Tropical Animal Health and Production (2021) 53:169

https://doi.org/10.1007/s11250-021-02618-1

REGULAR ARTICLES

Intake, digestibility, milk yield and composition, and ingestive

behavior of cows supplemented with byproducts from biodiesel
industry
Marcus Vinícius Gonçalves Lima 1 & Aureliano José Vieira Pires 2 & Fabiano Ferreira da Silva 2 &
Fábio Andrade Teixeira 2 & Bruna Rafaela de Carvalho Silva Castro Nogueira 2 & Leone Campos Rocha 3 &
Gleidson Pereira da Silva 2 & Weudes Rodrigues Andrade 2 & Gleidson Giordano Pinto de Carvalho 4

Received: 19 November 2019 / Accepted: 8 February 2021

# The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature 2021

Abstract
This trial aimed to analyze the effects of including three byproducts from the biodiesel industry on the intake, digestibility, milk
yield and composition, and feeding behavior of lactating cows. Eight crossbred Holstein-Zebu lactating cows with average body
weight 525 ± 18.5 kg and average milk yield of 8 ± 1.45 kg day–1 were assigned to four treatments (diets) in a double-Latin square
design, as follows: a diet based on corn- and soybean meal-based concentrate and three diets with 20% inclusion of byproducts
from the biodiesel industry (cottonseed cake, sunflower meal, and castor bean cake) on a total dry matter basis. The cows were
housed in individual covered stalls with concrete floor equipped with individual concrete troughs for feeding and automatic
drinkers, and fed diets containing 60% sugarcane and 40% concentrate. The inclusion of the byproducts in the diet changed the
intake, digestibility of some nutritional components, milk yield and composition, and feeding behavior of lactating cows. The use
of cottonseed cake and sunflower meal in the diet increased milk yield, and fat-corrected milk yield; while the use of castor bean
cake reduced the intake, digestibility of dry matter and total digestible nutrients, milk yield, and fat-corrected milk yield. The
inclusion of byproducts from the biodiesel industry in the diets did not change the fat, lactose, total solids, and solids-not-fat of
milk. Therefore, the cottonseed cake and sunflower meal can be included at up to 20% of the total diet.

Keywords Alternative feed . Feeding behavior . Milk yield . Protein sources

Introduction et al. 2008). However, environmental laws have become in-

creasingly stringent with the processing of plant materials and
Biodiesel production from vegetable oils is a sustainable way their waste (Oliveira et al. 2013; Moreira et al. 2014).
to meet worldwide energy demands since these oils are de- In tropical regions, it is necessary to use nutritional
rived from renewable sources (Ribeiro et al. 2018). The sources to balance the ration, due to the low nutritional
Brazilian climate is favorable to a wide diversity of raw ma- quality, mainly in the protein content. The search for
terials used for production, e.g., soybean, cottonseed, palm, alternative feeds to replace traditional ingredients while
sunflower, forage turnip, canola, castor, and jatropha (Abdalla maintaining the nutritional value of the diet has become
an important strategy (Silva et al. 2016). Protein sources
derived from biodiesel production emerge as a promis-
* Marcus Vinícius Gonçalves Lima ing option given the large volume generated and the
marcusviniciusagro@hotmail.com
versatility of their use.
In 2016, cotton, sunflower, and castor production in Brazil
1
Mato Grosso State University, Tangará da Serra, MT 78300-000, exceeded 4300, 246, and 60,000 tons, with average yields of
Brazil 1900, 1721, and 1000 kg ha–1, respectively (MAPA. 2017).
2
Southwest Bahia State University, Itapetinga, BA 45700-000, Brazil The respective meal and/or cake yields were approximately
3
São Paulo State University, Botucatu, SP 18618-686, Brazil 1368, 990, and 550 kg ha–1, and these protein sources have
4
Bahia Federal University, Salvador, BA 40110-903, Brazil been used as feedstuffs in diets for lactating cows (Pereira
et al. 2010; Broderick et al. 2013; Brito et al. 2015).
 169 Page 2 of 11 Trop Anim Health Prod (2021) 53:169

