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The Journal of Veterinary Science. Photon 116 (2015) 457-464
https://sites.google.com/site/photonfoundationorganization/home/the-journal-of-veterinary-science
Original Research Article. ISJN: 1784-6372: Impact Index: 5.80
The Journal of Veterinary Science Ph ton
A herbal tool- AV/KPC/10 to check the sub clinical ketosis in cows
M.F.M.F.Siddiqui
a
, S.P.Waghmare
a
, S.G.Mode
a
, K.S.Pajai
a
, Adarsh
b*
, S.Maini
b
, K.Ravikanth
b
a
Teaching Veterinary Clinical Complex, PGIVAS, Akola, India
b
Clinical Research & Development, Ayurvet Limited, Baddi, India
Article history:
Received: 29 April, 2015
Accepted: 01 May, 2015
Available online: 03 July, 2015
Corresponding Author:
Adarsh.*
Trainee clinical research
Email: clinical (at) ayurvet.in
Siddiqui
M.F.M.F.
Hospital Registrar
Waghmare S. P.
Hospital Superintendent
Mode S.G.
Associate Professor
Pajai K.S.
Assistant Professor
Maini S.
Deputy Manager
Ravikanth K.
C.E.O
Abstract
Ketosis is a metabolic disorder that occurs in cattle
when energy demands exceed energy intake and
result in a negative energy balance. The current
study was designed to evaluate the efficacy of
herbal gel AV/KPC/10 (M/S Ayurvet Limited) for
control of sub clinical ketosis in cows. 18 animals
positive for sub clinical ketosis were randomly
divided into Group I (n=6), treated with AV/KPC/10
at dose 200 gm twice a day orally for first two days
followed by 100 gm once a day for next 3 days,
Group II (n=6), treated with Liquid Anabolite at dose
200ml twice a day orally for two days, followed by
100 ml twice a day for 3 days and Group III (n=6),
treated with Inj. Dextrose 25% at dose 500 ml
intravenously once in a day for three days. Total 12
healthy cows negative for sub clinical ketosis were
selected and divided randomly into two equal
groups, control Group IV (n=6), cows negative for
sub clinical ketosis were kept without any treatment
and treatment Group V (n=6), cows negative for sub
clinical ketosis were supplemented with AV/KPC/10.
Parameters viz. Ketosis diagnosis, biochemical
estimation and milk yield were recorded before
treatment (‘0’ day) and on 3
rd
, 7
th
and 30
th
day post
treatment. Statistical analysis of data revealed
efficacy of herbal gel AV/KPC/10 (M/S Ayurvet
Limited) in control and treatment of Ketosis in cattle.
Citation:
Siddiqui M. F. M. F., Waghmare S. P., Mode S.G., Pajai K.S.,
Adarsh, Maini S., Ravikanth, K., 2014. A herbal tool-
AV/KPC/10 to check the sub clinical ketosis in cows. The
Journal of Veterinary Science. Photon 116, 457-464.
All Rights Reserved with Photon.
Photon Ignitor: ISJN17846372D790703072015
1. Introduction
During calving nutritional requirements increase
rapidly with lactation. Ketosis, or acetonaemia, is a
multifactorial disorder of energy metabolism i.e.
metabolic disorder and is largely a management
influenced disease in dairy cows in early lactation
(Shpigel et al., 1996; Ingvartsen, 2006).
Hyperketonemia is associated with an increase of
oxidative stress levels (Youssef et al., 2010). Most
of the animals in developing countries are fed on
agriculture by-products and low quality crop
residues have low nutritive value, i.e. low contents
of metabolizable energy and crude protein (Vahora
et al., 2013). Normally ruminants have low levels
of blood glucose and during heavy milking when
glucose utilization is high, even slight fall is
enough to put the animal in a hypoglycemic state
(Dufflied, 2006). During the early post partum
period, milk production increases dramatically,
while energy intake may not be adequate to sustain
the higher production level. This results in negative
energy balance and cows metabolize fat to meet
their energy needs (Weber et al., 2012). During
early lactation incidence rates of ketosis between
about 40% and 60% (Duffield et al., 1998).
