The Effect of Morinda citrifolia on health and performance of ......Calf Health Scoring Criteria...
Transcript of The Effect of Morinda citrifolia on health and performance of ......Calf Health Scoring Criteria...
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The Effect of Morinda citrifolia on health and performance of neonates
Richard G. Godbee, Ph.D., PASProfessor, Clinical & Adjunct Faculty
University of Nevada, RenoBenjamin J. Darien, DVM, MS Dipl. ACVIM
Associate ProfessorSchool of Veterinary Medicine
University of Wisconsin, Madison
1
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Botanical Immunomodulators
Tomato (Lycopene)
Ginger (6-Gingerol)
Tumeric (Curcumin)
Metastasis Inhibitors Antioxidant Activity
Red Grapes (Resveratrol)
Anti-Inflammatory Activity
Capsicum Baccatum
Peruvian hot pepper
Blueberries
Cinnamon Pomegranate (Ellagic Acid)
Green tea
Analgesic Effects
Red Chili (Capsaicin)
Antibacterial Activity
CloveOregano
RosmarinusOfficinalis
Rosemary
2
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Morinda citrifolia(NONI)
• Noni
– Indian mulberry, great morinda, cheezefruit, mouse’s pineapple, yellow root, jumbie breadfruit, hog apple, pain killer,, nono,
– It is a large evergreen shrub or small tree to 6 m or more in height and 13 cm or more in stem
– The white tubular flowers are grouped in globose heads
AND… Recent studies show NONI can also function as an immunomodulator! 3
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• Hypothesis: Neonatal calves fed noni puree would demonstrate whole blood phagocytic capacity in Gram-negative and Gram-positive in vitro assays
Increased Health& Well-being
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Study Protocol• 2 groups (n = 9 in each)
– Calf pairs
– Control & Noni supplemented calves
• Inclusion criteria
– Normal physical examination and IgG 1200 mg/dl
• Noni puree per calf = 1 oz/day p.o., BID
• Day 0 blood samples 36-48 hours of age before the first feeding of puree (day 0) &d’s 3, 7, and 14
D 0(Start of
the study)
D3 D7 D14Day ofBirth
24 -48h
D0
(Start ofthe study)
D3 D7 D14Day of
24 -48h
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Study Methods• Grp 1 control calves; Grp 2 received 30mL of
noni puree q12h in milk replacer
• Daily health scores recorded
• Bactericidal assays determined % killing of E. coli and S. epidermidis.
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Bactericidal Assay
Estimate the ability of whole blood from control and noni puree Tx calves to engulf and kill Gram – and + bacteria
E. coli S. epidermidis
Bacteria only
Count colonies and determine percentage of killing
Serial dilutions
blood + E. coli or Staph. epidermidis
Serial dilutions
Over night incubation
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Bactericidal Assay
Bacterial control cfu/ ml – Sample cfu/ ml
Bacterial control cfu/ ml100 Percent bacteria killed
Formula used:
Example:
Bacterial control: 200 colonies
Sample: 100 colonies
200 - 100x 100 = 50 % bacteria killed
200
E. Coli on blood agar
Difference: Noni % kill – control % kill
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Results: Calf Health & Gram +• No calves were
removed from the study because of health reasons
• There were no differences between the groups for S. epidermidis bactericidal activity at any of the time points or between time points
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Results: E. coli
Significantly more bacterical killing on day 14 of E. Coli compared to control (p = 0.0215).
E. coli bacterial killing between noni puree supplemented calves and control calves increased over time.
(p = 0.0534).
Med
ian
Dif
feren
ce i
n P
ercen
t K
ill,
(N
on
i-C
on
tro
l)
D ay
Med
ian
Dif
feren
ce i
n P
ercen
t K
ill,
(N
on
i-C
on
tro
l)
D ay
Staph . ep iderm idis M edian Percent K illB .
E . coli M edian Percent K ill
*
A .
Med
ian
Dif
feren
ce i
n P
ercen
t K
ill,
(N
on
i-C
on
tro
l)
D ay
Med
ian
Dif
feren
ce i
n P
ercen
t K
ill,
(N
on
i-C
on
tro
l)
D ay
Staph . ep iderm idis M edian Percent K illB .
E . coli M edian Percent K ill
*
A .
0 3 Days 7 14Med
ian
Dif
fere
nce
in %
Kill
(N
on
i-C
on
tro
l)
*
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Conclusions• Given the role of polysaccharides as
immunomodulators, noni puree potentially may stimulate the acquired innate immune response by modulating colostral cells, neonatal mononuclear cells
– General mitogenic effect on select pattern-recognition receptor activation
• Subsequent trials was warranted to investigate a modulating effect on naïve T cell activation.