The inclusion of byproducts in diets for beef and dairy with 1% of urea:ammonium sulfate (9:1) on a fresh-matter
animals is a favorable option to reduce this generated “waste;” basis. The following diets (Table 1) were tested: corn- and
another advantage of this replacement is that they can be used soybean meal-based standard concentrate; inclusion of 20%
as energy and protein sources in the diet, lowering feed costs cottonseed cake; inclusion of 20% sunflower meal; and inclu-
without changing the level of milk and meat production sion of 20% castor bean cake, included on a dry matter basis
(Pedroso et al. 2009; Oliveira et al. 2012; Arruda et al. of the total diet.
2013). The satisfactory use of oilseeds as a biodiesel feedstock Feed was provided ad libitum twice daily and adjusted to
depends on their use as animal feed, which is economically allow refusals around 5–10% of the total supplied. Water was
competitive when the generated byproducts are successfully permanently available to the animals. Throughout the experi-
marketed in the animal feeding industry. ment, the feed supply was recorded daily. Cows were weighed
The availability of a large amount of byproducts and also at the beginning and end of the experimental period to esti-
the possibility of supplying different types of alternative pro- mate the nutrient intake as a percentage of body weight.
tein sources from industrial biodiesel production and the pos- During the collection period, from the 16th to the 19th
sible impacts on the feeding behavior of cows and the produc- experimental day, samples of roughage, concentrate, and re-
tion and composition of milk, to reduce feed costs, research fusals from each animal were collected daily, stored in plastic
must be carried out to seek the best level of inclusion, and also bags, and frozen at − 20 °C.
to enable the use of these byproducts with the most diverse
types of roughage, such as sugar cane, which is important in Castor bean cake detoxification and ricin
bulk. The study of these alternative sources has become of determination
fundamental importance to determine the intake and digest-
ibility of these feedstuffs. The castor bean cake detoxification was performed using 60 g
The hypothesis tested in this study is that byproducts de- of Ca(OH)2 kg−1 (Anandan et al. 2005). The determination of
rived from biodiesel production when included in 20% of the the ricin content in detoxified castor bean cake was conducted
total diet for lactating crossbred Holstein-Zebu cows, do not using a 0.5 M Tris-HCl buffer, pH 3.8 (Oliveira et al. 2010).
alter the intake or performance of these animals. In this con- The identification of the ricin fractions (A—approximately
text, the present study aimed to evaluate the inclusion of 36 kDa and B—approximately 29 kDa) was made by gel
byproducts derived from biodiesel production (cottonseed electrophoresis using a solution based on Coomassie
cake, sunflower meal, and castor bean cake) on the intake, Brilliant Blue G and molecular weight markers between 14
apparent digestibility, milk yield and composition, and feed- and 66 kDa (Sigma, Saint Louis, MO, USA) (Novaes et al.
ing behavior of lactating cows. 2020). The castor bean cake was considered detoxified with
the complete disappearance of the two ricin fractions (Fig. 1).

Material and methods Chemical analysis

Animals and diets Samples of diets, sugarcane, corn, soybean meal, cottonseed
cake, sunflower meal, and castor bean cake were pre-dried in a
The trial was conducted at the Laboratory of Forage and forced-air oven at 55 °C for 72 h to determine the nutritional
Pasture at the State University of Southwest of Bahia, content (Tables 2 and 3). The materials were then ground in a
Itapetinga Campus (BA, Brazil). Eight crossbred Holstein- Wiley mill with a 1.0-mm sieve for later analysis of dry matter
Zebu lactating cows in the third and fourth lactations, with (DM), crude protein (CP), ash, ether extract (EE), neutral de-
an average body weight of 525 ± 18.5 kg and an average milk tergent fiber (NDF), and acid detergent fiber (ADF) according
yield of 8 ± 1.45 kg day–1, were used in a double-Latin square to methodologies described by Detmann et al. (2012). Neutral
design (4 × 4). Each cow was weighed at the end of each detergent fiber corrected for ash and nitrogen compounds
experimental period. The trial consisted of four 19-day (NDFap) was determined according to Licitra et al. (1996).
experimental periods, represented by 15 days for adap- Total carbohydrates (TC) were calculated using the following
tation to the diets and 4 days for data collection. Cows equation: TC = 100 − (% CP + % ASH + % EE), according to
were housed in individual covered stalls with concrete Sniffen et al. (1992).
floor, equipped with individual concrete troughs for The contents of non-fiber carbohydrates corrected for ash
feeding and automatic drinkers. and protein (NFCap) were calculated by the following equa-
Animals were fed diets containing 60% sugarcane and 40% tion: NFCap = (100 − %NDFap − %CP – %EE − %ash), as
concentrate. Diets were formulated to provide an average dai- proposed by Hall (2003). Total digestible nutrients (TDN)
ly weight gain of 0.2 kg day–1 and average daily milk produc- were calculated using NDF and NFC corrected for ash and
tion of 12 kg day–1 (NRC 2001). Sugarcane was corrected protein: TDN (%) = CPD + NDFapD + NFCapD + (2.25 X
Trop Anim Health Prod (2021) 53:169 Page 3 of 11 169

