Small Ruminant Research 96 (2011) 178–184

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Small Ruminant Research

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Metabolic adaptations to pregnancy and lactation in German

Blackheaded Mutton and Finn sheep ewes with different
susceptibilities to pregnancy toxaemia
R. Duehlmeier a,∗ , I. Fluegge a , B. Schwert a , N. Parvizi b , M. Ganter a
a
Clinic for Swine, Small Ruminants, Forensic Medicine and Ambulatory Service, University of Veterinary Medicine Hannover, Foundation,
Bischofsholer Damm 15, D-30173 Hannover, Germany
b
Friedrich-Loeffler-Institute (FLI) Federal Research Institute for Animal Health, Institute of Farm Animal Genetics, Höltystrasse 10,
D-31535 Neustadt, Germany

a r t i c l e i n f o a b s t r a c t

Article history: The metabolic adaptations for pregnancy and lactation were evaluated in ewes with a higher
Received 29 March 2010 and a lower risk of being affected by pregnancy toxaemia. Plasma concentrations of glucose,
Received in revised form
insulin, non-esterified fatty acids (NEFA) and ␤-hydroxybutyrate (␤-HB) were determined
29 November 2010
in 7 pregnant, 4.5- to 6.5-year-old German Blackheaded Mutton ewes (higher risk = HR)
Accepted 8 December 2010
Available online 6 January 2011 and 5 pregnant, 2.5-year-old Finn sheep ewes (lower risk = LR) 8 and 2 weeks ante par-
tum (mid- and late pregnancy), peripartal, as well as 2 weeks post partum (lactation) and
2 weeks after weaning. The maternal body weight was monitored at all the reproduc-
Keywords:
Sheep tive stages mentioned. To compare the reproductive performance and the rearing success
Pregnancy toxaemia of the groups, the number and the body weight of the lambs born alive and the ADG of
Glucose the suckling lambs were recorded. In contrast to the ewes of the HR group, the LR sheep
Insulin showed a significant body weight increment of 49% during late, compared to mid pregnancy,
Non-esterified fatty acids and maintained their body weight after parturition. The reproductive performance and the
␤-Hydroxybutyrate rearing success did not differ between HR and LR groups (lambs born alive per dam: HR:
2.0 ± 0.2, LR: 2.3 ± 0.7; ADG of the lambs: HR: 265 ± 19 g, LR: 249 ± 23 g). In both groups, the
plasma glucose and insulin concentrations were significantly lower during late gestation,
compared to lactation. In the HR ewes, the insulin values were also lower in late, compared
to mid-pregnancy. Blood NEFA values were identical during all gestational stages in the LR
sheep, while values were significantly increased during late pregnancy and lactation in the
HR ewes. The NEFA concentrations during late pregnancy were significantly higher in the
HR compared to the LR ewes. In both groups, the ␤-HB levels were elevated during late
pregnancy, peripartal and during lactation, compared to the dry period and mid-pregnancy
(HR ewes only). Significantly different glucose, insulin and ␤-HB plasma levels between HR
and LR sheep were recorded after parturition. In conclusion, the absence of maternal body
weight gain, the decreased insulin secretion and the increased lipolysis at the end of preg-
nancy indicate that late pregnancy requires more metabolic adaptations in the HR, than in
the LR ewes. This high demand may contribute to the development of pregnancy toxaemia.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction

Pregnancy toxaemia in sheep is a metabolic disorder

∗ Corresponding author. Tel.: +49 511 856 7263; fax: +49 511 856 7684. characterised by ketonaemia and ketonuria, commonly
E-mail address: Reinhard.Duehlmeier@tiho-hannover.de affecting older ewes carrying multiple foetuses during late
(R. Duehlmeier). pregnancy (Sargison et al., 1994). The most common clini-

