Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
Markers for estimating total intake of penned Holstein x
Zebu heifers
SILVA, Janaina Januário da2; SALIBA, Eloísa Oliveira Simões3; BORGES, Iran3; GONÇALVES, Lúcio
Carlos3; RODRIGUÉZ, Norberto Mario3; AROEIRA, Luiz Januário Magalhães4; SILVA, André Guimarães
Maciel5; COSTA, Francisco José do Nascimento6
1
Part of PhD Thesis of first author, financed by FAPEMIG
Federal University of Mato Grosso, College of Agronomy and Veterinary Medicine, Basic Sciences and Animal
production Department, Cuiabá, Mato Grosso, Brasil.
3
Federal University of Minas Gerais, School of Veterinary Sciences, Animal Sciences Department, Belo Horizonte, Minas
Gerais, Brasil.
4
National Center of Research on Dairy Cattle, Forage Sector, Juiz de Fora, Minas Gerais, Brasil.
5
Federal University of Pará, Veterinary Medicine College, Department of Animal Production, Castanhal, Pará, Brasil.
6
Center of Superior Teaching of Juiz de Fora, Minas Gerais, Brasil
*
corresponding address: [email protected]
2
ABSTRACT
The purpose of the study was to evaluate the efficiency of internal markers: indigestible fibers
(iADF and iNDF), Klason lignin and external markers: Cromic oxide and modified enriched
hydroxyphenylpropane (LIPE®) for estimating total intake of Holstein x Zebu heifers. The animals
were kept in pens and assigned four different diets: Elephant Grass (Pennisetum Purpureum
Schum.) silage; Elephant grass silage and commercial concentrate; chopped sugar cane and urea;
chopped sugar cane, urea and commercial concentrate. The chromic oxide marker underestimated
the intake regardless of the diet, and its estimative differed both from reference intake (measured in
the trough) and from other marker’s. LIPE®) is suggested as a replacement of Chromium Oxide,
since its estimates did not differ from the reference measured intake, in all diets. Klason lignin has
shown to be more adequate in estimating intake on heifers fed diets based on Grass silage, than on
heifers fed sugarcane. For heifers fed sugarcane and urea, Klason lignin underestimated intake
(3.75kg DM/day) compared to reference measure in the trough (4,05kg DM/day). For heifers fed
sugarcane, urea, and commercial concentrate, this marker estimated 3.09kg DM/day, while
reference method indicated intake of 4,90kg DM/day. Indigestible fiber markers (iADF and iNDF)
were adequate to estimate intake regardless of diet. These results show that markers have different
effects according to the forage fed to animals.
Keywords: external markers, internal markers, dairy heifers, fecal output
INTRODUCTION
Several problems are reported with the use of Chromium Oxide (Cr2O3) as an external
marker to measure fecal output, such as incomplete mixing with ruminal digesta, passage
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
rate through the rumen being faster than fiber material, possibilities of buildup in some
parts of the digestive tract. Despite that, the marker is still the one that is most used to this
purpose in experiments with ruminants, specially for its low cost and relative ease of
administration and analysis (PAIXÃO et al., 2007).
Internal markers have also been employed in estimating fecal output, such as: Acid
Insoluble Ash (AIA), Acid Detergent Insoluble Ash (ADIA) Acid Detergent Insoluble
Lignin (ADIL), Klason Lignin and Neutral (iNDF) or Acid (iADF) Detergent Fiber. The
recovery of indigestible fractions of feeds is the base of utilization of internal markers,
conveniently applied in studies of this nature (BARROS et al., 2007).
Limitations inherent to markers, external as well as internal, triggered the search
for markers that are more precise, cheaper and preferably with a more simple analysis
method. When it comes to convenience, LIPE® (purified and enriched lignin) deserves a
highlight . That was the name given to the modified hydroxyphenylpropane, developed at
the Federal University of Minas Gerais. It is an alternative technique that demands a
smaller animal adaptation period, is low cost and is recovered in the feces nearly entirely,
something that is considered an essential quality of a good marker. (SALIBA et al., 2004).
