IDENTIFICATION AND CONFIRMATION OF EARLY PREGNANCY ASSOCIATED URINARY METABOLITES IN MURRAH BUFFALO
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Date
2019
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Publisher
ICAR-NDRI, KARNAL
Abstract
Evaluation of urinary metabolite dynamics pertaining to day 0 and day 42
post-insemination in healthy Murrah buffaloes (Bubalus bubalis). Metabolite
analysis was performed on urine samples of pregnant, non-pregnant, nonpregnant
inseminated heifers derived from three groups of artificially inseminated,
thirty (n=30) healthy cyclic Murrah buffalo heifers. Urinary metabolites were
analyzed in these samples corresponding to Day 0, 10, 18, 35 and day 42 of
pregnancy by nuclear magnetic resonance (NMR) and the results were further
validated by gas chromatography-mass spectrometry (GC-MS) analysis. Current
experiments revealed and confirmed 32 metabolites in urine of both pregnant,
non-pregnant buffaloes urine with global changes during early pregnancy by
multivariate statistical analysis such as principal components analysis (PCA)
model and heatmap. Significant increase in urinary concentration of some 2-
amino-3-(1H-indol-2-yl)propanoic acid derived metabolite was observed on day 42
as compared to day 0 of pregnancy. Similarly, ethylene glycol, melatonin and
fumarate appeared on specific days of early pregnancy. Contrary to this, decrease
in concentration of 1-methyl-(2S)-2-amino-3-(1H-imidazol-5-yl)propanoic acid and
3-indoxysulfate were observed with progression of pregnancy from day 0 to 42.
Multivariate analysis of the analysed metabolites generated distinct differential
pattern with progression of pregnancy at early stage. Pathway analysis of the
corresponding metabolites suggested high impact correlation with establishment
and succession of pregnancy in buffaloes. Urinary metabolites measured during
early pregnancy in buffaloheifers depicted close association of the metabolites
with physiological processes involved in successful conception and progression of
pregnancy. This study opens new avenues for use of urinary metabolites in
pregnancy diagnosis in bovine especially buffaloes.
Key words: Murrah buffalo; Urinary metabolites; Early pregnancy; NMR; GC-MS