Trop Anim Health Prod (2015) 47:627–631

DOI 10.1007/s11250-015-0757-z

SHORT COMMUNICATIONS

Reproductive performance, metabolic and hormonal profiles

of Santa Inês ewes in winter and summer
under tropical conditions
Mario Felipe Alvarez Balaro &
Felipe Zandonadi Brandão &
Anna Beatriz Veltri Peneiras & Eunice Oba &
Jeferson Ferreira da Fonseca &
Nádia Regina Pereira Almosny &
Elyzabeth da Cruz Cardoso

Received: 16 July 2014 / Accepted: 6 January 2015 / Published online: 18 January 2015
# Springer Science+Business Media Dordrecht 2015

Abstract The study aimed to evaluate the reproductive, met-
abolic and hormonal profiles of Santa Inês ewes during winter
and summer in the tropical climate of Rio de Janeiro State,
Brazil. The ewes (n=16) were prepared for induction of
synchronised oestrus with a short-term hormonal treatment,
and follicular dynamics were assessed with ultrasonography.
Blood samples for biochemical and hormonal analyses were
collected on days zero, six and eight of the protocol at 7, 13
and 19 h. Reproductive behaviour did not differ (P>0.05)
between seasons. Based on ultrasonography data, the ovula-
tion rate was 100 % in summer and 81.3 % in winter. Most of
the ewes showed double ovulation in summer, and the diam-
eter of the largest and second largest follicles differed
(P<0.05) between seasons. Among the biochemical parame-
ters, plasma glucose, plasma urea and serum albumin differed
(P<0.05) between the study periods. Regarding the hormonal
profile, higher serum triiodothyronine, thyroxine and prolactin
M. F. A. Balaro (*) : F. Z. Brandão : A. B. V. Peneiras :
N. R. P. Almosny : E. da Cruz Cardoso
Departamento de Patologia e Clínica Veterinária, Faculdade de
Veterinária, Universidade Federal Fluminense, Rua Vital Brasil, 64,
Santa Rosa, Niterói, Rio de Janeiro 24320-340, Brazil
e-mail: mariobalaro@hotmail.com
E. Oba
Departamento de Reprodução Animal e Radiologia Veterinária,
Escola de Veterinária, Universidade do Estado de São Paulo, Distrito
de Rubião Junior, s/n, Botucatu, São Paulo 18618-970, Brazil
J. F. da Fonseca
Caprinos e Ovinos, Empresa Brasileira de Pesquisa Agropecuária,
Rodovia MG 133, km 42, Coronel Pacheco, Minas
Gerais 36155-000, Brazil
concentrations (P<0.05) were observed in summer. The aver-
age progesterone concentrations demonstrated low reproduc-
tive seasonality. In conclusion, the reproductive performance
of the ewes was superior in summer, and higher levels of
plasma glucose and serum albumin during this season may
have played a role in the observed ovulation rates. The higher
levels of serum prolactin and serum thyroxine in summer
demonstrated that these compounds were not involved in the
regulatory mechanisms underlying the onset of anoestrus in
these Santa Inês ewes under tropical conditions.
Keywords Biochemical . Ewe . Hormones . Oestrous .
Seasonality . Tropical
Introduction
The animal adaptation to the environmental conditions is one
of the key factors that affect the success of the tropical live-
stock production system (Bishop 2012). In Brazil, the sheep
population increased between 2006 and 2012 by more than
20 % to an estimated 18 million head (OIE WAHID 2012).
Santa Inês sheep, a naturalised hair sheep breed, is known for
its production of low-fat meat and good maternal traits; this
breed provides an opportunity for crossbreeding with
specialised meat breeds, enhancing production efficiency
(Paim et al. 2013). Regarding their reproductive parameters,
data are limited from a limited number of animals to oestrus
behaviour and ovulation time (Cardoso Neto et al. 2012;
628
Cavalcanti et al. 2012). Field observations have shown that
Santa Inês ewes may exhibit reproductive activity throughout
the year if receive adequate nutrition outside of breeding sea-
son (Sasa et al. 2011). The metabolic profile in animal repro-
duction may assist in the diagnosis of the energy-protein im-
balances, and on elucidation of other biological responses
(Amaral et al 2009). Triiodothyronine (T3), thyroxine (T4)
and prolactin (PRL) have a hormonal role on the seasonality
and reproductive performance in sheep (Arunakumari et al.
2007; Lehman et al. 2010; Nother et al. 2011). Furthermore,
progesterone (P4) determination has been used to monitor
cyclic activity (Sasa et al. 2011).
Clear information on reproductive behaviour based in the
metabolic-hormonal profiles of breeds with economic and
productive relevance in tropical zones is required. Therefore,
this study aimed to analyse the reproductive performance after
