Original Article
Ovarian response and conception rate following
oestrus synchronization using three protocols in
Egyptian buffalo heifers
Hassan A. Hussein*1; Ragab H. Mohamed2; Mohamed Hossam3; Axel Wehrend4
1
Theriogenology Department, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt; 2Theriogenology Department, Faculty of Veterinary Medicine,
Aswan University, Aswan, Egypt; 3Ard-El-khair Buffalo Farm, Abnob, Assiut, Egypt; 4Clinic for Obstetrics, Gynaecology and Andrology of Large and Small Animals,
Justus Liebig University, Giessen, Germany
Keywords
Reproduction, CIDR, Ovsynch, PGF2α
Summary
Objective: The aim of this study was to monitor the ovarian response
and conception rate following estrous synchronization using CIDR,
Ovsynch and double prostaglandin F2α protocols in Egyptian buffalo
heifers. Material and methods: A total of 80 cyclic buffalo heifers
were divided into four equal groups: CIDR (intravaginal progesterone
releasing device, EAZI-BREEDTM CIDR®), Ovsynch (GnRH, PGF2α, GnRH
injections), PGF (double PGF2α doses) and control. Timed artificial in-
semination (TAI) was performed in all heifers. All animals were exa-
mined using ultrasound and blood samples were collected for mea-
surement of progesterone. Results: A new follicular wave occurred
earlier in the Ovsynch and PGF groups than in the CIDR group
(p < 0.05). The mean diameter of the ovulatory follicle was smaller in
the CIDR group than in the Ovsynch and PGF groups (p < 0.05). The
ovulation rate was 100% in the CIDR group, 75% in the Ovsynch
group and 70% in the PGF group. In the control group a lower preg-
nancy rate (20%) was determined in than in the CIDR (35%), Ovsynch
(40%) and PGF (35%) groups. Progesterone concentrations were nu-
merically higher in pregnant heifers of the CIDR group but the dif-
ference was non-significantly compared to the Ovsynch, PGF and con-
trol groups (p > 0.05). Conclusion and clinical relevance: EAZI-
BREEDTM CIDR®, Ovsynch-based TAI and PGF protocols were effective
in synchronizing oestrus and resulted in nearly similar pregnancy rates
in Egyptian buffalo heifers.
Correspondence to
Hassan A. Hussein
Theriogenology Department
Faculty of Veterinary Medicine
Assiut University
Assiut 71526
Egypt
Email: hassansabour69@yahoo.com
Introduction
Buffaloes are an integral part of the agricultural economy and live-
stock production in Egypt. The total number of buffaloes in Egypt
reaches about 3.950 millions, of which 42% are dairy cows, 6%
© Schattauer 2016
287
Schlüsselworter
Reproduktion, CIDR, Ovsynch, PGF2α
Zusammenfassung
Ziel dieser Studie war, bei ägyptischen Büffelfärsen die Reaktion der
Ovarien und die Konzeptionsrate nach Östrussynchronisation mit
CIDR, Ovsynch und zweimaliger PGF2α-Gabe vergleichend zu erfassen.
Material und Methoden: Insgesamt 80 zyklische Büffelfärsen wur-
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den in vier gleich große Gruppen eingeteilt: CIDR-Gruppe (intrava-
ginale progesteronfreisetzende Spange, EAZI-BREEDTM CIDR®), Ov-
synch-Gruppe (GnRH, PGF2α, GnRH-Injektion), PGF-Gruppe (zweima-
lige Gabe von PGF2α) und Kontrollgruppe. Alle Tiere wurden termin-
orientiert besamt. Bei allen Tieren erfolgten eine ultrasonographische
Untersuchung und eine Blutentnahme zur Progesteronbestimmung.
Ergebnisse: Eine neue Follikelwelle trat in der Ovsynch- und PGF-
Gruppe früher auf als in der CIDR-Gruppe (p < 0,05). Der mittlere
Durchmesser des ovulatorischen Follikel war in der CIDR-Gruppe
kleiner als in der Ovsynch- und PGF-Gruppe (p < 0,05). Die Ovulations-
rate betrug in der CIDR-Gruppe 100%, in der Ovsynch-Gruppe 75%
und in der PGF-Gruppe 70%. In der Kontrollgruppe ergab sich eine nie-
drigere Trächtigkeitsrate (20%) als in der CIDR- (35%), Ovsynch- (40%)
und PGF-Gruppe (35%). Die Progesteronkonzentration lag in der CIDR-
Gruppe höher als in der Ovsynch-, PGF- und Kontrollgruppe, doch war
der Unterschied nicht signifikant (p > 0,05). Schlussfolgerung und
klinische Relevanz: CIDR-, Ovsynch- und PGF-Protokolle erwiesen
sich als wirksam in der Brunstsynchronisation von ägyptischen Büffel-
färsen und führten zu fast gleich hohen Trächtigkeitsraten.
Ovarielle Reaktion und Konzeptionsrate nach Östrussynchronisation mittels
drei verschiedener Protokolle bei ägyptischen Büffelfärsen
Tierärztl Prax 2016; 44 (G): 287–294
http://dx.doi.org/10.15653/TPG-160214
Received: February 20, 2016
Accepted after revision: June 28, 2016
Epub ahead of print: September 6, 2016
buffalo bulls, 32% heifers less than 2 years old and 20% male calves
less than 2 years old. The aggregate share of buffalo milk from all
types of production systems is about 81% of the total milk produc-
tion in Egypt (10). In buffalo herds, late attainment of puberty,
lower intensity of oestrus expression, seasonality of calving and
Tierärztliche Praxis Großtiere 5/2016
288 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers
long subsequent postpartum anoestrus periods are all factors that
can affect reproductive efficiency much more than in cows (5, 20,
41). Thus, difficulty in oestrus detection is one of the main causes
for low reproductive performance in herds where reproductive
biotechnologies such as artificial insemination have been applied.
In these cases, the low incidence of homosexual behaviour during
oestrus and the wide variation in the duration of oestrus are as-
pects that limit the use of this technology.
Recently, oestrus synchronization and the synchronization of
ovulation have been studied with the aim of making this a viable
