Professional Documents
Culture Documents
CHRISTOPHER K. REYNOLDS
Ruminant Nutrition Laboratory
US Department of Agriculture, ARS
Beltsville, MD 20705
NEFA release from adipose tissue can occur liquid protein supplement, 4.7% soybean meal,
even prior to copious milk secretion. Recently, 2.3% tallow, 1.6% sodium phosphate, .5%
our laboratory (2) demonstrated that hepatic magnesium oxide, and .5% urea (OM basis).
TG content at 1 d postpartum was significantly Feed and orts from each cow were sampled
reduced in cows that were force-fed prepartum three times weekly. Three weekly samples
via rumen cannulas. Plasma glucose at 2 d were composited into one sample for each
prepartum also was elevated in force-fed cows week. Forage and concentrate DM were deter-
and was negatively correlated with liver TG at mined by oven-drying at 60·C for calculation
d I postpartum. of DM!. Cows received a l-L oral drench of
Propylene glycol (pG) is a glucogenic com- PG (Phoenix Pharmaceutical, Inc., S1. Joseph,
pound used to treat ketosis postpartum. The MO) or water once daily at 1230 h from 7 d
majority of PG escapes the rumen intact; a prior to expected calving until parturition.
portion is metabolized to propionate (7). Blood and plasma samples were as
Propylene glycol escaping rumen fermentation described by Skaar et al. (29) and were taken
is converted to glucose by the liver, primarily via the coccygeal vein on d 16 and 17 prior to
via the lactaldehyde pathway and subsequent expected calving. Blood was also sampled
oxidation to lactate (18). Administration of PG daily from 7 d prepartum to 3 d postpartum
to postpartum cows reduced plasma concentra- and on d 7, 14, and 21 postpartum. During
tions of NEFA and ketones (27). Addition of treatment, blood samples were taken 90 min
PG to the ration, compared with dietary starch, postdrenching. On d 5 prior to expected calv-
produced a greater molar percentage of rumen ing, additional blood was sampled at IS-min
propionate and elicited a greater insulin re- intervals postdrenching for 2.5 h. One cow
sponse (9). The objective of our study was to assigned to the control group calved before
determine whether prepartum administration of multiple sampling. Liver was sampled by
PG could reduce periparturient fatty liver. puncture biopsy at 17 d prior to expected
calving and at 1 and 21 d postpartum. All
plasma samples were analyzed in random or-
MATERIALS AND METHODS der. Plasma was analyzed for glucose and
NEFA (29). Plasma was deproteinized (29) fol-
Experimental De81gn end Sampling Protocol lowing storage at -20·C and then immediately
analyzed for BHBA (11). Plasma insulin was
Twenty-four multiparous Holstein cows, analyzed by radioimmunoassay (RIA) with a
pregnant and nonlactating, were initially paired modification to the kit procedure (Coat-a-
according to calving date and assigned either Count®; Diagnostic Products Co., Los An-
to a control (n = 11) or PG treatment (n = 13) geles, CA) to account for relatively low insulin
group. However, one cow assigned to the con- concentrations in bovine plasma. Zero calibra-
trol group was mistakenly treated, leaving an tor (100 JoLI) was added to 200 JoLI of each
unbr!lanced design. Cows began the trial at 21 standard; the standard concentration was
d p:i.or to expected calving and were fed diluted by a factor of .66. Plasma (300 JoLI) was
alfaL"a-bromegrass hay for ad libitum intake then used in the determination of each un-
and 1.8 kgld of concentrate separately to meet known sample. Insulin RIA intraassay varia-
NRC (20) dry cow requirements. The prepar- tion was 7.9%, and interassay variation was
tum concentrate contained 34% soybean meal, 6.8%. Glucagon was analyzed by RIA accord-
18.3% cracked corn, 18.3% soybean hulls, ing to the method of Reynolds et al. (25),
8.7% corn gluten meal, 8.7% blood meal, 8.7% except that Unger 30 K antibody was used.
meat and bone meal, 1.8% trace-mineralized Glucagon RIA intraassay variation was 8.2%,
salt, 1.3% vitamins A, D, and E (2,643,171 IUI and interassay variation was 6.0%. Liver tissue
kg of vitamin A, 881,057 IU/kg of vitamin D, was assayed for total lipid and TG (29).
