Nutraceutical Properties of Human and Bovine Colostrum in Health

Nutraceutical Properties of Human and
Bovine Colostrum in Health and Disease
Harpal S. Buttar , D.V.M., M.Sc., Ph.D., FICN
Senior Scientist, Therapeutic Products Directorate,
Health Canada (Retd.)
Adjunct Prof., Dept. of Pathol. & Lab. Medicine,
University of Ottawa, School of Medicine, Canada
18th World Congress on Clinical Nutrition,
Ubon Ratchathani, Thailand, Dec. 1-3, 2014
COLOSTRUM
 Colostrum - first mammary secretion that all
mammals provide for their newborns during the first
24-48 hours after parturition.
 It creates the foundation of life-long immunity.
 Colostrum contains various antibodies, and has lower
fat and higher protein content than ordinary milk.
COLOSTRUM
 Colostrum has a nutrient profile and immunological
composition that differs substantially from mature
milk.
 Colostrum contains macro- and micronutrients,
including vitamins and minerals. It contains higher
amounts of oligosaccharides, growth factors,
antimicrobial compounds, and immune regulators.
The nutrients in colostrum
contribute to the maturing
and development of the GI
tract in newborns.
Colostrum starts helping
GI
tract
to
heal
immediately by lowering
pathogens, virus, bacteria,
fungus, yeast, mold etc.) in
the GI tract
 Colostrum has been known for
centuries for its health benefits.
English
Marathi
Colostrum
Chiik
(Kharwas)
 Bovine Colostrum > potency than
Human colostrum.
Hindi
Tamil
Telugu
Konkani
Gujarati
Khiis
Seem paal
Junnu
Posu/Geen
Bari
Punjabi
Bauli
 Humans can rely on bovine
colostrum for their health benefit.
 Another purported use of colostrum
in the early 1950s was for treatment
of rheumatoid arthritis.
Some Physiological Effects of Colostrum
 May increase lean muscle,
 May decrease body fat levels,
 May improve sprint performance,
 May enhance anaerobic performance and buffers lactic
acid build up,
 May improve the recovery between bouts of exercise,
 May enhance endurance performance,
 Can improve immune function,
 Growth factors in colostrum may enhance differentiation
of bone marrow stem cells.
Benefits of Colostrum Supplements
 Because of the rapid gut cell turn over rate (cell replication),
colostrum supplements may help in curing mal-absorption of
nutrients due to GI tract problems such as chronic inflammatory
gut disorder, leaky-gut syndrome.
 Several investigators have reported that the immune and growth
factors present in human and bovine colostrum may not only
benefit the newborn, but may also help the adult humans.
 Since colostrum is a food rather than a drug, its adverse effects
are considered to be minimal, provided the product is obtained
from a reliable source and is prepared using good manufacturing
procedure.
Colostrum
Ig G1, G2,
A,M
PRP
GF I, II
Lactoferrin
Leptin
Different immunoglobulins present in bovine colostrum
Immunoglobulin
Quantity
IgG1
35.0 (mg/ml)
IgG2
16.0 (mg/ml)
IgA
1.7 (mg/ml)
IgM
4.3 (mg/ml)
Lactoferrin
0.8 (mg/ml)
Biological Activities of Colostrum
1
•Antimicrobial
2
•Antioxidant
3
•Anti-inflammatory
4
•Cardio-protective
Antimicrobial activity
 Antimicrobial activity of colostrum was tested
using both Gram positive & Gram negative
strains.
 Pour plate method was adopted, according to the
Indian pharmacopoeia 2010.
 MIC of colostrum was calculated by serial dilution
in triplicate and it was found to be around
100µg/ml.
Antimicrobial activity of Colostrum
Minimum inhibitory concentration of colostrum was found to
be 100 µg/ml against:
• E. coli
• S. aureus
• P. vulgaris
• E. aerogenes
• S. typhi
From the result obtained, it can be concluded that
Colostrum is a potent antimicrobial agent.
Anti-inflammatory Actions of Colostrum
• Lactoferrin contained in colostrum has strong antioxidant
characteristics which seem to act by two separate
mechanisms:
(i) binding catalytic iron generated during cell destruction,
(ii) and binding bacterial lipopolysaccharide (LPS) which
causes a decrease in LPS bioactivity, such as increasing
neutrophil O2 production.
•
Colostrum lactoferrin also inhibits the production of
inflammatory cytokines.
• Proline rich peptides (PRP) in colostrum also act as an
oxidative stress inhibitor.
Anti-inflammatory Actions of Colostrum
• Colostrum has anti-inflammatory activity.
• Tumor necrosis factor-α (TNF-α) present in colostrum
bind with the cytokine receptors which control the
inflammatory cytokine cascade, effectively turning off
inflammation at its source.