a short-term hormonal treatment in Santa Inês ewes associated
with their metabolic and hormonal profile in two distinct sea-
sons, winter and summer, under the tropical conditions in Rio
de Janeiro, Brazil.
Materials and methods
The study was performed at the Farm School of the Veterinary
Faculty at Universidade Federal Fluminense, located in Rio de
Janeiro, Brazil (latitude 22°27′45″ south). The climate is trop-
ical Aw according to Koppen’s classification. This research
was reviewed and approved by the Animal Care Committee
of the Universidade Federal Fluminense, and was conducted
under the ethical principles of the Brazilian Society of Labo-
ratory Animal Science.
The experiment was conducted in July 2011 (cold and dry
season—winter) and January 2012 (hot and rainy season—
summer). Sixteen nulliparous Santa Inês ewes (21±2.7 months
old/38.7±6.8 kg live weight) were kept in a semi-intensive
system with access to pasture (Panicum sp. and Brachiaria
sp.) and fold. Chopped elephant grass (Pennisetum
purpureum) was offered twice daily, and 300 g/animal of con-
centrate (17 % crude protein) was offered daily. The animals
received water and mineral salt ad libitum.
Intravaginal devices impregnated with 60 mg of
medroxyprogesterone acetate (Progespon®, Schering Plough,
São Paulo, Brazil) were used for 6 days. One day before
sponge removal, 300 IU of eCG (Folligon®, MSD Animal
Health, São Paulo, Brazil) and 37.5 μg of d-clorprostenol
(Prolise®, Tecnopec LTDA, São Paulo, Brazil) were adminis-
tered intramuscularly.
Ovarian ultrasonography was performed every 24 h while
the implant was in place. After sponge removal, the measure-
ments were performed every 12 h until confirmation of ovu-
lation. The exams were performed using an ultrasound ma-
chine (Aquila Vet®, Pie Medical Equipment BV, Maastricht,
Trop Anim Health Prod (2015) 47:627–631
Netherlands) at 7.5 MHz, coupled to a linear rectal transducer.
Diameters, position and characteristics of the ovarian struc-
tures were recorded and drawn using individual chips. A ram
with lateral deviation of the penis and a marker vest was used
to detect oestrus duration in the ewes after sponge removal.
On days zero (sponge insertion), six (sponge removal) and
eight (expected ovulation), blood was collected by jugular
venipuncture at 7, 13 and 19 h into three vacuum tubes (BD
Vacutainer®, United Kingdom) containing (a) sodium
fluoride to obtain plasma glucose, (b) 10 % EDTA to obtain
plasma, and (c) gel clotting activator to obtain serum. After
collection, the samples were centrifuged (2600×g) to obtain
the plasma/serum and were separated into plastic microtubes,
labelled and stored in a freezer at −20 °C for later laboratory
analysis. The plasma analyses included glucose, urea and pro-
gesterone. Serum analyses included triglycerides, total blood
proteins, albumin, prolactin, triiodothyronine and thyroxine.
Biochemical analyses were performed by spectrophotometric
techniques using commercial kits (Labtest System Clinical
Diagnostics) in a colourimetric spectrophotometer (BioPlus®,
São Paulo, Brazil). The hormone assays were performed by
radioimmunoassay using commercial reagents (COAT-A-
COAT®, Diagnostic Products Company, Los Angeles, USA).
Descriptive statistics (winter × summer) to the reproduc-
tive, hormonal and biochemical data were calculated. The
results were analysed with the Statistical Analysis System
programme (SAEG® 9.0, Federal University of Viçosa,
Viçosa, Brazil). Parametric variables were subjected to analy-
sis of variance (ANOVA), and the means were compared
using SNK test (P<0.05). Non-parametric variables were
assessed by chi-square test (P<0.05).
Results
The reproductive behaviour did not differ between winter and
summer (Table 1). With the exception of the ovulation rate,
the data obtained by ultrasonographic evaluation differed be-
tween seasons (Table 1).
All biochemical parameters from metabolic profile
(Table 2) during the study period were within the range for
the species (Kaneko et al. 2008). Higher concentrations of
glucose, urea, albumin, T3, T4 and PRL were found in sum-
mer. However, P4 did not change between seasons (Table 2).
Discussion
Although comparisons of reproductive parameters are diffi-
cult due to differences in breed, age, nutritional status, and
hormonal protocol, previous studies performed under similar
conditions to those used in the current study have reported
similar averages (Cardoso Neto et al. 2012; Cavalcanti et al.
Trop Anim Health Prod (2015) 47:627–631 629

