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Yassin M. Alsaleh
Supervised by:
Dr.Sameer Al Abdi
Lactobacillus journal club
Lactobacillus journal club
Introduction
• Infantile colic: paroxysmal, excessive
crying without an identifiable cause in an
otherwise healthy newborn infant.
Infantile colic
• Wessel’s criteria :
• The classic definition is
based on the rule of threes:
• crying that lasts for ≥3 hrs
per day.
• for ≥ 3 days per week.
• for a minimum of 3 weeks.
Infantile colic
• It usually occurs in infants < 3 months.
• Colic affects 3% to 28% of infants.
• causing considerable stress and concern
for parents.
Infantile colic
• The etiology:
• swallowed air
• Overfeeding .
• some foods.
• The role of an aberrant intestinal
microflora .
• Increased presence of hydrogen gas
produced by anaerobic Gram-negative
bacteria.

• Recently, coliform bacteria, particularly
Escherichia coli, were found to be more
abundant in the feces of colicky infants.
Infantile colic
• Treatments:
• no treatment consistently provides
satisfactory relief.

• Carminatives before feedings are
ineffective in preventing the attacks.
Old
treatments
Carminatives
Probiotics
• Probiotics are live microorganisms
thought to be healthy for the host
organism.
• Probiotics are commonly consumed as
part of fermented foods with specially
added active live cultures; such as in
yogurt, or as dietary supplements.
Probiotics
• Mechanism of action:
• improving intestinal microbial balance,
thus inhibiting pathogens and toxin
producing bacteria.
•
•
•
•
•

uses for probiotics

Managing lactose intolerance
Prevention of colon cancer
Lowering cholesterol
Lowering blood pressure
Improving immune function and
preventing infections
• Antibiotic-associated diarrhea
• Irritable bowel syndrome and colitis
• constipation.
• in a prospective randomized study,
supplementation with the probiotic
Lactobacillus reuteri ATCC 55 730
improved colicky symptoms in breastfed
infants within 1 week of treatment
compared with treatment with simethicone.
OBJECTIVES
• To test the efficacy of Lactobacillus reuteri
on infantile colic .
• to evaluate its relationship to the gut
microbiota.
Lactobacillus journal club
METHODS
• This randomized, controlled, double blind
Study.
• In general pediatricians and OPD at the
Department of Pediatrics, University of
Turin (Regina Margherita Children
Hospital)
METHODS
• between March 2008 and August 2009.
• All infants were diagnosed according to
the modified Wessel’s criteria.
• All were born at term, AGA and aged 2 to
16 weeks at recruitment.
Inclusion criteria
• Only exclusively breastfed infants were
enrolled.
• At enrollment, mothers were encouraged
to avoid cow’s milk in their diet.
Exclusion criteria
• clinical evidence of chronic illness or GIT
disorders.
• any intake of probiotics and/or antibiotics
in the week preceding recruitment.
• any formula-feeding.
• no acid blockers were used in any of the
infants who completed this study.
outcomes
• Primary outcome was defined as a reduction of average
crying time to < 3 hours a day on day 21.
• Secondary outcome was defined as the number of
responders in each group on days 7, 14, and 21.
• Responders were those who experienced a decrease in
the daily average crying time of 50% from baseline.
METHODS
• In addition, the intestinal microflora of the
infants was analyzed to determine the
effect of the by using (FISH).
Lactobacillus journal club
METHODS
• Colicky infants were randomly assigned to
groups by using a computer generated
randomization list created by an
independent departmental statistician.
• written informed consent was obtained
from parents .
METHODS
• Active study product consisted of a
suspension of freeze-dried L reuteri DSM
17 938 in a mixture of sunflower oil and
medium-chain triglyceride oil .
• The placebo was identical in appearance
and taste but without the live bacteria.
METHODS
• Both formulations were administered in 5
drops, once a day, 30 minutes before the
feed in the morning, for 21 days.
METHODS
• At enrollment (day 0), history taken .
• Medical examinations were performed.
• growth parameters were recorded at baseline
and day 21.