Nowroozi Asl et al., (2011) studied that 97 percent
of the cows experienced at least one episode of
subclinical ketosis during the sixth week post
partum period there was a statistical relation
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between the prevalence of sub clinical ketosis and
milk production. Ketosis leads to heavy economic
losses. Geishauser et al. (2011) estimated cost of
sub clinical ketosis per case was $78 CAD. A study
by Varga (2004) summarized ketosis costs at $140
for treatment, and the total cost at $2,520 per year
for a 120-cow farm. Economic losses are
considerable due to decreased milk production, cost
of treatment and occasionally from deaths or
disposal of affected cows. The wide ranges of
treatments have been recommended for ketosis.
Most of these are intended to alleviate the
hypoglycemia and increase the glycogen content of
liver, thereby, restoring the carbohydrate
metabolism and aims to provide an energy boost to
overcome negative energy balance for optimizing
the milk production.
1.1 Objective of Research
The current study was designed to evaluate the
efficacy of herbal gel AV/KPC/10 (M/S Ayurvet
Limited) in the treatment of sub clinical ketosis in
cows.
2. Materials and Methods
Total 87 lactating cows in early mid lactation were
screened for ketosis by Modified Rothera’s test
(Kelly, 1974) and Keto-Diastix Strip test in urine.
18 animals positive for sub clinical ketosis were
selected and divided randomly into three equal
groups, comprising of 6 cows in each group. Group
I, cows positive for sub clinical ketosis were
treated with AV/KPC/10 @ 200 gm twice a day
orally for first two days followed by 100 gm once a
day for next 3 days. Group II, cows positive for sub
clinical ketosis were treated with Liquid Anabolite
@ 200ml twice a day orally for first two days,
followed by 100 ml twice a day for 3 day. Group
III, positive for sub clinical ketosis were treated
with Inj. Dextrose 25% @ 500 ml intravenously
once in a day for three days. Total 12 healthy cows
negative for sub clinical ketosis were selected and
divided randomly into two equal groups,
comprising of 6 cows in each group. Control Group
IV, cows negative for sub clinical ketosis were kept
without any treatment and treatment Group V,
cows negative for sub clinical ketosis were
supplemented with AV/KPC/10 @ 200 gm twice a
day orally for first two days followed by 100 gm
once a day for next 3 days. Biochemical parameters
viz. plasma glucose (mg/dl), serum triglyceride
(mg/dl), serum calcium (mg/dl), serum phosphorus
(mg/dl) were determined on ‘0’ day (before
treatment) and on 3
rd
, 7
th
and 30
th
day post
treatment. The milk yield was recorded before
treatment (‘0’ day) and daily after treatment during
the experimental period.
2.1 Statistical analysis
The data collected during the present study with
respect to different parameters was analyzed
statistically by two ways factorial design using web
based agricultural statistics software package
(WASP), ICAR Research Complex, Ela, Old Goa.
3. Results and Discussion
3.1 Detection of ketosis in cows (Ketone
bodies)
In Group I, Group II and Group III cows were
positive for sub clinical ketosis on day ‘0’ (before
treatment) diagnosed by Modified Rothra’s Test
and Keto-Diastix strip test.
3.1.1 Modified Rothra’s Test
In Modified Rothra’s Test, a mixture of reagent
was taken up to half of the test tube. Urine sample
was added slowly through the wall of test tube. The
positive test showed purple colour formation in
urine. The results are recorded as follows.
Slight purple colour +
Moderate purple colour ++
Dark purple colour +++
3.1.2 Keto-Diastix strip test
In Keto-Diastix strip test, dipstick containing the
salt nitroprusside, which provide semi quantitative
results. This test is based on the development of
colours ranging from buff pink for a negative
reading to purple when acetoacetic acid reacts with
nitroprusside. The colour intensity varies with the
amount of acetoacetic acid present in urine. Read
the ketone test area by comparing with the colour
chart at exactly 40 seconds after wetting and
recorded the results. The Keto-Diastix® diagnostic
test can be read in five categories: 1) negative (0
mg/dl), 2) trace (5 mg/dl), 3) small (15 mg/dl), 4)
moderate (40 mg/dl), and 5) large (greater than 80
mg/dl of AcAc).