technique for buffalo herds. Progesterone and prostaglandin F2α
(PGF2α) were administered to synchronize oestrus in swamp buf-
faloes (15), Mediterranean buffaloes (18, 32), Murrah buffaloes
(33), Mediterranean x Murrah buffaloes (8), Egyptian buffaloes (2,
24, 26), and beef cattle (30). It was reported that a sequence of
GnRH and PGF2α could synchronize ovulation using fixed timed
artificial insemination (TAI). The CIDR device is successful in
synchronizing oestrus and resulted in higher pregnancy rates in
cattle (16). The use of CIDR in reproductive management to in-
crease fertility is gaining popularity because of the short treatment
period (7 days) and the reduced incidence of persistent follicles
(1). In Egypt, there are no data on the use of this protocol in
swamp buffaloes (45). Despite the number of studies that have
been published in the last few years on oestrus synchronization in
buffaloes (12, 18, 19), not many studies have described follicular
dynamics during these hormonal protocols; this knowledge is of
great importance for TAI (7). Furthermore, limited work has been
carried out on the application of oestrus synchronization in buffa-
lo heifers (14, 32, 36).
This study evaluated the possibility of controlling oestrus and
ovulation synchronization in cyclic buffalo heifers and compared
three treatments (Eazi-BreedTM CIDR®, Ovsynch and PGF2α re-
gimes) used to facilitate artificial insemination (AI) and improve
conception rates in Egyptian buffalo heifers in Upper Egypt.
Materials and methods
Animals and management
A total of 80 apparently healthy Egyptian female buffalo heifers
(Bubalus bubalis) were included in this study. The mean age
± SEM was 28.7 ± 0.9 months and mean weight was 365 ± 2.5 kg;
body condition scores were ≥ 3.5 (scale: 1 = thin, 5 = fat [9]). The
animals were housed in groups in open yards with shelters that
correspond to 50% of the total area in the commercial buffalo farm
(Abnoob, Assiut province). Buffalo heifers were fed forage dry
matter (Egyptian clover, alfalfa) and a concentrate mixture. Wheat
straw was available ad libitum. These rations provided 16% crude
protein (CP) and 67% total digestible nutrient (TDN). The ex-
periment was conducted between January and April 2013
(57.29 ± 2.3% relative humidity and 18.45 ± 0.9 °C maximum at-
mospheric temperatures). Before starting the synchronization pro-
gram, the reproductive tracts of all females were examined trans-
Tierärztliche Praxis Großtiere 5/2016
rectally and by ultrasound to record ovarian and uterine variables.
All heifers were cyclic and none exhibited congenital anomalies or
clinical endometritis.
Experimental design and treatments
The 80 heifers were allocated to one of four groups. Heifers in the
first group (CIDR, n = 20, two replicates) received a controlled in-
ternal progesterone releasing device (EAZI-BREEDTM CIDR®,
Pfizer, USA) at random stages of the oestrous cycle (designated as
day 0). Thereafter, heifers in this group received 25 mg PGF2α in-
tramuscularly (Dinoprost, Lutalyse®, Pfizer, Pharmacia and Up-
john Company, NY, USA) on day 6 of the CIDR insert. The inserts
were removed on day 7. The animals were artificially inseminated
48 and 72 hours after CIDR removal using thawed semen from the
same origin.
Heifers in the second group (Ovsynch, n = 20, two replicates)
were synchronized using a 10 µg GnRH treatment on day 0 (Bu-
serelin, Receptal®, Intervet International B.V., Boxmeer, Holland).
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Seven days later, 25 mg PGF2α (5 ml) were administered intramus-
cularly and 48 hours later (day 9) the animals received a second in-
tramuscular dose of 10 µg GnRH. All animals were artificially in-
seminated 16–28 hours after the second GnRH treatment.
Heifers in the third group (PGF, n = 20, two replicates) were
synchronized using double PGF2α injections. All animals received
25 mg PGF2α intramuscularly on day 0. Animals exhibiting oes-
trous symptoms were artificially inseminated. The other animals
were injected with a second PGF2α dose on day 11 and insemi-
nation took place 3 days after the second treatment.
Heifers in the fourth group (control, n = 20) were inseminated
during natural oestrus; these animals served only as a reference for
the pregnancy rates. Follicular dynamic and ovulation rate could
not be recorded in this group.
Ultrasound examination
All heifers were examined rectally and by ultrasound (Pie Medical,
100 LC, Maastricht, the Netherlands) with a sonar device con-
nected to a 6/8 MHz changeable transducer. Examinations were
performed once daily from day 0 to day 8 in both CIDR and Ov-
synch groups and from day 0 to 11 in the PGF group, then every
12 hours until ovulation was observed. The number, diameter and
relative position of all follicles were recorded. Follicles were cat-
egorized as either small (3–5 mm), medium (5–8 mm) or large
(≥ 9 mm). Corpora lutea diameters were also measured and
sketched separately for each heifer. Ovulation was considered to
have occurred when a large, growing antral follicle that had been
observed for several days was no longer detectable (22). If the large
antral follicle persisted for a long time without ovulation and with
increasing wall thickness, it was considered luteinized. The cor-
pus luteum diameter was measured at the widest part; the mean
of two measurements was used if the corpus luteum was elon-
gated. Regression of the corpus luteum was considered to have
© Schattauer 2016
 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers 289

