Uso Da Creatinina Urinária Como Marcador Nutricional e de Volume Urinário em Ovinos Alimentados Com Forragem Tropical Ou Temperada
Uso Da Creatinina Urinária Como Marcador Nutricional e de Volume Urinário em Ovinos Alimentados Com Forragem Tropical Ou Temperada
Uso Da Creatinina Urinária Como Marcador Nutricional e de Volume Urinário em Ovinos Alimentados Com Forragem Tropical Ou Temperada
1590/1678-4162-7759
D.B. David1, C.H.E.C. Poli2, J.V. Savian2, G.A. Amaral1, E.B. Azevedo3, F. Jochims2
ABSTRACT
To test the accuracy of creatinine as a marker for estimating urinary volume and its use as a nutritional index,
the possible interference of forage intake and forage quality over creatinine excretion was evaluated. For this,
sheep were fed different levels of pearl millet (Pennisetum americanum (L) Leeke) or Italian ryegrass (Lolium
multiflorum Lam). The experiment consisted of a compilation of digestibility trials (n=6) with pearl millet or
Italian ryegrass in completely randomized designs with four replications and four forage levels: 1.5, 2.0, 2.5%
(kg dry matter (DM)/ 100 kg of live weight (LW)). The trials were repeated at different periods to evaluate how
stable the average metabolic excretion of creatinine is. In each trial, total urine collection was performed
individually during a period of 24 hours for five consecutive days and subsequently analyzed by colorimetry
for creatinine and purine derivatives. The creatinine excretion was not affected (P>0.05) by forage offer or
forage type, but there were period effects (P=0.0001). The average creatinine excretion for both forages was
0.21mmol/kg PV0,75. Linear regressions between the purine derivatives:creatinine index with total excretion of
purine derivatives were detected for pearl millet (P<0.0001, R= 0.64) and Italian ryegrass (P=0.02, R=0.20).
These results demonstrate that creatinine excretion is independent of the type and availability of forage and can
be a marker for urinary volume prediction and nutritional measures under grazing systems.
RESUMO
Para testar a preciso da creatinina como marcador para estimativas de volume urinrio e ndice nutricional,
foram avaliadas a possvel influncia do consumo e a qualidade da forragem sobre esse marcador. Para isso,
ovinos foram alimentados com diferentes nveis de milheto (Pennisetum americanum (L) Leeke) ou azevm
(Lolium multiflorum Lam). O experimento consistiu de uma compilao de ensaios de digestibilidade (n=6)
com milheto ou azevm, em um desenho experimental de blocos completamente ao acaso, com quatro
repeties e quatro nveis de forragem: 1,5; 2,0; 2,5% (kg de matria seca (MS)/ 100kg de peso vivo (PV)). Os
ensaios foram repetidos em diferentes perodos, com ambas as forragens, se para avaliar a estabilidade da
excreo mdia de creatinina metablica. Em cada ensaio, foi coletado o volume total de urina
individualmente, durante perodos de 24 horas, por cinco dias consecutivos. Posteriormente, esses ensaios
foram analisados por colorimetria para creatinina e derivados de purina. A excreo de creatinina no foi
afetada (P>0,05) pelo consumo de forragem ou pelo tipo de forragem, mas foi influenciada pelo perodo
(P=0,0001). A excreo mdia de creatinina para ambas as forragens foi 0,21mmol/kg PV 0,75. Regresses
lineares entre os ndices derivados de purina:creatinina com a excreo total de derivados de purina foram
detectadas para milheto (P<0,0001; R=0,64) e azevm (P=0,02; R=0,20). Os resultados demonstraram que
a excreo de creatinina independente do tipo e do consumo de forragem e pode ser usada como marcador
preditivo do volume urinrio e do status nutricional em sistemas de pastejo.
1010 Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015
Uso da creatinina urinria...
