Application of sensitive enzymeimmunoassays for oxytocin and prolactin determination in blood plasma of yaks ( Poephagus grunniens L.) during milk let down and cyclicity

Abstract

Highly sensitive and specific enzymeimmunoassays for oxytocin and prolactin determination in yak plasma using the biotin–streptavidin amplification system and the second antibody coating technique were validated and applied for determining their profiles during milk let down and cyclicity in yaks. Oxytocin EIA was conducted taking duplicate 200 μl of unknown plasma samples and standards per well. The lowest detection limit was 0.2 pg/well, which corresponded to 1 pg/ml plasma. Prolactin EIA was carried out directly in 50 μl of yak plasma. The sensitivity of EIA procedure was 5 pg/well prolactin, which corresponded to 0.1 ng/ml plasma. Mean plasma prolactin concentrations although high at estrus were not statistically different ( P > 0.05) from the hormone concentrations on other days. Mean plasma prolactin concentrations during non-breeding season were significantly higher ( P < 0.001) than that recorded in breeding season. Oxytocin and prolactin profiles were also obtained in two yaks before, during and after milking. A sharp release of oxytocin and prolactin shortly after udder stimulation was observed. High levels of oxytocin and prolactin were maintained during milking, falling sharply thereafter.