Pre-IACUC Approval

1.
Antibiotic
Group
N=5
High
Fat
Diet
Group
N=5
Control
Group
N=5
Diet:
Provided
ad
libitum
3
Kg-­‐
T.D.
088061i
(Harlan
Teklad,
Madison)
Purified
Control
Diet
(10%
fat
kcal)
Irradiated
Pellets
Sterilized
Water
Provided
ad
libitum
Diet:
Provided
ad
libitum
3
Kg-­‐
T.D.
088061i
(Harlan
Teklad,
Madison)
Purified
Control
Diet
(10%
fat
kcal)
Irradiated
Pellets
Sterilized
Water
Supplemented
with
Ampicillin
(sodium
salt)
(1g/L)
(>95%
Purity)
Provided
ad
libitum
Diet:
Provided
ad
libitum
3
Kg-­‐
T.D.
06414i
(Harlan
Teklad,
Madison)
Matched
Adjusted
Calories
(60%
fat
kcal)
Irradiated
Pellets
Sterilized
Water
Provided
ad
libitum
Gavage:
Total
Volume
10ml/kg
Body
Weight
Non-­‐Pharmaceutical
(Cayman
Chemical)
Broad-­‐
Spectrum
Antibiotic
Concoction
Dosage:
Vancomycin
(hydrochloride)
(100mg/kg
BW)
(>98%Purity)
Neomycin
(sulfate)
(100mg/kg
BW)
(>90%
Purity)
Metronidazol
(100mg/kg
BW)
(>98%
Purity)
Amphotericin
B
Antifungal
(1mg/kg
BW)
(>95%
Purity)
Combinational
Bactericidal
Activity
Against
Full
Spectrum
and
Notably
Dual
Activity
Against
Both:
Gram
Positive
(Ampicillin
and
Vancomycin)
And
Gram
Negative
Aerobic
and
Facultative
Strains
(Ampicillin
and
Neomycin)
Gavage
Twice
Daily
for
14
Days
Gavage:
Total
Volume
10ml/kg
Body
Weight
Saline/Water
Gavage
Twice
Daily
for
14
Days
Total
Volume-­‐
10ml/kg
Body
Weight
Gavage:
Total
Volume
10ml/kg
Body
Weight
Saline/Water
Gavage
Twice
Daily
for
14
days
Volume-­‐
10ml/kg
Body
Weight
Info:
Diet-­‐Induced
Obesity
Model
Diet-­‐Induced
Dysbiosis
Increased
Colonic
Permeability
Increased
Sleep
Duration
Aberrant
Microbial
Signals
Circadian
Disruption
Obese
Info:
“Germ
Free-­‐Like”
Phenotype**
Microbial
Depletion
Antibiotic-­‐Induced
Dysbiosis
Absent
Microbial
Signals
Circadian
Disruption
Increased
Colonic
Permeability
Decreased
Sleep
Duration
Lean
Info:
Conventionally
Raised
Intestinal
Microbial
Composition
Standard
Colonic
Permeability
Normal
Microbial
Signals
Normal
Host-­‐Microbial
Standard
Circadian
Function
Lean
Means
and
Standard
Deviation
of
Mouse
Sleep
Pattern
Correlations
%
TST
54.8
(6.21)
%PS/TST
9.60
(2.60)
SWS
per
length
(min)
5.79
(0.58)
PS
per
length
(min)
2.32
(0.23)
Sleep
Epoch
Length
(min)
7.77
(1.39)
Sleep
Cycle
Length
(min)
12.3
(1.78)
Ratio
day/night
sleep
1.43
(0.41)
No.
Sleep/Day
(111)
(18.5)
20
Mice:
C57CL/6
Male
9/10-­‐week
old