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Utilizing Vaginal Cytology and Gelatin Zymography for Pregnancy Diagnosis in Pigs

Utilizing Vaginal Cytology and Gelatin Zymography for Pregnancy Diagnosis in Pigs

Adsadawut Sanannam1, Worawatt Hanthongkul2, Yu-Jing Liao3, Kunlayaphat Wuthijaree4, Pattaraporn Tatsapong4, Amornrat Wanangkarn4,5, Anurak Khieokhajonkhet4,5, Niran Aeksiri4,5, Sureeporn Saengwong6, Wilasinee Inyawilert4,5*

1Department of Animal Science, Faculty of Sciences and Agricultural Technology, Rajamangala University of Technology Lanna, Phitsanulok, Thailand; 2Phitsanulok artificial insemination and biotechnology research center, Phitsanulok, Thailand; 3Genetics and Physiology Division, Taiwan Livestock Research Institute, Ministry of Agriculture, Tainan, Taiwan; 4Department of Agricultural Science, Faculty of Agriculture Natural Resources and Environment, Naresuan University, Phitsanulok, Thailand; 5The Center for Agricultural Biotechnology, Naresuan University, Phitsanulok, Thailand; 6Division of Animal Science, School of Agriculture and Natural Resources, University of Phayao, Phayao, Thailand.

 
*Correspondence | Wilasinee Inyawilert, Department of Agricultural Science, Faculty of Agriculture Natural Resources and Environment, Naresuan University, Phitsanulok, Thailand; Email: [email protected]

ABSTRACT

Timely identification of non-pregnant sows and gilts in the early stages can greatly enhance reproductive efficiency and financial success in pig farming. Vaginal cytology is a reliable and direct technique used to identify the estrus phase and can distinguish animals that are returning to the estrus stage. Furthermore, matrix metalloproteinases (MMPs) are involved in altering the extracellular matrix (ECM) throughout pregnancy and parturition. The aim of this study was to examine changes in the characteristics of vaginal cells and alterations in the expression of matrix metalloproteinases (MMPs) using gelatin zymography in pregnant pigs. Eight sexually mature individuals were identified as being in early pregnancy using the vaginal smear technique within 18 to 26 days following artificial insemination. The findings indicated an elevated leukocyte count in pregnant pigs between days 18 and 26. However, a small number of epithelial cells that were either nucleated or cornified were observed during the entire pregnancy period. Additionally, the MMP-2 and MMP-9 expression was analyzed using gelatin zymography in four pregnant pigs on day 60 and a non-pregnant pig on the day it returned to estrus after artificial insemination. Each sample was standardized by dividing the density of the band using Image J Software. Pregnant pigs showed significantly higher levels of MMP-9 compared to non-pregnant pigs (p≤0.05). While the level of MMP-2 expression was greater in pregnant pigs, there was no statistically significant differences in MMP-2 levels seen between pregnant and non-pregnant pigs (p≥0.05). Thus, vaginal cytology is a direct and efficient technique for detecting pregnancy in pigs. Furthermore, further investigation is necessary to evaluate MMP-9 as a potential biomarker for confirming pregnancy in pigs, with the objective of enhancing the precision of pregnancy detection in female pigs and resolving notable obstacles in the swine breeding industry.
 
Keywords | Vaginal cytology, Gelatin zymography, Pregnancy, Pig

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Advances in Animal and Veterinary Sciences

November

Vol. 12, Iss. 11, pp. 2062-2300

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