• Parents were asked to fill in a structured diary to
record:
– the daily crying time (minutes),
– stool characteristics and frequency,
– any adverse effects
Statistical Analysis
• The sample size was calculated to find a
clinically relevant difference in the
reduction in daily average crying .
• 20 patients were needed per group.
• Data are shown as mean or median and
interquartile range (IQR)
• differences were considered to be
significant when P<0.05.
• All reported P values are 2-sided.
Lactobacillus journal club
Lactobacillus journal club
Patient
Enrollment
RESULTS
• There were no significant differences
between the groups regarding:
• type of delivery.
• Gender.
• age on entry.
• Family history of gastrointestinal diseases
• Atopy.
• growth parameters
RESULTS
RESULTS
• At enrollment, there was no difference in
median crying time (minutes/day) between
the groups:
• 370 (IQR: 120) vs 300 (IQR: 150) in the L
reuteri and placebo groups, respectively
(P=0.127).
RESULTS
daily crying time
• In probiotic group there was a significant
reduction in daily crying time at the end of
the study (day 21) compared with placebo:
• 35 (IQR: 85) vs 90.0 (IQR: 148)
minutes/day, respectively (P=0 .022)
placebo

lactobacillus

min/day
Number of subject who cry for
3hrs
• By day 21, the number of subjects that
had crying times 180 minutes (3hrs) was
significantly lower in the L reuteri group
compared with the placebo group (4 vs
12, respectively; P= 0.009
placebo

lactobacillus

infant
number of responders
• There was a significantly higher number of
responders in the probiotic group
compared with placebo on days 7 (20 vs
8; P =0.006), 14 (24 vs 13; P =0.007), and
21 (24 vs 15; P =0.036)
Lactobacillus journal club
Microbiologic Analysis of
Fecal Cultures
• Fecal counts of
Lactobacillus, bifidobacteria,and
Clostridium butyricum species were similar
between the groups.
• the levels of E coli were higher in the L
reuteri group at the beginning of the study
reduction in E coli
• There was a significant reduction in E coli
in the probiotic group compared with the
placebo group (-6.55 x107 [IQR: 4.87 x108]
vs 4.30 x 105 [IQR: 4.35 x
107], respectively P =0.001) .
Lactobacilli level increament
• Lactobacilli were found to be significantly
increased in the L reuteri versus placebo
group (4.07x105 [IQR: 4.98x106] vs 0 [IQR:
3.27x104]( P=0.002).
GIT function
• Gastrointestinal function was similar
between the groups
Adverse events
• reported during the study were:

– rhinitis (L reuteri group, n 1).
–eczema (placebo group, n1).
– fever (placebo group, n 1).
–otalgy (placebo group,n 1).
–GER (placebo group, n 1).
placebo

lactobacillus
Lactobacillus journal club
Lactobacillus journal club
PICO
• Population: colicky infant
aged 2 to 16 weeks
• Intervention: probiotic
• Control: placebo.
• Out come: improvement of
colic.
Relevance
1. Does the study address a common
problem in your practice?
YES
2. Does the study address an important
outcome to you or to your patient? (DOE
vs. POEM).
YES
3. Assuming that the study conclusion is true
would it lead to a change in your practice?
YES
Validity
1. Was the assignment of patients to
treatment randomized?
Yes
2- Was the assignment concealed?
Yes
Validity
3- Were patients analyzed in the groups to
which they were randomized (intention to
treat analysis)?
YES
• Was follow-up complete& long enough?
YES
Validity
3. Were the groups similar at the start of the
trial? Baseline prognostic factors
(demographics, co-morditity, disease
severity, other known confounders)
balanced?
YES
4. Were patients, their clinicians, and study
personnel 'blind' to treatment?
YES
Validity
5. Aside from the experimental intervention,
were the groups treated equally?
• Co-intervention?
• Contamination?
• Compliance?

yes
Validity
6. Were all clinically important outcomes
considered?

YES
Results clinical significance
• Precision of the effect:
• Confidence intervals?
•

no
Calculation
colic
Y
treatment Lactobacillus 1
placebo

7
8

N
a

24 (96%) b

25

n1

c

18 (71%) d

25

n0

42

50

• Absolute Risk Reduction = |EER-CER| =
• a/n1 - c/n0 =0.04 - 0.28 = -0.24 = 24%
• NNT number need to treat = 1/absolute
risk reduction = 1/0.24=4.1
Lactobacillus journal club
Applicability
1.

Can you do the Intervention exactly as it is described
in the paper
YES

2. Is your Patient is similar to the population of the study?
YES
3. Are the likely treatment benefits worth the potential
harms and costs?
???????
product
Lactobacillus journal club
CONCLUSION
• L. reuteri DSM 17 938 at a dose of 108
colony-forming units per day in early
breastfed infants improved symptoms of
infantile colic .
• well tolerated and safe.
• Gut microbiota changes induced by the
probiotic could be involved in the observed
clinical improvement.
Points
• those who received placebo thrived and
had reduced crying time by the end of the
study.

