Animal (2018), 12:2, pp 312–321 © The Animal Consortium 2017

doi:10.1017/S175173111700163X
animal

Effects of maternal nutrient restriction during early or

mid-gestation without realimentation on maternal physiology
and foetal growth and development in beef cattle
R. K. Taylor, C. T. LeMaster, K. S. Mangrum, R. E. Ricks and N. M. Long†
Department of Animal and Veterinary Sciences, Clemson University, Clemson, South Carolina 29634, USA

(Received 26 October 2016; Accepted 12 June 2017; First published online 12 July 2017)

The aim of this study is to determine the effects of early and mid-gestation nutrient restriction on maternal metabolites and foetal
growth. Primiparous Angus cows were synchronized and inseminated with semen from one sire. Dietary treatments were: control
to gain 1 kg/week (CON) or 0.55% maintenance energy and CP requirements (nutrient restricted; NR). A subset of dams was fed
NR (n = 8) or CON (n = 8) from days 30 to 110 of gestation. Another group was fed CON ( n = 8), days 30 to 190; NR ( n = 7),
days 30 to 110 followed by CON days 110 to 190; or CON, ( n = 7) days 30 to 110 followed by NR days 110 to 190. Cows were
harvested at days 110 or 190 of gestation, when foetal measurements and samples were collected. Cows that were NR during
days 30 to 110 or 110 to 190 of gestation lost significant BW and body condition score ( P < 0.001), this was associated with
reduced plasma glucose during NR ( P < 0.002). Foetal weights, empty foetal weights, abdominal and thoracic circumferences were
all reduced ( P < 0.03) in day 110 NR animals. Foetal perirenal adipose as a percentage of empty foetal weight was increased
( P = 0.01) in NR day 110 female foetuses compared with CON foetus. Maternal serum triglycerides at day 110 of gestation were
decreased ( P < 0.05) in NR dams, whereas foetal serum triglycerides were increased ( P < 0.05) in response to maternal NR. Foetal
weights tended to be reduced ( P = 0.08) in NR/CON and CON/NR v. CON/CON cattle at day 190 of gestation. Empty foetal
weights, abdominal and thoracic circumferences were reduced ( P ⩽ 0.03) in NR/CON and CON/NR v. CON/CON cattle. Brain
weight as a percentage of empty foetal weight was increased ( P < 0.001) in NR/CON and CON/NR v. CON/CON cattle. Foetal
perirenal adipose as a percentage of empty foetal weight was increased ( P = 0.003) in NR/CON and CON/NR v. CON/CON cattle.
Maternal serum triglycerides at day 190 of gestation were decreased ( P < 0.05) in association with maternal NR. Foetal serum
triglycerides at day 190 of gestation were increased ( P < 0.05) in response to maternal NR during early gestation but
decreased by NR in mid gestation compared with CON foetuses. The data show that maternal nutrient restriction during early or
mid-gestation cause’s asymmetrical foetal growth restriction, regardless if the restriction is preceded or followed by a period of
non-restriction.

Keywords: bos taurus, foetal growth, foetal programming, maternal metabolites, maternal undernutrition

Implications
It has been previously highlighted that nutrient restriction
during early gestation causes reduced foetal growth and
development. Mid-gestation nutrient restriction has now
been shown to cause the same damaging effects to the same
extent, even when no nutritional stress is incurred during
early gestation. This could explain why experiments on mid
gestation nutrient restriction can find many of the same
results as early gestation nutrient restriction.
†
E-mail: nlong2@clemson.edu
Introduction
Because most beef cattle are maintained in pastoral production
systems, they are subject to seasonal changes in nutrient
availability that correspond to changes in forage quantity and
quality. Beef cattle experiencing nutritional stress in utero
can have reduced skeletal growth, which can affect carcass
merit and meat production (Long et al., 2012b). This could be
especially problematic in heifers used as breeding stock as it has
been shown that nutrient restriction during early gestation leads
to reduced antral follicle counts in heifer offspring at 12 months
of age (Mossa et al., 2013). Any stimulus or insult that alters
foetal growth during gestation can lead to permanent negative