Preliminary studies have shown a recovery rate of LIPE® ranging between 97.9 and 99.3%
for rabbits, 95.9% for sheep, 94.6 to 102.6% on swine and 96% on equines, with dry matter
intake estimated by LIPE® statistically identical to the actual measured intake
(RODRIGUÉZ et al., 2007).
Due to the lack of consensus among results of experiments using external and
internal markers, and the promising potential of LIPE®, this study had as an objective to
evaluate the efficiency of indigestible fibers (iADF and iNDF) and Klason Lignin as
internal markers, as well as Chromium Oxide and the modified enriched
hydroxyphenylpropane (LIPE®) for estimating total intake in crossbred Holstein x Zebu
heifers, kept in pens and fed four different diets.
Material and Methods
The experiment was conducted in the Experimental Field of Embrapa Dairy Cattle,
in the county of Coronel Pacheco – MG at geographical coordinates 21° 33'22"S e 43°
6'15"W and altitude of 410m. The regional climate is, according to the Köppen
classification, a CWA type (mesothermical).
An experimental design in random blocks (animals) under a factorial arrangement
(four diets and five markers, compared to measured intake at the trough and five days for
feces collection). A total of 20 crossbred Holstein x Zebu heifers were assigned to
individual stalls, with cement troughs and automatic drinking fountain of the clamshell
type. Fifteen days were dedicated to adapting the animals to the diets and the facilities. The
20 heifers were randomly sorted into four groups of five, and for each group, a different
diet was fed. The experimental diets were: Elephant Grass silage (Pennisetum purpureum
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
Schum.) with 25.9% DM; Elephant Grass silage with concentrate, at a 80:20 ratio, dry
matter based; chopped sugarcane (28.5% DM) with a mix of urea and ammonium sulfate
(9:1 ratio, dry matter based); chopped sugarcane with urea and ammonium sulfate (9:1) and
commercial concentrate (80:20 ratio, dry matter based), presented at Table 1.
Table 1: Bromatological composition of experimental diets
% DM
DM
(%)
CP
EE
ADF
NDF
Elephant Grass silage
20.00
5.05
1.78
34.59
Elephant Grass silage +
commercial concentrate
21.46
6.90
2.30
Sugarcane + 9:12 urea
and ammonium sulfate
25.8
11.89
Sugarcane, 9:12 urea +
ammonium sulfate +
commercial concentrate
27.76
Commercial concentrate
90.00
Diets
KLF
CE
(Mcal/kg DM)
IVDDM
(%)
55.75
11.59
4.24
45.68
31.83
53.07
12.16
3.94
48.05
0.80
48.93
66.48
13.96
3.95
43.20
12.13
2.19
34.10
55.22
12.43
4.06
44.63
16.00
4.70
6.40
17.50
-
4.60
90.00
DM = dry matter; CP = crude protein; EE = ethereal extract, ADF = acid detergent fiber; NDF = neutral
detergent fiber; KLF = Klason lignin fraction, CE = crude energy, IVDDM = in vitro digestibility of dry
matter; 2Urea and ammonium sulfate 9:1 ratios and 80:20 of Elephant grass silage or sugarcane and
commercial concentrate were established based on natural matter.
The choice for low quality diets was made due to fact that the majority of
experiments researched that have marker evaluation as their objective are using high quality
diets (based on corn silage, highly energetic concentrate, protein supplements, etc.). so
information regarding low quality diets is scarce.
Diets with commercial concentrate were supplied as a total mixture, homogenized
in the trough before heifer’s access to it. Samples of the complete diets and orts were
weighed and stored for posterior laboratory analysis. The determination of dry matter,
crude protein and ethereal extract were made according to AOAC International (CUNIFF,
1995) procedures. Neutral detergent fiber and acid detergent fiber were determined
according to method proposed by Robertson & Van Soest (1981). The digestibility of
experimental diets was determined using the in vitro method described by Tilley & Terry
(1963), and the results were applied to the calculus of dry matter intake of heifers.