0921-4488/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.smallrumres.2010.12.002
 R. Duehlmeier et al. / Small Ruminant Research 96 (2011) 178–184
cal signs of pregnancy toxaemia are weakness, depression,
mental dullness, disorientation, anorexia, blindness, and
finally recumbency and death after 3–10 days (Rook, 2000).
Due to non-specific clinical symptoms, the presence
of pregnancy toxaemia in ewes can only be determined
by elevated plasma ketone body concentrations, i.e. of
␤-hydroxybutyrate (␤-HB). Plasma ␤-HB levels generally
higher than 1.6 mmol/L indicate severe undernutrition
(Andrews, 1997). In ewes suffering from pregnancy tox-
aemia, the plasma ␤-HB concentrations usually exceed
3.0 mmol/L (Sargison et al., 1994).
Prognosis of pregnancy toxaemia is generally very poor.
Even if treated intensively, about 40% of the affected ewes
die and in about 20% of the cases of clinical pregnancy
toxaemia, the offspring die before, or immediately after
parturition (Henze et al., 1998).
The aetiopathogenesis of pregnancy toxaemia has still
not been clarified completely. The affection resulting from
an impaired energy supply during late pregnancy is cur-
rently the most common hypothesis. Almost 80% of the
foetal growth takes place in the final 6 weeks of pregnancy,
with 30–40% of the maternal glucose supply being utilised
by the foetal–placental unit (Rook, 2000). Therefore, the
higher the number of lambs carried by the ewe, the higher
the foetal glucose demands (Sargison et al., 1994). The
maternal glucose undersupply is promoted by the fact that
placental glucose transport takes place via the insulin inde-
pendent glucose transporter 1 (GLUT1) – while the glucose
uptake in the maternal skeletal muscle and adipose tissue
is mediated by the insulin dependent GLUT4 (Anderson
et al., 2001). Thus, the physiologically impaired insulin
sensitivity of ewes during late pregnancy reinforces the
disturbance of the maternal glucose supply. To compensate
for the lack of glucose, maternal triglycerides are mobilised,
resulting in increased plasma levels of non-esterified fatty
acids (NEFA). Due to the incomplete NEFA break down, the
plasma ␤-HB concentrations increase. The elevated ␤-HB
levels inhibit the hepatic gluconeogenesis, and thus fur-
ther increase maternal hypoglycaemia (Schlumbohm and
Harmeyer, 2004).
Although this hypothesis appears conclusive, there are
certain characteristics of pregnancy toxaemia which can-
not be explained by energy deficiency, as the primary
cause of this disease. The first of these phenomena is the
fact that pregnancy toxaemia occurs predominantly dur-
ing late pregnancy, and not during peak lactation – which
in sheep, as in dairy cows, is the gestational stage of the
highest energy demand (Baird, 1981). Furthermore, in a
preliminary investigation, only 40% of the ewes suffering
from pregnancy toxaemia demonstrated hypoglycaemia
– normoglycaemia being observed in 40% and hypergly-
caemia in 20% of the sheep (Henze et al., 1998). Not all
cases of normo- and hyperglycaemia reported in this study
could be attributed to foetal death – which has been
identified as a reason for hyperglycaemia in sheep with
pregnancy toxaemia (Wastney et al., 1983). Finally, it is still
unclear why there are individual (Marteniuk and Herdt,
1988) and breed-dependent differences in susceptibility.
For example, German Blackheaded Mutton ewes have been
recognised as being highly susceptible to pregnancy tox-
aemia (Bickhardt et al., 1998). On the other hand, no cases
179
of pregnancy toxaemia have been documented in purebred
Finnish Landrace ewes, although this breed is characterised
by a high reproductive performance (Oltenacu and Boylan,
1981).
The aim of this investigation was thus to identify possi-
ble differences regarding the metabolic adaptations during
late pregnancy and lactation in ewes with presumably dif-
ferent risks of being affected by pregnancy toxaemia.
2. Materials and methods
The animal tests included in this study were reported to the district
government of Hannover, Lower Saxony, Germany according to the Ger-
man Animal Welfare Legislation (reference number: 33-42502–05/928).
According to the major predisposing factors to pregnancy toxaemia (poly-
tocous pregnancy, breed, higher age and inappropriate feeding), two
groups of experimental ewes were created. The group with the presum-
ably higher risk (HR) for pregnancy toxaemia consisted of 7 pregnant, 4.5-
to 6.5-year-old German Blackheaded Mutton ewes. The control group with
the lower susceptibility (LR) for pregnancy toxaemia consisted of 5 preg-
nant, 2.5-year-old Finn sheep ewes. All animals were in excellent physical
condition and healthy, according to clinical and haematological evaluation