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effects on the offspring’s physiology and development (Baker
et al., 1969). Even minor events like transportation of the
dam can lead to alteration of the hypothalamic-pituitary axis
development in the foetus, inhibiting correct regulation of
growth and reproduction in the postnatal animal (Lay et al.,
1997). Foetuses that undergo intrauterine growth retardation
(IUGR) often exhibit asymmetrical growth patterns, which
suggests that the foetus compartmentalizes nutrients and blood
flow to the organs and structures that are most pertinent to
development (Anthony et al., 2003). Early gestation is a critical
period for foetal organ and tissue development (Ford, 1995).
It has been shown that young cows respond differently than
older cows when exposed to nutrient restriction and this leads to
differences in foetal growth and development (Long et al., 2009).
It has also been shown that nutrient restriction during early
gestation will result in calves that are similar in BW at birth but
have organs and tissues that are functionally compromised
(Long et al., 2009). The objective of this study was to determine
the severity of asymmetrical foetal growth and changes in
maternal and foetal metabolites caused by maternal nutrient
restriction during early or mid-gestation and to what extent, if
any, the effects of foetal growth restriction could be alleviated
by feeding the dam slightly over maintenance requirements
during the second trimester of gestation in young cows. Our
hypothesis is that mid-gestation nutritional stress will alter foetal
development similar to early gestation nutrient restriction.
Material and methods
Animals and sample collection
All animal procedures were approved by Clemson University
Animal Care and Use Committee (AUP no. 2013-062).
Primiparous Angus-cross cows (2.5 to 3.5 years of age) were
initially synchronized. On day 0, cows received an easy breed
(CIDR) (Zoetis Animal Health, New York, NY, USA) and
gonadotropin releasing hormone (100 µg (i.m.); Zoetis Animal
Health). On day 7, CIDRs were removed and cows received
prostaglandin F2α (25 mg (i.m.); Zoetis Animal Health). Cows
were observed for oestrus twice daily for 30 min and
inseminated by one technician with male sexed (day 190
foetuses) or conventional (day 110 foetuses) semen from
one Angus sire 12 h after observed oestrus. From the start of
synchronization to day 30 of gestation, cattle were group fed in
excess of requirements (National Research Council (NRC),
1996). At 30 days post breeding, singleton pregnancy was
determined via transrectal ultrasonography (Aloka 500-V with
7.5-MHz probe; Corometrics Medical Systems, Wallingford, CT,
USA). Blood samples were collected from pregnant animals via
caudal venipuncture into 9 ml Lithium-Heparin vacutainers
(Becton, Dickinson and Company, Franklin Lakes, NJ, USA) and
placed immediately on ice. Samples were centrifuged within 2 h
of collection at 2000 × g for 20 min at 4°C. After centrifuging,
the plasma was collected and stored at −20°C until analysis.
Animals were blocked by body condition score (BCS) (Wagner
et al., 1988) into treatment groups. Throughout the experiment,
all animals were fed a total mixed ration (30% soya bean hulls,
20% cottonseed hulls, 17.5% peanut hulls, 17% corn screening,
Nutrient restriction in early and mid-gestation
12.5% corn gluten feed, 1.5% salt, 0.5% calcium, 0.5% trace
mineral, 0.5% vitamin premix: 1.72 Mcal net energy of
maintenance (NEm)/kg, 8.9% CP, 1.4% ether extract on a DM
basis, 94.1% DM) at one of two levels. Diets were control (CON),
fed to gain 1 kg of BW/week and exceed CP requirements or
nutrient restricted (NR) fed 0.55% of maintenance energy and
CP recommendations based on BW (NRC, 1996). A subset of
animals was fed either NR (n = 8) or CON (n = 8) from days 30
to 110 of gestation. The remaining animals were fed CON
(n = 8) days 30 to 190; NR (n = 7) days 30 to 110 followed by
CON days 110 to 190; or CON (n = 7) days 30 to 110 followed
by NR days 110 to 190.
Cattle were maintained together on a dry lot with free
access to water and were fed individually once daily. Animal
BW was collected weekly and used to adjust feed allowances
throughout the experiment; BCS and blood samples were
collected every 2 weeks. All blood samples were collected
and processed as previously described.
Final maternal BW, BCS and blood samples were collected
before transport to commercial abattoir. Blood samples were
collected as previously described into 9 ml sodium heparin,
serum and Ethylenediaminetetraacetic acid (EDTA) vacutainers.
The plasma tubes were processed as previously described. Serum
samples were allowed to clot for ~60 min at room temperature
and then refrigerated overnight before centrifuging and stored
the same way as plasma samples. All animals were harvested at
the same commercial abattoir. Cows were harvested and the
gravid uterus was removed. At harvest, foetal gender was
determined. Within the day 110 dams there were two female
and six male foetuses per treatment and within the day 190
dams only male foetuses were recovered and used for this study.
Foetal blood samples were collected from the umbilical cord into
10 ml sodium heparin, serum and EDTA vacutainers, handled
as previously described, and stored at −80°C. Foetal weight,
crown rump length, abdominal circumference and thoracic
circumference were recorded. Foetal brain, heart, liver, lungs,
kidneys, adrenals, perirenal adipose, viscera and longissimus
dorsi (LD) muscles were collected and weights were recorded.
The foetal heart was dissected into right and left atria, right and
left ventricle, and septum; each component was individually
weighed. Thickness was measured in three locations and aver-
aged. The eviscerated foetus was weighed to obtain an empty
foetal weight (EFW).
Histology
Two samples of foetal perirenal adipose tissue were collected
and stored in 10% formalin for histology. All samples were
embedded in paraffin and stored at room temperature. Four,
5 µm, sections, five sections apart were collected for evalua-
tion. Tissues were deparaffinized then stained using standard
haematoxylin–eosin Y staining procedures. Two randomly
chosen fields were captured digitally at 10 × magnification
per section for a total of eight fields per animal. Cell diameter
was evaluated on fifty randomly chosen cells per field for a
total of 400 adipocytes per animal. The histological measure of
cell diameter has been previously validated (Long et al.,
2012b). Only adipocyte diameters for day 190 foetuses are
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reported due to finding preadipocytes and no filled adipocytes to be normally distributed (P > 0.17). All data for days 110
for day 110 foetuses. and 190 cows, initial BW and BCS along with changes in
each, were analysed as an ANOVA analysis using the GLM
procedure of SAS. Maternal plasma glucose concentrations
Hormone and metabolite quantification
for days 110 and 190 animals was analysed separately as a
Animals were blocked by treatment and gestation group into
repeated measures analysis using the MIXED procedure of
assay plates. All day 190 samples (n = 22) were run separately
SAS with treatment, day and their interaction in the model
from day 110 samples (n = 16). Within each gestation group,
statement. For all maternal day 110 date, foetal gender
all treatments were represented on an assay block. All colori-
was initially included in the model but was found to be
metric reagents and standards were obtained from Pointe
non-significant (P > 0.67) and was therefore removed from
Scientific (Canton, MI, USA). Plasma glucose concentrations
the final model. For day 110 animals, foetal weights and
were determined by colorimetric assay previously validated in
measurements, plasma glucose, serum triglycerides and
our lab (Long and Schafer, 2013) on both maternal and foetal
serum cholesterol in dams and foetuses and maternal serum
plasma for all animals. Intra-assay and inter-assay CV of the
cortisol at harvest were analysed using the MIXED procedure
glucose assays were 3.5% and 3.1%, respectively. Serum
of SAS with treatment, gender and their interaction in the
triglyceride concentrations were determined by colorimetric
model statement. For day 190 animals, foetal weights and
assay previously validated in our lab (Long et al., 2014) on all
measurements, plasma glucose, serum triglycerides, serum
samples. Intra-assay and inter-assay CV were 2.4% and 3.1%,
cholesterol in dams and foetuses and maternal serum cortisol
respectively. Serum cholesterol concentrations were deter-
at harvest, and adipocyte diameter were analysed as an
mined by colorimetric assay previously validated in our lab
ANOVA analysis using the GLM procedure of SAS with
(Long et al., 2014) for all samples. Intra-assay and inter-assay
treatment in the model. A tendency was defined as P ⩽ 0.10
CV were 2.7% and 3.7%, respectively. All assays were read at
and a significant difference was defined as P ⩽ 0.05
their respective wavelengths on a Synergy HT 96 well plate
Spectrophotometers (BioTek Instruments, Winooski, VT, USA).
Maternal serum cortisol concentrations were quantified in
duplicate in a single assay via a previously validated RIA (Long Results
and Schafer, 2013) with a sensitivity of 0.25 ug/dl (Coat-A-
Maternal BW, BCS, metabolites and foetal metabolites
Count cortisol RIA; Siemens Medical Solutions Diagnostics, Los
Maternal BW and BCS changes are shown in Table 1. At day
Angeles, CA, USA) with an intraassay CV of 5.3%.
30 of gestation cow BW and BCS were similar (P > 0.35) for
all cows. At day 110, NR cattle had a decrease in both BW
Statistics (P < 0.0001) and BCS (P < 0.0001) from days 30 to 110 when
All data were tested for normality using the Shapiro–Wilk compared with day 110 CON cattle. In day 190 cows,
test using the UNIVARIATE procedure of SAS and was found NR/CON cattle had a decrease in both BW (P < 0.0001) and