occurred when its diameter decreased markedly and this was la-
ter confirmed by hormonal analysis (progesterone concentration
< 1 ng/ml).
The following ovarian characteristics were identified and com-
pared between groups: emergence of the new follicular wave;
emergence is defined as the first day in which the dominant follicle
is retrospectively identified and has a diameter of 3–5 mm (21);
the interval between the beginning of treatment and the emer-
gence of a new follicular wave; the diameter of the ovulatory fol-
licles; ovulation, defined as occurring on the day when the large
follicle (≥ 10 mm) was no longer observed. Pregnancy was diag-
nosed sonographically 40 days after AI. The pregnancy rate per
TAI was defined as the percentage of cows that were confirmed
pregnant at the single pregnancy diagnosis after one TAI (37).
Blood sampling
In animals of the treatment groups blood samples were collected
from the jugular veins into non-heparinised tubes twice weekly
inter- and intra-run precision coefficients of variation of 4.5% and
2.6%, respectively.
Statistical analysis
The data were presented as mean ± SEM. Statistical analysis was
carried out using the statistical package for social sciences for
Windows (43). Differences among groups were evaluated by one-
way ANOVA for the replicated parameters. Changes in the proges-
terone concentration were tested with repeated measure analysis.
Means were compared by the least significance difference at 5%
level of probability.
Results
At the beginning of the experiment, 50 heifers had corpora lutea
and 35 heifers had a follicle ≥ 10 mm (▶ Table 1, only the data of
the treatment groups are presented). All heifers in the CIDR group