After treatment with AV/KPC/10, Anabolite and
Dextrose 25% injection all the cows in respective
groups were found negative for sub clinical ketosis
on day 7th, 14th and 30th day. In Control Group
IV, initially on day 0 all the cows were negative for
sub clinical ketosis. During subsequent period 4
animals were found positive for sub clinical ketosis
on 30
th
day. This means in untreated group 75%
new cases of ketosis were observed. In Group V
all the animals were negative for ketosis on day ‘0’
and no incidence of ketosis was observed on the
day 3
rd
, 7
th
and 30th day after AV/KPC/10
supplementation. Which means AV/KPC/10 was
efficacious to control the new incidences of ketosis.
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3.2 Biochemical studies
3.2.1 Plasma Glucose (mg/dl)
In ketotic cows during early lactation period
glucose supply is not sufficient for milk production
(Padilla et al., 2005). Group I treated with
AV/KPC/10 showed significant
(P<0.05)improvement in plasma glucose level in
comparison to groups treated with liquid
Anaboliteand Injection Dextrose 25%. In
AV/KPC/10 treated group I cows, on day 3
rd
(53.85
mg/dl) a significant (P<0.05) increase in plasma
glucose level was observed in comparison of day 0
(43.05 mg/dl) and level was maintained till 30
th
day. The overall mean plasma glucose level
(mg/dl) in AV/KPC/10 treated group (I) was 51.71
mg/dl. Increase in plasma glucose concentration is
well in comparison with other supplemented
groups. In Anabolite treatment group (II) and
dextrose injection treatment (III) group, the overall
mean plasma glucose values were 50.51 mg/dl and
49.94 mg/dl respectively (table 1). When treating
cows for negative energy balance, it is essential to
meet the need for glucose so that the ketogenic
process in the liver are reversed (Sezer et al.,
2010). The improvement in plasma glucose level
was because the supplementation may have
facilitated cellular uptake of glucose, suppresses
fatty acid mobilization and stimulate hepatic
gluconeogenesis (Radostits et al., 2000).
Table 1: Mean values of plasma glucose (mg/dl) in different groups before treatment and at different intervals after
treatment
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group I 43.05
a
±0.77
53.85
efgh
±0.92 54.10
fgh
±0.57 55.83
hi
±0.42 51.71
b
±1.10
Group II 44.93
a
±0.81
50.98
bc
±0.30 52.93
cdefg
±0.50 53.18
defg
±0.49 50.51
a
±0.54
Group III 44.57
a
±0.80
51.03
bcd
±0.60 52.31
cdef
±0.64 51.83
cde
±0.37 49.94
a
±0.71
Group IV 56.90
i
±0.63
54.98
ghi
±0.54 53.70
efgh
±1.01 49.00
b
±1.16 53.64
c
±0.73
Pooled Mean (B) 47.36
a
±1.21
52.71
b
±0.47 53.26
b
±0.36 52.46
b
±0.61
Similar superscript shows non significant differences
The analysis of variances indicated significant
(P<0.05) rise in plasma glucose on 3
rd
(52.68
mg/dl), 7
th
(57.77 mg/dl), and 30
th
(56.38 mg/dl)
day (mean, 55.56 mg/dl) post AV/KPC/10
supplementation in group V (table 2). AV/KPC/10
supplementation has significantly maintained the
plasma glucose concentration. In control group IV
i.e. unsupplemented group there was a decrease in
plasma glucose levels from 3rd (54.98), 7th (53.70)
and 30th (49.00) day (Mean 53.64 mg/dl).