Table 1 Maternal BW (kg) and body condition score (BCS) changes throughout gestation for days 110 and 190 animals1
Items CON1 NR1 SEM P value

Day 110 animals

n 8 8
Day 30 of gestation BW 573.8 553.9 54.2 0.891
Days 30 to 110 BW 11.9a −53.5b 3.2 <0.0001
Day 30 of gestation BCS2 5.8 5.7 0.6 0.796
Days 30 to 110 0.1a −1.2b 0.2 <0.0001
CON1 NR/CON1 CON/NR1
Day 190 animals
n 8 7 7
Day 30 of gestation BW 537.4 537.1 522.0 22.6 0.866
Days 30 to 114 BW 11.7a −50.2b 12.9a 4.6 <0.0001
Days 114 to 190 BW 14.5a 19.3a −77.3b 4.7 <0.0001
Day 30 of gestation BCS2 5.3 5.6 5.2 0.2 0.351
Days 30 to 114 BCS 0.1a −1.1b 0.1a 0.2 0.0008
Days 114 to 190 BCS 0.0a 0.3a −1.3b 0.2 <0.0001
Means differ P ⩽ 0.05.
a,b
1
CON = control cows fed to gain 1 kg of BW/week and exceed CP recommendations. NR = nutrient restricted cows fed 0.55 × of NRC NEm and
CP recommendations, NR/CON and CON/NR cows were fed respective treatments from days 30 to 110 of gestation and the other treatment from
days 110 to 190 of gestation.
2
Wagner et al. (1988).

