Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis

Abstract Background Reproductive efficiency is paramount in the dairy industry, where early pregnancy detection of dairy cows will allow to detect the non-pregnant animals early, thus enabling to re-synchronize them and getting them pregnant leading to decrease in calving interval, which, in turn, i...

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Main Authors: Malshani De Silva, Devinda Wickramasingha, Nirodha Abayalath, Basil Alexander, Missaka Wijayagunawardane, Kai Fai Lee, Suranga Kodithuwakku
Format: Article
Language:English
Published: BMC 2024-12-01
Series:BMC Veterinary Research
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Online Access:https://doi.org/10.1186/s12917-024-04438-5
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author Malshani De Silva
Devinda Wickramasingha
Nirodha Abayalath
Basil Alexander
Missaka Wijayagunawardane
Kai Fai Lee
Suranga Kodithuwakku
author_facet Malshani De Silva
Devinda Wickramasingha
Nirodha Abayalath
Basil Alexander
Missaka Wijayagunawardane
Kai Fai Lee
Suranga Kodithuwakku
author_sort Malshani De Silva
collection DOAJ
description Abstract Background Reproductive efficiency is paramount in the dairy industry, where early pregnancy detection of dairy cows will allow to detect the non-pregnant animals early, thus enabling to re-synchronize them and getting them pregnant leading to decrease in calving interval, which, in turn, is critical for maximizing productivity and economic gain. The objective of this study was to evaluate the colour Doppler ultrasonography (CDUS) and peripheral blood leukocytes (PBLs)-based pregnancy-associated biomarker mRNAs expression for the earliest detection of pregnancy status in the dairy cows at post insemination. Intensively managed animals were ovulation synchronized and subjected to timed artificial insemination (TAI). On day 20, corpus luteum blood flow (CLBF) was evaluated using CDUS in 30 cows. The percentage of the incoming blood flow (as an area) of the corpus luteum (CL) was determined using an image analysis software. On day 35, the same operator performed a final pregnancy diagnosis using transrectal ultrasonography to confirm the pregnancy. Blood samples were collected on day 20 and 28 after TAI for biomarkers analysis. The mRNA expression levels of ISG15, MX1, MX2, and PAG9 genes in PBLs were determined by quantitative polymerase chain reaction (qPCR). Results The identified CLBF cutoff point resulted 100% sensitivity and negative predictive value (NPV) in determining non-pregnant status on day 20 in the cows. Overall, MX2 and ISG15 mRNAs showed the most significant (P < 0.05) expression levels in pregnant animals on day 20 and 28 compared to non-pregnant animals. Among them, MX2 showed the highest expression levels on both days, ascertaining it as the better candidate biomarker for the earliest identification of pregnancy. Conclusions The CDUS-based CLBF analysis on day 20 after TAI can be potentially used for the early identification of non-pregnancy status in dairy cows and MX2 could be a potential mRNA candidate for the identification of pregnancy in cows. Further studies should be conducted in large scale to validate these findings due to the small sample number used in the current study.
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spelling doaj-art-05be1ad6a0694866b828ca6cb15e7e122025-08-20T02:39:40ZengBMCBMC Veterinary Research1746-61482024-12-0120111110.1186/s12917-024-04438-5Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysisMalshani De Silva0Devinda Wickramasingha1Nirodha Abayalath2Basil Alexander3Missaka Wijayagunawardane4Kai Fai Lee5Suranga Kodithuwakku6Department of Animal Science, Faculty of Agriculture, University of PeradeniyaDepartment of Farm Animal Production and Health, Faculty of Veterinary Medicine and Animal Science, University of PeradeniyaDepartment of Animal Science, Faculty of Agriculture, University of PeradeniyaDepartment of Farm Animal Production and Health, Faculty of Veterinary Medicine and Animal Science, University of PeradeniyaDepartment of Animal Science, Faculty of Agriculture, University of PeradeniyaDepartment of Obstetrics and Gynaecology, LKS Faculty of Medicine, The University of Hong KongDepartment of Animal Science, Faculty of Agriculture, University of PeradeniyaAbstract Background Reproductive efficiency is paramount in the dairy industry, where early pregnancy detection of dairy cows will allow to detect the non-pregnant animals early, thus enabling to re-synchronize them and getting them pregnant leading to decrease in calving interval, which, in turn, is critical for maximizing productivity and economic gain. The objective of this study was to evaluate the colour Doppler ultrasonography (CDUS) and peripheral blood leukocytes (PBLs)-based pregnancy-associated biomarker mRNAs expression for the earliest detection of pregnancy status in the dairy cows at post insemination. Intensively managed animals were ovulation synchronized and subjected to timed artificial insemination (TAI). On day 20, corpus luteum blood flow (CLBF) was evaluated using CDUS in 30 cows. The percentage of the incoming blood flow (as an area) of the corpus luteum (CL) was determined using an image analysis software. On day 35, the same operator performed a final pregnancy diagnosis using transrectal ultrasonography to confirm the pregnancy. Blood samples were collected on day 20 and 28 after TAI for biomarkers analysis. The mRNA expression levels of ISG15, MX1, MX2, and PAG9 genes in PBLs were determined by quantitative polymerase chain reaction (qPCR). Results The identified CLBF cutoff point resulted 100% sensitivity and negative predictive value (NPV) in determining non-pregnant status on day 20 in the cows. Overall, MX2 and ISG15 mRNAs showed the most significant (P < 0.05) expression levels in pregnant animals on day 20 and 28 compared to non-pregnant animals. Among them, MX2 showed the highest expression levels on both days, ascertaining it as the better candidate biomarker for the earliest identification of pregnancy. Conclusions The CDUS-based CLBF analysis on day 20 after TAI can be potentially used for the early identification of non-pregnancy status in dairy cows and MX2 could be a potential mRNA candidate for the identification of pregnancy in cows. Further studies should be conducted in large scale to validate these findings due to the small sample number used in the current study.https://doi.org/10.1186/s12917-024-04438-5Colour Doppler ultrasonographyCorpus luteum blood flowDairy cowsEarly pregnancy diagnosisGene expression
spellingShingle Malshani De Silva
Devinda Wickramasingha
Nirodha Abayalath
Basil Alexander
Missaka Wijayagunawardane
Kai Fai Lee
Suranga Kodithuwakku
Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
BMC Veterinary Research
Colour Doppler ultrasonography
Corpus luteum blood flow
Dairy cows
Early pregnancy diagnosis
Gene expression
title Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
title_full Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
title_fullStr Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
title_full_unstemmed Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
title_short Early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour Doppler ultrasonography and mRNA analysis
title_sort early pregnancy diagnosis in cows using corpus luteum blood flow analysis based on colour doppler ultrasonography and mrna analysis
topic Colour Doppler ultrasonography
Corpus luteum blood flow
Dairy cows
Early pregnancy diagnosis
Gene expression
url https://doi.org/10.1186/s12917-024-04438-5
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