2023.08.03.125
Files > Volume 8 > Vol 8 No 3 2023 > Diyala 3

Comparison of the three methods for pregnancy detection in Iraqi Sheep

A. M. F. Al-Mohammedy1
and Y. T. Abdul-Rahaman1,*
1 College of Veterinary Medicine, University of Fallujah, Iraq.
* Correspondence:
[email protected].
Available from.
http://dx.doi.org/10.21931/RB/2023.08.03.125
ABSTRACT
Twenty Iraqi ewes aged between 2-3
years were used in the current study. All animals were raised in an animal
field belonging to the Fallujah in the Saqlawiyah area from August 2022 to
February 2023. It was fed naturally, and on one diet and T1 Control, T2 was
given hCG- COX 100 IU/ ewe injection, T3 was given Biotin 10 mg/ ewe/ daily
oral, and T4 was given Biotin 10 mg/ ewe/ daily oral + hCG- COX 100 IU/ ewe
injection. After mating, blood samples for progesterone and Sheep Pregnancy
Associated Plasma Protein B (PAPPB) measurements were collected on Days 48 and
79. The study's results showed accuracy when diagnosing pregnancy by the PAPPB
method, which was 100% during the 48th and 79th day of pregnancy—followed by
the method of diagnosis by ultrasonography. Hence, the progesterone assay
method. At the same time, specificity was superior in the PAPPB method. The
accuracy of pregnancy diagnosis between PAPPB and ultrasonography on day 48 of
pregnancy was equal. While there were no results during the diagnosis using
progesterone assay. In conclusion, the pregnancy diagnosis by PAPPB is
considered one of the most accurate and modern methods.
Keywords:
Pregnancy
detection; progesterone; ultrasonography; PAPPB; Iraqi Sheep.
INTRODUCTION
Livestock contributes a large percentage to the
agricultural sector, around 50% to most countries. This wealth is essential in
the national income of many countries, and livestock is meant to be raised
cattle, sheep, goats, buffaloes, camels, horses, poultry and fish. Concerning
sheep, they have an essential role in the economies of many countries, past and
present, and are mainly raised for the production of meat, wool, milk, and
leather3.
Early diagnosis of pregnancy in sheep has
an essential role in the economy of countries to reduce increased expenses on
the herd and exclude sick animals 4. The method of diagnosing progesterone is one of the primary methods for
diagnosing pregnancy in animals, including sheep, as the high concentration of
the hormone in the blood or milk is considered an indication of the presence of
pregnancy due to the presence of the corpus luteum.
Nevertheless, sometimes, the diagnosis cannot be
considered correct in this way due to early embryonic death and the survival of
the corpus luteum in the secretion of progesterone. Therefore, the diagnosis of
pregnancy by ultrasonography method was essential in detecting pregnant females
on the 21st day of pregnancy, but this method requires an
experienced person and a high cost. The PAPPB method is a modern and accurate
method for detecting pregnancy in sheep at an early stage of pregnancy through
placenta formation. Early detection of pregnancy reduces the expenses, costs
and effort of the farmer while excluding sick animals, increasing the
productivity and fertility of the herd1,2,11,15,16,17,20. This field
study aimed to compare the accuracy of progesterone, ultrasonography and PAPPB
test for early pregnancy detection in Iraqi Sheep.
MATERIALS
AND METHODS
Twenty Iraqi ewes aged
between 2-3 years with body weight between 34-48 kg were used in the current
study. All animals were raised in an animal field belonging to the Fallujah in
the Saqlawiyah area between August 2022 and February 2023. The animals were
isolated for 30 days before the study began and were examined by the
ultrasonography apparatus to ensure they were not pregnant. The animals were
divided randomly into 4 equal groups. It was fed naturally and on one diet and
the first group (T1) Control, the second group (T2) was
given hCG- COX 100 IU/ ewe injection, the third group (T3) was given
Biotin 10 mg/ ewe/
daily oral (30 days) and the fourth group (T4)
was given Biotin 10 mg/
ewe/ daily oral (30 days) + hCG- COX 100 IU/ ewe injection.
Estrous synchronization is unified by placing vaginal sponges for 13 days.