and 881 IU/kg of vitamin E), and .3% mag- Milk production was recorded daily through
nesium oxide (OM basis). Following calving, 21 d postpartum. Milk samples were taken at
cows were fed 50% alfalfa silage and 50% a.m. and p.m. milkings on d 13, 14,20, and 21
concentrate (OM basis) as a TMR for ad libi- postpartum and analyzed for fat and protein
tum intake. The postpartum concentrate con- percentages and somatic cells by infrared spec-
tained 77.4% shell corn, 7.0% molasses, 6.0% trophotometry (Wisconsin DHI Cooperative
Journal of Dairy Science Vol. 76. No. 10. 1993
EFFECT OF PROPYLENE GLYCOL ON FATTY LIVER 2933
Laboratory, Appleton, WI). Body weight and Period represented day for OMI and milk
body condition score (6), averaged from two production and week for milk composition and
evaluators, were recorded once prior to treat- FCM.
ment. Plasma metabolites and hormones were ana-
lyzed independently within sample period (i.e.,
Statistical Analysis pre- or postpartum) according to the following
=
model: Yijk p. + b + Ti + Cj + Pk + TPik +
Total liver lipid and TG were analyzed in- c;jk where Yijk =
the dependent variable, p. =
dependently within sample period, i.e., 1 or 21 the overall mean of the population, b = the
d postpartum, according to the following regression coefficient for the plasma variable
model: Yi = p. + b + Ti + ej where Yi = the averaged between measurements on d 16 and
dependent variable, p. =the overall mean of the 17 prior to expected calving for cow j, Ti = the
population, b = the regression coefficient for average effect of treatment i, Cj = the average
the liver variable measured at d 17 prior to effect of cow j, Pic = the average effect of
expected calving, Ti = the average effect of period k, and eijk =
the unexplained residual
treatment i, and c; = the unexplained residual element assumed to be independent and nor-
element assumed to be independent and nor- mally distributed. Period represented day for
mally distributed. daily plasma measurements and 15-min inter-
The OMI, analyzed independently within val for multiple plasma samples. Models were
sample period (i.e., pre- or postpartum), milk originally constructed to contain pair (i.e.,
production, and milk composition were ana- block), but the term was dropped because of
lyzed according to the following model: Yijk = events resulting in an uneven experimental de-
p. + Ti + Cj + Pic + TPik. + ejjlc where Yijlc =the sign. Variation between cows was used as the
dependent variable, p. =the overall mean of the error term when treatment effects on OMI,
population, Ti =the average effect of treatment milk production and composition, and all
i, Cj = the average effect of cow j, Pic = the plasma and liver measurements were tested.
average effect of period k, and c;jlc = the Differences were significant at P < .05, indi-
unexplained residual element assumed to be cated a tendency at P = .05 to .15, and were
independent and normally distributed. Analysis nonsignifIcant at P > .15.
of covariance using fIrst lactation 305-d ma-
ture equivalent milk production as the regres- RESULTS AND DISCUSSION
sion coeffIcient was not successful in demon-
strating differences in milk production. The Average BW and body condition score,
regression coeffIcient was not included in the measured prior to treatment, were not different
fInal statistical model for milk production. between treatment groups (Table I). Cows in
..
control group and 10 ± 3.6 d (range = 5 to 15
C.
7
d) for the PG group. Multiple blood samples
were obtained on d 6 ± 1.5 (range = 3 to 8)
.
E
ii:
6
prepartum for the control group and d 7 ± 3.8
(range = 1 to 12) prepartum for the PG group.
5D+....,.....-r-....,-.,........,.-..,..-r-..,.-..,.--.
Fisher et al. (8) reported a depression in -25 -20 -15 -10 -5 0 5 10 15 20 25
forage OMI when PG was fed at 9% of the Day relative to parturition
concentrate mixture (460 mlId). However, to
minimize the potential for confounding effects Figure 2. Plasma glucose concentration of cows given
a drench of 1 L of water (0) or propylene i!ycol (e) once
of PG and OMI, we delivered PG as an oral daily begiMing at an average of 8 ± 2.5 d (X ± SO; range,
drench. Both groups experienced a moderate 2 to II d) prepartum for cows assigned to the control
depression in feed intake immediately prepar- group and 10 ± 3.6 d (range, 5 to 15 d) prepartum for cows
tum, but the depression was similar between treated with propylene glycol through parturition (unad-
justed means). A prepartum treatment effect (P < .01)
treatments (Figure I). occurred. Postpartum plasma glucose concentrations were
Plasma glucose was elevated by PG not significantly different between groups. The pre- and
throughout treatment (Figure 2). Plasma glu- postpartum standard errors were 2.1 and 1.9 mg/dl. respec-
cose concentrations peaked at 75 min post- tively.