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CD4+ and CD8+T Cell Response In Neonatal Calves Fed Morinda
Citrifolia (Noni)
V.J. Brooks1*, R.G. Godbee2, S.F. Peek1, B.J. Darien1
1University of Wisconsin, Madison2University of Nevada, Reno
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Hypothesis
• Colostral leukocytes & cytokines play a role in enhancing neonatal immunity
• Colostral IL-1, TNF-α, IFN-γ, play a role in enhancing neonate immunity by up-regulation of IL-2r on PBMC
• Ho #1: That supplementation of Noni pure would enhance T cell expression of CD25
• Ho #2: Noni induces cytokine expression in colostral leukocytes
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Protocol• 2 groups (n = 10 in each)
– Calf pairs
– Control & noni supplemented calves
– Daily calf health score evaluations
• Noni puree per calf = 30 mL po, q12h for 14 days
• Day 0, 3, 7 and 14 samples
• T cell activation and proliferation
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Lymphocyte Activation Assay• Objective– Evaluate IL-2 receptor (CD25) expression as index of
lymphocyte activation
PBMC NoMitogen
Con A PHA
Fix and label cells
72 hr
2-color flowcytometry
CD4+
CD25CD8+
CD25 T cell
CD25
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Lymphocyte Blastogenesis Assay• Objective
– Measure lymphocyte proliferation in response to mitogen stimulation
PBMC
48 h
18 h
NoMitogen
Con A PHA
Pulse with 3H Thymidine
Harvest and count
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CD4+T Cell Activation in Neonatal Calves Fed (Noni) puree
Significant increase in % activation(CD25r) of CD4+ T cells on day 3 after ConA stimulation
0
20
40
60
80
100
0 3 7 14
Control Noni Puree
*
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CD8+ T Cell Activation in Neonatal Calves Fed (Noni) puree
Significant increase in % activation of CD8+ T cells on dayAfter ConA stimulation
Significant effect over time for CD8+ T cells
0
10
20
30
40
50
60
70
80
90
0 3 7 14
Control Noni Puree
*
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Results• No significant differences
– Activation of γδ TCR+ cells
– PHA activation response
– Proliferation
• Increase in CD25 % expression on day 3– CD4+ T cells
– CD8+ T cells
• Significant effect over time for CD8+ T cells
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Mode of Action: Noni Immune Modulating Effect
More prolific CD4+
clonal expansion
More potent attack on invader
Noni CD25 expression in CD4+ and CD8+ T-Cells
IL-2 release by CD4+ cells
CD4+
CellsCD4+
cell activation
number of CD25 receptors on CD8+ cells
+
CD8
APC
CD4Clonal Expansion
CD4
CD4
Un-Supplemented Calves
IL-2
IL-2IL-2
Pathogen
CD25 Receptor
APC
CD4Clonal Expansion
CD4
CD4
CD4
CD8
Noni Supplemented Calves
Pathogen
IL-2
IL-2 IL-2
CD25 Receptor
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A Clinical Trial Evaluating MorindaMaxTM
Calf® Immune-supplement on Health and Performance of Pre-weaned, Holstein, Heifer
Calves.
V.J. Brooks1*, R.G. Godbee2, S.F. Peek1, B.J. Darien1
1University of Wisconsin, Madison2University of Nevada, Reno
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Central Hypothesis• Neonatal calves fed noni puree would be
healthier and consequently wean earlier and increase profitability of the producer
IncreasedHealth
AndWell-being
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Clinical Trial
• Large scale calf-raising farm in Central Wisconsin
• Endemic Salmonellosis
• MorindaMax™ Nutritional Support for Calves was fed in a controlled blinded study
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Clinical Trial• 226 calves enrolled
– Total Protein ≥ 5.0 g/dL
– IgG ≥ 1200 mg/dL
– Birth weight 70-120 lbs
– Alive 48 hrs
– 10 calves dropped
• Trial A (n=106) followed by Trial B (n=110)
• Calves fed 0, 15 (A) or 30 (B) mL of MorindaMax po, q12h for 21 days
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Outputs• Daily health scores
• Weaning age
• Average daily gain
• Medical treatments
• Economic impacts
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Scoring example
0 0
0
1 2
3
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Calf Health Scoring Criteria
Score Temperatur
e
Cough Nasal
Discharge
Eyes or ears Fecal
Score
0 100-100.9° F
(37.8-38.3°
C)
None Normal serous
discharge
Normal Normal
1 101-101.9° F
(38.3-38.8°
C)
Induce
single cough
Small amount of
unilateral, cloudy
discharge
Small amount
of ocular
discharge
Semi-
formed,
pasty
2 102-102.9° F
(38.9-39.4°
C)
Induced
repeated
cough or
occasional
spontaneous
cough
Bilateral, cloudy
or excessive
mucus discharge
Moderate
ocular
discharge for
both eyes or
slight ear
drop
Loose but
enough
consistency
to stay on
bedding
3 103° F
( 39.4° C)
Repeated
spontaneous
coughing
Copious,
bilateral,
mucopurulent
nasal discharge
Head tilt or
both ears
dropped
Watery,
sifts
through
bedding
Combined Respiratory Score
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Treatment protocol
Health Score Treatment Cost*
Fecal = 2 Electrolytes $0.03
Fecal = 3 Electrolytes (2x) $0.06
Fecal = 2 + blood Electrolytes + Gentamicin $0.03 + 0.26
Fecal = 3 + blood Electrolytes (2x) + Gentamicin $0.06 + 0.26
Total respiratory
score of ≥ 5
1st Ceftiofur $3.20
2nd Tulathromycin $7.56
3rd Trimethoprim Sulfa $1.40
Temperature ≥
39.4°C (≥ 103°F)
Flunixin meglumine $0.50
Incidence of Disease:Fecal score ≥ 2 Total respiratory score of ≥ 5Temperature ≥ 39.4° C
* Cost of treatment per calf per day
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Economic Analysis• Cost figures were attained from the Economic Costs and Labor
Efficiencies Associated with Raising Dairy Herd Replacements on Wisconsin Dairy Farms and Custom Heifer Raising Operations report (2007) as analyzed by the Intuitive Cost of Production Analysis (ICPA) computer model.