• They include patient at age of 2 weeks
and (16 weeks) 4 months
• This study was funded by BioGaia AB
(Stockholm, Sweden).
Appendages
Lactobacillus journal club
Lactobacillus reuteri
• Lactobacillus reuteri is a Gram-positive
bacterium that naturally inhabits the gut of
mammals and birds.
History
• Discovery
• As early as the turn of the 20th century, L.
reuteri was recorded in scientific classifications
of lactic acid bacteria[1], though at this time it was
mistakenly grouped as a member of
Lactobacillus fermentum. In the 1960s, further
work by German microbiologist Gerhard Reuter
– for whom the species eventually would be
named – began to distinguish L. reuteri from L.
fermentum. Reuter re-classified the species as
"Lactobacillus fermentum biotype II".[2]
History
• L. reuteri was eventually identified as a
distinct species in 1980 by Kandler et al.[3]
This group found significant differences
between L. reuteri and other biotypes of L.
fermentum, and thus proposed that it be
given formal species identity. They chose
the species name "reuteri", after
discoverer Gerhard Reuter, and L. reuteri
has since been recognized as a separate
species within the Lactobacillus genus.
level of evidence
GLOSSARY
• Randomization: Ideally, a process that
ensures every member of a population has
an equal chance to be included in the
study's sample.
• Randomized Controlled Trial (RCT): A true
experiment, in which the researcher
randomly assigns some patients to at least
one maneuver (treatment) and other
patients to a placebo, or usual treatment.
GLOSSARY
• Intention-to-treat Analysis: analyzed for
according to the groups for which they were
originally assigned .
Interquartile range
• also called the midspread or middle fifty,
is a measure of statistical dispersion,
being equal to the difference between the
third and first quartiles.
• the distance between the 75th percentile
and the 25th percentile. The IQR is
essentially the range of the middle 50% of
the data. Because it uses the middle 50%,
the IQR is not affected by outliers or
extreme values.
Interquartile range
• Example
• Compute the interquartile range for the
sorted Cotinine data:
• 18, 33, 58, 67, 73, 93, 147
• The 25th and 75th percentiles are the
.25*(7+1) and .75*(7+1) = 2nd and 6th
observations, respectively.
• IQR = 93-33 = 60.
Lactobacillus journal club