For every heifer group, the following average live weights were observed: 268.5;
267,83; 267,50 and 273.33kg, respectively for heifers fed grass silage; grass silage and
concentrate; sugarcane and urea; sugarcane, urea and concentrate.
Since total fecal collection was impossible, fecal output (kg DM/day was estimated
using only the external markers (iADF, iNDF and Klason lignin) and internal (LIPE® and
Chromium Oxide). The external markers were administered following the scheme of two
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
doses, at 9am and 2pm, for Chromium Oxide, and only the 9am dosage for the LIPE®.
Capsules of 0.25g of LIPE® were supplied, and the Chromium Oxide was wrapped in
bullet-form paper bags, containing 5g of the marker. Both markers were orally introduced
using a bullet launcher. Internal markers iADF and iNDF were obtained after incubating the
feeds, orts and feces, grinded, put through a 5mm mesh sieve and allocated in 9 x 12cm
nylon bags, with 50µm porosity, on the rumen of three fistulated cows for 144 hours. The
basal diet of these cows was chopped sugarcane and 9:1 urea and ammonium sulfate. After
incubation, the bags were washed and dried in a ventilated oven at 55°C for 72 hours.
Determination of the markers were made sequentially according to Van Soest et. al.
(1991) using Ankon 200 apparatus Ankon Technology Corp., Fairport, NY, USA).
The Klason fraction of lignin was determined using gravimetric method, adding
sulfuric acid 72% to the samples of offered feeds, trough orts and feces, according to Van
Soest (1994).
Feces sampling followed protocol established for Chromium Oxide, being the 6 first
days the adaptation period and the 5 last days for sampling, straight from the rectum of the
animals, in the morning and afternoon, in a total of 11 days of administering the marker.
For LIPE®, however, 3 days were dedicated to adaptation (coinciding with the 3 last days
of adaptation to the chromium oxide) followed by 5 days of sampling. For the internal
markers, five days of weighing of the offered diet and orts were assigned, for comparison
purposes. Feces samples were dried in regulated and ventilated oven at 55°C for 72 hours
and processed in a Willey type grinder with a 1mm mesh sieve. An Atomic Absorption
Flame Spectrophotometer (Perkin Elmer 3110) was used, at the Nutrition Laboratory of EV
– UFMG, for dosing the chrome, according to methodology proposed by Berchielli et al.
(2000). Part of the grinded feces samples were mixed with Potassium bromide (KBr) at
1%, in order to read the concentrations of LIPE® with infrared spectroscopy. Fecal output
was calculated using the logarithmic ratio of the spectral bands between wavelengths λ1
(1050 ηm) and λ2 (1650ηm) (SALIBA et al., 2002).
The general equation to determine fecal output (kg DM/day) using both external and
internal markers was based on the ratio of the amount of the marker ingested by the animal
and its concentration on the feces. (Fecal Output = amount of marker ingested/ marker
concentration in feces)
The intake was obtained with the ratio of the fecal output, obtained with the
markers, and the inverse of digestibility, according to the equation: Intake (kg DM/day) =
Fecal output / (1 – Digestibility) (Prigge et al., 1981).
The mathematical model used for the ANOVA was:
Yijk = µ+ B + Di + Ij + DCk + DIij + DDCik + IDCjk + Eijk
Where:
Yijk = variable observed for diet I, using marker j, on day k;
µ = overall average
B = blocks (animals)
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
Di = effect of diet i
Ij = effect of marker j
DCk = effect of sampling day k
DIij = effect of the interaction diet x marker
DDCik = effect of the interaction diet x sampling day
IDCjk = effect of the interaction marker x sampling day
Eijk = random error
Results were analyzed using procedure PROC GLM of SAS (1996), and differences
between treatments were compared using the Tukey test with 5% probability.