at the onset of the investigation. During the course of the study one of the
German Blackheaded Mutton ewes died due to mastitis.
Irrespective of the breed and body weight, the ewes all received a diet
consisting of 500 g hay and 3 kg grass silage, daily. During the last four
weeks of pregnancy and during lactation, the animals were additionally
fed 300 g concentrate (barley 23%, oat 8%, wheat 35%, peas 12%, sugar beet
pulp 11%, extracted soybean meal 3%, mineral premix 3%, and calcium
carbonate 3%) per day. The resulting daily energy intake per sheep was
approximately 21.1 MJ of metabolisable energy (ME). The recommended
daily energy requirements 2 weeks before parturition were 22 MJ ME (HR
ewes) and 18.7 MJ ME (LR ewes), respectively (Kamphues et al., 2004).
According to these calculations the ewes in the HR group were fed to meet
their energy requirements, while the LR sheep were slightly overnour-
ished. During the post partum period, 100 g of extracted soya bean meal
per lamb/day was added to this diet. Water was available ad libitum for
all animals. Ewes were housed in groups of 4 in a barn fitted with a slatted
floor, throughout the entire experiment at the Friedrich-Loeffler-Institute,
Neustadt, Germany.
Suckling lambs remained with the dams, until weaning. Lambs were
weaned at the age of 10 weeks (lambs of the HR ewes) and 8 weeks (lambs
of the LR sheep), respectively. In the case of Finn sheep bearing more than
two lambs, the third and fourth lambs were removed from their mothers
and reared on a sheep milk replacer. Hay and a concentrate were offered
to the lambs after the 3rd and the 10th day post natum, respectively.
In total, each animal was evaluated 5 times at different stages of preg-
nancy and lactation. Blood sampling was performed 8 and 2 weeks ante
partum (a.p.), peripartal (p.p.), 2 weeks post partum (2 weeks p.p.) and
2 weeks post weaning (p.w.). The exact times of blood collection in the
HR group were 56.4 ± 7.5 days a.p., 10.6 ± 2.3 days a.p. and 2.7 ± 1.6 days
a.p., as well as 17.0 ± 2.2 and 84.9 ± 7.4 days post partum. In the LR ewes,
blood samples were collected 51.3 ± 2.1 days a.p., 15.3 ± 4.5 days a.p. and
2.0 ± 3.4 days a.p., as well as 15.3 ± 0.5 and 69.3 ± 1.9 days post partum.
In all cases blood sampling took place following overnight fasting. Imme-
diately after the blood collection, the samples were chilled on ice and
centrifuged for 15 min at 4000G at 4 ◦ C within 1 h. Plasma was separated
and stored at −20 ◦ C until used for biochemical analyses.
All experimental animals were weighed at each blood sampling
period. To evaluate the reproductive performance, the live weight of the
offspring was recorded immediately after birth, and the percentage birth
weight of the lambs in relation to the body weight of the dams (relative
offspring weight) was calculated. The lambs were weighed at weaning to
calculate the ADG and thus the rearing success of the dams.
Plasma samples were analysed for glucose, insulin, NEFA and ␤-
HB concentrations. Plasma concentrations of glucose were assayed
using the hexokinase method of the commercially available Gluco-quant
Glucose/HK® test kit. The precision of the test was characterised by an
intra assay coefficient of variance (CV) of 4.5% and an inter assay CV of
6.0%. Plasma insulin levels were determined using the Coat-A-Count®
Insulin RIA kit, with human insulin serving as the standard in antibody-
coated tubes. The intra assay CV of the plasma insulin determination
180 R. Duehlmeier et al. / Small Ruminant Research 96 (2011) 178–184
was 1.4%, and the inter assay CV 5.1%. The ovine plasma NEFA con-
centrations were analysed using the enzymatic colour test kit NEFA C
ACS-ACOD® , as described by McMullen et al. (2005). The intra assay CV
and the inter assay CV of the plasma NEFA determination were 1.8% and
6.8%, respectively. The recovery of diluted pool plasma ranged between
90.9 ± 0.4% and 98.1 ± 2.1%. Linearity of the determination for plasma
NEFA concentrations was between 0.26 and 2.29 mmol/L (the calculated
coefficient of regression in this range being 0.9999). As a consequence of
this result, samples with NEFA values of 2.0 mmol/L or higher were diluted
1:2 and measured again. The plasma ␤-HB concentrations were assayed
using the d-␤-HB-dehydrogenase method, as described by Williamson
and Mellanby (1970). The intra assay CV and the inter assay CV of this
procedure were 12.6% and 15.6%, respectively.
Results were expressed as the mean and standard error of the
mean (SE). Statistical analyses were carried out using SigmaStat® 3.0
(SPSS Inc.). Data were checked for normal distribution, using the
Kolmogoroff–Smirnoff test. Some ␤-HB values were out of the range of
the normal distribution (values differed from the mean of all other sheep,
