Animal Feed Science and Technology 226 (2017) 56–64

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Animal Feed Science and Technology

journal homepage: www.elsevier.com/locate/anifeedsci

Optimizing the use of spineless cactus in the diets of cattle:

Total and partial digestibility, fiber dynamics and ruminal
parameters
Michelle C.B. Siqueira a , Marcelo de A. Ferreira a , João Paulo I. dos S. Monnerat a ,
Janaina de L. Silva a,∗ , Cleber T.F. Costa b , Maria G. da Conceição a ,
Rafael de P.X. de Andrade c , Leonardo J.A. Barros a , Tobias T. de B. Melo a
a
Department of Animal Science, Universidade Federal Rural de Pernambuco, Recife, Brazil
b
Department of Agriculture, Instituto Federal do Sertão Pernambucano, Floresta, Brazil
c
Studies Institute of the Humid Tropics, Universidade Federal do Sul e Sudeste do Pará, Xinguara, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: The effects of the inclusion of spineless cactus (0; 147; 294; 441 and 588 g kg−1 ) in replace-
Received 11 March 2016 ment of Tifton hay on intake, total and partial (ruminal and intestinal) digestibility obtained
Received in revised form 2 December 2016 from samples of digesta collected in reticulum and omasum, fiber dynamics, and rumen
Accepted 12 December 2016
parameters were evaluated. Five rumen fistulated crossbred steers with an average body
weight of 380 ± 5.3 kg were assigned to a 5 × 5 Latin square design. The nutrient intake
Keywords: showed a quadratic effect, with a maximum intake of dry matter (8.89 kg d−1 ; P = 0.012)
Drought
and digestible organic matter (5.75 kg d−1 ; P = 0.013) estimated with 339 and 418 g kg−1 of
Energy
inclusion, respectively. The total (P < 0.01) and ruminal (P < 0.05) digestibility of dry matter
Fiber
Rumen metabolism (DM), organic matter (OM), crude protein (CP), and degradation rate (Kd) of DM increased
Volatile fatty acids linearly. There was no effect on the total and ruminal digestibility, ingestion rate (Ki), pas-
sage rate (Kp) and Kd of neutral detergent fiber (NDF; P > 0.05). The maximum pool of NDF
(2.46 kg; P < 0.05) was estimated with 201 g kg−1 of spineless cactus inclusion. Except for the
proportion of volatile fatty acids that did not differ (P > 0.05) with the inclusion of spineless
cactus, there was a linear decrease (P = 0.001) of rumen pH, and a quadratic effect (P < 0.001)
of ammonia nitrogen. It is recommended to include 418 g kg−1 of spineless cactus on a DM
basis as a replacement of Tifton hay in the diet of cattle to maximize energy intake.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction

Climate changes have caused frequent and prolonged droughts, with losses to ruminants reared in degraded pastures
or supplemented with low-quality roughage (Ben Salem, 2010). Under these conditions, the farmer depends on roughage
importing (hay or silage) from other regions, burdening the production system.

Abbreviations: DM, dry matter; OM, organic matter; CP, crude protein; EE, ether extract; aNDFom, neutral detergent fiber assayed with a heat stable
amylase and corrected for ash and nitrogenous compounds; iNDF, indigestible neutral detergent fibre; NFC, non-fibre carbohydrates; DOM, digestible
organic matter; BW, body weight; Ki, ingestion rate; Kp, passage rate; Kd, digestion rate; Kpi, passage rate of iNDF; RAN, rumen ammonia-nitrogen.
∗ Corresponding author.
E-mail addresses: michelle.siqueira2@gmail.com (M.C.B. Siqueira), marcelo.aferreira@ufrpe.br (M.d.A. Ferreira), joao.monnerat@dz.ufrpe.br (J.P.I.d.S.
Monnerat), silva janainalima@yahoo.com.br (J.d.L. Silva), clebertcosta@gmail.com (C.T.F. Costa), gabizoo2283@hotmail.com (M.G. da Conceição),
rafaeldepaula@zootecnista.com.br (R.d.P.X. de Andrade), leonardobarros92@hotmail.com (L.J.A. Barros), tobiaszootec@gmail.com (T.T.d.B. Melo).

http://dx.doi.org/10.1016/j.anifeedsci.2016.12.006
0377-8401/© 2016 Elsevier B.V. All rights reserved.
 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64 57

Fig. 1. Estimate of ruminal pH on the collection times (T) for the respective dietary spineless cactus (SC) levels (g kg−1 ).

