Milk Metabolomics Reveals Potential Biomarkers for Early Prediction of Pregnancy in Buffaloes Having Undergone Artificial Insemination

This study aimed to identify potential biomarkers for early pregnancy diagnosis in buffaloes subjected to artificial insemination (AI). The study was carried out on 10 pregnant and 10 non-pregnant buffaloes that were synchronized by Ovsynch-Timed Artificial Insemination Program and have undergone the first AI. Furthermore, milk samples were individually collected ten days before AI (the start of the synchronization treatment), on the day of AI, day 7 and 18 after AI, and were analyzed by LC–MS. Statistical analysis was carried out by using Mass Profile Professional (Agilent Technologies, Santa Clara, CA, USA). Metabolomic analysis revealed the presence of several metabolites differentially expressed between pregnant and non-pregnant buffaloes. Among these, a total of five metabolites were identified by comparison with an online database and a standard compound as acetylcarnitine (3-Acetoxy-4-(trimethylammonio)butanoate), arginine-succinic acid hydrate, 5′-O-{[3-({4-[(3aminopropyl)amino]butyl}amino)propyl]carbamoyl}-2′-deoxyadenosine, N-(1-Hydroxy-2-hexadecanyl)pentadecanamide, and N-[2,3-Bis(dodecyloxy)propyl]-L-lysinamide). Interestingly, acetylcarnitine was dominant in milk samples collected from non-pregnant buffaloes. The results obtained from milk metabolic profile and hierarchical clustering analysis revealed significant differences between pregnant and non-pregnant buffaloes, as well as in the metabolite expression. Overall, the findings indicate the potential of milk metabolomics as a powerful tool to identify biomarkers of early pregnancy in buffalo undergoing AI.

Download Full-text

Evaluation of synchronization protocols and methods of early pregnancy diagnosis in dairy cattle

South African Journal of Animal Science ◽

10.4314/sajas.v50i6.4 ◽

2021 ◽

Vol 50 (6) ◽

Author(s):

A.H. Shahzad ◽

A. Sattar ◽

N. Ahmad ◽

I. Ahmad ◽

M.S. Yousaf ◽

...

Keyword(s):

Pregnancy Rate ◽

Early Pregnancy ◽

Artificial Insemination ◽

Pregnancy Loss ◽

Holstein Cows ◽

Plasma Progesterone ◽

Pregnancy Diagnosis ◽

Milk Samples ◽

Timed Artificial Insemination ◽

In Pregnancy

The studies aimed to evaluate the pregnancy rate (PR) for timed artificial insemination (TAI) after G7G-Ovsynch, modified G7G-Ovsynch (MG7G-Ovsynch) and Ovsynch protocols and to assess the accuracy of using pregnancy-associated glycoproteins (PAGs) and plasma progesterone (P4) in pregnancy diagnosis compared with ultrasonography (US). In study 1, Holstein cows (n = 37) were bred by TAI following the G7G-Ovsynch protocol (n = 19) or MG7G-Ovsynch (n = 18). Pregnancy was evaluated by US at days 31, 59, and 87 after breeding. The PR was not different for the G7G-Ovsynch and MG7G-Ovsynch. Blood and milk samples were collected on day 3 after insemination and then weekly through day 59 post TAI in cows diagnosed as not pregnant on day 31 and through day 87 in pregnant cows. PAGs were measured using ELISA and P4 by radioimmunoassay (RIA). In the second study, Holstein cows (n = 212) were bred by TAI following G7G-Ovsynch protocol (n = 110) or standard Ovsynch (n = 102). Cows were subjected to pregnancy diagnosis on days 30, 60, and 90. A subset (n = 15 in each group) was subjected to blood and milk samples on days 30, 45, 60, 75, and 90 to measure PAGs and P4. In study 2, PR was not significantly different between synchronization protocols on days 30, 60, and 90. Pregnancy loss averaged 15% between day 30 and day 90. The use of PAGs and P4 proved equally effective in diagnosis of pregnancy. Thus, G7G-Ovsynch was deemed the protocol of choice in postpartum cows, and PAGs assayed in milk or plasma could be used to diagnose pregnancy.

Download Full-text

Pregnancy rates and peripheral progesterone levels following Ovsynch or CIDR ovulation synchronization/timed artificial insemination protocols in postpartum dairy cows

Canadian Journal of Animal Science ◽

10.4141/cjas07150 ◽

2008 ◽

Vol 88 (3) ◽

pp. 457-461

Author(s):

M. Aali ◽

T. Pretheeban ◽

G. Giritharan ◽

R. Rajamahendran

Keyword(s):

Dairy Cows ◽

Key Words ◽

Pregnancy Rate ◽

Artificial Insemination ◽

Sampling Period ◽

Pregnancy Rates ◽

Key Words Pregnancy ◽

Milk Samples ◽

Timed Artificial Insemination ◽

Rectal Palpation

Peripheral progesterone (P4) concentrations as well as pregnancy rates (PR) between cows treated with Ovsynch or CIDR ovulation synchronization/TAI protocols were compared. Two hundred and twenty-seven postpartum lactating Holstein cows were randomly assigned to Ovsynch (n = 111) or CIDR (n = 116) ovulation synchronization/TAI protocols. Pregnancy was diagnosed by ultrasonography at day 35 after TAI and later confirmed by rectal palpation at day 60 post TAI. Milk samples were taken from the beginning of treatment (day -10) until day 35 after TAI (day 0) for P4 determination. Pregnancy rates for Ovsynch and CIDR groups of cows were 31 and 41%, respectively. The PR of ovulation synchronized cows for Ovsynch and CIDR groups were 49 and 69%, respectively. Progesterone levels during the sampling period were similar (P > 0.05) between Ovsynch and CIDR treated cows. Key words: Pregnancy rate, progesterone, Ovsynch, CIDR, cows

Download Full-text