by more than 2-fold the standard deviation). These values were logarith-
mised to achieve a normal distribution. To evaluate possible differences
between the data of different stages of pregnancy and lactation (8 weeks
a.p., 2 weeks a.p., p.p., 2 weeks. p.p., 2 weeks p.w.) within each group of
the experimental animals, a one-way analysis of variance (ANOVA) with
repeated measures was performed. The Tukey test was subsequently per-
formed if the ANOVA yielded significant differences regarding the main
effects. Data of both groups at the different gestational stages were com-
pared, using the Student’s T-test. To evaluate any possible relationships
between the biochemical data, the correlations between selected param-
eters (insulin-glucose, NEFA-␤-HB, NEFA-insulin and ␤-HB-insulin) were
analysed using the Pearson product moment correlation. Significance was
accepted at a level of P < 0.05 in all analyses.
3. Results
The sheep in the HR group on average weighed
95.0 ± 4.4 kg at the beginning of the study, and maintained
their body weight until 2 weeks before lambing. There-
after the HR ewes lost significant body weight (Fig. 1). Thus,
throughout the entire trial, a body weight loss of approx-
imately 12.5% was recorded in the HR sheep. The ewes in
the LR group had an initial body weight of 44.1 ± 4.3 kg and
developed a significant weight gain of about 49% until 2
weeks before parturition. In contrast to the ewes in the HR
group, the sheep in the LR group maintained their body
weight after parturition (Fig. 1).
HR
110 A
A LR
B
B B
Body weight [kg]
90
70 a
ab ab
ab
50 b
Parturition
30
-8 -6 -4 -2 0 2 4 6 8 10
Weeks relative to parturition
Fig. 1. Mean (±SE) body weight during different reproductive stages of
ewes with a high risk (HR: dot) and low risk (LR: circle) of being affected
by pregnancy toxaemia. Different superscripts (HR: A, B; LR: a, b) indicate
significant (P < 0.05) differences between the reproductive stages.
4 HR LR a
ab A ab
A
AB ab
Glucose [mmol/L]
AB
3 b
B
2
1
0
8 W a.p. 2 W a.p. p.p. 2 W p.p. 2 W p.w.
Reproductive stage
Fig. 2. Mean (±SE) plasma glucose concentrations during different repro-
ductive stages in ewes with a high risk (HR: ) and low risk (LR: ) of being
affected by pregnancy toxaemia. Different superscripts (HR: A, B; LR: a, b)
indicate significant (P < 0.05) differences between the reproductive stages.
*Significant (P < 0.05) differences between HR and LR ewes.
The mean number of lambs born alive per ewe did
not differ significantly between both experimental groups
(HR: 2.0 ± 0.2; LR: 2.6 ± 0.7). The mean offspring weight
(sum of the body weights of all lambs born per ewe) was
10.0 ± 0.8 kg in the HR group and 7.1 ± 1.3 kg in the LR
group. The relative offspring weight was 11.8 ± 1.0% in the
HR ewes and 12.7 ± 2.5% in the LR group. There was no
significant difference with regard to the total and to the
relative offspring weights between both groups. Suckling
lambs of the HR ewes recorded an ADG of 265 ± 19 g, which
did not differ significantly from that recorded in the lambs
of the LR group (249 ± 23 g). During the suckling period in
the HR group, 2 out of 14 lambs born alive died, while in
the LR group, 4 out of 13 lambs did not survive. In the latter
case, 3 of the dead lambs had been born to the same ewe,
and 2 of these lambs had been reared artificially.
In both groups the lowest plasma glucose concentra-
tions were measured 2 weeks a.p. (HR ewes: 1.97 ± 0.23;
LR ewes: 2.45 ± 0.25 mmol/L), while the highest glucose
levels were recorded 2 weeks p.p. (Fig. 2). One differ-
ence between both groups was that only in the HR
ewes the plasma glucose values during late pregnancy
(2 weeks a.p.) significantly (P < 0.05) underestimated the
levels measured at parturition (p.p.). The second differ-
ence between both groups was that, 2 weeks p.w., the HR
ewes exhibited significantly (P < 0.05) lower glucose lev-
els (2.62 ± 0.21 mmol/L), than the ewes in the LR group
(3.27 ± 0.07 mmol/L).
The plasma insulin levels recorded during the dif-
ferent stages of pregnancy and lactation were similar
to the glucose concentrations in both groups. However,
the differences between individuals regarding the plasma
insulin concentration were higher than those for the
plasma glucose values (Fig. 3). In general, the plasma
insulin concentrations tended to be higher in the LR
ewes (10.92 ± 5.32 to 22.8 ± 7.73 mIU/L), compared to the
HR ewes (2.79 ± 0.52 to 7.56 ± 1.52 mIU/L). Significant
(P < 0.05) differences in this regard being determined dur-
ing lactation (2 weeks p.p.) and the dry period (2 weeks
p.w.). A second important result was that in the HR group,
 R. Duehlmeier et al. / Small Ruminant Research 96 (2011) 178–184 181