Fig. 2. Estimate of rumen ammonia nitrogen (RAN) concentration on the collection times (T).

Spineless cactus use can reduce concentrate supplementation effective cost in the dry season due to being able to with-
stand prolonged droughts and having high production potential in these regions (Nefzaoui et al., 2014). Currently, spineless
cactus has the cost of US$0.13 kg−1 dry matter (DM) to the producer. It is cheaper if compared to other roughages, such as
corn silage and Tifton hay, which cost US$0.27 kg−1 DM.
Among attributes, spineless cactus has low crude protein (40–50 g kg−1 dry matter (DM)) and neutral detergent fiber (NDF;
170–280 g kg−1 DM) contents, but high non-fibrous carbohydrate content (NFC; 640–710 g kg−1 ) (Nefzaoui and Ben Salem,
2002; Ferreira et al., 2012) and a rapid NDF degradation rate (associated with low lignin content). Due to this composition,
spineless cactus presence in the diet associated with protein and fiber sources promotes nutrient degradability increase
(Batista et al., 2009).
The use of non-protein nitrogen compounds (urea) aims to raise the protein content of the diet, and its use is justified by
its low cost if compared to true protein sources (Felix et al., 2014). According to Ben Salem and Abidi (2009), total replacement
of concentrated feeds (such as corn or barley) by spineless cactus can be performed without causing any negative effect in
ruminants. However, it is suggested that forage replacement (such as hay, silage or straw) does not exceed 50% dry matter,
since digestion and animal performance may be impaired. Thus, it was hypothesized that providing more digestible feeds
(rich in NFC), such as spineless cactus associated with urea, replacing the diet roughage (Tifton hay), could interfere with
cattle ruminal metabolism.
Therefore, this study aimed to evaluate the effects of spineless cactus inclusion replacing Tifton hay on intake, total and
partial (ruminal and intestinal) digestibility of nutrients, fiber dynamics and ruminal parameters in cattle. In addition, to
identify the ideal ratio of Tifton hay:spineless cactus that would maximize the use of this forage in cattle systems Figs 1 and 2.
58 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64

Table 1
Chemical composition of ingredients used in the experimental diets (g kg−1 dry matter).

Item Ingredients

Tifton hay Spineless cactus Ground corn Soybean meal

Dry mattera 839 139 859 888

Organic matter 914 860 987 931
Crude protein 94,6 41,0 88,2 508,2
Ether extract 19,4 15,7 48,4 16,5
aNDF(n) 654 277 76,8 92,8
iNDF 229 94.7 23.3 21.1
Non-fibre carbohydrates 147 523 774 312

aNDF(n) – neutral detergent fiber assayed with a heat stable amylase and corrected for ash and nitrogenous compounds; iNDF – indigestible neutral
detergent fibre.
a
g kg−1 as fed.

Table 2
Proportion of ingredients and chemical composition of the experimental diets.

Item Inclusion of spineless cactus in replacement of Tifton hay (%)

0 14.7 29.4 44.1 58.8

Ingredients (g kg−1 )
Tifton hay 690 540 390 240 90.0
Spineless cactus 0 147 294 441 588
Ground corn 155 155 155 155 155
Soybean meal 140 140 140 140 140
Urea/ASa 0 3.00 6.00 9.00 12.0
Mineralb 15.0 15.0 15.0 15.0 15.0

Diet composition (g kg−1 of DM)

DM 852 466 326 252 209
OM 929 922 914 906 899
CP 150 150 150 149 149
EE 23.1 22.5 21.9 21.3 20.7
aNDF(n) 476 419 361 304 247
iNDF 165 145 124 103 82.9
NFC 280 329 376 422 471

DM – dry matter; OM – organic matter; CP – crude protein; EE – ether extract; aNDF(n) – neutral detergent fibre assayed with a heat stable amylase and
corrected for ash and nitrogenous compounds; iNDF – indigestible neutral detergent fibre; NFC – non-fibrous carbohydrates.
a
9 parts of urea and 1 part of ammonium sulphate (AS).
b
Nutrients per kg of product: Ca (min.) – 98 g, Ca (max.) – 113 g, P – 45 g, S – 40 g, Mg – 44 g, K – 61.5 g, Na – 114.5 g, Co – 48.5 mg, Cu – 516 mg, I – 30 mg,
Mn – 760 mg, Se – 9 mg, Zn – 2516 mg, F – 450 mg.