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BCS (P = 0.0008) from days 30 to 114 when compared with
CON/NR and CON cattle. A decrease in BW (P < 0.0001) and
BCS (P < 0.0001) occurred in CON/NR cows from days 114 to
190 when compared with NR/CON and CON cattle.
Monthly plasma glucose concentrations of day 110 cows
(Figure 1a) exhibited a treatment × day effect (P = 0.002),
with the plasma concentration of NR group decreased
(P < 0.05) compared with CON cows at day 56 that was
maintained until harvest. Maternal plasma glucose concentra-
tions at harvest of day 110 cows (Figure 2a) were decreased
(P = 0.027) in dams on the NR diet compared with dams on
the CON diet. Foetal plasma glucose concentrations at harvest
on day 110 (Figure 2a) were decreased (P = 0.014) in foetuses
from dams on NR diet v. foetuses from dams on CON diet.
Monthly plasma glucose concentrations of day 190 cows
(Figure 1b) exhibited a treatment × day effect (P = 0.0001).
Cows that were nutritionally restricted on days 30 to 110 had
decreased (P < 0.05) plasma glucose compared with animals
on the CON diet during early gestation. On day 114, both NR
treatments had decreased (P < 0.05) plasma glucose compared
with CON cows. Cows that were NR during mid-gestation had
Figure 1 Plasma glucose concentrations of control cows (fed to gain
1 kg of BW/week and exceed CP recommendations, CON) and nutrient
restricted (NR) cows (fed 0.55 × of NRC NEm and CP recommendations,
NR) cows either harvested at day 110 of gestation (a, treatment × day,
P = 0.002, *means differ within a time point P < 0.05) or day 190 of
gestation (b, treatment × day, P = 0.0001, *means differ between CON
and NR/CON; P < 0.05. #Means differ between CON and CON/NR;
P < 0.05). Data are presented as least square means ± SEM.
Nutrient restriction in early and mid-gestation
decreased (P < 0.05) plasma glucose compared with CON
treatments during the same time period. Maternal plasma
glucose concentrations at harvest for day 190 cows (Figure 3a)
were decreased (P < 0.001) in NR dams, whereas plasma
glucose concentrations were similar between animals on the
CON diet before harvest. Foetal plasma glucose concentrations
at harvest on day 190 of gestation (Figure 3a) were decreased
(P = 0.049) in foetuses from NR dams, whereas plasma
glucose concentrations were similar between foetuses from
dams on CON diets before harvest.
Maternal serum triglyceride concentrations of day 110
cows of gestation (Figure 2b) were decreased (P = 0.003) in
NR dams compared with CON dams. Foetal serum triglycer-
ide concentrations at day 110 of gestation (Figure 2b)
were decreased (P = 0.033) in foetuses from CON dams
compared with NR dams. Maternal serum triglyceride
concentrations at day 190 of gestation (Figure 3b) were
decreased (P = 0.008) in NR dams compared with CON
dams. Foetal serum triglyceride concentrations at day 190 of
gestation (Figure 3b) were decreased (P < 0.001) in CON/NR
v. CON foetuses, however NR/CON foetuses had increased
(P < 0.05) serum triglyceride concentrations.
Maternal and foetal serum cholesterol concentrations at
day 110 of gestation (Figure 2c) were similar among
experimental groups (P = 0.984 and 0.285, respectively).
Maternal serum cholesterol concentrations at day 190 of
gestation (Figure 3c) tended to be decreased (P = 0.098)
in NR dams compared with dams on CON. Foetal serum
concentrations at day 190 of gestation (Figure 3c) were
decreased (P = 0.019) in foetuses from NR dams compared
with foetuses from dams on CON.
Maternal serum cortisol concentrations at day 110 of
gestation was increased in NR cows compared with CON cows
(1.13 ± 0.27 v. 0.56 ± 0.27, least square means (LSM) and SEM,
respectively P = 0.04). At day 190 of gestation maternal serum
cortisol concentration were greater (P = 0.003) in CON/NR
cows compared with CON and NR/CON cows (2.13 ± 0.24,
0.86 ± 0.26 and 0.89 ± 0.26 LSM and SEM, respectively).
Foetal measures at day 110 of gestation
Foetal size and weights at day 110 of gestation are shown in
Table 2. Foetal weights showed a treatment by gender inter-
action (P = 0.01) with NR male foetuses having a reduced
foetal weight compared with the other foetal groups. Empty
foetal weights and crown rump length and thoracic cir-
cumference was reduced (P = 0.01, P = 0.05 and P = 0.003,
respectively) in NR foetuses compared with CON foetuses.
Abdominal circumference tended to be reduced (P = 0.08) in
NR compared with CON foetuses. Liver weights tended to be
decreased (P = 0.09) in NR foetuses v. CON foetuses. Total LD
muscle weights was decreased (P = 0.03) in NR foetuses v.
CON foetuses. Brain weight had a tendency (P = 0.07) to be
decreased in NR female foetuses v. male NR and CON foetuses
and female CON foetuses. Total heart, left atrium, right
ventricle and septum weight were unaffected by treatment,
whereas the right atrium weight had a tendency (P = 0.09) to
be reduced in NR v. CON foetuses. Left ventricle weight was
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Taylor, LeMaster, Mangrum, Ricks and Long

Figure 2 Plasma glucose (a), serum triglycerides (b), and serum cholesterol (c) concentrations from control (cows fed to gain 1 kg of BW/week and exceed
CP recommendations, n = 8) and nutrient restricted NR (NR cows fed 0.55 × of NRC NEm and CP recommendations, n = 8) foetuses and dams at day
110 of gestation. Data are presented as least square means ± SEM. a,bMeans differ P < 0.05.

