
a model for evaluation of mucosal and systemic immune responses and memory formation elicited by transformed
Lactobacilli. Res. Microbiology, 141, 955-962.
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Andrew Maltez Thomas & Nicola Segata BMC Biology 17: 48 (2019)
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De putrefactie van de dikke darm
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Het humane microbioom heeft 1-10 maal
zoveel bacteriën als lichaamscellen!
• Er leven ongeveer 1014
(honderd-duizend-miljard!) micro-organismen in de mens
• Dat is 1-1½ kilogram per gezonde volwassene
• 100x meer bacteriële genen in vergelijking met humaan genoom
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Nieuwboer, Claassen et al 2019, in druk
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10 100 1000 100.000 100.000.000
3-4
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When children are born, they emerge fromthe relatively sterile environment of the uterusinto a world teeming with bacteria . . .
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Stunted microbiota and opportunistic pathogen colonization in caesarean-section birth
Yan Shao, Samuel C. Forster, Evdokia Tsaliki, Kevin Vervier, Angela Strang, Nandi Simpson, Nitin Kumar, Mark D. Stares, Alison Rodger, Peter Brocklehurst, Nigel Field & Trevor D. Lawley Nature (2019)
KEIZERSNEDE GEEFT MICROBIOTA ACHTERSTAND
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Stunted microbiota and opportunistic pathogen colonization in caesarean-section birth
Yan Shao, Samuel C. Forster, Evdokia Tsaliki, Kevin Vervier, Angela Strang, Nandi Simpson, Nitin Kumar, Mark D. Stares, Alison Rodger, Peter Brocklehurst, Nigel Field & Trevor D. Lawley Nature (2019)
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Mouth
105 – 107
polymorph germs stomach
102 – 103
Streptococcus
duodenum
< 104-5
Streptococcus
jenunum
105 – 106
Streptococcus
ileum
103 – 107
Streptococcus
Bacteroides
colon
109 – 1011
Bacteroides
Clostridum
Streptococcus
Bifidobacteria
Enterobacteria
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Darmvlokken (villi) geven oppervlakte vergroting
• De totale oppervlakte van de ‘uitgespreide darm’ komt daarmee op meer dan 300 vierkante meter
• Dat is een tennisveld
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Hier zit uw
BROODJE met uw
levende bacteriële
gasten!!
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Bacterieen moeten aanhechten aan darm eptiheel om te koloniseren: competitie
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MELKZUUR EN WATERSTOF PEROXIDE
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melkzuurbacteriën
melkzuur
azijnzuur
mierenzuur
(boterzuur indirect)
ethanol
diactyl
acetaldehyde
waterstofperoxide
zuurstofkatabolieten
suikerkatabolieten
koolstofdioxide
caproïnezuur
3-hydroxy vetzuren
vet- en aminozuurmetabolieten
eiwitachtige stoffen
phenyl-melkzuur
4-hydroxy-phenyl melkzuur
aromaten
heterocyclische molekulen
Reuterine
Reuteri-cycline
kleine peptiden
cyclische di-peptiden
anti-fungale-peptiden
bacteriocinen
bacteriocine-achtige molekulen
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Darmwand en gezond epitheel is een barriere
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Afbraak van de barriere functie -> opportunisten & ontsteking
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Lactobacilli versterken tight junctions
Occludin
Claudin 1
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Knol et al., 2002
0
10
20
30
40
50
60
70
80
study start after 4 weeks after 6 weeks
% B
ifid
ob
ac
teri
a o
f to
tal fa
ec
al
ba
cte
ria
(m
ea
n ±
SE
)
Human milk (n = 16)
Control (n=18)
Clinical Studies: Effects of the Prebiotic
GOS/FOS Mixture in Infants 4-12 Weeks
- Bifidogenicity -
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Knol et al., 2002
0
10
20
30
40
50
60
70
80
study start after 4 weeks after 6 weeks
% B
ifid
ob
ac
teri
a o
f to
tal fa
ec
al
ba
cte
ria
(m
ea
n ±
SE
)
Human milk (n = 16)
GOS/FOS (n=16)Control (n=18)
*
p<0.05, 6 wk vs. study start,
# GOS/FOS vs. control
Clinical Studies: Effects of the Prebiotic
GOS/FOS Mixture in Infants 4-12 Weeks
- Bifidogenicity -
#
*
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0
1
2
3
Control GOS/FOS
Acinetobacter