2. Material and methods

2.1. Experimental area

This study was carried out in the Department of Animal Science at the Federal Rural University of Pernambuco, located
in Recife, Pernambuco State, Brazil.

2.2. Animals and experimental design

The management and care of animals were performed in accordance with the guidelines and recommendations of the
Committee of Ethics on Animal Studies at the Federal Rural University of Pernambuco (License N◦ 009/2015), Recife, Brazil.
Five rumen fistulated steers (½ Holstein-Zebu) with an average initial body weight (BW) of 380 ± 5.3 kg were assigned in
a 5 × 5 Latin square design. The animals were weighed, identified and vermifuged prior to the experiment and housed in
individual pens fitted with feeders and waterers. The trial lasted 80 days, with five consecutive 16-day periods, and was
divided into a seven-day adaptation according to literature reference (Storry and Sutton, 1969; Menezes et al., 2011) and
nine-day sampling.

2.3. Experimental diets

The chemical composition of the ingredients and proportion of the ingredients in the concentrate mixture and the chem-
ical composition of the diets are shown in Tables 1 and 2. The experimental diets consisted of the inclusion of spineless
cactus (Nopalea cochenillifera Salm-Dyck) (0; 147; 294; 441 and 588 g kg−1 of DM basis) replacing Tifton hay (Cynodon spp.).
 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64 59

Urea and ammonium sulfate were added to the diets to adjust the crude protein (CP) content to 150 g kg−1 DM, to meet the
requirements of crossbred cattle (½ Holstein-Zebu), with an average daily gain of 1.5 kg (Valadares Filho et al., 2010).
The spineless cactus were cut and chopped daily and then provided to the animals. The evaluation of DM content of
spineless cactus was performed weekly to adjust the amount of feed allowed to the animals. The mixture of ingredients was
performed manually in the feeders, highlighting that the spineless cactus mucilage allowed a uniform aggregation of urea.

