REPRODUCTIVE PERFORMANCE OF ZEBU CATTLE IN MEXICO

2. SEASONAL INFLUENCE ON THE LEVELS OF PROGESTERONE,

ESTRADIOL-17l3, CORTISOL AND LH DURING THE ES-I-ROUS CYCLE

C. Lamothe-Zavaleta3, G. Fredriksson’ and A. Madej’

Departments of ‘Obstetrics and Gynaecology and ‘Clinical Chemistry

Faculty of Veterinary Medicine, Swedish University of Agricultural
Sciences, S-750 07 Uppsala, Sweden
3Department of Animal Reproduction, Faculty of Veterinary Medicine
University of Veracruz, Veracruz, Mexico

Received for publication: October 30, 1990

Accepted: September 30, 1991

ABSTRACT

Levels of progesterone, estradiol-1713, cortisol and LH were determined in 18

cyclic Zebu females during the rainy (n= 11) and the dry season (n =7). Four cows
studied during the rainy season stopped cycling and were considered to be
anestrous. Blood samples were collected from the animals once daily from Day 2
(day of estrus=Day 1) to Day 16, and three times daily from Day 17 to the day
before the expected onset of estrus, when the frequency was increased to every 3
hours until ovulation. Increases in concentrations of progesterone were more
pronounced in the rainy season (n = 7) than in the dry season (n = 7): on Days 4,
5 and 7 higher concentrations were found during the rainy season (P < 0.1, P < 0.05
and PcO.05 respectively). Decrease in the concentrations of progesterone started
on Day -4 before the next estrus during the rainy season and on Day -3 during the
dry season. The area under the curve of estradiol-17l3 from Day -3 to ovulation was
greater during the rainy period than during the dry period (P < 0.05). In all animals,
a peak of estradiol-17l3 was found around Days 4 to 7 of the cycle (n = 14). In five
animals, a second peak was detected around midcycle. A third peak was recorded
on Day 18 (n=S). Levels of cortisol were higher during the rainy than the dry
season (PC 0.05). The interval between the onset of estrus and the LH peak was
6.7 hours (n=5), while the interval between the LH peak and ovulation was 21.3
hours (n=8). In the dry period, three of the animals had significantly elevated LH
values the day before the LH surge.

Key words: Zebu cattle, season, estrous cycle, reproductive hormones, cortisol

Acknowledgments
Financial support for Carlos Lamothe-Zavaleta from the Swedish International
Programme on Animal Reproduction is gratefully acknowledged. The authors
wish to thank the Faculty of Veterinary Medicine, University of Veracruz,
Mexico, for the facilities provided and to its students and staff for their help. We
are indebted to NIDDK as well as NHPP. University of Maryland School of
Medicine, USA,for the generous donation of bLH.

DECEMBER 1991 VOL. 36 NO. 6 667

 INTRODUCTION

Even though cattle are basically nonseasonal breeders, there is evidence

suggesting that variations in ambient temperature, photoperiod, humidity and feed
supply contribute to seasonal variations regarding reproductive efficiency. High
environmental temperatures increase the secretion of adrenocorticotropin, which in
turn stimulates the secretion of progesterone from the adrenal gland (1). Moreover,
there is a decrease in the secretion of basal levels of LH as well as of the
preovnlatory surge of LH (2). In Zebu cattle, temperatures above 27°C increase the
length of the estrous cycle, reduce the duration and the intensity of estrus, and also
lead to a greater percentage of cows coming into estrus at night (3).

The endocrine regulation of reproductive functions has not been sufficiently

documented in Zebu cattle. Previous studies have reported that Zebu cows produce
less progesterone than Bos cows (4-7). Furthermore, it is known that the
corpus luteum is smaller in Bos than in Bos taut-us (8). Hansel et al. (9)
have reported that the luteal cells of Brahman cows produce less progesterone in
vitro than cells from European breeds. However, other studies report similar
progesterone values in Zebu cows and in Bos taunts breeds (10-12). No detailed
data for LH, estradiol-178 and cortisol during the estrous cycle are available for
Zebu cattle. Most data indicate that the reproductive efficiency in Zebu cows is
lower than in European breeds. Whether this is due to breed differences or to the
climate has not been fully clarified. It is, therefore, of importance to study hormonal
profiles to determine whether these profiles are similar to those reported for
European breeds (Bos taunts) under temperate conditions as well as to determine
the seasonal influence on reproductive hormonal levels in Zebu cows under tropical
conditions.