Klebsiella
Bacillus subtilis
Streptococcus haemolyticus
Enterobacter
Clostridium dificile
Proteus
Pseudomonas aeruginosa
Staphylococcus aureus
Staphylococcus haemolyticus
Staphylococcus epididermidis
% o
f to
tal
mic
rob
ial
cou
nts
(m
ean
)
Difference group
Mann-Whitney U-test:
Sum of all pathogens:
* p=0.01 vs. Control
*
Boehm et al., JPGN, 2003Slide from Yvan van den Plas
Reduction of pathogens in the gut after GOS&FOS
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% w/w (vers)
Witlof / Cichorei 15-20Knoflook 9 - 16Schorseneer 4 - 10Prei 3 - 10Ui 2 - 6Artisjok 2 - 6Asperge 2 - 3Tarwemeel 1 – 4Borstvoeding Mens 0.5 - 1.5Haver 0.5 - 1.5Banaan 0.3 - 0.7Zoogdier melk 0.01 - 0.1
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Puntarelle is soort roodlof
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From Holland witlof
designed © by slappo
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39
Directly linked to gut dysbiosis• Inflammatory Bowel Disease (IBD)
• Irritable Bowel Syndrome (IBS)
• Diarrhea
• Antibiotic associated diarrhea
• Travelers diarrhea
• Infantile diarrhea (Rotavirus)
• Caused by food pathogens
• Constipation
• Helicobacter pylori infections
Indirectly linked to gut dysbiosis• Allergies
• Cancer
• Cardiovascular disease
• Urinary tract infections
• Bacterial vaginosis
• Upper respiratory tract infections
Disturbance microbiota (=dysbiosis)
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The human microbiota
In health and disease
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.
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Fecal transplantation NOT SAFE but “Poopular”
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VALIDITY
BIAS
Meta-analysisof RCTs
Randomised controlled trial (RCT)
Observational studies(case-control, cohort)
Observational studies(case report, case series)
Experimental studies
Hierarchy of evidenceFor questions about the effectiveness of an intervention
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The forest plot
D’Souza AL. et al. BMJ 2002; 324: 361-4
LEFT
LESS EFFECT
RIGHT
MORE EFFECT
Line of no effect
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The overall evidence suggests a moderate protective effect of probiotics for preventing
AAD (NNTB 9, 95% CI 7 to 13). Using five criteria to evaluate the credibility of the
subgroup analysis on probiotic dose, the results indicate the subgroup effect based on
high dose probiotics (≥ 5 billion CFUs per day) was credible. Based on high-dose
probiotics, the NNTB to prevent one case of diarrhea is 6 (95% CI 5 to 9). The overall
certainty of the evidence for the primary endpoint, incidence of AAD, based on high dose
probiotics was moderate due to the minor issues with risk of bias and inconsistency
related to a diversity of probiotic agents used. Evidence also suggests that probiotics may
moderately reduce the duration of diarrhea, a reduction by almost one day. The benefit of
high dose probiotics (e.g. Lactobacillus rhamnosus or Saccharomyces boulardii) needs to
be confirmed by a large well-designed multi-centered randomized trial. It is premature to
draw firm conclusions about the efficacy and safety of 'other' probiotic agents as an
adjunct to antibiotics in children. Adverse event rates were low and no serious adverse
events were attributable to probiotics.
Guo Q, Goldenberg JZ, Humphrey C, El Dib R, Johnston BC. Probiotics for the prevention of pediatric antibiotic-
associated diarrhea. Cochrane Database of Systematic Reviews 2019
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Examples Atopic / Constitutional Eczema
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Epidemiologic Associations between Antibiotic Use
and Allergies; 14 STUDIES AND OVER 90
THOUSANDS PERSONS
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Year Designa Number b Residence Age Disease c Diagnosis d Sample Assaye Negative g Positive h Reference
29
31
32
32
28
microbiol Bifidobacterium Staphylococcus
21
30
22
23
24
25
26
Table 1. The Epidemiological Association Between Allergies and Microbiota Composition
gBacteria (or markers) are negatively associated with allergic subjects or positively associated with healthy control subjects
2003 C-C 30-68 Japan < 20 yr AD clinical feces
Aerobes, Coliforms, S.