Results and Discussion
Given the impossibility of doing the total collection of feces for the DM intake
calculation, estimates of fecal output (kg DM/day) were made using internal markers
(iADF, iNDF and Klason lignin) and external markers (LIPE® and Chromium Oxide),
observing the significant effect (P = 0.0001) of the marker according to the diet (Table 2).
Chromium Oxide marker underestimated (P<0,05) fecal output of the heifers,
compared to the other markers, regardless of the diet. Soares et al. (2004) have verified that
the fecal recovery of Chromium Oxide is incomplete, mainly due to variability of results
obtained with its analysis methodology. Besides, the variation of excretion between
animals, a variation throughout the day of its feces excretion, form of administration,
number of doses, method and time of sampling the marker in the feces, duration of
adaptation and sampling periods, incomplete mixture with the ruminal digesta, a passage
rate that is faster through he rumen than the fibrous material, all could have contributed for
the underestimation of fecal output using the Chromium Oxide. In a similar manner, Myers
et al. (2006), evaluating the excretion pattern of Titanium Dioxide and Chromium Oxide in
sheep, have verified that using two daily applications of the marker leads to reduction of the
total amplitude of variation, making the excretion profile of the Chromium Oxide more
stable and closer to the desired balance, as opposed to what occurs with a single dosage.
Detmann et al. (2004), when revising the use of markers, have arrived to the same
conclusion, and recommended the application of two daily dosages.
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
Table 2: Average estimates of fecal output (kg DM/day) of penned Holstein x Zebu heifers
Diet
Elephant Grass silage
Elephant Grass silage
+ commercial concentrate
Sugarcane + 9:12 urea
and ammonium sulfate
Sugarcane, 9:12 urea +
ammonium sulfate +
concentrate
Chromium
Oxide
LIPE®
Klason
lignin
iADF
iNDF
1,62±0,45b
2,71±0,5a
2,41±0,4a
2,69±0,2a
2,54±0,23a
1,78±0,15c
2,95±0,5a
2,28±0,0b
2,95±0,60a
2,74±0,69a
1,29±0,13b
2,24±0,3a
1,90±0,0a
1,90±0,11a
1,82±0,09a
1,29±0,09c
2,38±0,3a
2,17±0,1ab
1,91±0,07b
2,09±0,06ab
1
Averages and standard deviations followed by distinct letter in the same line differ by the Tukey test
(P<0,05); 2 LIPE®, iADF and iNDF are: purified and enriched hydroxyphenylpropane, indigestible fiber in
acid detergent and indigestible fiber in neutral detergent, respectively.
In this study, the use of two daily doses of Chromium Oxide did not avoid the
underestimation of the heifer’s fecal output, making evident the complex interaction of
factor that may lower the efficiency of this as an external marker. From an ethological
standpoint, applying 2 dosages in animals unaccustomed to experimental handling, such as
in this case, alters the intake, and therefore, the fecal output (ZEOULA et al., 2002).
The fecal output estimate provided by the internal markers have differed among
themselves (P<0,05) only when the diet based on Elephant grass silage an concentrate was
used. The indigestible fibers have given higher values (2.95 and 2.74kg DM/day, for iADF
and iNDF, respectively) than the ones obtained with Klason lignin (2.28kg DM/day)
Values obtained using LIPE® differed from the ones obtained using internal
markers for diets based on grass silage and concentrate (2.95 and 2.28kg DM/day, for
LIPE® and Klason lignin, respectively), and diets based on sugarcane, urea and concentrate
(2.38 and 1.91kg DM/day, for LIPE® and iADF, respectively).
In a study conducted by Freitas et al. (2002), the authors evaluated different
markers’s estimates of fecal output in crossbred bullocks with average weight similar to the
animals used in this study (271.7kg), and verified that the biggest estimate was obtained
using iNDF (2.99 kg DM/day), statistically similar to the one obtained with Chromium
Oxide (2.69kg DM/day). They concluded that, for conditions presented in their study, the
ivADF (in vitro) was the marker that did best estimating fecal output (2.39 kg DM/day), not
differing (P>0,05) from total collection.
