Reprod Dom Anim 2017; 1–5 wileyonlinelibrary.com/journal/rda | 1 © 2017 Blackwell Verlag GmbH Received: 30 September 2016 | Accepted: 27 December 2016 DOI: 10.1111/rda.12933 SHORT COMMUNICATION Urine levels of luteinizing hormone as predictor of the period of ovulaton for advantage of tmed-artfcial inseminaton in murrah bufalo (Bubalus bubalis) RM Selvam 1 | D Singh 2 | MA Akbarsha 1 | G Archunan 1 1 Centre for Pheromone Technology, Department of Animal Science, Bharathidasan University, Tiruchirappalli, India 2 Molecular Endocrinology Laboratory, Animal Biochemistry Division, Natonal Dairy Research Insttute, Karnal, India Correspondence Govindaraju Archunan, Bharathidasan University, Tiruchirappalli, India. Email: garchu56@yahoo.co.in Funding informaton Indian Council of Agricultural Research, Grant/ Award Number: NFBSFARA/4006. Contents Assessment of urine levels of luteinizing hormone (LH) for predictng the reproductve status of animals is in practce. The aim of this study was to predict the period of ovulaton based on the urine levels of LH for tmed-artfcial inseminaton to increase the concepton rate in bufaloes, which are naturally silent-oestrous animals. Level of LH in urine was assessed using ELISA, and a cut-of LH concentraton for predicton of ovulaton period was obtained using receiver operatng characteristc analysis. Artfcial inseminaton was performed before- and afer -positve predicton of ovulaton period adoptng this method, and the rates of concepton were assessed. Urine LH level of 105 mIU/ml (n = 14) was derived as a cut-of concentraton which predicts the ovulaton period. The bufaloes in the positvely predicted group (day 1 or 2) inseminated via intracervical route had an increase in the concepton rate (83.33%); however, the inseminaton in the before-positve-predicton group resulted in poor concepton rates (day 0; 16.66%) compared to that of the naturally inseminated group (day 0; 75.0%). In conclusion, the urinary LH would possibly be a fairly reliable predictor of the ovulaton period. The day when cut-of LH concentraton is obtained may be taken as the most favourable tme for artfcial inseminaton, so as to atain a much beter rate of concepton in the bufalo. 1 | INTRODUCTION Bufalo cows have gained considerable atenton over bovine cows for the economic advantage on meager quantty of roughage feed- ing, even in hot and humid climates. The reproductve efciency of this peculiar shy breeder is afected by factors such as silent heat and poor concepton rate, resultng in substantal decline of their popula- ton. Assisted reproductve techniques (ART) do not fnd optmal utl- ity in this species, and ART such as artfcial inseminaton (AI) may be practced accurately and precisely if the tming of ovulaton in relaton to oestrus and LH surge is accurately assessed (Mondal, Prakash, & Palta, 2007), otherwise it would result in poor concepton. Although oestrous-specifc pheromones and proteins have been reported in buf- faloes (Archunan, Rajanarayanan, & Karthikeyan, 2014; Muthukumar et al., 2014), no kit has yet been developed for oestrous detecton and ovulaton predicton. Reproductve endocrinology has received considerable atenton during the recent decades. The frequency of sampling is an import- ant factor for assessing gonadotropins like luteinizing hormone (LH) which is released in a pulsatle fashion (Fitzgerald, I’Anson, Legan, & Loy, 1985). However, the chance of missing the peak is high even with frequent blood samples collecton (Paul, 2003). The excretory fuids like urine refect the internal status of the animal and have been proposed to be a beter medium for assessing the reproduc- tve status non-invasively. Moreover, the fuctuatons occurring due to the pulsatle release of LH could be overcome by detectng LH in urine, as it is accumulated in the bladder for quite some tme before being voided by the animal (Khatab, Mustafa, & Taylor, 2005). Standard immunoassay techniques (French et al., 1999; Miller & Soules, 1996) and portable dip strips are available in the market for quanttatve detecton of urinary LH in several mammals to fx the tme for AI (Robeck et al., 2005).