aureus , BacteroidesfecesAD (FA)2 yrC-C1999
AllergiesSubjects
clinical and
SPT
Estonia +
Sweden27-35
propionic, butyric, i-butyric, i-
valeric, and valeric FAmolecular
microbiol
Gram-positive species
within aerobes, S.viridans ,
Clostridia j
Lactobacilli, Bifidobacteria,
CONS, Anaerobes,
Bacteroides
fecesclinicalAD and FA2-7 mo
C-C2000 fecesAD, asthma, FA12 moclinical and
SPTSweden25-47
fecesclinical and/or
SPTAD (FA)0-2 yr
Estonia +
Sweden44 (18)Pro2001
NR4-6C-C2001
i-caproic FAi
microbiol,
molecular,
genetic
Similarity (%) of bacterial FA
profileClostridia
Enterococci, Bifidobacteria,
Bacteroides
S. aureus , Lactobacilli,
Clostridiamicrobiol
Bifidobacterium
adolescentisBifidobacterium bifidummicrobiol
fecesSPT (clinical)atopy (+/-
AD/FA)0-12 moFinland76 (22)Pro (FH)2001
27-10 Lactobacilli/enterococci jmicrobiol,
geneticfeces
clinical and
SPT
NR
AD (FA)0-14 moFinland
12 mo
C-C2001
Bifidobacterium
adolescentisBifidobacterium bifidummicrobiolfeces
clinical and
SPTAD2-7 mo
C-C2002
7-6C-C2001
(Bifidobacteria) k
UK10-10 C. difficile-specific IgGmolecularbloodclinical and
SPTatopic wheeze
atopic wheeze
(AD)k
clinical and
SPTfeces genetic2005 C-C UK 3-5 yr
33-33 (8-
8)k
2005 fecesclinical and
SPT and/or IgE
AD, asthma,
allergic rhinitis5 yrEstonia
AD
bFor C-C, 'number of cases-number of controls'; for Pro, 'total subjects (allergic subjects)'
microbiol ClostridiaBifidobacteria
Lactic acid bacteria,
enterococci
Bifidobacterium spp.,
Clostridium spp.
microbiol,
genetic
19-19C-C
2006
Microflora Associations with Allergic Disease fExperiments
aC-C = Case-Control; Pro = Prospective; FH = inclusion based on Family History of allergy
~3 yrSingapore21-28C-C clinical feces
kWithin the study of 33 pairs of atopic, wheezy cases and controls, only the subset of 8 atopic dermatitis cases exhibited significant differences with their controls
cAD = atopic dermatitis or eczema; FA = food allergy; (FA) = food allergens included in skin-prick test
dSPT = positive skin-prick test; IgE measured by ELISA
jAssociations made between disease severity and bacterial counts
hBacteria (or markers) are positively associated with allergic subjects or negatively associated with healthy control subjects
emicrobiol = various microbiological culture techniques
ii-caproic fatty acid is "associated with the presence of C. difficile"
fSignificant associations (p<0.05) listed between allergic disease and microflora, generally in terms of microbial counts and/or the frequency of colonizaition with microbe (associations made with
characteristic bacterial fatty acids (FA), or with IgG antibodies specific to bacteria as indicated)
Epidemiologic Associations between Fecal
Microbiota Composition and Allergies
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Identification and antibiotic resistance of isolates from probiotic productsT. Temmerman. Ghent, Belgium
55 European probiotic products 25 dairy products
20 dried food supplements
Antiobiotic resistance
antibiotic % resistance
Kanamycin 79 %
Vancomycin 65 %
Tetracycline 26 %
Pencillin G 23 %
Erythromycin 16 %
Chloramphenicol 11 %
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Examples Atopic / Constitutional Eczema
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Hygiene hypothesis: relevant microbial
exposures?
• Childhood infections (Hep A?): NO
• Domestic hygiene (bathing, soaps,
detergents, antibacterial kitchen cutting
boards etc.): NO
• “Old Friends” (helminths, mites, saprophytic
mycobacteria, lactobacilli): YES?
• Slide stolen from prof. van Eden
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With increased food safety our food has also became more and more sterile
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Xenia Warrior Princess
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Friedman & Zeiger (2005) J Allergy Clin Immunol 115: 1238-48
Breastfeeding reduces chance on allergy
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Allergisch meisje of?
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Allergisch voor meisjes?