2.4. Experimental procedures and sampling

The diets were supplied ad libitum, allowing approximately 100 g kg−1 DM of orts. The animals were fed twice a day,
at 06:00 a.m. (60%) and 6:00 p.m. (40%). Forage provided and orts were sampled daily during the collection period and
subjected to partial drying in a forced ventilation oven set at 60 ◦ C for 72 h. The ingredients that comprised the concentrate
were sampled directly from the feed mill silos on the days that they were mixed. All samples were processed in a Wiley mill
to pass through a 2-mm screen sieve. After that each sample was homogenized and divided in two portions. Half of each
sample was processed again in the same mill to pass through a 1-mm screen sieve.
From the eighth to the tenth day of each period, the fecal dry matter output was estimated using the total collection of
feces through spontaneous defecation, used to estimate the total apparent digestibility of nutrients. Six collections of omasal
digesta were performed at 12-h intervals between day 11 and day 13. On day 11, samples were collected at 10:00 a.m. and
10:00 p.m. On day 12, they were collected at 8:00 a.m. and 8:00 p.m. On day 13, they were collected at 6:00 a.m. and 6:00 p.m.
Also, approximately 200 g of feces was collected from each animal between day 9 and day 11, and the indigestible neutral
detergent fiber (iNDF) content was used to estimate partial digestibility, ruminal pool, and passage rate. Fecal samples
were partially dried in a forced ventilation oven at 60 ◦ C. Subsequently, these samples were milled to a size of 1–2 mm in a
Wiley-type laboratory mill.
Briefly, to collect the omasal digesta, the technique reported by Huhtanen et al. (1997) was adapted as follows: the
collection of omasal digesta was performed by introducing the end of a collection plastic tube (1.2 m long by 14 mm i.d.)
into the rumen and passing it into the reticulum-omasal orifice until the first part of the tube passed into the orifice, where
it was secured by hand during the collection period. The other end of the collection tube was fitted to one of the openings
of a kitassato flask, and a vacuum pump was attached to the other opening. The vacuum pump was subsequently turned
on to begin a collection, and the digesta was collected through the tube via suction until it reached the kitassato flask.
Approximately 800 mL of digesta was obtained per collection and frozen (−20 ◦ C).
To estimate the omasal flow, a double marker system was employed in which cobalt-EDTA (6 g d−1 divided into four
doses were infused into the rumen of each animal at 12:00 a.m., 6:00 a.m., 12:00 p.m. and 6:00 p.m., beginning three days
before omasal digesta sampling) was used as the liquid phase and a small particle marker and indigestible neutral detergent
fiber (iNDF) were used as particulate phase (France and Siddons, 1986).
After collection, the omasal digesta sample was frozen (−20 ◦ C) until processing. At the end of each experimental period,
the samples were thawed at room temperature and filtered through a 100-␮m nylon filter with pores covering 44% of the sur-
face to generate two phases: the filtrate, which corresponded to the liquid phase and small particles, and the residue, which
corresponded to the large particle phase. Subsequently, these samples were oven dried (60 ◦ C) and ground as previously
described for forage and orts samples.
On day 14, the rumen was completely emptied four hours after the morning diet was provided to determine the rates
of indigestion and ruminal pool for each diet using the technique described by Allen and Linton (2007). On day 16, the
rumen was emptied immediately before morning feeding. After emptying the rumen, the total weight of the digesta was
determined, followed by filtering through four layers of cheesecloth to separate the solid and liquid phases. A representative
sample of both phases was collected and frozen (−20 ◦ C) for subsequent evaluation of DM, neutral detergent fiber (NDF) and
iNDF contents. After sampling, the phases were again mixed, and the remaining digesta returned to the rumen.
At the end of the sampling period, the feces, orts, ingredients of the diets, and omasal digesta and ruminal content samples
were oven dried (60 ◦ C) and ground. At the end of each period, a composite sample was prepared from the orts, feces, and
omasal digesta based on the air-dried weight of the samples for each animal, and the samples were then properly identified
and stored in plastic containers for further analysis.
Ruminal fluid was collected from steers on three days (day 11, 12 and day 13) at 6:00 a.m., 8:00 a.m., 10:00 a.m., and
12:00 p.m. on the same day. A total of 250 mL was collected from the anterior dorsal, anterior ventral, medium ventral,
posterior dorsal regions following ventral locations within the rumen using a 50-mL syringe screwed to a plastic tube
ending with a probe covered with a fine metal mesh. The rumen fluid was collected through the cannula and then filtered
through four layers of cheesecloth within 1 min and measured using a digital pH meter (Model k39-0014P-kasvi, Taiwan,
CHN). Ruminal fluid was acidified to a pH of 2 with 50% sulfuric acid, and subsamples (40 mL) were frozen at -20 ◦ C for the
later determination of volatile fatty acids (VFA) and rumen ammonia nitrogen (RAN) concentrations. The analysis of VFA
was performed using a gas chromatograph (Model CG-Master, Ciola and Gregori, Brazil) equipped with a flame ionization
detector and auto-injector and fitted with a GC column (30 m × 0.250 mm, 0.25 ␮m; Chromosorb WAW).
60 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64

Table 3
Mean values for the intake of nutrients in cattle.

Item Inclusion of spineless cactus (%) SEM Effect (P value)

0 14.7 29.4 44.1 58.8 Linear Quadratic

Intake (kg d−1 )

DM 7.56 8.02 9.08 8.85 8.01 0.500 0.145 0.012
OM 6.90 7.22 8.07 7.79 6.97 0.450 0.484 0.013
CP 1.21 1.25 1.38 1.30 1.20 0.070 0.881 0.018
aNDF(n) 3.44 3.14 3.10 2.72 2.04 0.180 <0.001 0.068
NFC 2.06 2.70 3.54 3.77 3.80 0.200 <0.001 0.079
DOM 4.75 5.12 5.72 5.83 5.52 0.290 0.119 0.034

Intake (g kg−1 BW)

DM 19.6 20.4 23.2 23.0 20.8 1.180 0.103 0.019

SEM – standard error of the mean; DM – dry matter; OM – organic matter; CP – crude protein; aNDF(n) – neutral detergent fibre assayed with a heat stable
amylase and corrected for ash and nitrogenous compounds; NFC – non-fibrous carbohydrates; DOM – digestible organic matter; BW – body weight.

