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Department of Animal Nutrition, Madras Veterinary College, Chennai 600 007, India
ABSTRACT
Rapid hydrolysis of urea in the rumen is the principle cause for urea (ammo-
nia) toxicity. Efforts were directed to retard urea hydrolysis by supplementing
zinc at gradual levels viz, 0, 5.0,10.0, 15.0 and 20.0 pprn in one ml solution to
35 mg of urea in one ml solution, and in one ml distilled water with 40 ml
buffered rumen liquor in 100 ml syringe fitted with butyl rubber cap and
incubated at 39OC. The 35 mg of urea per 43ml of liquor in the in vitro batch
culture is equivalent to toxic dose of 100 mg per 100ml of rumen liquor in adult
cattle. The incubation was carried out in an anaerobic environment at pH of
6.8. Five replications were conducted twice, resulting in ten replications in
each treatment. The residual urea retained in each tube at 0, 1, 2 and 3 hour
intervals from the respective aliquot (1ml) was measured at wavelength of 520
nm. Highest residual urea (P<0.01)was observed in 10 pprn zinc supplemen-
tation over the rest of the treatments imposed across incubating hours. The
residual urea (mg/dL) at the end of 1,2 and 3 hours of incubation were 28.99f
1.04, 18.33f 0.04 and 15.45+ 0.18 respectively at 10 pprn of zinc compared to
+
18.95+ 0.38,10.00+0.16 and 7.48 0.12 in control (0 pprn). The result divulged
that 10 pprn zinc was able to effectively retard the urea hydrolysis up to 3
hours, reflecting its effect on the extent of duration. Though the results demon-
strated the superiority of 10 pprn zinc treatment over the rest of the treatments
in retarding urea hydrolysis, yet another experiment was conducted to further
improve the precision at a pH of 7.4 that is considered to be favorable
environment for ammonia toxicity. The second trial was conducted following
the same procedure except that of the level of zinc supplemented. In this exper-
iment, zinc was supplemented at 0, 7.5, 10.0 and 12.5 pprn to 35 mg of urea
with 40 ml of buffered rumen liquor incubated for 0,1,2 and 3 hours. The intial
pH was brought to 7.4 by addition of a suitable quantity of soda bicarbonate to
simulate a conducive environment for free ammonia production. The results of
this experiment further strengthened the previous experiment's results with a
significantly (P<0.01) higher residual urea in 10 pprn against the rest of the
treatments. Thus it can be concluded that supplementation of 10 pprn zinc
delayed the hydrolysis of urea.
the present study. Similar reports on the than the unsupplemented group. How-
negligible quantity of zinc in rumen liquor ever, at the end of the second hour incuba-
has been reported by Arelovich et al. tion, the 15 pprn of zinc had significantly
(2000). (P<0.01) lower residual urea compared to
10 pprn zinc, but it was comparable to 5
EXPERIMENT 1: Studies on the effect of ppm. At the end of the third hour of
zinc at 0, 5.0, 10.0, 15.0 and 20.0 pprn in incubation, the highest residual urea was
retarding urea hydrolysis at pH 6.8 observed in the 10-ppm zinc, which was
(Normal rumen pH) significantly (P<0.01) higher than 5 or 15
The residual urea concentration pprn zinc. However, the latter was in turn
ranged from 7.48 to 51.81 mg/dl across significantly (P<0.01) higher then 20 pprn
treatments and periods studied (Table 1). zinc. Thus, it is evident that 10 pprn of zinc
The values reported in this study con- was effective in reducing urea hydrolysis.
curred with Arelovich et al. (2000), who Similar observations were made by
recorded 14.77 to 40.34 mg /dl of residual Arelovich et al. (2000), who reported that
urea. supplementation with zinc at 10 to 15 pprn
No significant difference was concentration inhibited in vitro urea
observed among various levels of zinc at hydrolysis. Lowered and controlled urea
the start of the experiment (0 h). At the end disappearance is desired, as it would
of the first hour, the highest (P<0.01) avoid ammonia spikes resulting in urea
amount of residual urea was recorded in toxicity. Further excess of ruminal ammo-
10 pprn and 15 pprn compared to 5, and 20 nia can decrease N retention, reduce
pprn respectively, which in turn had productivity (Doyle, 1987) and is proven
significantly (P<0.01)higher residual urea to be toxic (Froslie, 1977).
Table 1. The effect of zinc at various levels (pprn) on the residual urea (mg/dl) at three
hourly intervals (Mean f SE)
Note:
a Mean of five observations
Means bearing different superscripts within a column differ significantly (P<0.01)for
each treatment
a NS - Non significant
Effect of Zinc Supplementation on Urea Hydrolysis 97
The mechanism by which zinc exerts (1995) who showed that zinc intake to the
influence to inhibit urease secretion by the equivalent of 13.95 pprn improved urea
microorganism has been attributed to the utilisation and nitrogen balance in sheep.
fact that urease produced by the rumen The percentage of urea disappear-
bacteria is considered to be a metal ance as altered by various levels of zinc at
inhibitory intracellular enzyme. It has cumulative hour intervals is presented in
been postulated that zinc ion forms an Figure 1.
active complex with the enzyme to inhibit The percentage of urea disappear-
its activity (Jones, Macheod and ance at cumulative hours was calculated to
Blackwood, 1964). The gradual decline in evaluate the extent of the effect of zinc in
retarding urea hydrolysis by higher con- preventing urea toxicity. It was observed
centrations of zinc could be attributed to that 10 pprn zinc continued to retard urea
its toxic effect on rumen microbes hydrolysis until the end of 3 hours.
(Martinez and Church, 1970) or its modify- Similarly, zinc at 5 or 15 pprn was able to
ing effect on enzymatic action (Hare Singh retard urea disappearance better than 0
and Cole, 1988) and 20 ppm. The graph reveals that zinc at
Hence, it is concluded that zinc at 10 10 pprn was able to reduce urea disap-
pprn attains the effective concentration to pearance to the extent of 18 per cent over
achieve the desired effect of regulating control in the first hour, which is the cru-
ammonia spikes from urea. The results of cial hour for ammonia toxicity. Rodriguez
this study concurred with Rodriguez et al. et al. (1993) reported that zinc at 600 pprn
5 10 15
Zinc (ppm)
Figure 1. The effect of zinc at 0, 5, 10, 15 and 20 pprn on the percentage of urea
disappearance at cumulative hour intervals in experiment 1
98 Kathirvelan C O Balakrishnan V