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Rautava S. J. Allergy Clin Immunol 2002: 109 pp 119-121
& reviews by E. Isolauri
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Now in its 15th year the probiotic
use by pregnant mothers +
breastfeeding study still shows a
45% reduction in children’s clinical
manifestations of IgE+ve allergy
over placebo
Breastfeeding remains key!!!!!!
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Yazdanbakhsh et al. 2002
The Africa Paradox and the hygiënehypothesis
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‘Old friends trigger regulation through innate dendritic cell receptors’
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Scout senses
no danger
(sees “old
friends”): info to
officer “steady
state of
tolerance”
“Hold your fire”
Allergy: Scout
senses danger &
no old friends:
info to officer and
command to fire
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The influence on metabolism
• This explains the differences in metabolic mechanisms• These insights are a base to understand the Mechanism Of Action (MOA) of future probiotic
treatment• Probiotic microorganism might manipulate these mechanisms
– Reducing effect on weight and weight gain
Probiotics & Obesity Marcel Mekkes © 1-6-2012 74
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HARA HACHI BU
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78
Darmmicrobiota bepalen gedrag: Angst zit in je darm en niet in je brein!
Cell 155:1451-1463; 2013
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Gezonde studenten: probiotica gedurende weken →minder gevoelig voor negatieve gedachten typisch voor depressie→ ondersteunende/preventieve therapie bij PDS?
CXCR
IL-RBCR
TCR PRRImmune receptors
Endocrine receptors
ProlactineGrowth
hormone
Cortisol Sex
hormones
OpioidSAdrenalin
Melatonin
cytokines
hormones
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a) Gastrointestinal
symptom rating scale
b) Parent global
impressions-III
c) Childhood autism
rating scale
d) Social
responsiveness scale
e) Aberrant behavior
checklist
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The history of probiotics: the untold story.
Ozen M, Dinleyici EC
Probiotic, a word derived from Latin, means 'for life'. A long time before
the awareness of probiotic microorganisms, fermented products, such as
beer, bread, wine, kefir, kumis and cheese had been very frequently used
for nutritional and therapeutic purposes.
Yoghurt is most likely resulted from a fermentation process within the
animal skin bags used for transportation of water and milk in regions
with low humidity and high temperatures (Middle Asia and Middle East).
The history of probiotics goes parallel with the evolution of human race
and, thanks to the sophisticated techniques at the moment, can be traced
back to the ancient times, nearly 10,000 years ago.
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Cheese
Salami and pickles
cured ham
Yogurt
Sauerkraut
Vinegar
Beer
Sourdough
bread
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ADJUVANT
• From Latin “adjuvare” meaning to support
or to help.
• In the immunesystem this help is non-
specific, so “general help” a sort of “code
red” alarm
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Strain dependent intrinsic adjuvanticity
0 10 20 30 40
***
**
NaHCO3
L. murines
L. casei
L. gasseri
L. plant. 14917
L. plant. NCIB
L. fermentum
L. brevis
L. reuteri
Anti-TNP (hapten) respons in arbitraire eenheden
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concept of probiotic in medicine
“biotherapy”
1935
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DEALING WITH THE REMAINING
CONTROVERSIES OF PROBIOTIC
SAFETY
M. van den Nieuwboer and E. Claassen. Beneficial Microbes
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PROBIOTICS REDUCE COMPLICATIONS
5
- Confidential -
REDUCED COMPLICATIONS - Probiotic intake is associated
with a reduction in symptoms and AEs compared to the
placebo. Examples are:
✓ In the prevention of Clostridium difficile associated diarrhoea, probiotics reduce the
risk of AEs by 17%
✓ Probiotic intake for preventing necrotizing enterocolitis is associated with fewer
infections and gastrointestinal symptoms and 50% reduced mortality rate
✓ Lower rates of respiratory and gastrointestinal infections in healthy infants as a
result of probiotic intake
✓ Reduced incidence of catheter related bloodstream infections in mechanically
ventilated individuals
✓ Lower incidence of complications after surgery with probiotic intake.
▪ It is difficult for an investigator to discriminate whether
an AE is a result of the patient condition or from the
investigated product.
▪ Effects of probiotics can be widespread, exerting a subtle
positive influence extending far beyond the gut
▪ Experienced AEs in placebo and probiotic groups are
often a result of the health condition of the individual
Due to heterogeneity and publication bias, the evidence for this reduction in adverse
events is low. In order to make these general conclusions, additional data is required.