MATERIAL AND METHODS

Eighteen cyclic Zebu females, 3 to 8 years of age, with a quiet temperament,

were used in the study. The cattle came from the herd belonging to the Faculty of
Veterinary0 Medicine at the University of Veracruz in Mexico. Veracruz is located
on the Gulf of Mexico (at a latitude of 19”12’ north and a longitude of 96”08’ west).
It has a humid subtropical climate. However, from October to May there are
recurrent winds from the north and rains from June to October, with an annual
precipitation of 1677 mm, of which 87% falls during the rainy season; the mean
daily temperature during the dry and rainy season is 22.3”C and 27.4”C, respectively
(13).

The experiment was carried out between September 10 and October 17 1988
during the rainy season (n = 7); between March 22 and April 16 1989 during the
dry season (n = 7); and between August 5 and September 2, 1989 during the rainy
season (n = 4).

The cattle were placed in small fenced pastures with an area of 2 to 3 hectares.
They were grazed on estrella de Africa pasture (m) during the

898 DECEMBER 1991 VOL. 36 NO. 6

 THERIOGENOLOGY

rainy seasons, and were fed with “estrella de Africa” hay and concentrates during
the dry season. Mineral supplementation was given during both seasons.

Sexual behavior

Signs of estrus were observed during one estrous cycle in each animal.
Observation was done three times daily (sunrise, noon and sunset) when estrus was
not expected and then continuously (24 hours/day) starting with signs of proestrous.
Estrus was checked with the aid of penis-deviated bulls. The cow was considered
to be in estrus from the first time she aIlowed a teaser to mount until she refused
to be mounted, or when she had not been mounted by the bull or by another cow
for more than 1 hour.

Clinical examination

Palpation per rectum to determine follicular growth and corpus luteum

development was performed every 4 days until Day 16 of the cycle, when the
animals were palpated daily and checked for external signs ofproestrous until the
following estrus. Palpation per rectum was done every 4 hours to determine the
time of ovulation starting 4 hours after the end of estrus.

Weather report

Reports of temperature, humidity and speed of the wind and rain were obtained
every day from the weather bureau located at the international airport in Veracruz,
located 2 km from the experimental center.

Sample collection

Blood samples were drawn by puncture of the jugular vein into vacutainer tubes.
Blood was collected bled at the same time once daily from Day 2 (day of estrus =
Day 1) to Day 16 of the cycle, when the frequency of samplings was increased to
three times daily. One day before the expected onset of estrus, the frequency was
increased to every 3 hours until ovulation. Since the cows were kept on pasture,
cannulation was not possible as the cannulas would be removed immediately by the
animals. Within 15 minutes after collection, the samples were centrifuged, and
plasma was removed and stored at -20°C until analysis.

Hormone analysis

Analysis of progesterone was performed on one sample per day from Day 2 of
the cycle until the next ovulation. The levels were determined by radioimmunoassay
using an antiserum to 1 la-hydroxyprogesterone-hemi-succinate-bovine serum
albumin (14). The plasma was extracted by petroleum ether with an extraction
recovery of 80%. Dextran-coated charcoal was used for the separation of the free
and antibody-bound hormone. The antiserum cross-reacted < 1% with progestagens,
estrogens, androgens and corticoids except for deoxycorticosterone (3.8%) and 58

DECEMBER 1991 VOL. 36 NO. 6 899

THERIOGENOLOGY

pregnane-3,20-dione (11%). The practical detection limit was 0.5 mnol/l for analysis
of 250 ~1 of plasma. The intra-assay coefficient of variation varied between 8.5 and
11.5% for different ranges of the standard curve and the inter-assay coefficient of
variation ranged from 10.8 to 11.3%.