40 HR LR 8 HR LR

a A a a
30 7 AB
Insulin [mIU/L]

-HB [log µmol/L]

AB
ab a
ab
BC
20 b
ab 6
ab C
b
A A
10 AB 5
AB
B

0 4
8 W a.p. 2 W a.p. p.p. 2 W p.p. 2 W p.w. 8 W a.p. 2 W a.p. p.p. 2 W p.p. 2 W p.w.
Reproductive stage Reproductive stage

Fig. 3. Mean (±SE) plasma insulin concentrations during different repro-
ductive stages in ewes with a high risk (HR: ) and low risk (LR: ) of being
affected by pregnancy toxaemia. Different superscripts (HR: A, B; LR: a, b)
indicate significant (P < 0.05) differences between the reproductive stages.
*Significant (P < 0.05) differences between HR and LR ewes.
Fig. 5. Mean (±SE) logarithmised plasma concentrations of ␤-
hydroxybutyrate (␤-HB) during different reproductive stages in
ewes with a high risk (HR: ) and low risk (LR: ) of being affected
by pregnancy toxaemia. Different superscripts (HR: A, B, C; LR: a, b)
indicate significant (P < 0.05) differences between the reproductive
stages. *Significant (P < 0.05) differences between HR and LR ewes.

in contrast to the LR ewes, the plasma insulin concentra-
tion decreased significantly during late, compared to mid
pregnancy.
The NEFA concentrations at the different gestational
stages in the HR ewes and the LR group were as set out
in Fig. 4. Significant differences between the reproductive
stages were recorded only in the HR ewes. The ewes in
this group produced significantly (P < 0.05) higher plasma
NEFA values 2 weeks a.p., compared to mid-pregnancy
and the dry period. Furthermore, the plasma NEFA levels
during parturition and during lactation were significantly
(P < 0.05) higher than after weaning. When comparing both
experimental groups, the HR ewes showed significantly
higher (P < 0.05) plasma NEFA concentrations 2 weeks a.p.
(1.36 ± 0.19 mmol/L), compared to the sheep in the LR
group (0.80 ± 0.09 mmol/L). The opposite was recorded
8 weeks a.p., when the HR sheep recorded significantly
(P < 0.05) lower plasma NEFA levels, than the LR ewes.
2.0
A AB
1.5
NEFA [mmol/L]
As shown in Fig. 5, the blood ␤-HB concentrations
in the HR ewes paralleled the NEFA concentrations i.e.
the logarithmised ␤-HB levels were significantly (P < 0.05)
higher during late pregnancy, at parturition and during
lactation. In the LR group, significant (P < 0.05) elevations
in the logarithmised plasma ␤-HB values were recorded
during late pregnancy and lactation – this contrasting to
the blood NEFA concentrations in this group. Different ␤-
HB levels between both groups were recorded 2 weeks
p.p., when the ␤-HB concentrations in the HR ewes were
significantly (P < 0.05) higher than those in the LR sheep
(P < 0.05). A further striking result was that the ␤-HB levels,
especially 2 weeks a.p., were characterised by high indi-
vidual differences (HR sheep: 0.35–4.60 mmol/L; LR ewes:
0.42–2.79 mmol/L).
Positive correlations between the plasma glucose and
insulin concentrations were recorded in HR (r = 0.449,
P < 0.01), as well as in LR sheep (r = 0.398, P < 0.05) –
independent of the gestational stage. Similarly, a positive
correlation between plasma NEFA and logarithmised ␤-HB
HR LR levels was observed in both groups (HR sheep: r = 0.794,
P < 0.01; LR sheep: r = 0.524, P < 0.01). In the LR ewes, a neg-
ative correlation between insulin and NEFA concentrations
(r = −0.469, P < 0.05) was recorded. These relationships