Table 2: Mean values of plasma glucose (mg/dl) in different groups of healthy animals before supplementation and at
different intervals after supplementation
Intervals Before Treatment After Treatment Pooled Mean
(A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group IV 56.90
de
±0.63
54.98
bcd
±0.54 53.70
bc
±1.01 49.00
a
±1.16 53.64
a
±0.73
Group V 52.68
b
±0.88
57.77
e
±0.40 56.38
cde
±0.53 55.42
cde
±0.34 55.56
b
±0.47
Pooled Mean (B) 54.79
b
±0.82
56.37
b
±0.52 55.04
b
±0.68 52.21
a
±1.13
Similar superscript shows non significant differences
3.2.2 Serum Triglycerides (mg/dl)
A significant (P<0.05) decrease in serum
triglycerides levels was observed in all treatment
groups from day 0 to day 30
th
. In group I, the
triglycerides level was 40.37 mg/dl on day 0 which
was significantly reduced to 34.12 mg/dl on day 3
rd
after AV/KPC/10 treatment and decreasing trend
continued till day 7
th
(29.28 mg/dl) and 30
th
(26.18
mg/dl). The overall mean serum triglyceride levels
(mg/dl) in group I treated with AV/KPC/10 was
32.49 mg/dl and was in well comparison with
group II treated with liquid Anabolite (29.29
mg/dl) and group III treated with Injection
Dextrose 25% was 31.57 mg/dl (table 3). Overall
finding indicated that all the treatment found
equally effective in improving triglyceride level in
sub clinical ketotic cows within the experimental
period.
Significant (P<0.05) reduction in serum
triglyceride levels was observed in AV/KPC/10
supplemented Group V (pooled mean, 22.77 mg/dl)
in comparison untreated group IV (pooled mean,
25.06 mg/dl) (table 4). The overall results indicated
the effectiveness of AV/KPC/10 in restoring serum
triglyceride level within treatment regimen.
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Table 4: Mean values of serum triglyceride (mg/dl) in different groups of ketotic cows before treatment and at different
intervals after treatment
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group IV 18.58
a
±1.14
22.58
abc
±0.85 27.25
cd
±1.24 31.82
d
±1.31
25.06
b
±1.16
Group V 25.05
bc
±1.32
23.30
acb
±1.15 22.38
abc
±1.88
20.37
ab
±1.42 22.77
a
±0.77
Pooled Mean (B) 21.82
a
±1.28
22.94
ab
±0.69 24.82
bc
±1.30 26.09
c
±1.96
Similar superscript shows non significant differences
3.2.3 Serum Calcium (mg/dl)
Several workers reported, low level of serum
calcium in sub clinical ketotic cows, which might
be due to increase loss of base in urine to
compensate ketosis induced acidosis (Duffield,
2006). The lower level of serum calcium level in
subclinical ketosis can also be attributed to high
concentration of β-hydroxybutarate (BHBA),
which impairs the absorption and utilization of
calcium in dairy cows during the early lactation
period (Zhang et al., 2009).
The mean values of serum Calcium (mg/dl) in
Group I, group II, group III and group IV of
cowson day ‘0’ (before treatment) and on 3
rd
, 7
th
and 30
th
day after treatment are presented in Table
5. Serum calcium level was improved in
AV/KPC/10 supplemented group I from day 0
(7.85 mg/dl) to day 3
rd
(7.95 mg/dl) and
improvement was also observed on day 7
th
(8.23
mg/dl) till day 30
th
(8.32 mg/dl). Although overall
result indicates that the non significant increase in
serum calcium level was at par in animals treated
with AV/KPC/10, Liquid Anabolite and Inj
Dextrose 25% but the supplement products
maintained the serum calcium level. Even in cows
negative for ketosis i.e. in group IV and V, the
mean serum calcium level varied non significantly
(P<0.05) (table 6).