Table 1 Percent composition of

diet ingredients, % in DM Item Standard concentrate Cottonseed cake Sunflower meal Castor bean cake

Sugarcane 60.0 60.0 60.0 60.0

Ground corn 22.1 12.4 14.0 16.1
Soybean meal 15.3 5.10 3.70 1.00
Cottonseed cake - 20.0 - -
Sunflower meal - - 20.0 -
Castor bean cake - - - 20.0
Mineral supplement1 2.66 2.52 2.35 2.90
100.00 100.00 100.00 100.00

Standard concentrate: corn- and soybean meal-based; Cottonseed cake: 50% inclusion in the concentrate (20% in
the total diet); Sunflower meal: 50% inclusion in the concentrate (20% in the total diet); Castor bean cake: 50%
inclusion in the concentrate (20% in the total diet)
1
Provides per kg of product: calcium—187 g, phosphorus—85 g, sulfur—18 g, magnesium—15 g, sodium—90
g, copper—1350 mg, cobalt—80 mg, iron—1450 mg, iodine —90 mg, manganese—1700 mg, selenium—22
mg, zinc—5800 mg, fluorine (max) —850 mg

EED). The estimated contents of TDN were calculated using Additionally, DM and NDFap were also expressed as % body
equations described by NRC (2001). weight (BW) and DM as metabolic weight (in g kg–0.75).
The intakes of DM, CP, EE, NDFap, indigestible NDF
(iNDF), NFCap, and TDN were determined in kg day–1. Apparent nutrient digestibility

To estimate the apparent digestibility of nutrients, feces sam-

ples were collected during the last 4 days of the experimental
period in alternating shifts (alternating morning or afternoon).
The feces were collected directly from the rectum. For the
estimation of fecal output, iNDF was used as an internal mark-
er. The iNDF content of the feedstuffs was determined by in
situ incubation for 240 h (Casali et al. 2008) followed by
analysis of NDF. The fecal excretion values were obtained
as the ratio of intake to the fecal concentration of iNDF. The
fecal dry matter (FDM) excretion was estimated according to
the equation: FDM (g/day) = [quantity of the indicator pro-
vided (g)/concentration of the indicator at feces (%)] × 100.

Milk yield and composition

Cows were mechanically milked once daily, at 06 h00, and

their milk yield (MY) was determined considering the results
obtained on four consecutive milking sessions, between the
16th and 19th days of each experimental period.
Milk yield was corrected to 3.5% fat (FCMY) by the fol-
lowing equation: FCMY = (0.432 + 0.1625 × F) × kg of milk,
where F = % milk fat (Sklan et al. 1992). Milk samples col-
lected on the 15th day were refrigerated in a plastic container
at 4 °C with bronopol (2-bromo-2-nitropropane 1,3-diol), at
the ratio of 10 mg bronopol to 50 mL milk, and immediately
sent to the Milk Clinic at ESALQ-USP, for analyses of
protein, fat, lactose, total solids (TS), solids-not-fat
(SNF), urea nitrogen (UN), casein (CAS), and percent-
Fig. 1 Polyacrylamide gel electrophoresis of castor bean cake treated or age of casein in protein (PCAS). The near-infrared ra-
untreated with Ca(OH)2 diation method was applied for these analyses, using a
 169 Page 4 of 11 Trop Anim Health Prod (2021) 53:169