After
mating, blood samples for progesterone and Sheep Pregnancy Associated Plasma
Protein B (PAPPB) measurements were collected on Days 48 and 79. The blood was
centrifuged at 4000 rpm for 15 minutes. The plasma was collected in sterilized
plastic tubes and kept at -20°C. Progesterone concentrations were measured by COBAS (E-411). The kit provided by Roche, Inc.,
Switzerland, and Sheep Pregnancy Associated
Plasma Protein B (PAPPB) concentrations were measured using ELISA technology—the kit provided by Sunlong
Biotech Co., Ltd, China. The indices were determined as Positive predictive
value: 100 × a/a + b; Negative predictive value: 100 × c / c + d; Sensitivity:
100 × a/a + d; Specificity: 100 × c / c + b; Accuracy (%) = (a +c/ a + b + c +
d) ×100.
RESULTS
Between days 48 PM (16 ewes) and 79 PM (20 ewes),
were declared pregnant using the progesterone assay. The sensitivity of
progesterone assay for detecting pregnant ewes increased from 76.47 to 100%
during the experimental period. The sensitivity of progesterone assay for
detecting pregnant ewes decreased accuracy (65%) at day 48 PM and increased
accuracy (85%) at day 79 PM. The specificity of the progesterone test in
detecting non-pregnant ewes on days 48 and 79 was equal (0%). A similar trend
was observed for positive predictive values, which decreased progressively from
81.25% at day 48 PM, which increased again at day 79 PM to 85%. On the
contrary, the negative predictive value of this test for the detection of
non-pregnant females was equal at day 48 PM and day 79 PM. Better results were
obtained for detecting pregnant and non-pregnant ewes using the ultrasonography
technique Fig (1). The sensitivity for detecting pregnant ewe was high (94.12%)
at day 79 PM and moderately accurate (76.47%) at day 48 PM Fig (2). The
specificity (%) for detecting non-pregnant ewe obviously from 66.67% at day 79
PM Fig (3), and 100% during day 48 PM. The positive predictive value (%) for
detecting the pregnant ewe was increased (100%) at day 48 PM and lower at day
79 PM (94.12%). In contrast, negative predicting values were lower (42.86%) at
day 48 PM and higher (66.12%) at day 79 PM. Accuracy decreased (80%) at day 48
PM and increased (90%) at day 79 PM. The results of the current study showed
the compatibility between accuracy, specificity, sensitivity, and predictive
value of positive and negative tests during the test periods days 48 and 79
using the PAPPB method (100%) (Table 1).
Figure 1. Ultrasonogram of the ewe before treatment
Figure 2.
Ultrasonogram of the ewes at day 48 PM
Figure 3.
Ultrasonogram of the ewes at day 79 PM
Table 1. Early pregnancy detection of ewes using three different methods
DISCUSSION
The plasma progesterone assay is
often the method for early pregnancy detection. This method is accurate and can
be done for the second week of pregnancy15.
Karen et al.11 found
that sensitivity (100%), specificity (95.4%), positive (81.6) and negative
(100%) predictive values for progesterone at day 18 after AI.
Ganaie et al.7 noted
that the accuracy of detecting pregnancy at 100 % and non-pregnancy at 83.3%
was the same for days 15-30 and 31-45 of pregnancy, And the accuracy of 98%,
prediction values of 97.7 and 100% for positive and negative detection, at days
18.23±0. 78. The researcher also showed that the determination of progesterone
hormone by ELISA on the 18th day after fertilization is considered one of the
reliable methods for diagnosing pregnancy in Corriedale ewes.
The difference in results may be
due to the difference in the sensitivity of the progesterone test.
Affects: irregularity of the estrous
cycle, early embryonic deaths, ovarian and ovarian pathological conditions like
hydrometra, pyometra and luteal cysts as the result of hormone concentration8.
The results of the current study
did not agree with this as it was found that sensitivity (95.2%), specificity
(100%), positive predictive value (100%), negative predictive value (80%) and
accuracy (96%) and 19, noted that sensitivity 91.30%,
specificity100%, positive predictive value 100%, negative predictive value
ration 83.3% for pregnancy examination findings by transabdominal
ultrasonography on days 34- 35.
Ultrasonography is used to detect
pregnancy in a variety of domestic species, such as ewes. The use of
ultrasonography has proven effective for the examination of ovarian follicles,
corpora lutea and pregnancy. Genetic factors and nutrition affect the process
of diagnosing pregnancy in ewes by ultrasound, and the placenta and its
development6 determine the gestational age.