• Colostrum also contains interleukin-1, the IL-1 receptor
protein, which also acts to inactivate the inflammatory
activity of IL-1.
• Colostrum may be beneficial in chronic inflammatory
diseases, such as various forms of arthritis and Crohn’s
disease, which has been linked to gut inflammation.
Athletic Benefits of Colostrum Supplements
 Research suggests that to enhance supporting
performance, a minimum dose of 20 mg/day is
required, and this may take up to 8-weeks to have a
beneficial effect. It is not clear whether lower doses
(e.g. 10 g/day) would also be effective.
Note: Colostrum appears to be well tolerated. However, some
people may experience gastrointestinal problems such as
flatulence and nausea.
Biochemical Components of Colostrum
Components
1) Nutritional components
• Vitamins (A, B12 and
E)
• Minerals
• Amino Acids
• Essential fatty acids
Uses
Promote Health, Vitality,
and Growth of
the Newborn
Biochemical Components of Colostrum
Components
2) Immune Factors
• Proline-Rich Polypeptide
(PRP)
• Immunoglobulins (A, D, E, G
and M)
• Lactoferrin
Uses
Regulate the thymus gland
IgG neutralizes toxins and
microbes
IgM destroys bacteria
IgE and IgD are highly antiviral
A nti-viral, anti-inflammatory and
anti-bacterial iron-binding protein
with therapeutic effects in cancer,
HIV etc.
Biochemical Components of Colostrum
Components
3) Growth Factors
• Epithelial growth factor
(EGF)
• Insulin-like growth factor-I
and II (IGF-1 and IGF-II)
• Fibroblast growth factor
(FgF)
• Platelet-derived growth
factor (PDGF)
• Transforming growth factors
A (TgA) and B (TgB)
• Growth hormone (GH)
Uses
Help in enhancing cell and
tissue growth by stimulating
DNA formation
Comparison of Human, Cow, Buffalo & Goat Colostrum
Constituents
Human
Cow
Buffalo
Goat
Fat
3-5%
6.7%
7.56%-
4.1%
11.31%
Protein
0.8-0.9%
14.9%
4.3%
3.4%
Lactose
6.9-7.2%
2.5%
4.7%
4.7%
• Fat content in Buffalo Colostrum is higher than other species.
• Protein in Cow Colostrum is higher than other species.
• Lactose content in Human Colostrum is higher than other
species.
Comparison of water soluble vitamin and mineral contents in
Human, Cow, Buffalo & Goat Colostrum
Water Soluble
Vitamin
Human (mg/100 Cow (µg/ml)
Buffalo (µg/ml)
ml)
Goat
(µg/ml)
Constituents
Niacin
0.02
0.34
-
Approximately 0.6 µg/ml water soluble
vitamins are present.
Thiamine
0.017
0.90
-
Riboflavin
0.04
4.55
3.4
Vitamin B12
0.03
0.60
1.59
Pyridoxal
-
0.15
-
Pyridoxamine
-
0.21
-
Pyridoxine
-
0.04
3.25
Fat soluble vitamins present in bovine colostrum
Fat soluble vitamins
Quantity
Retinol (vitamin A)
4.9 (µg/g)
Tocopherol (vitamin E)
2.9 (µg/g)
Beta-carotene
0.7 (µg/g)
Mineral Constituents
Human (mg/100 ml)
Cow (mg/kg)
Buffalo (mM)
Goat (g/kg)
33
4716
47.1
0.65
Phosphorus
13-16
4452
27.7
0.36
Magnesium
4
733
7.3
-
Sodium
50
1058
20.3
1.44
Potassium
74
2845
28.7
3.38
Zinc
0.53
38
147-728
-
Iron
0.15
5.3
42-152
-
Copper
0.04
0.3
7
-
Sulphur
-
2595
15700
0.2
Manganese
-
0.1
38.2
-
Calcium
The level of water soluble Vitamin content and Mineral constituent is much
higher in Cow Colostrum.
Manufacturers of colostrum-derived nutraceuticals
Types
Colostrum powder
Colostrum capsule
manufacturer
manufacturer
Cow colostrum
APS BioGroup
APS BioGroup
Buffalo colostrum
Biostrum Nutritech
Biostrum Nutritech
Pvt. Ltd
Pvt. Ltd.
Goat colostrum
Mt. Capra Whole food Mt. Capra Whole food
Nutritionals
Nutritionals
Conclusions
 Colostrum acts as an anti-microbial, anti-oxidant, antiinflammatory and cardio-protective agent.
 Colostrum concentrate was safe in athletes at 20 mg/day
dose.
 Well designed placebo-controlled clinical trials are
required to access the safety and efficacy of colostrum
products.
 Further studies are needed to understand the underlying
mechanisms by which colostrum acts as anti-microbial,
anti-inflammatory and cardio-protective agent.
Acknowledgement
I would like to acknowledge the collaboration of Dr.