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Lactobacillus journal club

  • 1. Yassin M. Alsaleh Supervised by: Dr.Sameer Al Abdi
  • 4. Introduction • Infantile colic: paroxysmal, excessive crying without an identifiable cause in an otherwise healthy newborn infant.
  • 5. Infantile colic • Wessel’s criteria : • The classic definition is based on the rule of threes: • crying that lasts for ≥3 hrs per day. • for ≥ 3 days per week. • for a minimum of 3 weeks.
  • 6. Infantile colic • It usually occurs in infants < 3 months. • Colic affects 3% to 28% of infants. • causing considerable stress and concern for parents.
  • 8. • swallowed air • Overfeeding . • some foods. • The role of an aberrant intestinal microflora .
  • 9. • Increased presence of hydrogen gas produced by anaerobic Gram-negative bacteria. • Recently, coliform bacteria, particularly Escherichia coli, were found to be more abundant in the feces of colicky infants.
  • 10. Infantile colic • Treatments: • no treatment consistently provides satisfactory relief. • Carminatives before feedings are ineffective in preventing the attacks.
  • 13. Probiotics • Probiotics are live microorganisms thought to be healthy for the host organism. • Probiotics are commonly consumed as part of fermented foods with specially added active live cultures; such as in yogurt, or as dietary supplements.
  • 14. Probiotics • Mechanism of action: • improving intestinal microbial balance, thus inhibiting pathogens and toxin producing bacteria.
  • 15. • • • • • uses for probiotics Managing lactose intolerance Prevention of colon cancer Lowering cholesterol Lowering blood pressure Improving immune function and preventing infections • Antibiotic-associated diarrhea • Irritable bowel syndrome and colitis • constipation.
  • 16. • in a prospective randomized study, supplementation with the probiotic Lactobacillus reuteri ATCC 55 730 improved colicky symptoms in breastfed infants within 1 week of treatment compared with treatment with simethicone.
  • 17. OBJECTIVES • To test the efficacy of Lactobacillus reuteri on infantile colic . • to evaluate its relationship to the gut microbiota.
  • 19. METHODS • This randomized, controlled, double blind Study. • In general pediatricians and OPD at the Department of Pediatrics, University of Turin (Regina Margherita Children Hospital)
  • 20. METHODS • between March 2008 and August 2009. • All infants were diagnosed according to the modified Wessel’s criteria. • All were born at term, AGA and aged 2 to 16 weeks at recruitment.
  • 21. Inclusion criteria • Only exclusively breastfed infants were enrolled. • At enrollment, mothers were encouraged to avoid cow’s milk in their diet.
  • 22. Exclusion criteria • clinical evidence of chronic illness or GIT disorders. • any intake of probiotics and/or antibiotics in the week preceding recruitment. • any formula-feeding. • no acid blockers were used in any of the infants who completed this study.
  • 23. outcomes • Primary outcome was defined as a reduction of average crying time to < 3 hours a day on day 21. • Secondary outcome was defined as the number of responders in each group on days 7, 14, and 21. • Responders were those who experienced a decrease in the daily average crying time of 50% from baseline.
  • 24. METHODS • In addition, the intestinal microflora of the infants was analyzed to determine the effect of the by using (FISH).
  • 26. METHODS • Colicky infants were randomly assigned to groups by using a computer generated randomization list created by an independent departmental statistician. • written informed consent was obtained from parents .
  • 27. METHODS • Active study product consisted of a suspension of freeze-dried L reuteri DSM 17 938 in a mixture of sunflower oil and medium-chain triglyceride oil . • The placebo was identical in appearance and taste but without the live bacteria.
  • 28. METHODS • Both formulations were administered in 5 drops, once a day, 30 minutes before the feed in the morning, for 21 days.
  • 29. METHODS • At enrollment (day 0), history taken . • Medical examinations were performed. • growth parameters were recorded at baseline and day 21. • Parents were asked to fill in a structured diary to record: – the daily crying time (minutes), – stool characteristics and frequency, – any adverse effects
  • 30. Statistical Analysis • The sample size was calculated to find a clinically relevant difference in the reduction in daily average crying . • 20 patients were needed per group. • Data are shown as mean or median and interquartile range (IQR) • differences were considered to be significant when P<0.05. • All reported P values are 2-sided.
  • 34. RESULTS • There were no significant differences between the groups regarding: • type of delivery. • Gender. • age on entry. • Family history of gastrointestinal diseases • Atopy. • growth parameters
  • 36. RESULTS • At enrollment, there was no difference in median crying time (minutes/day) between the groups: • 370 (IQR: 120) vs 300 (IQR: 150) in the L reuteri and placebo groups, respectively (P=0.127).
  • 38. daily crying time • In probiotic group there was a significant reduction in daily crying time at the end of the study (day 21) compared with placebo: • 35 (IQR: 85) vs 90.0 (IQR: 148) minutes/day, respectively (P=0 .022) placebo lactobacillus min/day
  • 39. Number of subject who cry for 3hrs • By day 21, the number of subjects that had crying times 180 minutes (3hrs) was significantly lower in the L reuteri group compared with the placebo group (4 vs 12, respectively; P= 0.009 placebo lactobacillus infant