decreased (P = 0.05) in female CON foetuses v. female NR
foetuses and male CON and NR foetuses. Both ventricles wall
and septum wall thickness was increased (P ⩽ 0.05) in NR
foetuses compared with CON foetuses. Total weight of kidneys,
lungs and adrenal glands (P = 0.68, P = 0.14, P = 0.12,
respectively) were unaffected by maternal treatment.
Foetal organ weights as a percentage of EFW at day 110 of
gestation are shown in Table 3. Brain weight as a percentage
of EFW was increased (treatment × gender interaction
P = 0.01) in NR male foetuses v. CON male foetuses, with
no differences in female foetuses. Total lung weight as a
percentage of EFW was increased (P = 0.03) in female foe-
tuses v. male foetuses. Foetal heart weight as a percentage
of EFW tended (P = 0.06) to be increased in female v. male
foetuses. Right atrium weight as a percentage of EFW had a
tendency (P = 0.09) to be increased in CON foetuses v. NR
foetuses. Total kidney weight as a percentage of EFW were
increased (P = 0.04) in female foetuses v. male foetuses.
Perirenal adipose tissue weight as a percentage of EFW
was increased (P = 0.04) in NR foetuses compared with
CON foetuses. Foetal left and right ventricles, left atrium,
septum, liver, adrenal, LD muscles and viscera, as a percen-
tage of EFW were unaffected by either maternal diet or foetal
gender.
Foetal measures at day 190 of gestation
Foetal measurements and foetal organ weights at day 190 of
gestation are shown in Table 4. Foetal weights tended to be
reduced (P = 0.08) in NR/CON and CON/NR v. CON/CON
foetuses and EFWs were reduced (P = 0.03) in NR/CON
and CON/NR v. CON/CON foetuses. Abdominal and thoracic
circumference were reduced (P = 0.02 and P = 0.03,
respectively) in NR/CON and CON/NR foetuses v. CON/CON
foetuses, however crown rump length was unaffected
(P = 0.32). Foetal liver weight was reduced (P = 0.05) in
NR/CON and CON/NR v. CON/CON foetuses. Total LD muscle
weight was decreased (P = 0.03) in NR/CON and CON/NR v.
CON/CON foetuses. Perirenal adipose weight tended to be
reduced (P = 0.09) in NR/CON and CON/NR compared with
CON/CON foetuses. Total heart weight tended to be reduced

316
 Nutrient restriction in early and mid-gestation

Figure 3 Plasma glucose (a), serum triglycerides (b) and serum cholesterol (c) concentrations from control (cows fed to gain 1 kg of BW/week and exceed
CP recommendations) and nutrient restricted (NR cows fed 0.55 × of NRC NEm and CP recommendations) foetuses and dams at day 190 of gestation.
n = 8, 7 and 7 for CON, NR/CON and CON/NR treatments, respectively. Data are presented as least square means ± SEM. a,b,cMeans differ P < 0.05 and e,
f
means differ P < 0.10.

(P = 0.08) in NR/CON and CON/NR v. CON/CON foetuses.
Left ventricular weights were reduced (P = 0.04) in NR/CON
and CON/NR v. CON/CON foetuses. Atria, right ventricle and
septum weights were similar between foetal groups. Left
ventricle thickness tended to be increased (P = 0.08) in
NR/CON and CON/NR v. CON/CON foetuses. Right ventricles
and septum thickness were both unaffected (P = 0.37 and
P = 0.44, respectively) by maternal treatments.
Foetal organ weights as a percentage of EFW at day 190 of
gestation are shown in Table 5. Brain weight as a percentage
of EFW was increased (P = 0.01) in NR/CON and CON/NR v.
CON/CON foetuses. Total heart weight as a percentage of
EFW was increased (P = 0.04) in NR/CON and CON/NR v.
CON/CON foetuses. Left and right ventricle weight as a per-
centage of EFW was increased (P < 0.01) in NR/CON and
CON/NR v. CON/CON foetuses. Septum weight as a percen-
tage of EFW was also increased (P = 0.04) in NR/CON and
CON/NR v. CON/CON foetuses. Foetal perirenal adipose as a
percentage of EFW was increased (P < 0.01) in NR/CON and
CON/NR v. CON/CON foetuses. Adipocyte diameter of peri-
renal adipose tissue at d 190 of gestation were similar
(P = 0.86) regardless of treatment (CON = 23.9 ± 0.6; CON/
NR = 24.4 ± 0.7; NR/CON 24.3 ± 0.7 µm).
Discussion
To the authors’ knowledge, this is the first study with
maternal nutrient restriction during early or mid-gestation
in young cattle without realimentation with foetal endpoints.
Previous studies have shown in multiple mammal species
that maternal nutrient restriction during early or mid-
gestation can lead to IUGR exhibited by asynchronous
organ growth (i.e. enlarged hearts and brains) (McMillen
et al., 2001; Vonnahme et al., 2003; Platz and Newman
2008; Long et al., 2009). However, there have been little
comparisons of early versed mid-gestation IUGR in the
same experimental conditions in cattle. This study revealed
asynchronous organ growth in foetuses from NR dams