drenching (Figure 3). The elevation of plasma
glucose agrees with that of other reports (9,
27), indicating that PG is an effective gluco- cally in both groups at parturition (Figure 2),
genic agent. Plasma glucose increased dramati- which agrees with results of Schwalm and
Schultz (28). Increased plasma glucose concen-
tration could be a result of increased gluconeo-
2 genesis, glycogenolysis, or both, stimulated by
catecholamines and glucocorticoids (21) at par-
turition.
2 Plasma insulin concentrations were signifi-
cantly elevated by 15 min following PG ad-
ministration (Figure 4). Insulin concentration
peaked prior to peak glucose concentration.
:iii Significant amounts of PG may have been
c
metabolized to propionate, which stimulates
pancreatic insulin secretion (14), or PG or in-
termediates of PG metabolism might be direct
insulin secretagogues. Insulin concentrations
5+-....,.-..,......,.-,......,.-..,.-...,.-..,.-..,.--. were depressed on the day of calving (Figure
-25 -20 -15 -10 -5 0 5 10 15 20 25 5), in contrast to results of Kunz et al. (17),
Day relative to parturition who reported an insulin surge at calving. Our
Figure 1. The OM! of cows given a drench of 1 L of blood samples were taken at 24-h intervals;
water (0) or propylene glycol (e) once daily beginning at therefore, an acute insulin increase might not
an average of 8 ± 2.5 d ex
± SO; range, 2 to II d) have been detected. Plasma insulin concentra-
prepartum for cows assigned to the control group and 10 ± tions were not different between treatment
3.6 d (range, 5 to 15 d) prepartum for cows treated with groups following parturition.
propylene glycol through parturition (unadjusted means).
Differences in DMI were not significant between groups
Prepartum plasma glucagon concentration
pre- or postpartum. The pre- and postpartum standard was not affected by prepartum PG administra-
errors were .5 and .9 kg/d, respectively. tion (Figure 6), but daily samples were ob-
8 .80
.70
7
;;
Q E
Q .60
E c:
.
oi
0
U
7
.5
'3
WI .50
.
::I
..a::..
'6> .5
6
E
.
E
WI
a::
.40
6 .30
5s+-......r--"'T""--r-T"""....,..-r-""T'"--.r--"'T"""""
o 15 30 45 60 75 90 105120 135150
.20
-25 ·20 ·15 ·10·5 0 5 10 15 20 25
Postdrenching, min Day relative to partu rillon
Figure 3. Plasma glucose concentration of cows given Figure 5. Plasma insulin concentration of cows given a
a dren<2!! of 1 L of water (0) or propylene glycol (e) on d 6 drench of 1 L of water (0) or propylene &!ycol (e) once
± 1.5 (X ± SO; range, 3 to 8) prepartum for cows assigned daily beginning at an average of 8 ± 2.5 d (X ± SO; range.
to the control group and d 7 ± 3.8 (range, I to 12) 2 to 11 d) prepartum for cows assigned to the control
prepartum for cows treated with propylene glycol (unad- group and 10 ± 3.6 d (range, 5 to 15 d) prepartum for cows
justed means). A treatment effect if' < .01) and a treatment treated with propylene glycol through parturition(un-
by time interaction (P < .05) occurred. The overall stan- adjusted means). A prepartum treatment effect (P < .001)
dard error was 1.2 mgldl. occurred. Postpartum plasma insulin concentrations were
not significantly different between groups. The pre- and
postpartum standard errors were .034 and .030 ng/mI.
respectively.
.30
.27
E
Q
c:
C
E
Q
c:
..
0
01
u
::I
.25
.s'3
.. ..
'6> .22
....