Analysis of estradiol-1713 was done on a single daily sample from Day 2 to Day
16 of the cycle. From Day 16 to the day of ovulation it was performed on three
samples per day. The levels were determined by enzymeimmunoassay (15) using an
antiserum against estradiol-1786-(0-carboxymethyl)-oxime-bovine serum albumin
conjugate (16). The plasma samples were extracted with peroxide-free diethyl ether
prepared by treatment with A&O,. The enzyme conjugate was prepared by coupling
estradiol-1786-(0-carboxymethyl)-oxime to horseradish peroxidase. The sensitivity
of the assay was 0.22 fmol/well (1.1 pmol/l). The amount of estradiol-178 causing
a 50% reduction of maximum binding was 4.4 fmol/well. The relative cross-
reactions of the antibody were 7% with oestrone sulphate and less than 0.01% with
cortisol, progesterone and testosterone. The intra-assay coefficients of variation
varied between 10.0 and 20.0%. The inter-assay coefficients of variation varied
between 5.9 and 27.1%.

Cortisol was analyzed using the same samples as for estradiol-178, i.e., one
sample daily from Day 2 to Day 16 of the cycle and three samples per day from
Day 16 to the day of ovulation. A Coat-A-Count, solid-phase [‘251]-
radioimmunoassay kit (Diagnostic Products Corporation, Los Angeles, CA, USA)
was used to determine plasma levels of cortisol. Standard curves, using processed
human serum, represented 0, 27.6, 138, 276, 552 and 1380 nmol/l of cortisol. The
detection limit of the assay was 5.5 nmol/l. The Coat-A-Count cortisol antiserum
is highly specific for cortisol with very low cross-reactivity to prednisone (3.1%) ll-
deoxycorticosterone (1.5%), corticosterone (1.4%) and tetrahydrocortisol (1.0%).
Other steroids tested either cross-reacted less than 1.0% or were not detectable by
the assay. The intra-assay coefficient of variation varied between 3.0 and 8.0%. The
inter-assay coefficient of variation varied between 4.7 and 6.3%.

The LH levels in plasma were measured in the samples collected during the day
before estrus until ovulation. The levels were determined by radioimmunoassay
(17), using an antiserum against bovine LH (a-bLH/R7) with a final dilution of 1:60
000. Bovine LH (USDA-bLH-B-5-AFP-5500) was used as the standard. Overnight
incubation, including a 2-hour delay in the addition of [lZI]-bLH, was carried out
at room temperature. The separation of free and antibody-bound hormone was
carried out by adding a second antibody coupled to Micro Sepharose beads
(Pharmacia AB, Uppsala, Sweden). A 0.5-ml suspension was dispensed into each
tube, and after incubation at room temperature for 30 minutes the contents of the
tubes were centrifuged for 20 min at 1500 x g. The intra-assay coefficient of
variation was below 8.0% and the inter-assay one varied from 6.7 to 8.2%.

Statistical Evaluation

Evaluation of the curve of progesterone was done by dividing the estrous cycle

Boo DECEMBER 1991 VOL. 36 NO. 6

 THERIOGENOLOGY

of each cow into four periods. Period 1 (from Day 2 to Day 6, after the first estrus);
Period 2 (from Day 7 to Day 12 after the first estrus); Period 3 (from Day -10 to
Day -5 before the second estrus); and Period 4 (from Day -4 to Day 1 of the
following estrous cycle). The area under the curve for each period was calculated
using an electronic integrator (Digiplan, Kontron Messgerate, FRG). The results
were analyzed as dependent variables using a one-way factorial analysis of variance
from Statgraphics (STSC, Inc., Rockville, MD, USA). The Student t-test was used
to compare the means.

The area under the estradiol-17B curve from Day -3 to ovulation was measured
and analyzed using the same methods as described above for progesterone. The
Scheffe test was used to compare the means.

To evaluate cortisol, the cycle was divided into two parts. The first part included
Day 2 to Day 16. The second part included Day 17 to the day of estrus. The results
were analyzed as dependent variables using a two-ways factorial analysis of variance
from Statgraphics. The Scheffe test was used to compare the means.

