Transcript Slayt 1 - Werfft

SS Ay1, I Kucukaslan2, D Kaya3, SB Mulazimoglu2, B Emre4, C Kacar3, H Kalender5, H Bollwein6, M Findik1, J Scholbach7, S Aslan2
1Veterinary
Faculty, Samsun, Turkey; 2Veterinary Faculty, Ankara, Turkey; 3Veterinary Faculty, Kars, Turkey; 4Veterinary Faculty, Urfa, Turkey; 5Veterinary Faculty,
Kırıkkale, Turkey; 6University of Veterinary Medicine, Hannover, Germany ;7Chameleon Software, Leipzig, Germany
INTRODUCTION
β-carotene functions independently of vitamin A in regulation of ovarian functions (Lotthammer, 1999). Supplementation to the ration or injection of β-carotene increase plasma concentration of βcarotene (Arechiga et al., 1998; Wang et al., 1988; Sales, 2007). β-carotene also has a positive effect on the size of corpus luteum (Aslan et al., 1998; Agaoglu et al., 2009). The stimulation of the
gene Connexin-43 and formation of Gap-junction by β-carotene shows its effect positive on functions of the corpus luteum (Arıkan and Rodway, 2000; Arıkan et al., 2002). Angio mod of the color
Doppler ultrasonography is a non-invasive and safe imaging method to display blood viscosity of corpus luteum (Myamoto et al., 2005; Honnes et al., 2009).
The aim of the present study was to determine the effect of administration of GnRH or GnRH+ β-carotene combinations on Luteal size (LS) and Luteal Blood Flow (LBF) in pregnant and non-pregnant
cows.
MATERIALS and METHODS
In the present study, 29 Holstein-Friesian cows were used in the postpartum 50-80 day. The animals were divided into two groups as β-carotene uninjected (β-Car (-); n=15) and β-carotene injected
(β-Car (+); n=14). All cows were synchronized with single dose of PGF2 (Enzaprost®; 5ml/cow, im, Ceva-Dif) and double artificial inseminations were performed at the time of 48 and 72 hours after
synchronization. Last insemination was accepted as a day 0. GnRH (Receptal®; 2.5ml/cow, im, Intervet) was injected in both groups at the days of synchronization (-3), 7 and 17. The injections of βCar (Carofertin®; 20ml/cow, im, AlvetraWerfft AG) were performed only β-Car (+) group unlike β-Car (-) group at the day -3, 7 and 17. Images of corpus luteum (CL) and blood samples were collected
to measure the luteal size (LS), luteal blood flow (LBF) and blood serum β-Car and progesterone concentrations at the days -3, 7, 10, 17, 27 and 37. The CL was displayed with color-doppler
ultrasonography (LOGIQ BookXP, Germany) with a 10.0 MHz linear-array transducer and at least six images were taken in each examination. In order to minimize variations between recordings, the
settings for the Power Doppler system were fixed and the same settings were used in all examinations. The images were taken when the largest CL was displayed from the route of transducer lateromedial and dorso-ventral. After the transfer of these images as a DICOM format to the computer, four images with the maximum number of color pixels within the sectional planes near the maximum
diameter of the CL were selected for calculating the mean blood flow. The colored area was measured using a software (Pixel Flux™, Version 1, Chameleon Software, Leipzig, Germany) and used as
a semi-quantitative parameter for LBF. Data were analyzed (Descriptive Statistic, Mann Whitney U-Test, Repeated Measures Define Factors-Test) using SPSS 16.0 (SPSS Inc., Chicago, USA).
Table 1. Study Dizayn
Synchronization
Groups
-3
Inseminations
-1
βC-(n=15) : PGF2
0
7
10
GnRH GnRH GnRH
βC+(n=15): +βC
+βC
17
27
37
DAYS
GnRH
+βC
+βC: days of β-car injections
RESULTS
There was significant increase in β-Car levels after injection of β-Car in β-Car(+) group
(Graphic 1). After inseminations, it was determined that there was significant difference
between β-Car(-) and β-Car(+) groups at all of the pregnancy days (except day 17)
(P<0.05; P<0.01) (Graphic 2). It was not found any difference in blood serum
Graphic 1. Changes of the blood serum β-car concentrations in
groups.