Table 2 Dry matter content and

chemical composition of Item Sugarcane Ground Soybean Cottonseed Sunflower Castor bean
sugarcane, corn, soybean meal, (corrected) corn meal cake meal cake
cottonseed cake, sunflower meal,
and castor bean cake DM, % 32.2 86.1 87.8 93.1 90.3 90.5
CP1 9.61 8.91 43.7 36.5 33.3 34.6
EE1 1.51 5.81 1.31 16.3 1.72 7.42
NDFap1 52.2 11.7 8.51 20.8 29.9 36.1
iNDF1 28.5 2.94 2.12 12.9 28.5 34.2
ADF1 37.6 2.15 5.32 25.3 29.3 25.3
TC1 88.6 83.7 48.6 41.7 59.1 46.0
NFCap1 37.1 71.9 40.0 20.8 29.1 9.95
Cellulose1 29.1 2.11 4.92 19.0 21.8 13.6
Hemicellulose1 18.8 11.6 23.8 10.0 15.8 14.9
Lignin1 8.52 0.25 0.30 6.35 7.52 14.5
Ash1 3.01 1.62 6.40 5.52 5.72 11.9
NDIP1 2.21 1.41 16.7 11.4 12.8 7.81
TDN1,2 48.0 87.5 80.3 73.7 54.9 37.8
1
Values as a percentage of DM
2
Estimated according to the NRC (2001)
DM, dry matter; CP, crude protein; EE, ether extract; NDFap, neutral detergent fiber corrected for ash and protein;
iNDF, indigestible neutral detergent fiber; ADF, acid detergent fiber; TC, total carbohydrates; NFCap, non-
fibrous carbohydrates corrected for ash and protein; NDIP, neutral detergent insoluble protein; TDN, total digest-
ible nutrients

Bentley 2002 apparatus. Somatic cell count (SCC) was Instruments Incorporated, Chaska, MN, USA), by flow
determined using a Somacount 500 device (Bentley cytometry (IDF/FIL 148A).

Table 3 Dry matter content and

chemical composition of the total Item Standard concentrate Cotton Sunflower meal Castor bean cake
diets cake

DM, % 54.7 55.4 55.0 54.5

CP1 13.8 14.6 13.4 14.0
EE1 4.21 4.22 4.42 4.71
NDFap1 33.8 36.1 38.1 36.4
iNDF1 18.9 21.6 23.6 27.5
ADF1 24.4 28.1 29.8 30.0
TC1 76.9 75.8 77.2 75.6
NFCap1 43.1 39.8 39.1 39.3
Cellulose1 19.0 21.6 21.9 21.5
Hemicellulose1 16.9 16.5 17.5 14.6
Lignin1 5.42 6.52 7.91 9.52
Ash1 6.12 5.81 5.91 6.12
NDIP1 2.95 3.92 4.41 3.62
TDN1,2 67.3 64.6 62.4 62.0
1
Values as a percentage of DM
2
Estimated according to NRC (2001)
DM dry matter; CP crude protein; EE ether extract; NDFap neutral detergent fiber corrected for ash and protein;
iNDF, indigestible neutral detergent fiber; ADF, acid detergent fiber; TC, total carbohydrates; NFCap, non-
fibrous carbohydrates corrected for ash and protein; NDIP, neutral detergent insoluble protein; TDN, total digest-
ible nutrients
Trop Anim Health Prod (2021) 53:169 Page 5 of 11 169