To identify significant values, LH and estradiol-17B peaks were calculated

using an iterative approach described previously by Zarco et al. (18). The mean and
standard deviation were calculated in all the samples, and any value greater than
two standard deviations above the mean was considered to be a significant increase.
The calculation was repeated after eliminating the values corresponding to points
identified as significant values. As a result of the elimination of peak values, the
mean value decreased, the standard deviation was reduced, and new points were
identified as significant elevations. The process was repeated until no new
significant elevations were detected.

The results, unless otherwise indicated, are expressed as mean ? standard error
of the mean (SEM).

RESULTS

Four of 18 animals stopped cycling (two from each rainy season) and were
considered to be anestrous. The remaining animals (n= 14) represent two groups
referred to as the rainy season group (n =7) and the dry season group (n=7). No
differences in the climatic conditions were found when rainy seasons from 1988 and
1989 were compared.

The area under the curve of progesterone of the first period was larger in the
rainy season than in the dry season (P~0.05). During the other three periods no
significant differences were found between seasons.

The data for all cycles (Figure 1) show that progesterone levels on Days 2 and
3 of the cycle averaged 0.6 nmol/l during both seasons. Increases in concentrations
started from Day 4 of the cycle in the rainy season and from Day 5 in the dry
season, reaching peak values (20 to 22 nmol/l) between Day 12 (second period) and

DECEMBER 1991 VOL. 36 NO. 6 901

THERIOGENOLOGY

20-
Rainy season -
Dry season ---
I
5 24-
0
E

= 20-
%
0
fi 16-
z
g 12-

i?
z *-
5
= 4-

2 4 6 8 10 12-10-8
-6 -4 -2 1
DAYS OF ESTROUS CYCLE
Figure 1. Levels of progesterone (x+SEM) in the rainy season (-; n=7) and
in the dry season (----; n=7) during the estrous cycle. * indicates
significantly different values.

Day -7 (third period) in the rainy season and peak values of 18 to 21.5 nmol/l
between Day 12 (second period) and Day -5 (third period) in the dry season. The
increase in concentrations of progesterone was more pronounced in the rainy than
in the dry season as well as on Day 4 (P < O.l), Day 5 (P < 0.05), and Day 7 (P < 0.05)
with the higher concentrations found during the rainy season. The decrease in
concentrations of progesterone started on Day -4 in the rainy season and on Day -
3 in the dry season. Values of progesterone during both seasons were below 1
nmol/l between Day -1 and the day of estrus (Day 1).

The area under the curve from Day -3 to the time of ovulation was larger (P
< 0.05) during the rainy period (n = 7) than during the dry period ( n .= 7) (1178
4 336 mm2 vs 753 * 175 mm’). The first peak of 19.5 pmol/l (range 12 to 39
pmol/l) of estradiol-1713 was found in all the animals around Days 4 to 7 of the
cycle (n= 14). In five of the animals, a second mean peak of 27.6 pmol/l (range 14
to 47 pmol/l) of estradiol-17l3 was detected around midcycle. A third peak was
recorded on Day 18, with a mean value of 27.4 pmol/l (range 10 to 35 pmol/l)

902 DECEMBER 1991 VOL. 36 NO. 6

 24- 80

18- 60

12- 40 0,
I/,
: ;;:
20 b
; 6 0
;;
; o-
O 7
0 2 4 6 8 10 12 14 16 10 20 1 3
LT z
: 24- -80 =
vl -0
3
' 18- -60 :
:
a I
12- -40 1

6- - 20

O- -0
2 4 6 8 101214'l6182022 2
DAYS OF ESTROUS CYCLE
Figure 2. Levels of progesterone (-) and estradiol-17B (----) in Cows 205 and
304 during the estrous cycle. Horizontal black bar indicates estrus.
Vertical arrow shows time of ovulation.

(n=S). Plasma levels of estradiol-17l3 rose from a mean basal concentration of 10.2
pmol/l (range 5.4 to 19.8 pmol/l) during the period of luteal regression to 48.8
pmol/l (range 25 to 70 pmol/l) around the onset of estrus. In most of the animals,
the peak coincided with the onset of estrus and only in one animal did the peak
occur after the onset of estrus. After that, levels of estradiol-170 decreased rapidly
to basal levels. Figure 2 depicts estradiol-1713 levels in Cow 205, representing the
animals which had estradioL17l3 peaks on Days 4 to 7, around midcycle and at the
onset of estrus, and in Cow 304, representing animals which had estradiol-17l3 peaks
on Days 4-7, Day 18 and at the onset of estrus.