Table 5: Mean values of serum calcium (mg/dl) in different groups of cows before treatment and at different intervals after
treatment
Intervals Before Treatment After Treatment Pooled Mean(A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group I 7.85
ab
±0.17
7.95
abc
±0.13 8.23
abcd
±0.18 8.32
bcd
±0.16
8.09
ab
±0.08
Group II 7.85
ab
±0.05
7.88
abc
±0.05
8.05
abc
±0.06
8.08
abc
±0.05 7.97
a
±0.03
Group III 8.18
abcd
±0.09
8.13
abcd
±0.12 8.10
a
bcd
±0.85 8.02
abc
±0.06
8.11
ab
±0.05
Group IV 8.65
d
±0.23
8.35
cd
±0.17 8.18
abcd
±0.16
7.77
a
±0.12 8.24
b
±0.10
Pooled Mean (B) 8.13
NS
±0.10
8.08
NS
±0.07 8.14
NS
±0.06 8.05
NS
±0.06
Similar superscript shows non significant differences
Table 6: Mean values of serum calcium level (mg/dl) in different groups of healthy cows before supplementation and at
different intervals after supplementation
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group IV 8.65
NS
±0.23
8.35
NS
±0.17 8.18
NS
±0.16 7.77
NS
±0.12
8.23
NS
±0.05
Group V 8.45
NS
±0.12
8.32
NS
±0.10 8.18
NS
±0.07 8.07
NS
±0.13
8.25
NS
±0.06
Pooled Mean (B) 8.55
b
±0.12
8.33
b
±0.09 8.18
ab
±0.08 7.92
a
±0.09
Similar superscript shows non significant differences
3.2.4 Serum Phosphorus (mg/dl)
An important hydrogen buffering system involves
phosphorus functions in the blood and is a critical
component of nucleic acids, adenosine triphosphate
(ATP), and adenosine monophosphate (AMP)
(Cunningham, 2002). The lower level of serum
phosphorus in ketosis might be due to
compensation of ketosis induced acidosis, which
triggers excretion of phosphorus via urine (Fatur,
1994).
Table 7: Mean values of serum phosphorus (mg/dl) in different groups of ketotic cows before treatment and at different
intervals after treatment
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group I 5.18
ab
±0.20
5.33
bc
±0.08 5.48
bc
±0.09 5.42
bc
±0.09
5.35
NS
±0.06
Group II 5.30
abc
±0.13
5.28
abc
±0.09 5.32
bc
±0.10 5.32
bc
±0.13 5.30
NS
±0.05
Group III 5.55
bc
±0.12
5.47
bc
±0.13 5.52
bc
±0.05 5.47
bc
±0.14
5.50
NS
±0.05
Group IV 5.65
c
±0.22
5.48
bc
±0.10
5.22
abc
±0.08 4.90
a
±0.07
5.31
NS
±0.08
Pooled Mean (B) 5.42
NS
±0.09
5.39
NS
±0.65 5.38
NS
±0.04 5.27
a
±0.07
Similar superscript shows non significant differences
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462
Serum Phosphorus level was improved
significantly (P<0.05) in AV/KPC/10
supplemented group from day 0 (5.18 mg/dl) to day
30
th
(5.42 mg/dl) (table 7). Although statistical
analysis revealed non significant variation
(P<0.05) in serum phosphorus levels between all
groups. According to Rollin et al., (2010)
phosphorus concentrations did not differ between
treatment groups even after injectable
butaphosphan and cyanocobalamin.
Serum Phosphorus concentration significantly
(P<0.05) improved in ketosis negative AV/KPC/10
supplemented group V cows (5.54 mg/dl) in
comparison to ketosis negative unsupplemented
group IV cows (5.31 mg/dl).
Table 8: Mean values of serum phosphorus level (mg/dl) in healthy cows before supplementation and at different intervals
after supplementation
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group IV 5.65
b
±0.22
5.48
b
±0.10 5.22
ab
±0.08 4.90
a
±0.07
5.31
a
±0.08
Group V 5.60
b
±0.16
5.53
b
±0.11 5.53
b
±0.14 5.48
b
±0.09
5.54
b
±0.06
Pooled Mean (B) 5.62
b
±0.13
5.51
ab
±0.07 5.37
ab
±0.09 5.19
a
±0.10
Similar superscript shows non significant differences
3.3 Milk Yield (Kg)
Milk yield is negatively correlated with acetone
concentration in milk (Miettinen, 1994).