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for 2 weeks prior to the program start and on days 0, 2, 4, 7, 9,
10 and 11 of the program. The blood samples were then taken
twice weekly from all groups until pregnancy diagnosis on day 40.
The samples were transported on ice to the laboratory within
20–30 minutes, centrifuged at 3000 rounds/minute for 20 minutes
and the harvested serum was stored at –20 °C until analyzed for
progesterone (P4). The P4 concentration was determined using a
direct ELISA technique. Kits were provided by Diagnostic System
Laboratory Co. (Bio Check, Foster City, CA 94404, USA) and use a
micro-well method. This kit has a sensitivity of 0.0625 ng/ml with
did not show ovulation during the insertion period but ovulation
occurred after the CIDR removal. After CIDR insertion in the
same group, 10 animals showed regression of the follicles (atresia),
five heifers had a persistent follicle and the other five showed no
reaction (neither ovulation nor atresia). In the Ovsynch group
after the first GnRH injection, ovulation was recorded in seven
heifers, follicular regression was noticed in eight heifers, while one
had a persistent follicle and the other four heifers showed no reac-
tion. In the PGF group after the first dose of PGF2α, ovulation was
recorded in seven heifers, follicular atresia was reported in eight

Table 1 Parameter Treatment group

Ovarian reaction in buffalo heifers after begin-
ning of treatment with CIDR, Ovsynch and PGF. CIDR Ovsynch PGF
Tab. 1 Heifers with a CL 14/20 (70%) 13/20 (65%) 12/20 (60%)
Eierstockreaktion von Büffelfärsen nach Beginn Heifers with a follicle ≥ 10 mm 8/20 (40%) 10/20 (50%) 8/20 (40%)
der Behandlung mit CIDR, Ovsynch und PGF
Heifers with ovulation of the DF after – 7 7
CIDR insertion, GnRH-1, PGF-1
Heifers with follicle atresia 10 8 8
Heifers with a persistent follicle 5 1 1
Heifers with no response (no ovulation, 5 4 4
no atresia)
Diameter of the dominant follicle on 11.2 ± 0.7a 11.7 ± 1.0a 10.9 ± 0.8a
day 0 (mm)
Diameter of the CL on day 0 (mm) 13.7 ± 0.3a 13.9 ± 1.0a 14.0 ± 0.6a
CL regression after CIDR insertion, 4/12 (33.3%) 7/12 (58.3%) 11/11 (100%)
GnRH-1, PGF-1

Treatment groups: CIDR = controlled internal drug release (EAZI-BREEDTM CIDR®), Ovsynch =
protocol using GnRH – PGF2α – GnRH, PGF = double prostaglandin F2α protocol
CL = corpus luteum, DF = dominant follicle, GnRH-1 = first dose of GnRH, PGF-1 = first dose of
PGF2α
Values with the same superscripts in the same row do not differ significantly.

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290 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers

Parameter Treatment group Table 2

Effects of treatment protocols with CIDR, Ov-
CIDR Ovsynch PGF synch and PGF on follicular dynamics, ovulation
Number of heifers with the emergence 12/20 (60%) 18/20 (90%) 16/20 (80%) rate and time of ovulation in buffalo heifers.
of a new follicular wave after treatment Tab. 2
Interval between beginning of the proto- 4.9 ± 0.7a 2.5 ± 0.6b 2.9 ± 0.5b Auswirkungen der Behandlungsprotokolle mit
col and emergence of the new follicular (4–5) (2–3) (2–3) CIDR, Ovsynch und PGF auf Follikeldynamik,
wave (range in days) Ovulationsrate und Ovulationszeitpunkt bei
Büffelfärsen
Diameter of the dominant follicle on 9.1 ± 0.6a 11.3 ± 0.9b 11.0 ± 0.7b
day 7 (mm)
Diameter of the CL on day 7 (mm) 14.9 ± 0.3a 11.1 ± 1.7b 12.4 ± 0.2c
CL regression on day 9 (n [%]) 10/10 (100%) 8/12 (66.7%) 0/7 (0.0%)
a b
Diameter of the ovulatory follicle (mm) 11.3 ± 0.3 14.9 ± 0.1 15.3 ± 0.4b
Follicular growth rate (mm/day) 0.73 ± 0.04a 1.2 ± 0.07b 1.43 ± 0.1b
Ovulation rate (n [%]) 20/20 (100%) 15/20 (75%) 14/20 (70%)
Interval to ovulation after CIDR removal 59.7 ± 12.9a 20.5 ± 5.4b 68.2 ± 6.4a
and last treatment (hours)

