Page 714 - WSAVA2018
P. 714

 25-28 September, 2018 | Singapore
WSVA8-0020
REPRODUCTION, PEDIATRICS
SEX DETERMINATION OF CANINE SPERMATOZOA BY SYBR® GREEN QUANTITATIVE PCR
S. Kaewmanee1, N. Boonwittaya2, J. Thiengtham1, A. Pinyopumintr3
1Faculty of Agriculture- Kasetsart University, Animal Science, Bangkok, Thailand
2Faculty of Veterinary Medicine- Kasetsart University, Kasetsart University Veterinary Teaching Hospital KUVTH, Bangkok, Thailand
3Faculty of Veterinary Medicine- Kasetsart University, Large Animal and Wildlife Clinical Sciences, Nakorn-Pathom, Thailand
WSVA8-0104
REPRODUCTION, PEDIATRICS
A COMPARATIVE STUDY OF FETAL HEAD DIAMETER MEASURED BY RADIOGRAPHY AND ULTRASONOGRAPHY IN DOGS AND CATS
C. Limmanont1,2, S. Ponglowhapan3, T. Sirinarumitr2,4, K. Sirinarumitr2,5
1Kasetsart University, Graduate school- Faculty of Veterinary Medicine, Bangkok, Thailand
2Kasetsart University, Center for Advanced Studies in Agriculture and Food- KU Institute for Advanced Studies, Bangkok, Thailand
3Chulalongkorn University, Department of Obstetrics Gynaecology and Reproduction- Faculty of Veterinary Medicine, Bangkok, Thailand
4Kasetsart University, Department of Pathology- Faculty of Veterinary Medicine, Bangkok, Thailand
5Kasetsart University, Department of Companion Animal Clinical Science- Faculty of Veterinary Medicine, Bangkok, Thailand
INTRODUCTION
Radiography and ultrasonography are both widespread methods used in veterinary obstetrics. In a ventro-dorsal radiographic view, measurements of fetal biparietal or head diameter (HD) and maternal pelvic diameter are commonly used to predict the risk of dystocia in dogs and cats due to relatively oversized fetus. However, HD measurement is easier to perform with ultrasound scan.
OBJECTIVES
To compare and find the correlations of fetal HD measured by radiography and ultrasonography in dogs and cats.
METHODS
Twenty seven dogs and seventeen cats were diagnosed in last trimester of pregnancy at Kasetsart University Veterinary Teaching Hospital during 2017-2018. Fetal
HD was measured by radiography and transabdominal ultrasonography on the same day. The correlations between two techniques and linear regressions of HD were statistically analyzed.
RESULTS
Radiographic measurements of fetal HD were larger than those measured by ultrasonography in both dogs (P<0.0001) and cats (P<0.0001). The correlations were significant (r = 0.85 in dogs and r = 0.94 in cats, p<0.0001). Linear regression formulas were y = 1.0716x + 0.1661 (R2 = 0.72) in dogs, and y = 0.9627x + 0.3148 (R2 = 0.88) in cats. (y= HD by radiography, x= HD by ultrasonography)
CONCLUSIONS
Fetal HD measurements were different between two modalities. The formulas above can be implied to estimate relatively oversized fetus.
    INTRODUCTION
Effective preselection of sex has been done in various species of livestock and domestic animals using the flow cytometry. A guaranteed high sorting accuracy is a key performance for the widespread use of sperm sexing. However, the sexing technique needed to be validated to ensure the accuracy of the technology.
OBJECTIVES
To be able to specifically produce either male or female offspring in the dog, we developed a technique to determine the sex of canine spermatozoa using SYBR® Green real-time quantitative PCR (qPCR).
METHODS
Two sets of primers, ZFX and SRY were designed specifically for X- and Y- chromosome canine genes respectively. Plasmid was inserted with ZFX and SRY gene fragment separately to create standard curves that ranged from 300 to 3,000,000 copies. In canine genome, both ZFX and SRY genes were exist as single copy number and commonly used as a DNA marker in sex determination.
RESULTS
Real-time qPCR of bovine spermatozoa DNA samples and cloned plasmid ZFX and SRY genes for creating standard samples were performed simultaneously.
There was no significant difference in percentages of between the theoretical ration (1:1) and unsexed X- and
Y- chromosome-bearing canine spermatozoa in any of the nine dogs. In addition, the mean purities of sorted sex chromosomes in spermatozoa of the nine dogs were 91% for the X chromosome fraction and 90% for the Y chromosome fraction.
CONCLUSIONS
This technique was a rapid and reliable technique in quantifying the sex ratio of X- and Y- chromosome bearing spermatozoa in semen sample.
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43RD WORLD SMALL ANIMAL VETERINARY ASSOCIATION CONGRESS AND 9TH FASAVA CONGRESS

























































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