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Table 2 Foetal measurements and selected organ weights (wt) of male and female foetuses on day 110 of gestation from control (CON) and nutrient
restricted (NR) cattle1
CON NR P value

Items Male Female Male Female SEM Treatment Gender Treatment × gender

n 6 2 6 2
Foetal wt (kg) 0.71a 0.62b 0.59b 0.63 b 0.02 0.03 0.20 0.01
Empty foetal wt (kg) 0.54a 0.49a 0.47b 0.46b 0.01 0.01 0.10 0.31
Crown rump length (cm) 26.6a 26.2a 25.3b 25.2b 0.04 0.05 0.64 0.76
Abdominal circumference (cm) 20.3d 19.5d 17.4e 17.1e 1.1 0.08 0.68 0.86
Thoracic circumference (cm) 18.8a 18.5a 17.2b 17.5b 0.3 0.003 0.91 0.41
Viscera wt (g) 53.8 53.5 45.5 49.8 7.8 0.50 0.82 0.79
Liver wt (g) 24.6d 21.5d 20.9e 21.0e 1.1 0.09 0.22 0.19
Total lung wt (g) 20.1 18.9 18.3 21.3 1.3 0.65 0.80 0.14
Total kidney wt (g) 5.6 5.7 4.9 5.9 0.3 0.68 0.40 0.53
Adrenal wt (g) 0.2 0.9 0.6 0.3 0.4 0.57 0.42 0.12
Brain wt (g) 12.4 12.6 12.9 11.1 0.4 0.34 0.15 0.07
Total LD muscle (g) 12.7a 10.6a 9.4b 10.1b 0.4 0.03 0.39 0.11
Heart wt (g) 5.5 5.2 4.7 5.2 0.4 0.45 0.81 0.50
Left atrium wt (g) 0.55 0.26 0.32 0.32 0.08 0.40 0.16 0.16
Right atrium wt (g) 0.45d 0.38d 0.27e 0.33e 0.05 0.09 0.87 0.32
Left ventricle wt (g) 1.36a 0.82b 1.05c 1.41a 0.14 0.50 0.65 0.05
Right ventricle wt (g) 0.94 0.09 1.03 1.04 0.21 0.59 0.96 0.91
Septum wt (g) 1.34a 0.80b 1.22a 1.10b 0.09 0.52 0.05 0.13
Left ventricle thickness (mm) 4.1a 3.1a 4.5b 4.9b 0.3 0.02 0.41 0.11
Right ventricle thickness (mm) 3.0a 3.0a 3.9b 3.4b 0.2 0.05 0.40 0.43
Avg. septum thickness (mm) 4.0a 3.7a 4.7b 4.7b 0.2 0.02 0.72 0.57
Perirenal adipose wt (g) 1.69 1.57 1.47 1.57 0.2 0.72 0.98 0.70
LD = longissimus dorsi.
a,b,c,d,e
Means differ P ⩽ 0.05.
1
CON cows fed to gain 1 kg of BW/week and exceed CP recommendations. NR cows fed 0.55× of NRC NEm and CP recommendations.

Table 3 Foetal organ weights (wt) as a percentage of empty foetal weight (EFW) at day 110 of gestation from control (CON) and nutrient restricted
(NR) cattle1
CON NR P-value

Items Female Male Female Male SEM Treatment Gender Treatment × gender

n 2 6 2 6
Brain wt (g/EFW, g) 0.026 0.023 0.024 0.027 0.001 0.20 0.75 0.01
Lung wt (g/EFW, g) 0.045 0.035 0.046 0.039 0.003 0.44 0.03 0.70
Heart wt (g/EFW, g) 0.012 0.010 0.011 0.010 0.001 0.71 0.06 0.50
Left ventricle wt (g/EFW, g) 0.0019 0.0025 0.0031 0.0022 0.0004 0.31 0.75 0.14
Right ventricle wt (g/EFW, g) 0.0023 0.0016 0.0023 0.0022 0.0004 0.52 0.43 0.45
Left atrium wt (g/EFW, g) 0.0010 0.0009 0.0007 0.0007 0.0002 0.31 0.91 0.91
Right atrium wt (g/EFW, g) 0.0009 0.0008 0.0007 0.0006 0.0002 0.09 0.34 0.99
Septum wt (g/EFW, g) 0.0021 0.0024 0.0025 0.0026 0.0002 0.30 0.55 0.75
Liver wt (g/EFW, g) 0.043 0.046 0.046 0.044 0.002 0.82 0.74 0.40
Total kidney wt (g/EFW, g) 0.0132 0.0101 0.0129 0.0107 0.0009 0.97 0.04 0.78
Adrenal wt (g/EFW, g) 0.0019 0.0005 0.0007 0.00012 0.0006 0.69 0.48 0.17
Total LD (g/EFW, g) 0.025 0.022 0.022 0.020 0.001 0.16 0.29 0.89
Viscera wt (g/EFW, g) 0.092 0.104 0.107 0.096 0.014 0.82 0.99 0.46
Perirenal adipose wt (g/EFW, g) 0.0018 0.0036 0.0034 0.0035 0.0003 0.04 0.13 0.14
LD = longissimus dorsi.
1
CON cows fed to gain 1 kg of BW/week and exceed CP recommendations. NR cows fed 0.55 × of NRC NEm and CP recommendations.