μg/dl
μg/dl
700
700
b**
600
b*
400
300
b*
b*
a
a
a
a
a
300
a
There was a positive correlation (r=0.882; P<0.01) between LS and serum β-Car levels
200
200
in β-Car(+) group (Table 2).
100
100
0
-3βC
7βC
βC (-)
Correlation
CL size / Ser β-Car levels
R2
-0.129
Day
17 βC
β-Car (-)
Table 2. Correlation (R2) between LS and blood serum β-car concentrations in groups at day -3 and 37.
Groups
10
27
CL size / Ser β-Car levels
0.881
a
a
b*
b**
a*
a*
a
b*
a
b*
0
-3 βC
37
7 βC
β-Car (+)
10
β-car (-)
17
Day
βC
27
37
β-car (+)
P*<0.05; P**<0.01
P*<0.05; P**<0.01
Graphic 3. Changes of the CL size during early pregnancy in
groups.
Graphic 4. Comparisons of CL size between groups during
pregnancy.
P
P>0.05
cm2
βC (+)
a**
500
400
a
a*
600
a
500
progesterone concentrations between groups (P>0.05).
Graphic 2. Comparisons of blood serum β-car concentrations
between groups.
P<0.01
5.5
b**
b*
a
5
b*
a,b
a,b
b
a*
The mean size of CL at day 17 was determined significantly different in day 17 from
a**
4.5
a**
a*
a
other days in β-Car(-) group (P**<0.01; P*<0.05). On the other hand, size of CL at days
10, 17 and 37 was found significantly bigger than days 0 and 37 in β-Car(+) group
Day
4
-3βC
7βC
10
17 βC
27
37
cm2
2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
β-car (-)
A gradually increase of luteal blood flow was detected in β-Car(+) group from day 7
(except day 37) (P<0.01). In β-Car(-) group, there was an increase in day 10 (P<0.01),
a stable level in following days and no significant increase between days 10 and 37
(Graphic 5) .
significantly higher than β-Car(-) group (P*<0.01; P**<0.01) (Graphic 6).
a*
Day
7 βC
β-car (+)
10
β-Car (-)
17 βC
27
37
β-Car (+)
P*<0.05; P**<0.01
Graphic 5. Luteal blood flow values between day 7 and 37 of
pregnancy
Graphic 6. Statistical difference of luteal blood flow between day
7 and 37 of pregnancy.
mm2
3
d**,*
d**
2.5
a*,**
a,b,c**
b*
a,b,c**
c*,**
a,c
1.5
a,c
a,c
a*
b**
1
0.5
The pregnancy rate was found to be higher in β-Car(+) group (71.4%) than in β-Car(-)
a**
β-car (-) =P*<0.05; P**<0.01
β-car (+)=P*<0.05; P**<0.01
2
As a result of β-Car injections, LBF in β-Car(+) group at days 7 and 27 was determined
b*
-3βC
(P**<0.01; P**<0.05) (Graphic 3). CL size at days 27 (P*<0.05) and 37 (P**<0.01) in βCar(+) group was found to be bigger than β-Car(-) group at same days (Graphic 4).
b**
Day
0
-3βC
7βC
group (53.3%).
10
β-Car (-)
17βC
27
37
mm2
2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
b**
b*
a**
a*
Day
-3βC
β-Car (+)
β-car (-) =a*:b*=P<0.05; b*:c*=P<0.05; a:b:c P**<0.01
β-car (+)=a:b:c=P<0.05; a:b:c:d=P<0.01
7 βC
10
β-Car (-)
17 βC
27
37
β-Car (+)
P*<0.05; P**<0.01
CONCLUSION
Injection of β-Car in addition to the GnRH injections in early pregnancy days after inseminations LS, LBF and blood serum β-Car concentrations have significantly increased
compared to β-Car(-) group. It was thought that this resulted in higher pregnancy rate in β-Car(+) group.