Treatment groups: CIDR = controlled internal drug release (EAZI-BREEDTM CIDR®), Ovsynch =

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protocol using GnRH – PGF2α – GnRH, PGF = double prostaglandin F2α protocol
CL = corpus luteum
Values with different superscripts in the same row differ significantly (p < 0,05).

heifers, while one heifer had a persistent follicle and the other four
heifers showed no reaction (▶ Table 1). The diameters of both fol-
licles and corpora lutea on day 0 (day of treatment begin) did not
differ significantly among groups (p > 0.05; ▶ Table 1). Regression
of the corpus luteum was recorded in four, seven and 11 heifers
Fig. 1 Time of oestrus occurrence in buffalo heifers in the treatment
groups CIDR (EAZI-BREEDTM CIDR®), Ovsynch (protocol using GnRH – PGF2α
– GnRH) and PGF (double prostaglandin F2α protocol).
Abb. 1 Zeitpunkt der Auftretens von Brunstsymptomen bei Büffelfärsen in
den Behandlungsgruppen CIDR (EAZI-BREEDTM CIDR®), Ovsynch (Applika-
tion von GnRH – PGF2α – GnRH) und PGF (zweimalige Applikation von
PGF2α)
after CIDR insertion, first GnRH dose and first dose of PGF2α, re-
spectively (▶ Table 1).
The proportion of heifers with emergence of the follicular
wave within 7 days after the beginning of treatment differed
among the CIDR (60.0%), Ovsynch (90.0%) and PGF (80.0%)
groups (▶ Table 2). The interval from beginning of treatment to
follicular wave emergence was longer in the CIDR group than in
the two other groups (p < 0.05). There was no significant differ-
ence in the time of follicular wave emergence between the Ov-
synch and PGF groups (p > 0.05, ▶ Table 2).
The diameter of the dominant follicle on day 7 was significantly
smaller in the CIDR group than in the Ovsynch and PGF groups
(p < 0.05). Mean CL diameter on day 7 was significantly different
(p < 0.05) between the three protocols. The mean diameters of
ovulatory follicles on days 9–11 were significantly smaller in the
CIDR group than in the Ovsynch and PGF groups (p < 0.05).
Ovulation was recorded in all heifers (100%) of the CIDR group,
75% of the Ovsynch group and 70% of the PGF group. The inter-
val between CIDR removal and last treatment until ovulation was
the shortest in the Ovsynch group (p < 0.05) but did not signifi-
cantly differ between the CIDR and PGF groups (▶ Table 2).
All heifers showed oestrus after removal of the CIDR; this rate
was higher than in the Ovsynch and PGF groups. The proportion of
cows with synchronized oestrus (by 48 hours after CIDR removal in
the CIDR group and by 18 hours after the last dose of GnRH in the
Ovsynch group) differed from the PGF group (▶Fig. 1).
The pregnancy rate was lower in the control group (20%) than
in the CIDR (35.0%), Ovsynch (40.0%) and PGF (35.0%) groups
(▶ Table 3).

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 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers 291

Table 3 Parameter Treatment group Control

Effects of treatment protocols with CIDR, Ov-
synch, PGF or natural oestrus (control group) on CIDR Ovsynch PGF
the pregnancy rate in buffalo heifers. Pregnancy rate 7/20 8/20 7/20 4/20
Tab. 3 (35%) (40%) (35%) (20%)
Auswirkungen der Behandlungsprotokolle mit CIDR,
Treatment groups: CIDR = controlled internal drug release (EAZI-BREEDTM CIDR®), Ovsynch =
Ovsynch und PGF oder des natürlichen Östrus (Kon-
protocol using GnRH – PGF2α – GnRH, PGF = double prostaglandin F2α protocol
trolltiere) auf die Trächtigkeitsrate bei Büffelfärsen

The progesterone level in the Ovsynch group increased grad-

ually up to the 6th day after the first GnRH treatment (p < 0.05).
The average P4 concentration in the CIDR group decreased
sharply after administration of PGF2α on days 6 and 7 after the
CIDR was removed (p < 0.05; ▶ Fig. 2). Progesterone concen-
trations in the pregnant heifers in all groups did not differ up to
the 3rd week after insemination. Between the 4th and 7th weeks
post insemination the progesterone concentrations in pregnant
heifers in the CIDR group were numerically higher but not signifi-

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cantly different (p > 0.05) from those in the Ovsynch, PGF and
control groups (▶ Fig. 3).