  • 40. number of responders • There was a significantly higher number of responders in the probiotic group compared with placebo on days 7 (20 vs 8; P =0.006), 14 (24 vs 13; P =0.007), and 21 (24 vs 15; P =0.036)
  • 42. Microbiologic Analysis of Fecal Cultures • Fecal counts of Lactobacillus, bifidobacteria,and Clostridium butyricum species were similar between the groups. • the levels of E coli were higher in the L reuteri group at the beginning of the study
  • 43. reduction in E coli • There was a significant reduction in E coli in the probiotic group compared with the placebo group (-6.55 x107 [IQR: 4.87 x108] vs 4.30 x 105 [IQR: 4.35 x 107], respectively P =0.001) .
  • 44. Lactobacilli level increament • Lactobacilli were found to be significantly increased in the L reuteri versus placebo group (4.07x105 [IQR: 4.98x106] vs 0 [IQR: 3.27x104]( P=0.002).
  • 45. GIT function • Gastrointestinal function was similar between the groups
  • 46. Adverse events • reported during the study were: – rhinitis (L reuteri group, n 1). –eczema (placebo group, n1). – fever (placebo group, n 1). –otalgy (placebo group,n 1). –GER (placebo group, n 1).
  • 50. PICO • Population: colicky infant aged 2 to 16 weeks • Intervention: probiotic • Control: placebo. • Out come: improvement of colic.
  • 51. Relevance 1. Does the study address a common problem in your practice? YES 2. Does the study address an important outcome to you or to your patient? (DOE vs. POEM). YES 3. Assuming that the study conclusion is true would it lead to a change in your practice? YES
  • 52. Validity 1. Was the assignment of patients to treatment randomized? Yes 2- Was the assignment concealed? Yes
  • 53. Validity 3- Were patients analyzed in the groups to which they were randomized (intention to treat analysis)? YES • Was follow-up complete& long enough? YES
  • 54. Validity 3. Were the groups similar at the start of the trial? Baseline prognostic factors (demographics, co-morditity, disease severity, other known confounders) balanced? YES 4. Were patients, their clinicians, and study personnel 'blind' to treatment? YES
  • 55. Validity 5. Aside from the experimental intervention, were the groups treated equally? • Co-intervention? • Contamination? • Compliance? yes
  • 56. Validity 6. Were all clinically important outcomes considered? YES
  • 57. Results clinical significance • Precision of the effect: • Confidence intervals? • no
  • 58. Calculation colic Y treatment Lactobacillus 1 placebo 7 8 N a 24 (96%) b 25 n1 c 18 (71%) d 25 n0 42 50 • Absolute Risk Reduction = |EER-CER| = • a/n1 - c/n0 =0.04 - 0.28 = -0.24 = 24% • NNT number need to treat = 1/absolute risk reduction = 1/0.24=4.1
  • 60. Applicability 1. Can you do the Intervention exactly as it is described in the paper YES 2. Is your Patient is similar to the population of the study? YES 3. Are the likely treatment benefits worth the potential harms and costs? ???????
  • 63. CONCLUSION • L. reuteri DSM 17 938 at a dose of 108 colony-forming units per day in early breastfed infants improved symptoms of infantile colic . • well tolerated and safe. • Gut microbiota changes induced by the probiotic could be involved in the observed clinical improvement.
  • 64. Points • those who received placebo thrived and had reduced crying time by the end of the study. • They include patient at age of 2 weeks and (16 weeks) 4 months • This study was funded by BioGaia AB (Stockholm, Sweden).
  • 67. Lactobacillus reuteri • Lactobacillus reuteri is a Gram-positive bacterium that naturally inhabits the gut of mammals and birds.
  • 68. History • Discovery • As early as the turn of the 20th century, L. reuteri was recorded in scientific classifications of lactic acid bacteria[1], though at this time it was mistakenly grouped as a member of Lactobacillus fermentum. In the 1960s, further work by German microbiologist Gerhard Reuter – for whom the species eventually would be named – began to distinguish L. reuteri from L. fermentum. Reuter re-classified the species as "Lactobacillus fermentum biotype II".[2]
  • 69. History • L. reuteri was eventually identified as a distinct species in 1980 by Kandler et al.[3] This group found significant differences between L. reuteri and other biotypes of L. fermentum, and thus proposed that it be given formal species identity. They chose the species name "reuteri", after discoverer Gerhard Reuter, and L. reuteri has since been recognized as a separate species within the Lactobacillus genus.
  • 71. GLOSSARY • Randomization: Ideally, a process that ensures every member of a population has an equal chance to be included in the study's sample. • Randomized Controlled Trial (RCT): A true experiment, in which the researcher randomly assigns some patients to at least one maneuver (treatment) and other patients to a placebo, or usual treatment.
  • 72. GLOSSARY • Intention-to-treat Analysis: analyzed for according to the groups for which they were originally assigned .
  • 73. Interquartile range • also called the midspread or middle fifty, is a measure of statistical dispersion, being equal to the difference between the third and first quartiles. • the distance between the 75th percentile and the 25th percentile. The IQR is essentially the range of the middle 50% of the data. Because it uses the middle 50%, the IQR is not affected by outliers or extreme values.
  • 74. Interquartile range • Example • Compute the interquartile range for the sorted Cotinine data: • 18, 33, 58, 67, 73, 93, 147 • The 25th and 75th percentiles are the .25*(7+1) and .75*(7+1) = 2nd and 6th observations, respectively. • IQR = 93-33 = 60.