The levels of cortisol at Day 2 to Day 16 of the cycle averaged 35.2 nmol/l in
the rainy season (n = 7) and was significantly higher (P < 0.05) than 14.1 nmol/l
in the dry season (n = 7; Figure 3). From day 17 to the day of estrus, values of
cortisol were again higher (P < 0.05) in the rainy season than in the dry season
(52.8 nmol/l vs 20.8 nmol/l). The differences in cortisol values between the two

DECEMBER 1991 VOL. 38 NO. 6 903

THERIOGENOLOGY

loo-

$, 60-
;z
E 40-
%
w
2z
20-

1 3 5 7 9 11 13 15 17 19 21 23
DAYS OF ESTROUS CYCLE

Figure 3. Levels of cortisol (xr SEM) in the rainy season (-* n=7) and in the
dry season (----; n=7). Blood samples were taken once daily during the
days preceding e) and more frequently later.

parts of the cycle (Days 2 to 16 vs Days 17 to 1) were significant (PcO.05) during

both seasons.

Levels of LH were analyzed in only eight animals. Due to infrequent blood

collection, the results of the LH from the remaining 11 cows were not included in
this portion of the study. The interval between the onset of estrus and recording of
the LH peak was 6.7 hours (n=5), and the interval between the LH peak and
ovulation was 21.3 hours (n=8). The mean value of the LH peak was 13.0 pg/l,
with a range of 4.8 to 19.4 pg/l (n=8). In two of the animals from the dry season,
the onset of estrus took place after the LH peak had occurred (6 and 3.2 hours).
In one of the cows the estradiol-178 peak occurred 3 hours after the onset of estrus
(Figure 4, Cow 205), while in another cow the estradiol-1713 peak was recorded
before the onset of estrus. Three of the animals in the dry period had significantly
elevated LH levels the day before the LH surge (Figure 4). Levels of cortisol and
the area under the estradiol-178 curve were not different between animals with and
without significant LH values the day before the preovulatory LH surge. However,
the previous LH peaks were preceded by an increase in estradiol-178 values.

DECEMBER 1991 VOL. 36 NO. 6

 THERIOGENOLOGY

21- -70
cow 205 4 4

15- -50

9- -30

= 3- -10
z o- iz
-0 VI
g 21- -70 2
ZD
E (3
g IS- -50 5;
I - r

: 9- -30 i
;3

; 3- -10 :
0
; O- -0 z
= 21- -70 (
-l
I
15 - - 50

9- - 30

3- -10
O- -0
24 6 12 18 24 6 12 18 24 6 12 10 24 6 12 18 HOURS
0.20 0.21 0.1 0.2

DAYS OF ESTROUS CYCLE

Figure 4. Leve!s of luteinizing hormone (-) and estradioL17D (----) during the
estrous cycle in Cows 205, 210 and 213. First vertical arrow indicates
onset of estrus. Second vertical arrow/s indicate ovulation/s.

Anestrous Cows

Levels of progesterone on Days 4, $6, 7 and 8 of the cycle were higher in the
cows that stopped cycling (n=4) than in those that did not in the dry season
(P<O.O5), but similar to those found in the rainy season. The peak values (23 to 28
nmol/l) tended to be higher than in the other two groups (PcO.1). The decrease
in concentrations of progesterone started on Day 17 of the cycle in two cows and
on Days 19 and 20 in the other two cows. In two of the cows, progesterone values
above 20 mnol/l reached values below 1 nmol/l in 1 day, when luteolysis occurred.
The other two cows had a decrease in progesterone similar to that in the cyclic
cows (i.e., values below 1 nmol/l were reached within 2 to 3 days.