Ginpreet Kaur, Assistant Professor at School of
Pharmacy &Technology Management, V. L. Mehta
Rd, Vile Parle (W) Mumbai, India.
“Leave your drugs in the chemist’s pot, if
you can heal the patient with food”
“Illness occurs when the body’s systems are
toxic and out-of-balance. One must treat the
body as a whole, rather than a series of
parts”
Hippocrates Father of Medicine
ca.460-370 B.C.
Antimicrobial plate of Enterobacter aerogenes
Enterobacter aerogenes
Code
Description
Centre
Positive
control
1
Negative
control
2
Mixture of
sample
3
Colostrum
(100 ppm)
4
Amoxicillin
(10 ppm)
Zone of inhibition of
colostrum = 12 mm
Antimicrobial plate of Proteus vulgaris
Proteus vulgaris
Code
Description
Centre
Positive
control
1
Negative
control
2
Mixture of
sample
3
Colostrum
(100 ppm)
4
Amoxicillin
(10 ppm)
Zone of inhibition of
colostrum = 12 mm
Antimicrobial plate of Escherichia coli
Escherichia coli
Code
Description
Centre
Positive
control
1
Negative
control
2
Mixture of
sample
3
Colostrum
(100 ppm)
4
Amoxicillin
(10 ppm)
Zone of inhibition of
colostrum = 13 mm
Antimicrobial plate of Salmonella typhi
Salmonella typhi
Code
Description
Centre
Positive
control
1
Negative
control
2
Mixture of
sample
3
Colostrum
(100 ppm)
4
Amoxicillin
(10 ppm)
Zone of inhibition of colostrum =
11 mm
Antimicrobial plate of Staphylococcus aureus
Staphylococcus aureus
Code
Description
Centre
Positive
control
1
Negative
control
2
Mixture of
sample
3
Colostrum
(100 ppm)
4
Amoxicillin
(10 ppm)
Zone of inhibition of colostrum =
11 mm
Antimicrobial activity of Colostrum
 Zone of inhibition was measured in millimeter after 24 hours of incubation at 37 0C.
Free radical scavenging activity of colostrum by DPPH
Free radical scavenging activity of colostrum by DPPH
According to DPPH free radical scavenging activity
assay, Colostrum powder showed moderate activity
(47.07 %) at 200 µg/ml in comparison with Ascorbic
acid (89.72%).
The activity of colostrum
concentration dependent.
was
found
to
From the results obtained, it can be concluded that
Colostrum has moderate antioxidant activity in
comparison with standard ascorbic acid.
be
Lipid peroxidation inhibitory activity
 Evaluation of lipid per oxidation inhibitory activity of
colostrum was performed on rat liver homogenate
using Thiobarbituric acid reacting substances
(TBARS) method.
 In acidic condition, a pink colored complex was
formed, absorbance of which was measured at 532 nm
spectrophotometrically.
 Colostrum showed dose-dependent increase in the
antioxidant activity. The average IC50 value was found
to be 180 ppm.
Lipid peroxidation inhibitory activity of colostrum by
TBARS
Sr.No
Concentration (ppm)
% Inhibition
1.
20
11.1
2.
40
22.4
3.
80
40.9
4.
160
46.4
5.
200
52.5
Lipid peroxidation inhibitory activity by TBARS
% Lipid peroxidation inhibition
60
50
% Inhibition
40
30
% Inhibition
20
10
0
0
50
100
150
200
Concentration (ppm)
 IC50 of colostrum was found to be 180 ppm
250
Anti-inflammatory activity
 Male Wistar rats were fasted for 12 h with
Before any treatment, the average volume
each animal was determined using
(Plethysmometer 520). Immediately after,
were administered.
free access to water.
of the right paw of
a plethysmometer
the test substances
 The control group received only the vehicle. Sixty minutes later,
paw edema was induced by subcutaneous injection of 0.1 ml
carrageenan (0.1%) into the plantar surface of the right paw.
 The volume of the paw of each rat was measured 1, 2, 3 and 4 h
after the injection of the control substance, inflammatory agent
or Sample Extract.
Evaluation of Anti-inflammatory Activity of Colostrum
Four groups (N = 6)
Group I
Group II
Group III
Group IV
• Control group
• Standard drug Diclofenac sodium (30 mg/kg)
• Test substance Colostrum (500 mg/kg)
• Test substance Colostrum (500mg/kg) + Diclofenac (10
mg/kg)
Anti-inflammatory activity results of colostrum
Time
(hrs)
Diclofenac
Colostrum
Colostrum + Diclofenac
0
46.25 ± 0.07***
35.62 ± 0.03***
21.67 ± 0.02***
1
64.56 ± 0.05***
50.76 ± 0.02***
37.49 ± 0.02***
2
77.33 ± 0.06***
58.80 ± 0.01***
49.38 ± 0.02***
3
76.23 ± 0.11***
67.94 ± 0.02***
59.53 ± 0.02***
4
67.38 ± 0.06***
56.65 ± 0.02***
48.83 ± 0.01***
Anti-inflammatory Activity Results
Anti-inflammatory Activity Results
The reduction was of the highest percent in Colostrum
alone group (67.94%) was observed at 3rd hr.