Studies have shown that the optimum
time for transabdominal or transrectal ultrasonography in ewes ranges from 12-
23 days PM may correctly predict approximately 85% of pregnant sheep and by 40
days using external trans-abdominal. The sex is determined between 60-69 days,
as the accuracy was (100%) for males but only (76%) for female fetuses. The
number of fetuses can be counted accurately from about 45 to 100 days of
gestation. So detection is usually undertaken between the 12- 13 weeks after
insemination5, 14. The diagnosis was made on day 16. The accuracy
was 50%, whereas at day 23 (72.73%) and day 30 (90.91%)12.
Age affects the sensitivity of the
test by ultrasonography, as it increases in young ewes and decreases in older
ewes on the 40th day of pregnancy; in addition, the examination method, whether
external or internal, affects the accuracy of the ultrasonography result17,18.
Embryo development can be divided
into three periods: First period, initial attachment of the blastocyst; second
period, The embryonic period extends from day 12 to day 34 in the ewes. In this
period, growth and differentiation occur, during which the significant tissues,
systems and organs are established and the significant features of the external
body form. The third period, the fetal period, extends from about day 34 of
gestation in ewes until birth. Growth and changes in the form of the fetus characterize
this period. Species, breed, and strain differences in fetal size are due to
differences in the rate of cell division, which is determined genetically.
Thus, there is close integration between the feed supply to the fetus
(environmental factors), the rate of cell division (genetic factors), and
hence, the rate of growth9.
The current study agreed with the analysis
of 13, showing that pregnancy can be diagnosed by the PAPPB method
on the 45th day of gestation in ewes.
PAPPB are significantly higher in
sheep-carrying twin than in those carrying singles because the PAPPB are
secreted by binucleate cells, which represent 15–20% of fetal placental cells,
and the total placental masses was higher areas of contact between the fetal
placenta and maternal caruncles than those in sheep carrying singles10.
CONCLUSIONS
In
conclusion, the pregnancy diagnosis by PAPPB is
considered one of the most accurate and modern methods, followed by
ultrasonography and progesterone assay. In sheep, pregnancy can be clearly and
accurately diagnosed by ultrasound on the 48th day of pregnancy.
REFERENCES
1. Akköse,
M. Evaluation of a bovine rapid visual PAG ELISA test and transabdominal
ultrasonography for early pregnancy diagnosis in Awassi Sheep. KSU Journal of
Agriculture & Nature. 2020, 23 (5): 1366-1372. doi:10.18016/ksutarimdoga.vi.668707
2. Alkan,
H.; Kivrak, M. B.; Satilmis, F.; Tekindal, M. A. and Dinc, D. A. Detection of
twin pregnancies in ewes by pregnancy-associated glycoprotein assay and
transabdominal ultrasonography. Domestic Animal Endocrinology. 2020, 72:
106399.doi: 10.1016/j.domaniend.2019.106399
3. Alkass,
J. E.; Hermiz, H. N. and Baper, M. I. Some aspects of reproductive efficiency
in Awassi ewes: A review. Iraqi Journal of Agricultural Sciences. 2021, 52(1):
20-27.doi: 10.36103/ijas.v52i1.1232
4. Andueza,
D.; Alabart, J. L.; Lahoz, B.; Muñoz, F. and Folch, J. Early pregnancy
diagnosis in sheep using near-infrared spectroscopy on blood plasma.
Theriogenology. 2014, 81 (3): 509- 513.doi: 10.1016/j.theriogenology.2013.10.016
5. Bragança,
G. M.; Balaro, M. F. A.; Fonseca, J. F.; Pinto, P. H. N.; Rosa, R. M.; Ribeiro,
L. S.; Almeida, M. S.; Souza Fabjan, J. M. G.; Garcia, A. R. and Brandao, F. Z.
Doppler ultrasound in the diagnosis of early pregnancy in sheep. Animal
Reproduction. 2016, 13 (3): 587.
6. Othman
Ghazi Najeeb Alani , Yassen Taha Abdul-Rahaman and Thafer Thabit Mohammed.
Effect Of Vêo® Premium and Vitamin C Supplementation on Lipid Profile Before
and During Pregnancy in Some Local Iraqi Ewes During Heat Stress. Iraqi Journal
of Science.2021, Vol. 62, No. 7, pp: 2122-2130.
7. Ahmad,
A. Sh., F.T. Al-Rawi, Y.T. Abdul-Rahaman, TH. T. Mohammed, K.I. Mahmud. Effect
of Tryptophan injection on the semen quality in Iraqi Shami buck. IOP
Conference Series: Earth and Environmental Science, 2021, 761(1), 012093.