Table 1 Reproductive performance (means±SE) of Santa Inês ewes in winter and summer under tropical conditions

Data Season

Winter Summer Average

Reproductive behaviour
Animals in oestrus (%) 87.5 (14/16) 93.8 (15/16) 90.6 (29/32)
Duration of oestrus (h) 27.2±2.1 31.8±3.8 29.6±2.3
Interval from sponge removal to oestrus onset (h) 33.4±2.1 35.6±2.8 34.5±1.8
Occurrence of ovulation from sponge removal (h) 64.9±2.0 62.8±1.7 64.1±1.3
Oestrus onset to ovulation (h) 31.5±2.1 26.8±3.2 29.2±2.0
Ultrasonography evaluation
Ovulation rate (%) 81.3 (13/16) 100.0 (16/16) 90.6 (29/32)
Average number of ovulations per ewe 1.1b ±0.1 1.9a ±0.1 1.5±0.1
Largest follicle diameter (mm) 5.4b ±0.2 6.0a ±0.2 5.7±0.1
Second largest follicle diameter (mm) 3.4b ±0.2 5.3a ±0.2 4.4±0.2
a, b
Significant difference between means by SNK test (P<0.05)

2012). It is noteworthy that regardless of the season, the mean
interval from sponge removal to ovulation varied little, even
without the use of GnRH and hCG to synchronise ovulation.
Ovulation was concentrated in the last hour of oestrus. This
data is also important in controlled breeding programmes or
when the oestrus onset in ewes is otherwise controlled. A total
of 94 % (15/16) of the ewes showed double ovulation in
summer against 92 % (12/13) of single ovulation in winter.
In fact, these higher ovulation rates in summer indicate higher
productivity per ewe and consequently, higher milk produc-
tion (kg/ewe) and meat (kg/weaned lamb/ewe) for the farmer.
Higher concentrations of plasma glucose, plasma urea and
serum albumin found in summer may be related to the better
pasture quality in this season and the supply of soluble
carbohydrates and protein to the ewes. According to Moreira
et al. (2012), the nitrogen and carbohydrate fractions are in-
fluenced by the time of year and the production cycle in trop-
ical pasture systems. The reason for this metabolic effect can
be explained by the adopted semi-intensive management sys-
tem. Even with controlled feeding, the pasture to which the
animals had access was subjected to seasonal influences.
Moreover, the seasonal differences observed in the reproduc-
tive parameters, such as the ovulation rate and follicular diam-
eter, may be explained by the metabolic profile found. Ac-
cording to Smith and Stewart (1990), although protein and
energy can independently influence ovulation rates, better re-
sults are obtained when both components are simultaneously
increased. Additionally, the state of follicular development at

Table 2 Metabolic and hormonal profile (means±SE) of Santa Inês ewes in winter and summer under tropical conditions