AB

a
a were confirmed only to a much lesser extent when correla-
tions were calculated for the different reproductive stages.
1.0 a a
Positive correlations (P < 0.05) between insulin and glucose
BC
in the LR ewes, 2 weeks a.p. (r = 0.910), between NEFA and
logarithmised ␤-HB values in both groups p.p. (HR sheep:
0.5
C
a r = 0.76; LR sheep: r = 0.915) and in the LR group 2 weeks
a.p. (r = 0.897) were recorded.
0
8 W a.p. 2 W a.p. p.p. 2 W p.p. 2 W p.w. 4. Discussion
Reproductive stage
In the present trial, late pregnancy and lactation were
Fig. 4. Mean (±SE) plasma concentrations of non-esterified fatty acids characterised by a decrease of body weight in the HR ewes
(NEFA) during different reproductive stages in ewes with a high risk and a body weight gain in the LR ewes. The large body
(HR: ) and low risk (LR: ) of being affected by pregnancy toxaemia.
weight gain in the LR sheep during late pregnancy may
Different superscripts (HR: A, B, C; LR: a) indicate significant (P < 0.05)
differences between the reproductive stages. *Significant (P < 0.05) differ-
be attributed to slight overfeeding. Nonetheless, it was
ences between HR and LR ewes. surprising that the HR ewes showed no increase in body
182 R. Duehlmeier et al. / Small Ruminant Research 96 (2011) 178–184
weight during late pregnancy, although the energy supply
was sufficient to meet the requirements for maintenance
and for 2 growing foetuses, with a predicted birth weight
of 5 kg each. Even Romney ewes which were fed in accor-
dance with their maintenance energy requirements, still
produced a weight gain between day 53 and 140 of preg-
nancy (Tanaka et al., 2008). Twin pregnant, 3- to 5-year-old
Clun forest sheep fed to meet the energy demands for main-
tenance and for the growing foetuses, recorded a weight
gain of 11% between day 110 and day 140 of pregnancy
(West, 1996).
According to the total and to the relative offspring
weights, as well as to the ADG of the suckling lambs in
both groups, the energy requirements for the developing
foetuses and the growing lambs seem to have been iden-
tical and are probably not responsible for the differences
in weight gain during late pregnancy in the HR and LR
groups.
Assuming identical energy requirements of the growing
foetuses and the suckling lambs, and a calculative suffi-
cient energy supply in both groups, the body weight loss of
the HR ewes during late pregnancy and lactation can only
be attributed to an inappropriate feed utilisation. This may
be due to an insufficient nutrient uptake in the digestive
tract, or to deficiencies regarding the energy partitioning
in the dam. In the ruminant intermediary metabolism too,
glucose is the most important energy source, and insulin
is the major regulator of energy partitioning (Hart, 1983).
Thus, to explain the body weight loss of the HR ewes during
pregnancy and lactation, beside the plasma NEFA and ␤-HB
values, blood glucose and insulin concentrations have to be
evaluated during the different gestational stages.
The blood glucose levels, 2 weeks a.p. in both groups,
were still markedly higher than that reported in some cases
of severe spontaneous pregnancy toxaemia (Sargison et al.,
1994; Van Saun, 2000). With regard to the reference values
established in the laboratory (Bickhardt and Konig, 1985),
plasma glucose concentrations 2 weeks before parturition
were in the range of the lower physiological limit. Investi-
gations concerning the effect of the different reproductive
stages on the plasma glucose values revealed inconsistent
data. In healthy Merino sheep, significantly lower plasma
glucose concentrations were reported during pregnancy,
than during lactation (Henze et al., 1994). Other research
with Sakiz ewes (Firat and Ozpinar, 2002) revealed no
changes in the plasma glucose levels between the differ-
ent reproductive stages. Everts and Kuiper (1983) indicated
increased blood glucose levels at the end of pregnancy
in healthy Ile de France × Finn sheep crossbreds. Ewes of
the same breed suffering from subclinical or clinical preg-
nancy toxaemia demonstrated decreased plasma glucose
values during late pregnancy. Thus, the lower plasma glu-