.5
.75
.E
WI
a::
E
a:: .50
.17$.j-...,r--.....--r-.,..."""'!"-"!'"--r::--:w:--:r:~
.25 ·20 ·15 ·10·5 0 5 10 15 20 25
Day relative to parturition
.250+--..-..,.-..-..,.-,..........- ...........-.:;::.......
o 15 30 45 60 75 90 105 120135150
Figure 6. Plasma glucagon concentration of cows
Postdrenchlng. min given a drench of 1 L of water (0) or propylene llli'col (e)
once daily beginning at an average of 8 ± 2.5 d (X ± SO;
Figure 4. Plasma insulin concentration of cows given a range. 2 to 11 d) prepartum for cows assigned to the
dren£!! of 1 L of water (0) or propylene glycol (e) on d 6 ± control group and 10 ± 3.6 d (range. 5 to 15 d) prepartum
1.5 (X ± SO; range, 3 to 8) prepartum for cows assigned to for cows treated with propylene glycol through parturition
the control group and d 7 ± 3.8 (range, 1 to 12) prepartum (unadjusted means). Pre- and postpartum plasma glucagon
for cows treated with propylene glycol (unadjusted means). concentrations were not significantly different between
A treatment effect (P < .001) and a treatment by time groups. A day effect occurred across pre- and postpartum
interaction if' < .001) occurred. The overall standard error periods (P < .001). The pre- and postpartum standard
was .047 ng/mI. errors were .006 and .009 ng/mI. respectively.
2
.25
~
E'" 2
.(
E III
g. :z: 1
..
.22 III
co
~::J .
E
Ii:
1
.
Q
E
5
:I
;;:
O+-...,.-,.-..,......,-.,-..,.-,.-............
-25 -20 -15 -10 -5 0 5 10 15 20 25
ro--..
Day relative 10 parturition
11""5--'3r"'"0--'4r"'"5"""60-""75-"'90-1"!0~5-1"'2~0-1~5"":"!15 0
.17 50101-""1
Figure 9. Plasma BHBA concentration of cows given a
Posldrenchlng, min drench of I L of water (0) or propylene &!ycol (e) once
daily beginning at an average of 8 ± 2.5 d (X ± SO; range.
Figure 7. Plasma glucagon concentration of cows 2 to 11 d) prepartum for cows assigned to the control
given a drench of 1 L of water (0) or propylene glycol (e) group and 10 ± 3.6 d (range. 5 to 15 d) prepartum for cows
on d 6 ± 1.5 (X ± SD; range. 3 to 8) prepartum for cows treated with propylene glycol through parturition (unad-
assigned to the control group and d 7 ± 3.8 (range. 1 to 12) justed means). A prepartum treatment effect (P < .001)
prepartum for cows treated with propylene glycol (unad- occurred. Postpartum plasma BHBA concentrations were
justed means). Plasma glucagon concentrations were not not significantly different between groups. The pre- and
significantly different between groups. The overall stan- postpartum standard errors were .7 and 2.1 mg/dl. respec-
dard error was .009 ng/ml. tively.
ControI2 SE SE p
gradual increase in plasma NEFA concentra- change markedly during elevated NEFA up-
tion prior to parturition, or the acute surge at take (3). Both groups experienced a smaller
parturition, to development of hepatic TG ac- increase in liver TG concentration between 1
cumulation is not known. Plasma NEFA con- and 21 d postpartum, which supports the con-
centration of cows treated with PG was signifi- cept that the majority of hepatic TG deposition
cantly lower than that of cows in the control occurs between 16 d prepartum and 1 d post-
group prior to parturition, most likely because partum (12).
of inhibition of adipose adenylate cyclase ac- Correlation analysis was performed to iden-
tivity and lipolysis by elevated plasma insulin tify prepartum parameters related to peripar-
(32). We have no explanation for the tendency turient hepatic TG accumulation (Table 3).
toward lower postpartum plasma NEFA con- Prepartum plasma NEFA concentration was
centration in cows receiving PG because treat- highly correlated with liver TG at 1 d postpar-
ments were terminated at calving. tum. The likely importance of maintaining
Prepartum plasma BHBA concentrations
were decreased by PG treatment (Figure 9).