The second highest reduction (59.53%) was observed in
Colostrum with Diclofenac combination at 3rd hr.
It was concluded that Colostrum powder has
moderate anti-inflammatory activity in
comparison with Diclofenac.
Cardio protective activity of colostrum
MYOCARDIAL INFARCTION
 The plaque deposited in the arteries hardens and when it cracks
clots are formed around the plaque.
 Artery blockage and cessation of blood supply leads to damage
of the heart muscles (myocardial infarction)
Cardioprotective activity of colostrum
Induction of myocardial infarction
Isoproterenol
(200 mg/kg)
Administered to
Albino Wistar
rats
subcutaneously
for 2 consecutive
days
Saline
Animal Groups
• Seven groups each group containing six rats (150-250 g body wt.)
Group I
Group II
Group III
Group IV
Group V
Group VI
Group VII
• Rats received 1.0 ml water as a vehicle. Group I rats were referred as control rats.
• Rats were administered the dose of isoproterenol
• Rats were dosed with the test agent colostrum 250 mg/kg alone.
• Rats were treated with colostrum 250 mg/kg plus 0.25mg/kg enalapril
• Rats received the test agent colostrum 500 mg/kg alone.
• Rats were treated with colostrum 500mg/kg + 0.25mg/kg enalapril
• Rats received dose of standard ACE inhibitor enalapril 0.25mg/kg alone.
Biochemical serum estimation of CK-MB and LDH
 The isoproterenol-induced myocardial infarcted control group
had the highest CK-MB and LDH values indicating cardiac
damage.
 Colostrum produced a dose-dependent decrease in the toxicity.
 There was a significant reduction in cardiotoxicity after the
combined administration of 500 mg/kg of colostrum and
0.25mg/kg enalapril on comparison with the isoproterenol
myocardial infarcted group (Group II rats).
 The increase in the CK-MB values and LDH indicate the extent
of isoproterenol-induced cardiotoxicity.
CKMB
CKMB
2500
2000
**
ckmb IU/l
***
1500
***
***
***
1000
500
0
Group I
Group II
Group III
Group IV
Group V
Group VI
Group VII
Treatment Groups
Bars represented as Mean ± SEM, one way ANOVA followed by
Dunnett’s multiple comparison test. Significantly differ from
control Group 1, ** p<0.01 and ***p< 0.001.
LDH
LDH
1800
1600
1400
LDH (IU/L)
1200
***
1000
***
800
***
***
***
600
400
200
0
Group I
Group II
Group III
Group IV
Group V
Group VI
Group VII
Treatment Groups
Bars represent as Mean ± SEM, one way ANOVA followed by
Dunnett’s multiple comparison test. Significantly differ from
control Group 1, ***p< 0.001.
Conclusion
 Colostrum was found to act as an antimicrobial, antioxidant,
anti inflammatory and cardio protective agent.
 Colostrum was safe till a 2g/kg dose.
 Further studies to understand the underlying mechanisms by
which Colostrum acts in inflammation and cardio protection
are required.
Summary and Conclusions
Acute toxicity, antimicrobial, antioxidant, anti
inflammatory and cardio protective effects of Colostrum
were evaluated.
Colostrum was safe upto 2.0 g/kg dose.
MIC of Colostrum was 100 µg/ml.
DPPH free radical scavenging activity assay - 47.07 % at
200µg/ml.
Lipid peroxidation inhibitory assay - IC50 value was
found to be 180 ppm.
Summary and Conclusions
Colostrum alone reduced inflammation (67.94%)
compared to colostrum + diclofenac (59.53%).
Reduction could be due to an antagonistic effect.
Colostrum + enalapril reduced cardiotoxicity more
than colostrum alone.
Acknowledgement
I would like to acknowledge the
collaboration of Dr. Ginpreet Kaur,
Assistant Professor at School of
Pharmacy &Technology Management,
V. L. Mehta Rd, Vile Parle (W)
Mumbai, India.
“Leave your drugs in the chemist’s pot, if you
can heal the patient with food”
“Illness occurs when the body’s systems are
toxic and out-of-balance. One must treat the
body as a whole, rather than a series of parts”
Hippocrates Father of Medicine
ca.460-370 B.C.
Thank you !!
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Thank you !!
OBJECTIVES
7)
NDC Questions regarding Findings and Recommendations
Raphael, 1509, School of Athens, Causarum cognitio (Seek Knowledge of
59
Causes) “A Visualisation of Knowledge”