2.5. Chemical analysis

Samples of feeds, orts, feces, ruminal content and omasal digesta were analyzed for dry matter (DM; method 934.01),
organic matter (OM; method 930.05), ash (method 942.05), crude protein (CP; method 968.06), and ether extract (EE; method
920.39), according to AOAC (2000). Dry matter was analyzed by the gravimetric difference between dry and wet sample
weights; OM by ashing at 600 ◦ C for at least 8 h; CP using the macro-Kjeldahl procedure and multiplied by a factor of 6.25;
and EE by Soxhlet extraction with petroleum ether. Analysis of NDF followed the method described by Mertens (2002), using
a heat-stable alpha amylase, without using sodium sulfite, and corrected for residual ash. The NDF was also corrected for
the nitrogenous compounds contents by using the method described by Licitra et al. (1996). The omasal digesta sample was
analyzed to estimate cobalt concentrations using an atomic absorption spectrophotometer. All of these chemical analyses
were performed in samples processed to pass through a 1-mm screen sieve.
The quantification of non-fibrous carbohydrates (NFC) content was performed according to Hall (2001) as follows:
NFC = 1000 – [(CP – CPu + U) + aNDFom + EE + ash]; where CPu = CP content from urea, U = urea content, and aNDFom = neutral
detergent fiber corrected for ash and nitrogenous compounds. The other terms were previously defined and all of them are
expressed as g kg−1 DM.
The iNDF content was analyzed using the fecal, omasal digesta, feed and orts samples processed at 2-mm screen sieve,
obtained by using in situ procedures with 288 h of rumen incubation in cattle, as described by Valente et al. (2011). On the
other hand, the omasal flow was calculated as described by France and Siddons (1986) using a double marker system in
which cobalt-EDTA was used as the liquid phase and small particles marker and iNDF was used as the large particles marker.
The ingestion rates (ki), passage (kp), degradation of NDF (kd) and iNDFi (kpi) were calculated by dividing the daily intake
and omasal flow by their respective rumen pools (Allen and Linton, 2007).

2.6. Statistical analysis

The variables studied were analyzed with the PROC MIXED option in SAS software (version 9.2), adopting 0.05 as the
critical level of probability for type I error according to the following model:

Y ijk =  + T i + aj + pk + εijk

where, Yijk is the dependent variable measured in animal j that was subjected to the i treatment in period k;  is the general
mean; Ti is the fixed effect of treatment i; aj is the random effect of animal j; pk is the random effect of period k; and εijk is
the unobserved random error assuming normal distribution.
After the analysis of the variance, the significance of the linear and quadratic effects obtained with the inclusion of
spineless cactus replacing Tifton hay was evaluated. A significance value of 0.05 was adopted as the critical value of the
probability of type I error. Ruminal pH, RAN, and VFAs were analyzed as the effects of repeated measures over time.

3. Results

The intake of DM (P = 0.012), OM (P = 0.013), CP (P = 0.018), and DOM (P = 0.034) showed a quadratic behavior with spine-
less cactus inclusion replacing Tifton hay (Table 3), with maximum values estimated of 8.89; 7.88; 1.35 and 5.75 kg d−1 ,
and spineless cactus maximal inclusion calculated to 339; 316; 299 and 418 g kg−1 , respectively. aNDFom intake linearly
decreased (P < 0.001) from 3.44 to 2.04 kg d−1 , while the NFC intake linearly increased (P < 0.001) from 2.06 to 3.80 kg d−1
with spineless cactus inclusion (Table 3).
Except to aNDFom (P > 0.05), the DM, OM and CP total (P < 0.001) and ruminal (P < 0.05) apparent digestibility linearly
increased with spineless cactus inclusion replacing Tifton hay (Table 4). The DM (P = 0.038) and aNDFom (P = 0.020) intestinal
digestibility linearly decreased; however, there was no effect (P > 0.05) for OM and CP
 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64 61

Table 4
Mean values for the total, ruminal and intestinal digestibility of nutrients.

Item Inclusion of spineless cactus (%) SEM Effect (P value)

0 14.7 29.4 44.1 58.8 Linear Quadratic

Total apparent digestibility (g kg−1 )

DM 691 715 713 751 788 12.7 <0.001 0.197
OM 684 710 708 748 789 12.7 <0.001 0.159
CP 765 767 756 792 832 11.7 <0.001 0.017
aNDF(n) 649 629 564 595 597 22.2 0.071 0.114

Ruminal digestibility (g kg−1 )

DM 318 356 413 503 588 40.4 <0.001 0.418
OM 465 500 538 618 672 35.7 <0.001 0.560
CP 300 289 294 334 473 53.2 0.0394 0.118
aNDF(n) 528 503 473 473 463 27.9 0.084 0.117

Intestinal digestibility (g kg−1 )

DM 539 553 505 500 471 25.5 0.038 0.663
OM 401 417 363 326 335 44.4 0.138 0.982
CP 662 670 654 688 660 26.7 0.875 0.820
aNDF(n) 121 126 91.0 62.0 62.0 21.6 0.020 0.949

SEM – standard error of the mean; DM – dry matter; OM – organic matter; CP – crude protein; aNDF(n) – neutral detergent fibre assayed with a heat stable
amylase and corrected for ash and nitrogenous compounds; NFC – non-fibrous carbohydrates.