In three of the cows, three estradiol-171.3 peaks were recorded during the cycle
(Days 4 to 6, midcycle and Day 17). However, in the fourth cow, a peak was

DECEMBER 1991 VOL. 36 NO. 6 905

THERIOGENOLOGY

detected on Day 8 but not during the rest of the cycle, when the values were always
lower than in the cyclic cows (Figure 5). During the days following the decrease in
the concentration of progesterone, values of estradiol-1713 increased and decreased
repeatedly, ranging between 21 and 38 pmol/l in three of the cows. The fourth cow
had lower levels of estradiol-1713 during this time (Figure 5).

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29

DAYS AFTER ESTRUS

Fig. 5. Levels of progesterone (- ) and estradioL17l3 (---) in Cow 302, which

never returned in estrus.

During Days 2 to 16 of the cycle as well as Day 17 and onwards, the mean
values of cortisol (19.1 and 28.4 nmol/l, respectively) were significantly lower than
those observed in cows from the rainy season (PcO.05) but similar to those from
the dry season.

DISCUSSION

Progesterone levels during the first period of the estrous cycle were higher in
the rainy season than in the dry season. However, during the remainder of the
luteal phase, concentrations of progesterone were similar during both seasons.
These results are similar to those of Abilay et al. (l), who found that Guernsey
heifers exposed to temperatures of 33.5”C produced more progesterone throughout
the first estrous cycle than the controls (182°C). However, during the following

906 DECEMBER 1991 VOL. 36 NO. 6

 THERIOGENOLOGY

cycles differences were only noticed when progesterone concentrations increased,

but not during the remainder of the luteal phase. It seems that when the animals
become accustomed to high temperatures, the production of progesterone is not as
high as at the beginning of the exposure. The higher values of progesterone during
the rainy season could be explained by the possibility that high environmental
temperatures may increase the concentration of progesterone, this production being
of adrenal origin (1,19).

The maximum values of progesterone in our study support those of previous

reports (lo-12), but are somewhat different from those reported by Vaca et al. (4)
and Oyedipe et al. (7). It is known that the incubation period and temperature of
whole blood, from collection to the separation of serum or plasma, significantly
affect the concentration of progesterone in cattle (20-22).

The estradiol-1713 peak around Days 4 to 6 of the cycle has already been
reported in the Bos taurus (23-26), with the second peak recorded around midcycle
(27). Several authors have postulated that two to three waves of follicular activity
occur during the bovine estrous cycle (28-31). During the immediate post-estrus
period, FSH increases in the absence of any increase of LH (25). Furthermore,
Schams et al. (23) found that FSH peaks occurred around Days 4, 8, 12 to 13, 17,
I8 and 20 of the cycle and that a wave-like pattern could be observed. The increase
in FSH may play a role in the recruitment of preantral follicles which are the
source of the increase in estradiol-17!3.

The preovulatory peak in estradiol-17l3 around the onset of estrus has also been
reported in the Bos taurus (24,26). The marked decrease in plasma estradiol-17L3
after the onset of estrus is believed to be due to the preovulatory gonadotropin
surge converting estrogen-active follicles to estrogen-inactive follicles, changing the
secretory activity of the granulosa cells.

During the period from Day -3 to ovulation, the amount of estradiol-1713

produced was higher during the rainy than the dry season. Lamothe-Zavaleta et al.
(3), working with Zebu cows in Mexico, found that proestrus is longer during the
rainy than the dry season. Furthermore, our findings show that concentrations of
cortisol were higher in the rainy season than in the dry season. It seems that the
higher temperatures during the rainy season were more stressful, and that the
animals responded by producing more cortisol. Since cortisol inhibits the response
of the pituitary gland to GnRH, a longer period with high values of estradiol-1713
might be necessary to stimulate the release of LH and the onset of estrus.

The levels of cortisol found in our present study are similar to those of
previous reports in Bos tam-us (1,19,26). The higher levels of cortisol in the rainy
season can be explained by the fact that the temperature and humidity in the rainy
season are higher than in the dry season. Similar results have been reported by
Abilay et al. (1). This also indicates that Bos indicus cattle are susceptible to
climatic/heat stress. It should, however, be emphasized that the higher levels of
cortisol in the rainy season were seen despite the difference in frequency of blood

DECEMBER 1991 VOL. 36 NO. 6 907

THERIOGENOLOGY

collection. Results of this and our previous study (3) indicate that the relatively long
estrous cycle in Zebu cows during rainy season may be attributable to the relatively
high cortisol levels. This is similar to the findings of Vighio and Liptrap (32), who
recently reported that administration of dexamethasone during the middle of the
luteal phase prolonged the duration of the estrous cycle in cow.

