Diurnal pattern of urinary and faecal nitrogen excretion by dairy cows fed ryegrass pasture twice daily indoors.

C Clark, G Waghorn, P Gregorini, S Woodward, D Clark DairyNZ, Hamilton, New Zealand 2011 Email: dave.clark@dairynz.co.nz Introduction The predominant component of the New Zealand dairy cows diet is ryegrass pasture, which generally has a nitrogen (N) content in excess of cow requirement. Much of the surplus is excreted as urinary N which concentrates N in urine patches and contributes to groundwater and atmospheric (nitrous oxide) pollution. Urine patch soil N accumulation, and N loss to the environment, is influenced by urine volume and N concentration (Di and Cameron, 2002). However, the diurnal variation in dairy cow urine volume, and N concentration, is poorly understood. Grazing ruminants have a natural circadian grazing pattern, but strict pasture allocation may mean dairy cows consume up to 70% of their daily allowance in the first 3-4 h post-allocation (Gregorini et al., 2009). We tested the hypothesis that there is diurnal variation in the concentration and quantity of urine N excreted by dairy cows. Materials and methods Six multiparous lactating Holstein-Friesian dairy cows (54640 kg BW; 22122 DIM) were housed in metabolism stalls and offered ryegrass ( Lolium perenne, L.) pasture ad libitum for 10 days in March 2010 with measurements taken on d 9 and 10. Half the daily DM requirement of pasture was cut and offered to each cow at 0840 h. Feed was removed at 1300 h and cows milked at 1530 h. Fresh pasture was cut and offered to cows at 1600 h. Feed was removed at 2400 h and replaced at 0600 h prior to being removed again at 0730 h. Pasture intake was measured, and samples collected daily and stored for analysis by near infrared spectroscopy (NIRS systems 6500, FeedTech). Crude protein, NDF and ADF were 15, 59 and 32 g/100 g DM respectively. Milk production was recorded, and samples collected twice daily (0730 and 1530 h). Urine and faeces weight were determined every 2 hours for 48 hours on d 9 and 10 and subsamples were taken on each occasion. Each 50mL urine sub-sample was immediately acidified with hydrochloric acid to a pH < 4 to prevent ammonia volatilisation and stored at -20 0C. Total Kjeldahl N and creatinine (Roche Diagnostics, kit number 11875418) were determined from these urine sub-samples. Each 50g faeces sub-sample was sealed in a zip-lock bag and stored at -200C for Kjeldahl N determination. Data were analysed as repeated measurements with a mixed model fitted using REML with Day and Time as fixed effects and Cow and Time within cow as random effects. An autoregressive of order 1 covariance structure was used to model the within cow measurements.
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Figure 1 Urine nitrogen excretion (g/ 2h) over 24 h (mean = 8.11; SED=2.72). Arrows indicate when pasture was offered

Results There was no effect of day (P > 0.05) on the variables analysed so the data were analysed as a mean of each 2 h period for the two days. Urine N concentration (g/100g) varied with time (P < 0.01) with a peak at approximately 4 h after pasture allocation. Urine volume (L/2h) and N excreted (g/2h) (Figure 1) varied with time (P < 0.01) with a peak < 1h after feeding commenced. There was no effect of time on faecal N concentration (mean 2.3 g/100g), faecal N excreted (mean 10.9 g N/2h) and faecal weight (mean 480 g DM/2 h). Urine creatinine varied with time (P < 0.01) ranging from 2,653 to 1,287 mol/L and creatinine also varied between cows from 7 to 12 mg/kg liveweight per d.

Conclusions The volume of urine and quantity of N excreted were more than doubled within an hour after feed was offered. Clark et al. (2010) reported dairy cow urination frequency increased during feeding compared with resting. In contrast, urine N concentration reached a peak four hours after pasture was offered. This delay in peak urine N concentration may be associated with proteolysis and microbial growth relative to feeding because Kolver et al. (1998) found rumen NH3 concentration peaked three hours, and blood urea five hours after pasture feeding. There is good evidence that the quantity and concentration of excreted urine N is related to the timing of pasture ingestion. The diurnal variation has implications for mitigating N loss from grazed dairy farms because urinary N concentration changed from 0.3 to 0.5% (w/w) within a day, contrary to an assumed value of 100g N/m2 used to describe the dairy cow urine patch soil N loading (Di and Cameron, 2002). If the high N loading events could be captured (see Clark et al. 2010) and spread evenly across a paddock, the amount of N loss from grazed dairy systems could be substantially reduced. Also, the timing of pasture allocation could be altered to prevent peaks and troughs of herbage intake. For instance, more frequent allocations of pasture may decrease peak urine N concentration, high N urine patch events and subsequent N loss to the environment. References Di, H.J., Cameron, K.C. 2002. Nutrient cycling in agroecosystems. 46, 237-256. Gregorini, P., Clark, C.E.F., Jago, J.G. et al. 2009. Journal of Dairy Science 92, 4572-4580. Clark, C.E.F., Levy, G., Beukes, P. et al. 2010. Proceedings of the European Association of Animal Production Annual Meeting. Heraklion, Greece p.328. Kolver, E., Muller, L.D., Varga, G.A. et al. 1998. Journal of Dairy Science 81, 2017-2028.

Urinenitrogenexcreted (g/2h)