Figures are based on:• Weaning ages (days)
• Disease incidences derived from health scores
• Treatment costs
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Results: Trial A
Significantly more calves weaned by 6 weeks of age that had received noni puree
p= 0.0385
Approx. 30% more 0
10
20
30
40
50
60
70
80
Control Half Dose Full Dose
Percent weaned by 6 weeks of age
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Results of Trial B• Significantly fewer
respiratory, fecal and total treatments
• In control calves:
– 2.5X Respiratory
– 2X Fecal
– 2.2X Resp + Fecal scores
0
10
20
30
40
50
60
70
Resp. Fecal Total
Control Half Dose Full Dose
Number of medical treatments
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Summary• Trial A
– 30% increase in calves weaned by 6 weeks of age
– Monetary savings of $13.56 per calf
• Trial B
– Significantly fewer respiratory, fecal and total treatments
– Monetary savings of $8.19 per calf
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Summary of Calf Studies
• Significant E. coli phagocytosis on day 14 compared to control
• E. coli bacterial killing increased over time
• Significant increase in % activation of CD4+ T cells on day 3
• Significant increase in % activation of CD8+ T cells on day 3
• Significant effect over time for CD8+ T cells
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Summary of Findings• Significantly more calves weaned by 6 weeks
of age that had received 30 mL noni puree a day
(p = 0.0385).
– Monetary savings of $13.56 per calf
• Significantly fewer respiratory, fecal and total treatments
– Monetary savings of $8.19 per cal
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Effect of MorindaMax on performance of calves with APT or FPT (field trial)
• 180 calves divided into 6 treatment groups.– TP> 5.2 Adequate Passive Transfer
• control
• 15ml po BID 10d
• 30ml po BID 10d
– TP<5.2 Failure Passive Transfer• control
• 15ml po BID 10d
• 30ml po BID 10d
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36
1.319
1.196
1.3531.327
1.351
1.471
0.000
0.200
0.400
0.600
0.800
1.000
1.200
1.400
1.600
>5.2 TP <5.2 TP
lb
Average daily gain
control 0.5oz BID 1.0 oz BID
Utah ‘08
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Summary
• Small improvement (2.6%) in average daily gain using MorindaMax BID for 10d in calves with APT.
• Feeding 30ml BID MorindaMax for 10d in calves with FPT resulted in a 23% increase in average daily gain.
• Feeding 15ml BID MorindaMax for 10d in calves with FPT resulted in a 11% increase in average daily gain.
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Animal Research - Piglets
• Improved gains early in nursery phase.
• Carry-over effect after receiving MorindaMax Nutritional Support for Piglets.
• Improved Gain:Feed.
• Improved average daily gain.
38
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5%39Kansas State University '07
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Prolonged Storage & ADFI
266
455
596
293
500
633
317
562
624
200
450
700
d 7 to 14 d 14 to 21 d 21 to 28
Control
Stored
Fresh
g
*Control vs M. citrifolia (P < 0.09)
**Storage effect (P < 0.002)
SE = 21 *
SE = 22 * **
SE = 31
40
Kansas State University '07
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M. citrifolia Concentration & ADG(d 0 to 7)
150
215
280
0% 0.75% 1.5% 3.0% 6.0%
197
223
198
228
177
g/d
SE 12
Quadratic (P < 0.03), Quartic effect (P < 0.07)
41Kansas State University '07
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M. citrifolia Concentration & G:F (d 0 to 7)
950
1100
1250
0% 0.75% 1.5% 3.0% 6.0%
1059
11631151
1135
1041
g/k SE 46
Quadratic effect (P < 0.08)
Kansas State University '0742
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Animal Research - Broilers
• Improved average daily gain.
• Improved Gain:Feed.
• Improved total weight gain.
43
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Effect of M. citrifolia on Broiler Chick Performance 1-42 Days of Age
2564
2661
2500
2520
2540
2560
2580
2600
2620
2640
2660
2680
control treated 0%
wt gain Kg
P<0.01
44Southern Poultry Research & UGA '05
3.6% improvement
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Effect of M. citrifolia on Broiler Chick Performance 1-42 Days of Age
1.77
1.74
1.725
1.73
1.735
1.74
1.745
1.75
1.755
1.76
1.765
1.77
1.775
control treated 0%
feed/gain
45Southern Poultry Research & UGA '05
2% improvement
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Thank You