cose concentrations during late gestation measured in the
present study may be the expression of subclinical preg-
nancy toxaemia. However, the low blood glucose levels
determined 2 weeks before lambing may be the conse-
quence of an increased glucose utilisation by the growing
foetus(es) (Rook, 2000), or of an impaired hepatic gluco-
neogenesis (Schlumbohm and Harmeyer, 2004). To finally
interpret the glucose data, other parameters of the energy
metabolism have to be taken into account.
The current results regarding the insulin concentrations
at different gestational stages are in part contradictory
to earlier investigations. In Finn × Dorset Horn crossbreds
(Vernon et al., 1981), as well as in Merino ewes (Regnault
et al., 2004), insulin values decreased significantly during
late pregnancy. This could be confirmed in the HR, but not
in the LR ewes. Additionally, Vernon et al. (1981) recorded
significantly lower blood insulin values on day 18 of lac-
tation, compared to late pregnancy, late lactation and the
dry period. In the current study increased insulin concen-
trations during lactation in both groups were recorded. This
contradiction remains unclear, as reproductivity and rear-
ing success and thus milk production were comparable in
both trials.
The positive correlation between plasma insulin and
glucose concentrations in both groups suggest that the
variation in blood insulin levels between the different
reproductive stages was the consequence of variations in
blood glucose levels, i.e. the higher the plasma glucose val-
ues the higher the glucose-stimulated pancreatic insulin
secretion. Another explanation for the decreased plasma
insulin levels during late pregnancy may be increased lipol-
ysis (Regnault et al., 2004) – chronically elevated plasma
NEFA values have been demonstrated to inhibit pancre-
atic insulin secretion in human diabetes type 2, and obesity
because of a “lipotoxicity” of the fatty acids in the pancre-
atic ␤-cells (Boden and Shulman, 2002).
In this trial, only ewes of the HR-group produced signif-
icantly increased plasma NEFA concentrations during late
pregnancy and early lactation. Furthermore, the NEFA con-
centrations measured 2 weeks before lambing in the HR
ewes were significantly higher, than in the LR group. Sum-
marising these data, late pregnancy resulted in increased
lipolysis in the HR group. In Dorset and Dorset Horn ewes,
elevated plasma NEFA levels were also recorded in preg-
nant, compared to non-pregnant ewes – when the sheep
received a 50% feed restriction for 5 days (Petterson et al.,
1994). In Shropshire ewes fed a diet containing 50% of
the energy requirements for maintenance during the last
6 weeks of pregnancy, elevated plasma NEFA values were
recorded 14 days before parturition (Tygesen et al., 2008). A
second investigation with an identical design revealed no
effect of energy restriction on blood NEFA levels (Husted
et al., 2008). It thus appears unlikely that the increased
NEFA values in the HR ewes in the current study may
hint at undernutrition during late pregnancy – especially
as the calculated daily energy intake of the HR ewes was
sufficient to meet the requirements. Furthermore, a subse-
quent study in Blackheaded Mutton and Finn sheep ewes
of the same age fed an individual diet meeting the energy
requirements also revealed an increase of the plasma NEFA
concentrations during late pregnancy in the Blackheaded
Mutton ewes (data not presented). Thus, a higher rate of
lipolysis appears to be specific in German Blackheaded
Muttons during late pregnancy. Regarding the insulin NEFA
relation, only in the LR ewes was a negative correlation
between these both parameters determined. This yields
some evidence for the following interpretations: (i) A NEFA
induced ␤-cell disturbance as a reason for reduced plasma
insulin levels during late pregnancy appears to be possible
in the LR group, but not in the HR ewes. (ii) The regulation
 R. Duehlmeier et al. / Small Ruminant Research 96 (2011) 178–184
of lipolysis and lipogenesis may have been less insulin-
sensitive in the ewes of the HR group.
Plasma ␤-HB concentrations have been recognised to
be a sensitive tool to detect maternal undernutrition and