Data Season Average Reference valuesc

Winter Summer

Serum triglycerides (mg/dL) 19.9±0.7 20.3±0.7 20.1±0.5 9–30

Plasma glucose (mg/dL) 62.9b ±0.9 66.5a ±0.8 64.8±0.6 50–80
Plasma urea (mg/dL) 32.9b ±0.7 34.8a ±0.5 33.9±0.5 17.1–42.8
Total blood proteins (g/dL) 7.4±0.1 7.2±0.0 7.3±0.0 6.0–7.9
Plasma albumin (g/dL) 2.8b ±0.0 3.2a ±0.0 3.0±0.0 2.4–3.0
Serum triiodothyronine (ng/mL) 1.0b ±0.1 1.2a ±0.0 1.1±0.0 –
Serum thyroxine (ng/mL) 52.2b ±1.8 64.5a ±3.8 58.4±2.2 –
Serum prolactin (ng/mL) 3.4b ±0.3 5.8a ±0.3 4.6±0.3 –
Plasma progesterone (ng/mL) 2.7±0.6 3.3±0.9 3.0±0.5 –
a, b
Significant difference between means by SNK test (P<0.05)
C
Reference values according to Kaneko et al. (2008)
630
time when maximum concentrations of glucose are available
can be one of the factors that determine the efficiency of the
ovulation rate (Somchit et al. 2007).
Higher T3 serum concentrations in summer may have in-
fluenced the observed reproductive parameters. Dijkstra et al.
(1996) reported that T3 interacted with FSH to promote pre-
antral follicle growth, and this interaction was mediated by
increased FSH-R expression and the action of growth and
differentiation factor-9 (GDF-9) produced by the oocyte.
Higher concentrations of T4 in summer appeared to be
more adaptively favourable for the metabolism profile and,
therefore, for the sheep reproductive performance. T4 is in-
volved in carbohydrate metabolism and promotes hepatic glu-
cose production acting as an energy balance indicator
(Achmadi and Terashima 1995). The Santa Inês ewes exhib-
ited T4 values that were similar to those of Corridale ewes
described by Starling et al. (2005). However, unlike the
Corridale ewes, which were in seasonal anoestrus during sum-
mer, the reproductive performance of the Santa Inês ewes
improved during this same season. According to Lehman
et al. (2010), the increase in the circulating concentration of
T4 could actively act in an inhibitory manner in the central
nervous system (CNS) during spring and summer, preventing
wool sheep breeds from resuming sexual activity. Therefore,
although T4 concentrations were similar between breeds, the
permissive role of the neuroendocrine effect on reproduction
appears to be distinct. Furthermore, Arunakumari et al. (2007)
reported that T4, when associated with FSH, increased follic-
ular growth rates, antrum formation and meiotic competence
after the in vitro culture of preantral follicles. Thus, although
T4 may act as an inhibitor in the CNS, its biological action in
the ovary can be different.
Seasonal prolactin values were consistent with those in other
studies. One of these studies, performed in wild sheep species
by Santiago-Moreno et al. (2005) demonstrated the role of pro-
lactin in the regulation of seasonal reproductive activity,
and that their high concentration in the summer could be related
to the entry into anoestrus. Nother et al. (2011) also showed that
variations in prolactin secretion were not affected by light treat-
ment. Even with higher physiological hormone concentrations
in spring/summer, the ewes were able to become pregnant nat-
urally. Indeed, the role of this hormone in the timing of entry
into anoestrus in tropical ewes is not well understood, and other
neuroendocrine mechanisms may be involved.
In both seasons, more than 50 % of the ewes had progester-
one concentrations above 1.0 ng/mL previous to the hormonal
treatment, confirming the low reproductive seasonality in Santa
Inês ewes (Sasa et al. 2011). These authors demonstrated that
only the use of a ram and the flushing effect were possible
methods to establish their cyclical activity. The practicality of
these techniques is associated with the reduced use of hormones
resulting in time and manual labour savings. Therefore, these
techniques may improve profitability for the sheep farmer.
Trop Anim Health Prod (2015) 47:627–631
Conclusions
Santa Inês ewes showed better reproductive performance in
summer than in winter. Higher levels of glucose, urea and
albumin during summer may be related to the feeding man-
agement due to a better pasture quality, and together with
higher triiodothyronine values, may have influenced the
higher ovulation rate found in this season. Progesterone values
demonstrated the low reproductive seasonality of the breed.
Higher levels of serum prolactin and thyroxine in summer
supported the hypothesis that these hormones played no reg-
ulatory roles in the beginning of the anoestrus season in Santa
Inês ewes under tropical conditions.
Acknowledgments The authors thank the Universidade Federal
Fluminense-Fopesq/PROPPi and FAPERJ (E-26/102.963/2012) for
funding the project. Eunice Oba, Felipe Zandonadi Brandão and Jeferson
Ferreira da Fonseca are fellows of the CNPq.
Conflict of interest The authors declare that they have no conflict of
interest.
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