Table 1. Chemical composition and morphological composition of the pearl millet and Italian ryegrass fed
to sheep in indoor trials
Pearl millet Italian ryegrass
Parameters
Mean Range Mean Range
Dry Matter (g/kg) 128 119136 227 170-283
OM1 (g/kg DM) 890 876901 915 897-932
CP2 (g/kg OM) 229 217241 201 146-256
NDFacp3 (g/kg OM) 548 500596 493 418-568
Leaf blades (g/kg DM) 396 175-617
Stem + sheath (g/kg DM) 321 304-338
Seed head (g/kg DM) 283 78-486
1
OM= organic matter; 2CP = crude protein; 3NDFacp = neutral detergent fibre corrected to ash and crude protein.
For urine collection each animal was fitted with Statistical analysis was performed using SAS
an abdominal urine collector to avoid contact version 9.2 statistical package (SAS Institute
with the feces. Urine was conveyed to a plastic Inc., Cary, NC, USA). The analysis of the
container with a lid containing 100mL of 20% experiment was performed by PROC GLM
sulfuric acid. After 24h, the total urine collected taking a completely randomized experimental
was measured, homogenized and 1% of the urine design with four treatments (offers) and four
was withdrawn and diluted three times with replicates per treatment. The effect of periods
distilled water to prevent possible crystallization (experiments) within the same forage was also
(Chen and Gomes, 1995). The samples were tested as a main effect. The comparison of means
adjusted to pH values below three using 20% was performed by Tukey test at a 5% error
sulfuric acid to prevent destruction of the probability. The regression between the PDC
bacterial purine bases and uric acid precipitation. index and intake of digestible organic matter
This was frozen at -20 C until analysis. Urine were evaluated by linear regression using the
samples were collected from only 84 animals of PROC REG procedure of SAS. Cook's distance
the 96 used in the experiments. Those excluded was calculated, excluding the data of those
were due to problems with abdominal collectors. animals that showed a daily excretion of
creatinine per kilogram metabolic weight that
Urinary creatinine was determined by the was very different from the average of the assay
colorimetric reaction with endpoint in alkaline within each level of fodder supply.
picrate solution, using commercial kits (Ref: 35,
Labtest, Lagoa Santa, MG, Brazil), while the RESULTS
purine derivatives (PD) were determined by
colorimetric analysis of allantoin and uric acid as Differences for forage intake (Table 2) between
described by Chen and Gomes (1995). Uric acid the levels of both offers were seen in the
was determined using a commercial kit (Ref: 73, experiments with both millet (P<0.0001) and
Labtest, Lagoa Santa, MG, Brazil), xanthine and ryegrass (P=0.0079).
hypoxanthine are converted to uric acid with
xanthine oxidase (Ref: x1875, Sigma-Aldrich In animals fed millet, urinary volume (Uv) and
Co.). daily excretion of purine derivatives followed the
For estimates of food intake and intestinal flow same pattern of response as dry matter intake
of microbial purines a purine (DMI), increasing with the increase of forage
derivatives:creatinine index (PDC) was supply. However, with ryegrass, this behavioral
calculated. This index was calculated from the pattern was not statistically different between
urinary concentrations (mmol/L) and creatinine treatments for Uv (P=0.18) and PD (P=0.21). The
purine derivatives multiplied by metabolic average volume of urine was higher for millet
weight as described by Chen et al. (2004): than ryegrass (2723 vs 2211 ml; Table 3) but
lower for the daily excretion of purine
derivatives (13.10 versus 18.71 mmol/day),
respectively.
Where creatinine and purine derivatives (PD) are
expressed in mmol/L and LW is the liveweight
of the animal (kg).
Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015 1011
David et al.