For the study of the total DM intake, it’s important to infer once again that the
heifers have received the concentrate included in the total mix of the diet, therefore, results
presented refer to total intake (kg DM per day and % live weight), composed of 1.3kg of
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
concentrate (considering that it was completely consumed), added to the voluminous
portion, that can be known by subtracting this value from the total. (Tables 3 and 4).
Table 3: Average intake (kg DM/day) of penned Holstein x Zebu heifers
Measured
Intake
Chromium
Oxide
LIPE®
Klason
lignin
iADF
iNDF
Elephant Grass
silage
4,85±0,95a
2,98±0,33b
5,21±0,22a
4,63±0,40a
4,85±0,13a
5,33±0,37a
Elephant Grass
silage+ commercial
concentrate
5,86±0,67a
3,42±0,28b
5,42±0,39a
5,27±0,18a
5,48±0,19a
6,09±0,27a
Sugarcane + 9:12
urea + ammonium
sulfate
4,05±0,79ab
2,26±0,55c
4,04±0,18ab
3,57±0,27b
4,05±0,22ab
4,61±0,21a
Sugarcane, 9:12 urea
+ ammonium sulfate
+ concentrate
4,90±0,83a
2,19±0,48c
4,19±0,31ab
3,90±0,55b
4,43±0,34ab
4,99±0,15a
Diet
1
Averages and standard deviations followed by distinct letters in the same line differ by the Tukey test
2
(P<0,05); 2 LIPE®, iADF and iNDF are: purified and enriched hydroxyphenylpropane, indigestible fiber in
acid detergent and indigestible fiber in neutral detergent, respectively.
In the same that it happened with the fecal output estimative, Chromium oxide
underestimated (P<0,05) the dry matter intake of the heifers, regardless of diet. Results
with chromium Oxide not only differed from the intake measured directly at the trough, but
also from the ones obtained with the other markers. However, results obtained with LIPE®
have shown that it can be a substitute external marker for the Chorium oxide, given that its
estimates did not differ from the reference method, for all diets.
The Klason lignin was the most adequate method to estimate the intake of heifers
fed grass silage, however, it underestimated in value (P>0,05) the intake (3.75kg DM/day)
of heifers receiving sugarcane and urea, and statistically (3.99kg DM/day) from the ones
receiving sugarcane, urea and concentrate.
Despite the practicality of the analysis methodology, the Klason lignin fraction may
include a variety of non-lignin components, such as polymers (aldehyde phenol), tannin
tannin-protein complexes, polymers from the Maillard reaction, cutine, and also
contaminants from the additives (FUKUSHIMA & SAVIOLI, 2001). The use of the
Klason lignin fraction, as a marker, is highly criticized also for its impurities, inability to
recover phenols, and mainly, the alteration of lignophenols resulting of the use of strong
sulfuric acid. Besides, there is interference of the relative distribution of the lignin for
particle sizes. The lignin of the feeds is associated to a much more cohesive and firm
material, while in the feces, its associated with a thinner one. During the filtering step in the
laboratory, for obtaining the Klason fraction, very small particles of feces may be lost
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
(SAVIOLI et al., 2000). Another cause for the loss of fecal lignin may also incur with the
attack of the sulfuric acid to the very fine particles, since these present a larger surface of
contact with the acid.
These results reinforce the presumed idea that the markers have different responses
according to the type of voluminous feed offered (BERCHIELLI et al., 2005), and
emphasize the importance of research that associate types of markers (external and internal)
with different forages, intending to establish the best contrasts.