regardless of which trimester the restriction took place. Other Prezotto et al., 2016), however the BW and BCS loss of
studies have utilized a bovine maternal nutrient restric- dams in this study was more significant than these other
tion (Long et al., 2009 and 2010; Camacho et al., 2014; early gestation maternal nutrient restriction, despite similar

318
 Nutrient restriction in early and mid-gestation

Table 4 Foetal measurements and selected organ weights (wt) on day 190 of gestation from control (CON), nutrient restricted
(NR)/CON and CON/NR cows1
Items CON NR/CON CON/NR SEM P value

n 8 7 7
Foetal wt (kg) 10.3c 9.1d 8.9d 0.5 0.08
Foetal empty carcass wt (kg) 8.5a 7.2b 7.2b 0.4 0.03
CRL (cm) 65.6 62.4 63.5 1.5 0.32
Abdominal circumference (cm) 47.3a 43.5b 43.9b 0.9 0.02
Thoracic circumference (cm) 45.6a 42.7b 42.5b 0.9 0.03
Brain wt (g) 106.5 97.3 100.9 4.6 0.37
Heart wt (g) 77.2 65.2 59.9 5.4 0.08
Left atrium wt (g) 4.9 5.1 5.7 0.6 0.65
Right atrium wt (g) 5.9 3.9 4.8 0.8 0.23
Left ventricle wt (g) 21.4a 17.9b 18.9b 0.9 0.04
Right ventricle wt (g) 19.7 16.7 16.9 1.5 0.28
Septum wt (g) 18.2 18.2 17.2 1.2 0.80
Avg. left ventricle thickness (mm) 10.3 11.2 11.4 0.4 0.08
Avg. right ventricle thickness (mm) 8.4 7.4 7.8 0.5 0.37
Avg. septum thickness (mm) 10.3 11.1 10.7 0.4 0.44
Liver wt (g) 315.2a 267.0b 271.0b 14.3 0.05
Lung wt (g) 238.3 218.2 226.1 20.3 0.78
Total kidney wt (g) 81.5 77.2 71.5 5.3 0.43
Adrenal wt (g) 1.6 1.7 1.2 0.2 0.21
Total LD muscle (g) 224.3a 185.5b 162.2b 14.4 0.03
Viscera wt (g) 595.1 557.8 503.3 40.6 0.29
Perirenal adipose wt (g) 34.3c 28.3d 30.8d 1.8 0.09
CRL = crown rump length; LD = longissimus dorsi.
a,b
Means differ P ⩽ 0.05, c,dmeans differ P < 0.1.
1
CON cows fed to gain 1 kg of BW/week and exceed CP recommendations. NR cows fed 0.55 × of NRC NEm and CP recommendations.

Table 5 Foetal organ weights (wt) as a percentage of empty foetal weight (EFW) on day 190 of gestation from control (CON),
nutrient restricted (NR)/CON and CON/NR cows1
Items CON NR/CON CON/NR SEM P value

n 8 7 7
Brain wt (g/EFW, g) 1.21a 1.38b 1.44b 0.03 <0.01
Lung wt (g/EFW, g) 2.83 2.98 3.08 0.21 0.69
Heart wt (g/EFW, g) 0.82a 0.92b 0.93b 0.03 0.04
Left ventricle wt (g/EFW, g) 0.24a 0.28b 0.28b 0.01 <0.01
Right ventricle wt (g/EFW, g) 0.21a 0.25b 0.2 b 0.01 <0.01
Left atrium wt (g/EFW, g) 0.06 0.07 0.08 0.01 0.16
Right atrium wt (g/EFW, g) 0.07 0.13 0.07 0.04 0.45
Septum wt (g/EFW, g) 0.21a 0.25b 0.24b 0.01 0.04
Liver wt (g/EFW, g) 3.71 3.74 3.84 0.11 0.79
Total kidney wt (g/EFW, g) 0.96 1.07 0.99 0.06 0.38
Adrenal wt (g/EFW, g) 0.019 0.023 0.016 0.002 0.13
Total LD muscle (g/EFW, g) 2.64 2.63 2.33 0.19 0.38
Viscera wt (g/EFW, g) 7.04 7.76 7.08 0.43 0.43
Perirenal adipose (g/EFW, g) 0.38a 0.43b 0.44b 0.01 <0.01
LD = longissimus dorsi.
a,b
Means differ P ⩽0.05.
1
CON cows fed to gain 1 kg of BW/week and exceed CP recommendations. NR cows fed 0.55 × of NRC NEm and CP recommendations

restriction levels because our maintenance energy and CP
recommendations calculations did not incorporate nutrition
requirements for pregnancy and were instead based on the
requirements of a non-pregnant, non-lactating cow. This was
done to get back to absolute maintenance energy and
protein basis that other researched have neglected as most
have fed their control animals over requirements. This
allowed us to get both an early and mid-gestation IUGR that
others were not able to obtain (Prezotto et al., 2016). This
was probably due to both our level of NR and also our use of