Table 5
Mean values for the pool sizes and rates of ingestion (Ki), passage (Kp) and digestion (Kd) of DM and NDF, and passage rate of iNDF (Kpi).

Item Inclusion of spineless cactus (%) SEM Effect (P value)

0 14.7 29.4 44.1 58.8 Linear Quadratic

Ruminal pool (kg)

DM 3.34 3.66 3.78 3.48 2.91 0.267 0.265 0.059
NDF 2.27 2.44 2.41 2.20 1.70 0.032 0.179 0.042
iNDF 1.22 1.29 1.33 1.22 1.04 0.103 0.202 0.118

DM (h−1 )
Ki 0.1002 0.0928 0.1033 0.1124 0.1163 0.0110 0.163 0.623
Kp 0.0703 0.0603 0.0605 0.0554 0.0466 0.0070 0.053 0.929
Kd 0.0298 0.0324 0.0428 0.0572 0.0696 0.0070 0.009 0.415

NDF (h−1 )
Ki 0.0800 0.0640 0.0656 0.0638 0.0602 0.0080 0.121 0.466
Kp 0.0381 0.0321 0.0346 0.0294 0.0263 0.0040 0.062 0.903
Kd 0.0419 0.0318 0.0310 0.0344 0.0338 0.0040 0.351 0.188
Kpi 0.0451 0.0350 0.0320 0.0273 0.0228 0.0040 0.029 0.584

SEM – standard error of the mean; DM – dry matter; NDF – neutral detergent fibre; iNDF – indigestible neutral detergent fibre.

Table 6
Mean values for ruminal parameters in crossbred steers.

Item Inclusion of spineless cactus (%) SEM Effect (P value)

0 14.7 29.4 44.1 58.8 Treat. Time Treat. x Time

L Q L Q

pH 6.45 6.25 6.23 6.03 6.05 0.180 0.001 0.538 <0.001 <0.001 0.292
RAN (mg dL−1 ) 24.3 25.8 20.2 23.4 22.5 2.26 0.404 0.632 0.015 <0.001 0.159

Volatile fatty acids (mmol L−1 )

Acetate 71.3 69.6 69.2 70.6 68.3 1.95 0.362 0.918 0.913 0.917 0.888
Propionate 18.9 19.3 19.6 18.3 19.2 1.07 0.899 0.864 0.313 0.431 0.870
Butyrate 9.80 11.1 11.3 11.1 12.5 1.06 0.075 0.975 0.175 0.273 0.803

SEM – Standard error of the mean; Treat – treatment; RAN – rumen ammonia nitrogen.

Except for DM and iNDF pool, which were unchanged (P > 0.05), there was a quadratic effect for the NDF (Table 5; P = 0.042),
presenting a pool with spineless cactus inclusion replacing Tifton hay, and the maximum NDF pool estimated of 2.46 kg was
for 201 g kg−1 spineless cactus inclusion. There was no effect (P > 0.05) of spineless cactus inclusion on DM ingestion rate
(Ki) and passage rate (Kp). However, a linear increase (P = 0.009) was observed in the Kd of DM (Table 5). The Ki, Kp, and
Kd of NDF were not altered (P > 0.05) by spineless cactus inclusion. However, a linear decrease (P = 0.029) was observed for
iNDF Kp.
62 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64

Ruminal pH linearly decreased (P = 0.001) with spineless cactus inclusion replacing Tifton hay (Table 6). There was no
interaction (P > 0.05) between collection times and spineless cactus inclusion replacing Tifton hay on pH and RAN concentra-
tions (Table 6). The estimates of ruminal pH as a function of sample collection times for the five levels of spineless cactus is
shown in Fig. 1, thus the regression model adjusted for the ruminal pH (Y) was: Y = 6.6614 – 0.00588435*SC – 0.1788*T + 0.02*
T2 (R2 = 0.78), wherein “SC” is the level of spineless cactus and “T,” the collection time after the feed supply. A minimum pH
of 6.26, 6.17, 6.08, 6.00, and 5.91 was estimated for the spineless cactus levels of 0; 147; 294; 441 and 588 g kg−1 at 4.47
hours after feeding.
The estimates of RAN as a function of sample collection times for the five levels of spineless cactus is shown in Fig. 2, thus
the regression model adjusted for the ruminal pH (Y) was: Y = 21.7679 + 5.8439*T – 1.1069*T2 (R2 = 0.74), wherein “T” is the
collection time after the feed supply. A quadratic effect (P < 0.001) was observed for RAN related to collection times, with a
maximum concentration of 29.4 mg dL-1 estimated at 2.64 hours after feeding (Table 6). There was no effect (P > 0.05) on
short chain volatile fatty acid (VFA) proportion with the inclusion of spineless cactus (Table 6).