Table 3: Average intake in live weight percentage (%LW) of penned Holstein x Zebu heifers
Measured
Intake
Chromium
Oxide
LIPE®
Klason
lignin
iADF
iNDF
Elephant Grass
silage
1,81±0,12a
1,11±0,22b
1,95±0,17a
1,73±0,26a
1,81±0,07a
1,99±0,09a
Elephant Grass
silage+ commercial
concentrate
2,19±0,09a
1,28±0,18b
2,02±0,28a
1,97±0,17a
2,04±0,10a
2,27±0,13a
Sugarcane + 9:12
urea + ammonium
sulfate
1,51±0,07ab
0,85±0,21c
1,48±0,19ab
1,31±0,18b
1,51±0,02ab
1,72±0,17a
Sugarcane, 9:12 urea
+ ammonium sulfate
+ concentrate
1,79±0,06a
0,80±0,25c
1,57±0,30ab
1,45±0,22b
1,62±0,05ab
1,82±0,04a
Diet
1
Averages and standard deviations followed by distinct letters in the same line differ by the Tukey test
2
(P<0,05); 2 LIPE®, iADF and iNDF are: purified and enriched hydroxyphenylpropane, indigestible fiber in
acid detergent and indigestible fiber in neutral detergent, respectively.
The Chromium Oxide was the least adequate marker to estimate dry matter intake
(% LW) of the heifers, underestimating (P<0,05) the reference intake measured at the
trough, regardless of diet. The DM intake estimates (%LW) obtained with the external
marker LIPE® and internal markers iADF and iNDF and Klason lignin did not differ
(P>0,05) from intake measured at the trough, therefore were considered adequate to
estimate dry matter intake of animals fed with both elephant grass silage and sugarcane.
The measuring of the intake directly at the trough has shown a low ingestion of the
heifers. That was also noted by Ítavo et al. (2002), using the internal markers iADF and
iNDF in steers fed coast-cross grass hay, registering intakes of 1.71%LW. In contrast,
steers fed Tifton 85 hay had an intake of 2.27%LW, suggesting the higher nutritional value
of the latter hay. In the same study, the authors concluded that iADF was the most adequate
marker to estimate the steer’s intake. In contrast, Cabral et al. (2008) have registered an
inefficiency of this marker compared to Chromium Oxide and iNDF, as it overestimated
fecal excretion and underestimated the apparent dry matter digestibility for crossbred bulls
with 350kg live weight.
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
When it comes to intake of diets based on sugarcane, it was observed a lower intake
than the one registered by Queiroz et al. (2001). Authors evaluated the ingestive behavior
of heifers averaging 220kg live weight and 14 months old, fed sugarcane and two different
concentrates, whose difference was in a higher or lower ruminal degradation of the protein
source – soybean meal or bloodmeal and corn gluten, respectively. The
voluminous:concentrate proportion was equivalent to 77.77:22.23 for the sugarcane and
soybean meal based diet and 83.39:16.61 for the sugarcane, blood meal and corn gluten
diet. The authors registered intakes of 9.10kg and 8.07kg DM/day, respectively,
corresponding to 4.13 and 3.67% live weight. The fecal output estimates and, consequently,
the dry matter intake of dairy heifers consuming diets of low nutritional value can be
obtained using the markers LIPE®, iADF and iNDF, instead of Chromium oxide. However,
the utilization of Klason lignin as a marker with diets of this kind is not recommended.
Using LIPE®, iADF and iNDF as substitutes to Chromium oxide also allows the reduction
of the sampling period from six to three days, minimizing the animal’s stress.
Considering the similarity of the estimates obtained with the markers and the ones
obtained with the measuring at the trough, for heifers fed diets based on grass silage, the
most adequate markers were iADF and Klason lignin. When concentrate was added to the
diets, the most precise markers were iNDF, followed by iADF. For heifers fed a sugarcane
and urea diet, markers iADF and LIPE® provided the closets results to actual intake, and
when concentrated was added to these diets, iNDF and iADF were the most adequate.
These results corroborate the evidences that the type of voluminous feed requires
the use of specific markers, that are most adequate to obtain the estimative.