Discussion
The emergence of a new follicular wave was observed in all treat-
ment groups; the highest percentage occurred in the Ovsynch
group, followed by the PGF group, while the lowest percentage was
observed in the CIDR group. A possible explanation for this result
is that progesterone levels in the CIDR group were higher than in
the other two groups; this in turn led to decreased numbers of
heifers showing emergence of a follicular wave. The emergence of
a new follicular wave appears to be critical for high pregnancy
rates in a progestin-based TAI protocol. This event is necessary to
prevent the development of persistent follicles and must take place
early enough to allow for sufficient growth prior to the induction
of ovulation (16, 27, 37, 39).
Interestingly, the mean diameter of the dominant follicles on
day 7 (day of CIDR removal and PGF2α injection in the Ovsynch
group) was significantly smaller in the CIDR group than in the
other groups. Differences are presumed to be the consequence of
dominant follicles emerging from a new follicular wave in the
CIDR group, where progesterone levels were high. Consequently,
the proportion of heifers with pre-ovulatory follicles > 13 mm on
day 9, a condition which may reduce pregnancy rates following Fig. 2 Progesterone concentration (mean ± SEM) from beginning of treat-
ment (day 0) to day 11 in serum of buffalo heifers in the treatment groups
TAI, was greater in the Ovsynch and PGF groups than in the
CIDR (EAZI-BREEDTM CIDR®), Ovsynch (protocol using GnRH – PGF2α –
CIDR group. Similar results have been reported in cattle (27, 28). GnRH) and PGF (double prostaglandin F2α protocol).
In this study, subsequent follicular development was synchron- Abb. 2 Progesteronkonzentration (Mittelwert ± SEM) von Beginn der Be-
ous among treatment groups of buffalo heifers. In the CIDR group, handlung (Tag 0) bis Tag 11 im Serum von Büffelfärsen in den Behandlungs-
all heifers experienced ovulations due to very low levels of proges- gruppen CIDR (EAZI-BREEDTM CIDR®), Ovsynch (Applikation von GnRH –
terone, especially following PGF2α treatment; hence, all dominant PGF2α – GnRH) und PGF (zweimalige Applikation von PGF2α)
follicles had undergone ovulation within 72 hours. Oestrus with
subsequent ovulation (indicated by the presence of a CL) in heifers