Lower BHBA concentrations may reflect
higher concentrations of insulin and lower con- TABLE 3. Correlation coefficients between prepartum
centrations of NEFA. Insulin may act to reduce plasma metabolites. hormones. DMI. or body condition
score and liver triglyceride (TO) concentrations at I and 21
NEFA supply to the liver, to inhibit hepatic d postpartum.
ketogenesis, to increase ketone utilization by
peripheral tissue (4), or trigger a combination Liver TG
of these effects. Postpartum plasma BHBA Prepartum 1 d 21 d
concentrations were similar between treat- parameter1 postpartum postpartum
ments. r p r P
At 1 d postpartum, total liver lipid and TG Glucose -.49 .02 -.36 .09
were significantly lower in cows treated with NEFA .46 .03 .11 .60
PG, and the differences were maintained BHBA .36 .08 .45 .03
Insulin -.49 .02 -.09 .69
through 21 d postpartum (Table 2). The differ- Glucagon .03 .91 -.07 .75
ence in liver TG content postpartum accompa- DMI -.25 .24 -.22 .31
nied a difference in plasma NEFA concentra- Body condition
tion between treatment groups. Changes in score2 .23 .29 .12 .56
liver TG are a more sensitive measure of lplasrna concentrations and DMI are expressed as
hepatic lipid accretion because they ignore means between d I and 7 prepartum.
phospholipid and cholesterol, which do not 2Scale ranged from 1 = thin to 5 = obese.
lOne liter of water (control) or propylene glycol (PG) given once daily as an oral drench beginning at an average of 8
d (control) or 10 d (PG) prepartum through parturition.
prepartum plasma glucose and insulin concen- accumulation. This reduction could be
trations in indicated by their strong negative achieved by improved energy balance in late
correlation with liver TG at 1 d postpartum, gestation through maintenance of prepartum
but not at 21 d postpartum. Plasma BHBA DMI, enhanced carbohydrate and insulin status
concentration and liver TG accumulation were through administration of glucogenic precur-
positively correlated at 1 and 21 d postpartum. sors during the last few days of gestation, or
Milk production and composition were not both. Increasing dietary nonstructural carbohy-
affected by prepartum PG administration (Ta- drates may have limited effectiveness, con-
ble 4). In a previous study (2), cows force-fed sidering the inherent decline in feed intake
via rumen cannulas had a lower liver TG con- immediately prepartum and complications that
tent at 1 d postpartum and produced more milk can arise from rumen acidosis. Administration
through 28 d postpartum. Longer postpartum of PG elevated prepartum plasma glucose con-
periods would be desirable for detection of centrations; however, the stimulation of insulin
treatment effects on milk production. Cows secretion probably more directly modulated
used in this experiment were in moderate body prepartum plasma NEFA concentration. Fur-
condition (Table 1). Production benefits from ther research is needed to evaluate the possible
PG administration may have been realized if role of insulin as a preventative agent in the
experimental cows had been in excessive body alleviation of periparturient fatty liver.
condition, possibly predisposing them to fatty
liver and ketosis (24). Differences between ACKNOWLEDGMENTS
groups in animal health were not observed.
The PG dose given in this study was in The care and feeding of the experimental
excess of the standard postpartum field recom- cows was coordinated by Leland Danz, whose
mendation of 230 to 410 mUd (15). The mini- contribution is greatly appreciated. The authors
mum effective prepartum dose of PG for the also thank Dawn Miksic for her assistance in
alleviation of fatty liver is unknown. A sum- the laboratory.
mary of data from our laboratory and other
reports (9, 26, 27) indicate that the antilipolytic REFERENCES
effect of PG increases linearly from 117 mUd
of PG to 1 Ud of PG (r2 = .86). I Bell.A. W. 1980. Lipid metabolism in the liver and
selected tissues and in the whole body of ruminant
animals. Prog. Lipid Res. 18:117.
CONCLUSIONS 2 Bertics. S. J.• R. R. Grummer. C. Cadomiga-Valino.
and E. E. Stoddard. 1992. Effect of prepartum dry
Prepartum PG administration reduced matter intake on liver triglyceride concentration and
hepatic TG accumulation by 32 and 42% at 1 early lactation. 1. Dairy Sci. 75:1914.
and 21 d postpartum, respectively. Evidence 3 Butler. S. M.• A. Faulkner. V. A. Zammit, and R. G.
Vernon. 1988. Fatty acid metabolism of the perfused
from this experiment suggests that reducing caudate lobe from livers of fed and fasted non-
prepartum plasma NEFA concentration is im- pregnant and fasted late pregnant ewes. Compo Bio-
portant for reducing periparturient hepatic TG chern. Pbysiol. B. Compo Biochem. 91:25.