References
BARROS, E.E.; FONTES, C.A.A.; DETMANN, E.; VIEIRA, R.A.M.; HENRIQUES,
L.T.; RIBEIRO, E.G. Avaliação do perfil nictemeral de excreção de indicadores internos e
de óxido crômico em ensaios de digestão em ruminantes. Revista Brasileira de Zootecnia,
v.36, n.6, 2007. Supl.
BERCHIELLI, T.T.; OLIVEIRA, S.G.; CARRILHO, E.N.V.M.; FEITOSA, J.V.; LOPES,
A.D. Comparação de indicadores para estimativas de produção fecal e de fluxo de digesta
em bovinos. Revista Brasileira de Zootecnia, v.34, n.3, p.987-996, 2005.
BERCHIELLI, T.T.; ANDRADE, P.; FURLAN, C.L. Avaliação de marcadores internos
em ensaios de digestibilidade. Revista Brasileira de Zootecnia, v.29, n.3, p.830-833, 2000.
CABRAL, L.S.; VALADARES FILHO, S.C.; DETMANN, E.; ZERVOUDAKIS, J.T.;
SOUZA, A.L.; VELOSO, R.G. Avaliação de indicadores na estimação da excreção fecal e
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
da digestibilidade em ruminantes. Revista Brasileira de Saúde e Produção Animal, v.9, n.1,
p.29-34, 2008.
CUNIFF, P. Official methods of analysis of AOAC International. 16.ed. Arlington: AOAC
International, 1995.
DETMANN, E.; VALADARES FILHO, S.C.; PAULINO, M.F.; ZERVOUDAKIS, J.T.;
CABRAL, L.S. Avaliação da técnica de indicadores na estimação do consumo por
ruminantes em pastejo. Cadernos Técnicos de Medicina Veterinária e Zootecnia, n.46,
p.40-57, 2004.
FREITAS, D.; BERCHIELLI, T.T.; SILVEIRA, R.N.; SOARES, J.P.G.; ERNANDES,
J.J.R.; PIRES, A.V. Produção fecal e fluxo duodenal de matéria seca e matéria orgânica
estimados por meio de indicadores. Revista Brasileira de Zootecnia, v.31, n.3, p.1521-1530,
2002.
FUKUSHIMA, R.S.; SAVIOLI, N.M.F. Correlação entre digestibilidade in vitro da parede
celular e três métodos analíticos para avaliação quantitativa da lignina. Revista Brasileira
de Zootecnia, v.30, n.2, p. 302-309, 2001.
ÍTAVO, L.C.V.; VALADARES FILHO, S.C.; SILVA, F.F.; VALADARES, R.F.D.;
CECON, P.R.; ÍATAVO, C.C.B.F.; MORAES, E.H.B.K.; PAULINO, P.V.R. Níveis de
concentrado e proteína bruta na dieta de bovinos nelore nas fases de recria e terminação:
Consumo e Digestibilidade. Revista Brasileira de Zootecnia, v.31, n.2, p.1033-1041, 2002.
Supl.
MYERS, W.D.; LUDDEN, P.A.; NAYIGIHUGU, V.; HESS, B.W. Excretion patterns of
titanium dioxide and chromic oxide in duodenal digesta and faeces of ewes. Small
Ruminant Research, v.63, p.135-141, 2006.
PAIXÃO, M.L.; VALADARES FILHO, S.C.; LEÃO, M.I.;CECON, P.R.; MARCONDES,
M.I.; SILVA, P.A.; PINA, D.S.; SOUZA, M.G. Variação diária na excreção de indicadores
interno (FDAi) e externo (Cr2O3), digestibilidade e parâmetros ruminais em bovinos
alimentados com dietas contendo uréia ou farelo de soja. Revista Brasileira de Zootecnia,
v.36, n.3, p. 739-747, 2007.
PRIGGE, E.C.; VARGA, G.A.; VICINI, J.L.; REID, R.L. Comparison of ytterbium
chloride and chromium sesquioxide as fecal indicators. Journal of Animal Science, v.53,
n.6, p. 1629-1633, 1981.