The interval between the onset of estrus and the LH surge is similar to that in
studies done on the Boss (23,26). However, Randel (33) reported that in
Brahman cows, the LH surge occurs 0.4 hours after the onset of estrus. A report on
the Bos taurus (23) indicates that the interval between the LH surge and ovulation
is longer in heifers than in cows (27.1 vs 23.0 hours). However, according to our
data it seems that this interval is shorter in the Bos indicus. This is also supported
by the findings of Randel (33). The levels of LH recorded in this study are lower
than those previously found both in Bos and Bos indicus cows (23,26,33-36).

In the present study, the onset of estrus occurred after the LH peak in two
animals. It is possible that the manifestation of sexual behavior in one of the
animals was delayed. Vaca et al. (37) suggested that behavioral manifestations of
estrus can be inhibited or delayed by stress. In Cow 205 the preovulatory LH peak
was only 4.8 pg/l but it lasted around 20 hours. The peak of estradiol-1713 occurred
3 hours after the onset of estrus, when the preovulatory LH surge had already been
released. After that, the concentration of estradiol-178 decreased rapidly. These
findings suggest that in this cow, due to the low levels of LH, the preovulatory LH
surge was not able to modify the secretion of the granulosa cells until the total
release of LH had occurred.

The factors that influenced the release of LH prior to the preovulatory LH

surge in three of the animals remain unknown. Levels of estradiol-178 and cortisol
were not different in cows with or without peaks of LH prior to the preovulatory
LH surge. However, small increases in estradiol-178 were previously detected,
suggesting irregular follicular growth.

Four of the animals selected during the rainy season stopped cycling. The higher
temperatures and humidity during this season could have a negative effect on estrus
manifestation. This has been reported earlier in Bos taunts by several authors
(2,38-40). The hormonal findings support this in the way that the progesterone
levels showed an even more pronounced alteration than in the other cows during
the rainy season compared to levels during the dry season. Changes of estradiol-170
levels are inconsistent around the time of expected estrus, a finding which is
reflected by poor and indecisive follicular development.

The physiological significance of the low concentration of plasma cortisol in the

anestrous cows is not clear. It is possible that depressed plasma cortisol levels
reflect inactivity of the 17-hydroxylating enzyme in the adrenal cortex that is
responsible for the synthesis of cortisol from progesterone, thereby enhancing the
accumulation of metabolites of progesterone (1). The cows that stopped cycling are
probably those which were the most susceptible to climatic stress during the rainy

908 DECEMBER 1661 VOL. 36 NO. 6

 THERIOGENOLOGY

season.

These hormonal findings support the results of a clinical study on reproductive

performance in Zebu cattle by Lamothe-Zavaleta et al. (3), in which it is suggested
that well-nourished Zebu cattle show a better reproductive performance during the
dry season than during the rainy season.

REFERENCES

1. Abilay, T.A., Johnson, H.D. and Madan, M. Influence of environmental heat

on peripheral plasma progesterone and cortisol during the bovine estrous
cycle. J. Dairy Sci. 3:1836-1840 (1975).

2. Tucker, H.A. Seasonal@ in cattle. Theriogenology 1753-59 (1982).

3. Lamothe-Zavaleta, C., Fredriksson, G. and Kindahl, H. Reproductive

performance of Zebu cattle in Mexico. 1. Sexual behavior and seasonal
influence on estrous cyclicity. Theriogenology 36: (1991)

4. Vaca, L.A., Galina, C., Fernandez-Baca, S., Escobar, J. and Ramirez, B.

Progesterone levels and relationship with the diagnosis of a corpus luteum by
rectal palpation during the estrous cycle in Zebu cows. Theriogenology
&):67-76 (1983).

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