Subclinical ketosis in early lactation reduces milk
production; the average total loss in milk yield per
cow has significant impact on economy (Rajala-
Schultz et al., 1999; McArt et al., 2011, Paavo et
al., 1993). The average milk yield (Kg) in Group I,
group II, group III and group IV of cows on day ‘0’
(before treatment) and on 3
rd
, 7
th
and 30
th
day after
treatment are presented in Table 9.
In group I treated with AV/KPC/10, the milk yield
increased by 12.25% (4.58 Kg) on day 3
rd
, 17.1%
(4.78 Kg) on day 7
th
, and 24.26% (5.07 Kg) on day
30
th
from day 0 (4.08 Kg). In group II the increase
in milk yield was 30.91% (5.42 Kg) and in group
III increase in milk yield was 19.52% (5.02 Kg)
from day 0 to 30
th
. On other hand in
unsupplemented group IV milk yield decreased by
2.69% (4.82 Kg) on day 0 to day 30
th
(4.69 Kg).
Overall the pooled mean showed no significant
(P<0.05) variation between various treatment
groups.
Table 9: Average Milk yield (Kg) in different groups of cows at different intervals
Intervals Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group I 4.08
a
±0.24 4.58
abcd
±0.25 4.78
bcde
±0.26 5.07
de
±0.30 4.63
NS
±0.14
Group II 4.14
ab
±0.26 4.61
abcd
±0.29 5.11
de
±0.22 5.42
e
±0.24 4.82
NS
±0.16
Group III 4.20
abc
±0.10 4.52
abcd
±0.08 4.82
cde
±0.15 5.02
de
±0.15 4.64
NS
±0.08
Group IV 4.82
cde
±0.25 4.85
de
±0.27 4.87
de
±0.26 4.69
abcd
±0.26 4.81
NS
±0.12
Pooled Mean (B) 4.31
a
±0.12 4.64
b
±0.11 4.89
bc
±0.10
5.05
c
±0.12
Similar superscript shows non significant differences
A 9.18 % increase in milk yield was observed in
AV/KPC/10 supplemented group V from day 0
(4.90 Kg) to day 30
th
(5.35 Kg) (table 10). The
statistical analysis pooled mean revealed significant
(p< 0.05) overall 7.9 % increase (P<0.05) in milk
yield in AV/KPC/10 supplemented group V (5.19
Kg) as compared to normal control group IV (4.81
Kg).
Table 10: Average Milk yield (Kg) in different groups of cows at different intervals before and after
supplementation.
Intervals
Before Treatment After Treatment Pooled Mean (A)
Groups ‘0’ day 3
rd
day 7
th
day 30
th
day
Group IV 4.82
ab
±0.25
4.85
de
±0.27 4.87
de
±0.26
4.69
abcd
±0.26 4.81
a
±0.12
Group V 4.90
a
b
±0.12 5.22
ab
±0.18
5.28
ab
±0.18
5.35
b
±0.18 5.19
b
±0.09
Pooled Mean (B) 4.86
a
±0.13 5.63
b
±0.16
5.07
ab
±0.16
5.02
ab
±0.18
Similar superscript shows non significant differences
Conclusion
The group of ketotic cows treated with AV/KPC/10
showed improvement in altered levels of plasma
glucose, serum triglycerides and serum phosphorus
on day 3
rd
till day 30
th
post treatment. All
AV/KPC/10 supplemented cows were found
negative for sub clinical ketosis on day 3
rd
, 7
th
and
day 30
th
after supplementation. A significant
increase in milk yield was observed after
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463
AV/KPC/10 supplementation. AV/KPC/10
supplementation was in well comparison with other
treatments.
Acknowledgements
The authors are thankful to Ayurvet Limited,
Baddi, India for providing necessary samples and
guidance and Post Graduate Institute of Veterinary
and Animal Sciences, MAFSU, Akola, for
providing the required facilities, guidance and
support. The authors are also grateful to authors /
editors / publishers of all those articles, journals
and books from where the literature for this article
has been reviewed and discussed.
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