319
Taylor, LeMaster, Mangrum, Ricks and Long
young cows as young cows are more susceptible to produ-
cing IUGR compared with older cows (Long et al., 2009).
Plasma glucose concentration was decreased in NR dams
and foetuses at both days 110 and 190 of gestation. This
data are consistent with that of sheep that has shown that
NR (50% of requirements) or overfeeding (150% of require-
ments) altered plasma glucose concentrations in ewes and
their foetuses during nutritional treatments (Ford et al.,
2007; Tuersunjiang et al., 2013). In contrast to sheep, Long
et al. (2009) showed that not all NR cattle and foetuses
would exhibit decreased plasma glucose concentrations.
Older cows have been shown to have an increased ability to
partition nutrients to maintain normal foetal growth during a
period of NR. This study also helps to support the fact that
younger cows do not partition nutrients to their foetus
(Long et al., 2009). We would have expected at least some
increased foetal growth in the NR/CON dams after they
were placed on the control ration but there appeared to be
no compensatory foetal growth even while maternal plasma
glucose was similar to the CON/CON cows during mid-
gestation.
Serum triglycerides were decreased in NR dams and foe-
tuses at days 110 and 190 of gestation. Similar decreases in
maternal and foetal serum triglycerides have been shown in
ewes that were NR during early gestation (Ma et al., 2011).
However, of interest is that foetuses that were exposed to NR
during the first trimester and then returned to the control diet
had increased serum triglyceride compared with foetuses
that had been on the control diet. This could indicate possible
selected nutrient partitioning in these NR/CON foetuses.
In obese ewes and their foetuses increased triglyceride
concentrations have been reported relative to their control
counterparts and this through as a reflection of their
increased nutritional status (Zhu et al., 2010). However, in
this present study, this increased serum triglyceride con-
centration in the NR/CON did not result in any changes in
foetal measurements. It should be noted that increased lipid
transport into foetus may not be beneficial, as it has been
shown that increased lipid transport into ovine foetuses
leads to increased development and altered function of
adipose tissue (Zhu et al., 2010; Long et al., 2012a).
Although the foetus can synthesize cholesterol on its
own, there is still placental transport of maternal cholesterol
to the foetal environment (Woollett, 2011). During early
gestation, when demand for growth is lower, cholesterol is
not as compulsory as is indicated by the similarities in serum
cholesterol concentration at harvest in either maternal
or foetal serum at day 110 of gestation. This effect of
similar cholesterol has been noted in foetuses from an early
gestation maternal NR sheep study (Ma et al., 2011). By
mid-gestation (days 110 to 190), as demand for cholesterol
increases in the foetus, NR animals were not able to meet
the demand, resulting in the decreased serum cholesterol
in NR day 190 dams and foetuses. In other species this
reduced cholesterol concentrations is associated with
reduced foetal growth rates (McConihay et al., 2000; Edison
et al., 2007).
320
In day 110 foetuses there were multiple measures that
exhibited a treatment × gender effect, in which foetuses that
were male and NR showed increased evidence of growth
restriction. Typically, male foetuses grow quicker and heavier
than female foetuses (Pedersen, 1980) and are therefore
more susceptible to nutrient restriction. The mechanism for
this gender-specific response in unknown, but it has been
shown that protein restriction during early gestation
leads to gender-specific changes in foetal-placental hemo-
dynamic (Hernandez-Medrano et al., 2015). Another possible
mechanism is a gender-specific response to the elevated
cortisol detected in out NR cows, it has been shown that
gender affects the response to glucocorticoids in ovine
foetuses (Miller et al., 2012).
Increased percentage of perirenal adipose tissue in NR
foetuses has been shown in other species. Increased perire-
nal adipose tissue depots have been reported in twin bearing
ewes on a 70% restricted diet during gestation (Edwards
et al., 2005). Twin foetuses from restricted ewes were
potentially more severely restricted than singleton foetuses
from restricted ewes because of increased nutrient parti-
tioning in utero. This increase in adipose percentage may
alter appetite regulation in the postnatal animal, caused by
changes in endocrine secretions from the adipose tissue
(Wang et al., 2008). This alteration of appetite and increased
adiposity leads to further metabolic disorders such as insulin
and glucose dysregulation, also observed in sheep that were
restricted during early (days 28 to 78) gestation (George
et al., 2012).
In conclusion, whereas it has been previously highlighted
that nutrient restriction during early gestation causes IUGR
and decreased foetal development, mid-gestation nutrient
restriction has now been shown to cause the same damaging
effects to the same extent, even with no nutritional stress
during early gestation. Realimentation of animals after a
period of nutrient restriction alleviates some of the damaging
results of nutrient restriction (Long et al., 2009), but without
realimentation this study shows that the effects of nutrient
restriction cannot be overcome even though many maternal
metabolites return to control levels, indicating a partitioning
of nutrients for the maternal system and not for the foetus in
these young cows.
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