8. Ishwar,
A. K. Pregnancy diagnosis in sheep and goats: a review. Small Ruminant Research.
1995, 17: 37- 44.
9. Jainudeen,
M. R. and Hafez, E. S. E. Gestation, prenatal physiology, and parturition. In:
Reproduction in farm animals. Hafez, B. & Hafez, E. S. E., Part. 2, Chapter
10, 7th ed., Lippincott Williams & Wilkins, U.S.A. 2000, P. 147.
10. Karen,
A.; Amiri, B. E.; Beckers, J. F.; Sulon, J.; Taverne, M. A. and Szenci, O. Comparison of accuracy of
transabdominal ultrasonography, progesterone and pregnancy-associated
glycoproteins tests for discrimination between single and multiple pregnancy in
sheep. Theriogenology. 2006, 66(2): 314- 322.doi:10.1016/j.theriogenology.2005.11.017
11. Tatar
, A.; Kasaeizadeh, . R. .; Shobeirinia, B. .; Abdali, . L. .; Baeelashaki, R. .
The Effect Of Using Different Levels Of Vitamin E On The Productive Traits Of
Broiler Chickens. JLSAR 2023, 4, 30-36
12. Metodiev,
N.; Dimov, D.; Ralchev, I. and Raicheva, E. Efficiency of transrectal
ultrasonography for pregnancy diagnosis of Ile de France ewes during the first
month of gestation. Bulgarian Journal of Veterinary Medicine. 2017, 20 (3):
276- 280.doi:10.15547/bjvm.944
13. Pickworth,
C. L.; Sheffield, J. and Poole, R. K. Pregnancy specific B concentration for
the prediction of lambing rate in sheep. Small Ruminant Research. 2020, 184:
106052.doi:10.1016/j.smallrumres.2020.106052
14. Zain,
H.; Tatar , A.; Alabi, O. M. .; Samiei Zafarghandi, M. . The Effect Of Using
Different Levels Of Vitamin E On The Antioxidants Status Of Broiler Chickens.
JLSAR 2023, 4, 37-44..
15. Purohit,
G. Methods of pregnancy diagnosis in domestic animals: The current status.
Webmed Central Reproduction. 2010, 1(12)WMC001305: 1- 28.
16. Rovani,
M. T.; Cezar, A. S.; Rigo, M. L.; Gasperin, B. G.; Júnior, J. E. D.; Torres, F.
D.; Gonçalves, P. B. D. and Ferreira, R. Evaluation of a bovine
pregnancy-associated glycoprotein enzyme-linked immunosorbent assay kit for
serological diagnosis of pregnancy in sheep. Ciência Rural, Santa Maria. 2016, 46
(2): 362- 367.doi:10.1590/0103-8478cr20150270
17. Tavana,
M.; Peighambarzadeh, S. Z. and Bardshiri, B. Evaluation of Pregnancy Detection
in Lori-Bakhtiari Sheep by Transrectal Ultrasonography. International Journal
of Advanced Biological and Biomedical Research. 2014, 2 (7): 2268-2270.
18. Taverne,
M. and Noakes, D. E. Pregnancy and its diagnosis. In: Reproduction and
Obstetrics. Noakes, D. E.; Parkinson, T. J. and England, G. C. W., Part. 1,
Chapter 5, 10th ed., Elsevier. 2019, P. 101 -102.
doi:10.1016/C2014-0-04782-X
19. Tekin,
T. C. and Köse, A. M. Investigation of the effectiveness of ultrasonography in
determining pregnancy and the number of fetuses on the 35th day of
pregnancy in Awassi sheep. Journal of Advances in VetBio Science and Techniques.
2022, 7(2): 143-152.doi:10.31797/vetbio.1091526
20. Yotov,
S. and Sinapov, B. Effect of GnRH administration on Pregnancy- Associated
Glycoproteins in dairy sheep with different reproductive status. Acta Scientiae
Veterinariae. 2023, 51 (1920): 1- 8.doi:10.22456/1679-9216.129955.
Received: 25 June 2023/ Accepted: 26 August 2023 /
Published:15 September 2023
Citation: Al-Mohammedy
A. M. F. , Abdul-Rahaman Y. T. Comparison
of the three methods for pregnancy detection in Iraqi Sheep. Revis Bionatura
2023;8 (3) 125 http://dx.doi.org/10.21931/RB/2023.08.03.125