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Health Maintenance of
Athletes with Probiotics
Marta Oliveira
School of Sport, Exercise & Health Sciences
Loughborough University, UK
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94
Percentage change in saliva IgA
-40
-20
0
20
40
60
80
Before
training
After 3
weeks
training
After
combat
course
After 1 week
recovery
PRO
PLA
▪ 47 army cadets
▪ Daily 100 ml L. casei DN-114 001 probiotic drink or placebo for 3 weeks training, followed by a 5-day combat course
▪ No difference in incidence of respiratory illness
▪ Significant saliva IgA decrease only in placebo group
EFFECT OF A PROBIOTIC ON RESPIRATORY INFECTIONS AND SALIVA IGA DURING INTENSE MILITARY TRAINING
Tiollier et al 2007 Military Med 172: 1006-1011
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• ‘Healthy’ Shift-workers Probiotics and resistance to infections
•Gastro-intestinal and respiratory
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• Groente en fruit zijn de basis van dagelijkse voeding
• Verwen je darm eet witlof, asperge, artisjok, ui, knoflook, prei
• Eet gefermenteerde producten
• Eet elke dag een handje ongezouten noten
• Eet ten minste 2x per week vette vis
• Eet matig vlees (gevogelte is beter dan rood) en altijd met groene groente
• Let op en beperk je suiker inname waar je maar kunt
• Drink water/koffie/thee in plaats van fris- of fruitdrank
• Wees zuinig met zetmeel (brood, pasta, rijst, aardappelen)
• Gebruik liever altijd volkoren graanproducten (met mate)
• Gebruik volvette melkproducten, vooral yoghurt, kwark en kaas
• Vermijd industrieel geproduceerde voedingsmiddelen
• Gebruik olijfolie als dressing en om te braden
Voedingsadvies
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Disclaimer for use and copyright
THIS SERIES OF SLIDES IS FOR
PERSONAL USE ONLY. ANY OTHER
APPLICATION MIGHT INFRINGE
ON COPYRIGHTS or other rights
of third parties (other than
myself). DO NOT COPY UNLESS
PERMITTED.
tel: +31 6 20443098
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http://www.biomaatschappij.nl/product/ons-microbioom/
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Phylogeny; ‘a reminder….’Human phylogeny & namegiving
PHYLUM CLASS ORDER FAMILY GENUS SPECIES
Slide stolen from prof dr Kleerebezem
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Phylogeny; ‘a reminder….’Automobile phylogeny & namegiving ☺
PHYLUM CLASS ORDER FAMILY GENUS SPECIES
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1980
2018
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Bacterial phylogeny; ‘a reminder….’The yoghurt workhorse S. thermophilus
PHYLUM CLASS ORDER FAMILY GENUS SPECIES
(OTU)STRAIN
N=>>26
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FROM 0 TO 1
MILLION CFU ARE
TYPICALLY LOW
DOSE
SUPERMARKET
PRODUCTS WITH
NO CLINICAL
DATA
WHATSOEVER
![Page 109: MY SLIDES ARE AVAILABLE THROUGH TODAYS ......My day job is starting companies for infectious disease diagnostics and vaccines in a One Health setting mainly for zoonoses (animal to](https://reader034.fdocuments.us/reader034/viewer/2022051901/5ff03e7639100674ae0ab9e8/html5/thumbnails/109.jpg)
FROM 0 TO 1
MILLION CFU ARE
TYPICALLY LOW
DOSE
SUPERMARKET
PRODUCTS WITH
NO CLINICAL
DATA
WHATSOEVER
FROM 1 MILLION
TO 10 BILLION CFU
ARE TYPICALLY
ARTISANAL OR
HOME MADE HIGH
DOSE PRODUCTS
WITH NO CLINICAL
DATA
WHATSOEVER
![Page 110: MY SLIDES ARE AVAILABLE THROUGH TODAYS ......My day job is starting companies for infectious disease diagnostics and vaccines in a One Health setting mainly for zoonoses (animal to](https://reader034.fdocuments.us/reader034/viewer/2022051901/5ff03e7639100674ae0ab9e8/html5/thumbnails/110.jpg)
NOTABLE EXCEPTIONS HIGH DOSE CLINICALLY TESTED SUPERMARKET PRODUCTS