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292 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers
Fig. 3 Progesterone concentration (mean ± SEM) post insemination in
pregnant buffalo heifers in the treatment groups CIDR (EAZI-BREEDTM
CIDR®), Ovsynch (protocol using GnRH – PGF2α – GnRH) and PGF (double
prostaglandin F2α protocol) and in the control group.
Abb. 3 Progesteronkonzentration (Mittelwert ± SEM) nach der Besamung
im Serum tragender Büffelfärsen in den Behandlungsgruppen CIDR (EAZI-
BREEDTM CIDR®), Ovsynch (Applikation von GnRH – PGF2α – GnRH) und PGF
(zweimalige Applikation von PGF2α) sowie in der Kontrollgruppe
following CIDR removal suggests that progesterone (P4) released
from the intravaginal CIDR in these animals was absorbed
through the vaginal wall and entered circulation (31, 42, 45). This
increased P4 concentration exerted negative feedback on the hy-
pothalamus and anterior pituitary, which in turn promoted storage
of GnRH, FSH and LH. Once P4 therapy had stopped (after CIDR
removal by day 7), PGF2α treatment led to a rapid decline in P4
concentrations. This decrease promoted the release of GnRH, as
the negative feedback of P4 is followed by the release of FSH and
LH, with subsequent controlled ovulation and good synchroni-
zation (3, 4, 46).
After the second dose of GnRH in the Ovsynch group, ovu-
lation was precisely synchronized and was observed within
18 hours in 16 of 20 buffalo heifers. The precision of synchrony of
oestrus depends on the synchrony of follicle development (35).
The diameters of the largest follicle in heifers that ovulated were
larger than those in animals that showed no ovulation. In the pre-
sent study, ovulation time in heifers was earlier than that reported
in the previous studies. In crossbred (Murrah x Mediterranean)
buffaloes, ovulation occurred an average of 26.5 and 24.4 hours
after the second GnRH or LH treatments, respectively (8, 33). In
cattle, ovulation occurred 24–32 hours after the second GnRH
treatment (38). This difference in response to the same treatment
in different studies may be related to parity and species.
Conclusion for practice
EAZI-BREEDTM CIDR®, Ovsynch-based timed artificial insemination
and PGF2α protocols were effective in synchronizing oestrus and re-
sulted in nearly similar pregnancy rates in Egyptian buffalo heifers.
This information may be useful for oestrus synchronization in buffalo
heifers.
Tierärztliche Praxis Großtiere 5/2016
Pregnancy rates in the present study were higher in the CIDR
and Ovsynch groups than in the control group. In crossbred buffa-
loes (Murrah X Mediterranean [8]) and Brazilian buffaloes (6),
conception rates after TAI were reported as 55.6–64.2%; these
rates are higher than we documented in the present study. In
contrast, a pregnancy rate of only 15% has been reported in
swamp buffalo heifers and of only 42.9% in buffalo cows (14). Pos-
sible reasons for this marked difference may include differences in
parity, breed, management or environmental conditions. The low
pregnancy rates we documented may result from the fact that with
the Ovsynch TAI protocol, the second injection of GnRH may in-
duce ovulation of the dominant follicle before development is
complete and therefore the quality of the follicle, oocyte and/or the
subsequent corpus luteum may be compromised and plasma con-
centrations of progesterone are too low to maintain pregnancy.
Reduced secretion of progesterone to below a critical threshold is
one cause for embryonic loss in cattle (25, 29, 40) and a similar ex-
planation has been proposed for buffaloes (11–13).
To control the timing of oestrus with exogenous hormones, it is
Downloaded by: LMU München. Copyrighted material.
necessary to ensure that either induced regression of the CL or ter-
mination of a progesterone treatment coincides with the selection
of the dominant follicle during the wave; this method ensures pre-
cise onset of oestrus and high fertility (25, 34). In the present study,
all heifers were inseminated approximately 72 hours after the se-
cond PGF2α dose in the PGF group, an interval that should be ap-
propriate (oestrus at 60 + 12 hours, according to the a.m./p.m.
rule). The resulting pregnancy rate was only 35% but when other
significant variables and interactions are considered, pregnancy
rates after TAI improved. 100% of the heifers in the CIDR group
exhibited oestrus but only 45% of the treated heifers became preg-
nant; the reason remains unclear. Early embryonic death is a pos-
sible factor but further study is needed. Similar results have been
reported in cattle (25) and buffaloes (11).
Progesterone levels in the control group were not comparable
because there was wide variation in the timing of ovulation (asyn-
chronous); in the PGF and Ovsynch groups, progesterone concen-
trations decreased to basal levels in response to the PGF2α dose
and then increased linearly. This pattern appears to be associated
with ovulation of the dominant follicle present at the time of
GnRH injection and subsequent CL formation (2, 25–27). In
contrast, progesterone levels were high in the CIDR up to day 7
after CIDR insertion and then decreased markedly after the PGF2α
dose (▶ Fig. 2). Progesterone concentrations during the late luteal
phase but before insemination have been shown to positively in-
fluence conception rates (44). In this study, progesterone concen-
trations during the period when the CIDR was in place may have
influenced pregnancy rates but there is no direct evidence to sup-
port this statement. Progesterone levels after conception were high
and similar in all groups up to the 3rd week of pregnancy. After the
3rd week of pregnancy, the level of progesterone in the pregnant
heifers in the CIDR group was higher than in the other groups;
this may be because some heifers ovulated, even if they were al-
ready pregnant. It was reported that the CL diameter and maternal
© Schattauer 2016
 H. A. Hussein et al.: Oestrus synchronization in Egyptian buffalo-heifers
serum P4 concentration of pregnant ewes were higher than in
non-pregnant ewes, and ewes bearing a single fetus had a lower P4
concentration than that found in ewes bearing twin fetuses (23).
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgements
The authors thank Pfizer Animal Health Inc., Egypt for partial fi-
nancial support. We also thank all workers in the Abnub buffalo
farm for their help with this work.
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Rezension len der Leitindikationen für die Anwendung

Zivilisationskrankheiten des Pferdes – Ganzheitliche Behandlung
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