4. Discussion

Increased dietary NFC on diets containing greater levels of spineless cactus was expected. Additionally, increasing the
level of cactus substitution of Tifton hay alters the digestibility of dietary dry matter in the treatments. High-digestible diets
support higher DM intake than higher fiber diets due to their lower fiber content and greater rate and extent of digestion.
In contrast, high fiber diets are digested slowly and remain in the rumen for longer periods, limiting rumen capacity.
The digestion process must be assessed as a whole, especially ruminal degradation and passage since the time required for
the particle to be removed from the rumen decreases when the use speed of potentially degradable compounds is increased
by microorganisms (Allen, 1996).
On the other hand, if the energy concentration of the diet is too high, DM intake is reduced due to chemicals produced
during fermentation. Also, excessive amounts of nutrients may reduce intake due to physiological regulations. According to
Mertens (1997), physiological intake regulation is related to nutritional status or energy balance, and diets with high energy
and low fiber content promote a limited intake by animals by meeting their energy requirements.
The DM intake increase observed until 339 g kg−1 spineless cactus inclusion replacing Tifton hay was mainly due to
aNDFom reduction (476 to 247 g kg−1 DM) and NFC increase (280 to 471 g kg−1 DM) in the diets (Table 2). Nevertheless,
above the 339 g kg−1 spineless cactus inclusion, intake reduction can be attributed to animal satiation by meeting their
energy demand since the maximum DOM intake was estimated at 418 g kg−1 spineless cactus inclusion (Table 3). Besides,
the inclusion of spineless cactus up to 250 g kg−1 provided the largest ruminal pool of DM and NDF compared to diets without
spineless cactus, which provided higher DM intake due to the increased digestion rate of this constituent (Table 5).
Furthermore, the change of DOM intake (5.72 to 5.83 kg d−1 ) occurred between 294 and 441 g kg−1 spineless cactus
inclusion, and there was a decrease from 418 g kg−1 inclusion, which was probably related to the higher energy supply. In
this diet, there was an estimated 412 g kg−1 of NFC, which corroborates with the level (400–440 g kg−1 ) recommended by the
NRC (2001), indicating the fulfillment of energy requirements. Cavalcanti et al. (2008), Pessoa et al. (2004), and Wanderley
et al. (2002) observed similar results, with intake decrease from 420 g kg−1 of NCF from spineless cactus inclusion. NRC
(2001) showed DM intake reduction in diets with more than 70% moisture. This fact was confirmed in this study, in which
intake reduction occurred when diets had 69.9% moisture and 339 g kg−1 spineless cactus inclusion.
The DM, OM and CP total and ruminal digestibility increase with spineless cactus inclusion as a replacement for Tifton
hay could be explained by better use and availability of nutrients for rumen microorganisms. According to Ferreira (2005),
non-fibrous carbohydrates that form the largest fraction of the total carbohydrates in spineless cactus are readily fermented
in the rumen.
In this case, spineless cactus constituents showed a high degradation rate, especially in DM, favoring rumen fermentative
capacity maximization due to high NFC contents. The DM ruminal degradation rate increase resulted in the increased intake.
When evaluating different spineless cactus cultivars, Batista et al. (2009) found high DM ruminal degradability (averaged in
701 g kg−1 ), and concluded that there was high non-structural carbohydrate content and low lignin content. Thus, the Kd of
DM increase as spineless cactus replaced Tifton hay was justified.
Cavalcanti et al. (2006) reported NDF digestibility reduction due to high readily fermentable carbohydrate levels in the
rumen while assessing the effect of the replacement of Tifton hay with spineless cactus. However, in the present study
the ruminal and total digestibility of aNDFom was not altered with increasing NFC degradation rate in diets with a higher
proportion of spineless cactus, possibly due to of effective fiber content allowed in the diets (Table 2), since the minimal
inclusion of NDF should be 250 g kg−1 (NRC, 2001). Batista et al. (2009) reported ruminal degradability value of spineless
NDF of 501 g kg−1 , and approximate average values were observed in the present study 488 g kg−1 (Table 4). According to
Church (1988), ruminal pH is one of the mechanisms that explain roughage nutrient intake and digestibility since it is directly
related to the end products of fermentation and microorganism growth rate.
Ruminal fermentation pattern of diets with spineless cactus seems to affect pH drop slightly with the presented values
of 6.45 to 6.05 (Table 6), for no addition and higher inclusion of spineless cactus, respectively. It was observed in diets with
spineless cactus inclusion of 588 g kg−1 DM that there was a pH variation with small decrease at 4 hours after feeding (5.91)
and, successive pH increase (6.0; Fig. 1) at 6 hours after feeding. Thus, the short period observed to pH reduction (Fig. 1)
was not enough to limit or inhibit the degradability of NDF, i.e., fiber degradation may be adversely affected when ruminal
 M.C.B. Siqueira et al. / Animal Feed Science and Technology 226 (2017) 56–64 63