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
QUEIROZ, A.C.; NEVES, J.S.; MIRANDA, L.F.; PEREIRA, J.C.; PEREIRA, E.S.;
DUTRA, A.R. Efeito do nível de fibra e da fonte de proteína sobre o comportamento
alimentar de novilhas mestiças Holandês x Zebu. Arquivo Brasileiro de Medicina
Veterinária e Zootecnia, v.53, n.1, p. 84-88, 2001. [ Links].
ROBERTSON, J.B.; VAN SOEST, P.J. The detergent system of analysis and its
application to human foods. In: JAMES, W.P.T.; THEANDER, O. (Ed.). The analysis of
dietary fiber in food. New York: Marcel Dekker, 1981. p.123-158.
RODRIGUÉZ, N.M.; SALIBA, E.O.S; GUIMARÃES-JÚNIOR, R. Uso de indicadores
para estimar consumo y digestibilidade de pasto. LIPE, lignina purificada y enriquecida.
Revista Colombiana de Ciências Pecuárias, v.20, n.4, p. 518-525, 2007.
SALIBA, E.O.S.; FERREIRA, W.M.; RODRIGUEZ, N.M.; GONÇALVES, L.C.;
BORGES, I. Lignin from eucalyptus as indicator for rabbits in digestibility trials. Tropical
and Subtropical Agroecosystens, v.3, n.1, p.107-109, 2004.
SALIBA, E.O.S.; GONÇALVES, L.C.; MORAIS, S.A.L.; PILÓ-VELOSO, D. Lignina
isolada da palha de milho utilizada com indicador em ensaios de digestibilidade: estudo
comparativo. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, v.54, n.1, 2002.
SAS INSTITUTE. The SAS system for Windows. Version 6.12. Cary, 1996.
SAVIOLI, N.M.F.; FUKUSHIMA, R.S.; LIMA, C.G.; GOMIDE, C.A. Rendimento e
comportamento espectrofotométrico da lignina extraída de preparações de parede celular,
fibra em detergente neutro ou fibra em detergente ácido. Revista Brasileira de Zootecnia,
v.29, n.4, p.988-996, 2000.
SOARES, J.P.G.; BERCHIELLI, T.T.; AROEIRA, L.J.M.; DERESZ, F.; VERNEQUE,
R.S. Estimativas de consumo do capim-elefante (Pennisetum purpureumSchum), fornecido
picado para vacas lactantes utilizando a técnica do óxido crômico. Revista Brasileira de
Zootecnia, v.33, n.3, p. 811-820, 2004.
TILLEY, J.A.; TERRY, R.A. A two stage technique for the in vitro digestion of forage
crops. Journal of British Grassland Society, v.18, n.1, p. 104-111, 1963.
VAN SOEST, P.J. Nutritional ecology of the ruminant. 2.ed. Ithaca: Cornell University,
1994. 476p.
Rev. Bras. Saúde Prod. An., v.11, n.3, p.838-848 jul/set, 2010 ISSN 1519 9940
http://www.rbspa.ufba.br
VAN SOEST, P.J.; ROBERTSON, J.B.; LEWIS, B.A. Methods for dietary fiber, neutral
detergent fiber, and nonstarch polyssacharides in relation to animal nutrition. Journal of
Dairy Science, v.74, n. 10, p. 3583-3597, 1991.
ZEOULA, L.M.; PRADO, I.N.; DIAN, P.H.M.; GERON, L.J.V.; CALDAS NETO,
S.F.M.; MAEDA, E.M.; PERON, P.D.P.; MARQUES, J.A.; FALCÃO, J.S. Recuperação
fecal de indicadores internos avaliados em ruminantes. Revista Brasileira de Zootecnia,
v.31, n.4, p.865-1874, 2002.
Receiving date: February 3rd 2009
Approval date: August 22nd 2010