pregnancy toxaemia in sheep. Healthy animals generally
develop ␤-HB plasma levels below 0.8 mmol/L, while ␤-HB
concentrations of 0.8 to 1.6 mmol/L could suggest mod-
erate undernutrition (Andrews, 1997). In sheep suffering
from pregnancy toxaemia ␤-HB plasma concentrations of
more than 3.0 mmol/L are usually measured (Sargison et al.,
1994).
In the present experiment, the median ␤-HB concen-
trations recorded were below 0.8 mmol/L at all stages of
gestation. On the other hand very high individual dif-
ferences regarding the ␤-HB values in both groups were
recorded 2 weeks before parturition – indicating individual
rates of ketogenesis. Furthermore, in ewes of both groups,
␤-HB levels were elevated 2 weeks a.p., p.p. and 2 weeks
p.p., compared to the dry period. This finding contradicts
earlier trials on Merino sheep which showed higher ␤-
HB concentrations during lactation than during pregnancy
(Henze et al., 1994). Sakiz ewes developed identical plasma
␤-HB values at different stages of pregnancy and dur-
ing early lactation (Firat and Ozpinar, 2002). The elevated
␤-HB concentrations during late pregnancy and lactation
recorded in the present study may however suggest the
high challenge of these gestational stages on the mater-
nal energy metabolism. The main difference between the
ewes of both groups was, that only in the HR sheep, did
the ␤-HB concentrations increase during late, compared to
mid-pregnancy. This again underlines the higher demand
for metabolic adaptation at the transition between early
and late pregnancy and lactation in the HR ewes.
Significant higher ␤-HB levels were determined in the
HR group, 2 weeks after parturition – compared to the LR
sheep. According to the similar ADG of the lambs in both
groups, an identical milk yield is suggested in the HR and
LR ewes. Thus, the increased ␤-HB blood values in HR ewes
during lactation were not expected. Finally, a positive cor-
relation between plasma NEFA and ␤-HB concentrations
was determined in both groups, which may present some
evidence that changes in ␤-HB values in the blood of sheep
depend on the rate of lipolysis and ketogenesis, rather than
on the rate of ketone body utilisation.
5. Conclusion
Summarising the clinical data (lack of body weight gain
during late pregnancy in the HR group) and the results of
the biochemical analyses (elevated lipolysis and ketoge-
nesis, reduced insulin secretion during late, compared to
mid-pregnancy in HR ewes), late pregnancy appears to be
a higher challenge to the energy metabolism in older Ger-
man Blackheaded Mutton ewes (HR group), compared with
the younger, slightly overnourished Finn sheep ewes (LR
group). As insulin has an important regulatory influence
on lipolysis and ketogenesis, a possible insulin resistance
in the HR sheep may explain the increased plasma NEFA
and ␤-HB concentrations during late pregnancy recorded
throughout the trial and should be further elucidated. The
need for metabolic adaptation in the HR ewes may also
183
be a possible reason for a higher risk of being affected by
pregnancy toxaemia. According to the design of the present
study, these higher metabolic adaptations could be due to
the breed, the feeding regime and the different ages of
the ewes in both groups. In humans, fuel utilisation and
basal and total energy expenditure are impaired in older,
rather than in younger individuals (Roberts and Rosenberg,
2006). An impaired total energy expenditure has also been
reported in older sheep (Toutain et al., 1977). To evaluate
whether the higher metabolic adaptations are attributable
to the breed, further investigations are needed, excluding
the effect of feeding and age of the experimental animals.
Acknowledgements
The authors wish to thank B. Möller and T. Hantscher
for their excellent care of the experimental animals and are
grateful to R. Wittig for technical assistance. Mrs. Frances C.
Sherwood-Brock English Editorial Office, University of Vet-
erinary Medicine Hannover, Foundation is acknowledged
for the careful English proof reading. This study was sup-
ported by the German Research Organisation (DFG) Grant
SA 160/17-2.
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