pH reaches values below 6.0 (Slyter, 1986). The maintenance of ruminal pH above 6.0 is ideal for the activity of fibrolytic
microorganisms’ since the bacterial adhesion process is severely inhibited by the initial pH below 5.5 (Mourino et al., 2001).
The observed increase in total and ruminal digestibility of CP for spineless cactus diets inclusion observed can be related to
a neutral detergent insoluble crude protein (NDIP) content of Tifton hay (406 g kg−1 of NDIP), which is greater than spineless
cactus (273 g kg−1 of NDIP; Table 1), thus allowing for more available N to the rumen. Other aspects observed are related to
increase in urea levels in diets with more spineless cactus inclusion (Table 2), necessary for promoting the same N content
for all experimental diets. Non-protein nitrogen compounds, such as urea, are degraded immediately, quickly releasing
NH3 in the rumen for microorganism use (Calomeni et al., 2015), allowing for higher nutrient degradation and increase
in digestibility. Studies have reported that urea ruminal infusion increased NH3-N concentration (Gabler and Heinrichs,
2003; Griswold et al., 2003). Nevertheless, NH3 -N concentration was not increased as urea increased in diets in the present
experiment, probably in function of higher NCF availability of diets with greater inclusion of spineless cactus that promoted
an increase in the energy supply as a function of the content of fast degradable carbohydrates. Thus, the diet provides
adequate synchrony between energy and protein.
According to Leng (1990), cattle DM intake is maximized when ruminal ammonia nitrogen (RAN) reaches values close
to 20 mg dL−1 in tropical conditions. In this study, the RAN remained at slightly higher concentrations (23.2 mg dL−1 ) than
suggested by Leng (1990). However, Mehrez and Ørskov (1977) suggested 23.0 mg dL−1 concentration for the maximum fer-
mentation rate. Therefore, experimental diets provided adequate RAN amounts for proper microorganism growth. Moreover,
the lack of effects on NH3-N concentration in the rumen and N digestibility in the animals fed spineless cactus suggests higher
utilization of nitrogen by ruminal microorganisms and demonstrates the high N recycling when animals are supplemented
with Tifton hay. This fact is important since that protein is the most expensive nutrient in diets for animals.
Although diets with increasing spineless cactus inclusion replacing Tifton hay provided high NFC levels, there was no effect
on the short chain volatile fatty acid proportion, probably due to ideal rumen fermentation condition promoted by the diets.
According to Batista et al. (2003), spineless cactus contains approximately 286 g kg−1 DM neutral detergent-soluble fiber,
providing a higher acetate to propionate ratio and a lower lactic acid content compared to sugar and starch fermentation.
Considering the energy intake as an important factor to be analyzed in the ruminant diet, it was utilized as criterion
to estimates the ideal inclusion of spineless cactus to maximum DOM. The maximum value estimated for DOM intake
was 5.75 g kg−1 for 418 g kg−1 of spineless cactus inclusion, by regression model: Y = 4.677254 + 0.051327X – 0.000614X2
(P = 0.034).

5. Conclusions

It is recommended to include 418 g kg−1 of spineless cactus on a DM basis as a replacement of Tifton hay in the diet of
cattle to maximize energy intake without causing adverse effects on ruminal fermentation. Thus, a Tifton hay to spineless
cactus ratio of 28:42 (70%) in diets with 30% concentrate is recommended for cattle.

Conflict of interest

The authors declare that there was no conflict of interest in carrying out this work.

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