Table 2. Urine volume and urinary excretion of purine derivatives and creatinine by sheep fed different
allowances of pearl millet and Italian ryegrass
Main effects
Forage allowance (%LW)
Parameters SE2 (Pr>F)
1.5 2.0 2.5 AL1 Period Treatment
Pearl millet
DMI3 (g/day) 345c 444bc 578b 878a 33.72 0.01 <0.0001
Uv4 (ml/day) 2042b 2231b 2879ab 3696a 153.01 0.03 0.0001
PD5 (mmol/d) 8.02b 9.88b 14.07ab 20.11a 1.07 0.002 <0.0001
Creatinine
mmol/L 1.32 1.26 1.07 0.90 0.07 0.09 0.1529
mmol/day 2.50 2.79 2.97 2.99 0.15 0.02 0.7052
mmol/kg
0.20 0.21 0.22 0.24 0.01 0.13 0.6615
LW0,75
Italian ryegrass
DMI3 (g/day) 458b 556ab 731a 759a 39.57 0.0350 0.0079
Uv4(ml/day) 1897 2291 1856 2835 184.51 0.0005 0.1852
PD5 (mmol/d) 14.43 12.85 16.28 30.61 3.34 0.87 0.2153
Creatinine
mmol/L 1.63 0.94 1.41 0.96 0.15 0.94 0.1355
mmol/day 2.90 2.13 2.17 2.72 0.28 0.0026 0.6833
mmol/kg
0.23 0.16 0.16 0.21 0.02 0.0001 0.5285
LW0,75
1
AL= ad libitum; 2SE= standard error; 3DMI= dry matter intake;4Uv= urine volume; 5PD= purine derivatives
Averages in the same row followed by letters differ by the Tukey test at 5%.
Table 3. Urine volume and urinary excretion of purine derivatives and creatinine by sheep fed different
forage types
Pearl Italian Forage type
Parameters Mean SE1
millet Ryegrass (Pr>F)
DMI2 (g/day) 565b 605a 578 26.20 <0.0001
Uv3 (ml/day) 2723a 2211b 2563 122.04 <0.0001
PD4 (mmol/d) 13.10b 18.71a 14.87 1.30 0.0011
Creatinine
mmol/L 1.13b 1.29a 1.18 0.06 0.05
mmol/day 2.81 2.59 2.74 0.14 0.80
mmol/kg LW0,75 0.22 0.20 0.21 0.01 0.66
1
SE= standard error; 2DMI= dry matter intake; 3Uv= urine volume; 4PD= purine derivatives
Averages in the same row followed by letters differ by the Tukey test at 5%.
The daily excretion of creatinine (mmol/kg The purine derivatives:creatinine index (PDC
LW0.75) was not affected by the level of forage index) and digestible organic matter intake
allowance or type of forage. However, it was not (DOMI, Fig. 1) were significant for both pearl
constant over time, varying significantly millet (P<0.0001) and ryegrass (P=0.02) with
(P=0.0001) between the digestibility trials estimates of DOMI closer to the actual for pearl
(periods) for ryegrass. On average for the two millet (R=0.64) than for ryegrass (R=0.20). An
forages the daily creatinine excretion was inclination angle was also higher for millet (3.14)
0.21mmol/kg LW0.75. compared to ryegrass (1.23).
1012 Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015
Uso da creatinina urinria...
Figure 1. Relationship between the purine derivatives:creatinine index (PDC index) and digestible organic
matter intake (DOMI) of sheep in different forage allowances of pearl millet () or Italian ryegrass ().
Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015 1013
David et al.
with caution, since the synthesis of purine FAICHNEY, G.J.; WELCH, R.J.; BROWN,
derivatives is not constant and can vary G.H. Prediction of the excretion of allantoin and
considerably according to the type of diet total purine derivatives by sheep from the
(Makkar, 2004). This is due to differences in the creatinine cefficient. J. Agr. Sci., v.125, p.425-
efficiency of microbial protein synthesis due to 428, 1995.
the greater or lesser capacity of the diet to meet
GEORGE, S.K.; DIPU, M.T.; MEHRA, U.R. et
the nutritional requirements of rumen
al. Influence of level of feed intake on
microorganisms. Different slopes for ryegrass
concentration of purine derivatives in urinary
and millet (Figure 1) suggest different rates of
spot samples and microbial nitrogen supply in
excretion of purine derivatives (directly related
crossbred bulls. Asian Austral. J. Anim., v.9,
to rumen microbial protein) per unit of
p.1291-1297, 2006.
consumption of digestible organic matter. Thus,
adopting the same PDC index for different HARPER, H.A.; RODWELL, V.W.; MAYES,
forages can lead to under or over estimation of P.A. Manual de qumica fisiolgica. 5.ed. So
DOMI depending on the efficiency with which it Paulo: Atheneu, 1982, 736 p.
is used in the synthesis of microbial protein.
HOVELL, F.D.DEB.; RSKOV, E.R.; KYLE,
D.J.; MACLEOD, N.A. Under nutrition in sheep:
CONCLUSIONS
nitrogen repletion by N-depleted sheep. Br. J.
Creatinine excretion is not affected by the level Nutr., v.57, p.7788, 1987.
of forage allowance, nor the type of forage, HOVELL, F.D.DEB.; RSKOV, E.R.;
suggesting the use of a single value for estimates MACLEOD, N.A.; MCDONALD, N.A. The
of urinary volume. The digestible organic matter effect of changes in the amount of energy infused
intake and excretion of purine derivatives present as volatile fatty acids on the nitrogen retention
a relationship with the PDC index, and the same and creatinine excretion of lambs wholly
can be used as a nutritional indicator. But this nourished by intragastric infusion. Br. J. Nutr.,
index is dependent on the efficiency of microbial v.50, p.331343, 1983.
synthesis and therefore may vary between
different forages. KOREN, A. Creatinine urine. Medical
encyclopedia, 2000. Available in:
REFERENCES <http://www.nl.nih.gov/medlineplus/ency/article/
003610.htm>. Accessed in: Dec.10, 2013.
CHEN, X.B.; GOMES, M.J. Estimation of KOZLOSKI, G.V.; FIORENTINI, G.; HRTER,
microbial protein supply to sheep and cattle C.J.; SANCHEZ, L.M.B. Uso da creatinina como
based on urinary excretion of purine derivatives: indicador da excreo urinria em ovinos. Cienc.
an overview of the technical details. Bucksburn Rural, v.35, p.98-102, 2005.
Aberdeen, UK: International Feed Resources
Unit Rowett Research Institute, 1995. LIU, Z.J.; MCMENIMAN, N.P. Effect of
nutrition level and diets on creatinine excretion
CHEN, X.B.; JAYASURIYA, M.C.N.; by sheep. Small Ruminant Res., v.63, p.265-273,
MAKKAR, H.P.S. Measurement and application 2006.
of purine derivatives:creatinine ratio in spot urine
samples of ruminants. In: MAKKAR, H.P.S., MAKKAR, H.P.S. Development, standardization
CHEN, X.B. (Eds.), Estimation of microbial and validation of nuclear based technologies for
protein supply in ruminants using urinary purine estimating microbial protein supply in ruminant
derivatives. Kluwer Academic Publishers, 2004. livestock for improving productivity. In:
p.167179. MAKKAR, H.P.S.; CHEN, X.B. (Eds.),
Estimation of microbial protein supply in
EASLEY, J.F.; MCCALL, J.T.; DAVIS, G.K.; ruminants using urinary purine derivatives.
SHIRLEY, R.L. Analytical methods for feeds Dordrecht: Kluwer Academic Publishers, 2004,
and tissues. Gainesville: University of Florida, p.113.
Nutrition Laboratory, Dept. of Animal Science,
1965. 81 p.
1014 Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015
Uso da creatinina urinria...
Arq. Bras. Med. Vet. Zootec., v.67, n.4, p.1009-1015, 2015 1015