Feeding behavior Results

On the 18th day of each experimental period, cows were vi- Cows fed diets containing standard concentrate, cottonseed
sually observed by trained people to evaluate their feeding cake, sunflower meal, and castor bean cake had average daily
behavior. Each animal was observed at 10-min intervals, for weight gains of 0.313, 0.263, 0.165, and 0.069 kg, respective-
24 h, to determine the times spent on the feeding, ruminating, ly, averaging 0.202 kg, which is close to the initially recom-
and idle activities. On the following day, the number of rumi- mended value of 0.200 kg. However, it should be emphasized
nation chews was counted, and the time spent ruminating each that the use of sunflower meal and castor bean cake provided
cud was obtained per animal, using a digital timer. gains of 17.5 and 65.5% below the recommended (0.200 kg),
The time spent and number of chews per ruminated cud respectively, reducing the final weight of the animals.
were obtained from the observations made during the rumina- DM intake was higher (P < 0.05) in cows fed diets con-
tion of three cuds, in three different periods of the day (10 to taining standard concentrate (17.3 kg day−1), cottonseed cake
12 h00, 14 to 16 h00, and 19 to 21 h00). During the nocturnal (18.2 kg day−1), or sunflower meal (16.9 kg day−1). DM in-
observation of the animals, the environment was maintained take decreased by 16.0% (14.7 kg day−1) when castor bean
under artificial lighting. cake was included in the diet (Table 4).
CP intake was higher (P < 0.05) in lactating cows fed diets
Statistical analysis containing cottonseed cake (2.5 kg day−1), this intake was
12% higher than the intake of cows fed with a standard con-
Data were tested by analysis of variance according to the centrate (2.2 kg day−1) and 28% higher when castor bean cake
following mathematical model, using the SAS 8.0 software (1.8 kg day−1) was provided in the diet.
(SAS. 1991): Cows fed cottonseed cake (6.5 kg day−1) and sunflower
meal (6.3 kg day−1) showed higher NDFap intake (P <
Y ijkl ¼ μ þ C i ðlÞ þ P j ðlÞ þ T k þ LS þ TSkl þ εij l; 0.05) compared to NDFap intake when fed with castor bean
cake (5.3 kg day−1). The average reduction in the intake of
where Yijkl = observation of cow i in period j subjected to NDFap when the cows fed castor bean cake was 17.2%.
treatment k in Latin square l; μ = overall effect of the mean; Ci The intake of NFCap was higher (P < 0.05) when the
(1) = effect of cow i within Latin square l (i = 1,2,3,4); Pj(l) =
lactating cows were fed diets containing standard concentrate
effect of period j within Latin square l (j = 1, 2, 3,4); Tk = effect (7.6 kg day−1) and cottonseed cake (7.5 kg day−1); this aver-
of treatment k (k = 1,2,3,4); LS = effect of Latin square l (l = age intake was 21.8% higher compared to cows fed castor
1,2); TSkl = effect of the interaction between treatment k and
bean cake (5.9 kg day−1) in the diet.
Latin square l; and εijkl = random error associated with each The average intake of TDN decreased by 23.8% when lactating
observation ijkl, eijkl~NID(0, ⌠2). Means were compared by cows fed diets containing castor bean cake (9.6 kg day−1)
Tukey’s test at 5% probability (SAS. 1991).

Table 4 Intake of nutritional

components by lactating dairy Variable Experimental diet
cows fed diets containing
byproducts derived from Standard concentrate Cotton cake Sunflower meal Castor bean cake P value
biodiesel production
Intake, kg day–1
DM 17.3a ± 2.7 18.3a ± 2.2 16.9a ± 1.9 14.7b ± 2.6 0.001
CP 2.2b ± 0.4 2.5a ± 0.3 2.1ab ± 0.5 1.8c ± 0.3 0.000
EE 0.7 ± 0.1 0.7 ± 0.1 0.7 ± 0.1 0.6 ± 0.1 0.439
NDFap 5.9ab ± 1.1 6.5a ± 0.7 6.3a ± 0.6 5.3b ± 1.0 0.002
iNDF 9.8 ± 2.3 10.7 ± 1.5 10.2 ± 1.5 9.8 ± 2.0 0.168
NFCap 7.6a ± 1.1 7.5a ± 1.1 6.8ab ± 3.9 5.9b ± 3.9 0.001
TDN 12.9a ± 2.0 13.0a ± 1.5 12.0a ± 1.7 9.6b ± 1.8 0.000
Intake, % BW
DM 3.2a ± 0.4 3.3a ± 0.3 3.1a ± 0.4 2.7b ± 0.3 0.001
NDFap 1.1ab ± 0.2 1.2a ± 0.1 1.2a ± 0.1 1.0b ± 0.1 0.001
Intake, g kg0.75
DM 154a ± 19.5 161a ± 15.4 151a ± 16.5 132b ± 17.0 0.001

Means followed by the same letter in the row do not differ by the Tukey test (5% probability)
 169 Page 6 of 11 Trop Anim Health Prod (2021) 53:169

compared to cows fed standard concentrate (12.9 kg day−1), cot- Table 6 lists the summary of the analysis of data on milk
tonseed cake (13.0 kg day−1), and sunflower meal (12.0 kg day−1). yield, milk yield corrected for 3.5% fat, and the milk compo-
The intake of DM and NDFap in % BW and the intake of sition. Use of standard concentrate and cottonseed cake in the
DM in g kg0.75 presented similar patterns (P < 0.05); these diet for lactating cows increased (P < 0.05) milk yield (10.2
were higher when cows fed diets containing standard concen- and 10.6 kg day−1, respectively), while milk yield corrected
trate (3.2 and 1.1% BW and 154.1 g kg0.75, respectively), for 3.5% fat was increased (12.0 kg day−1) when the diet was
cottonseed cake (3.3 and 1.2% BW and 161.1 g kg0.75, respec- included with cottonseed cake. Use of castor bean cake in the
tively) and sunflower meal (3.1 and 1.2% BW and 150.8 g lactating cow concentrate reduced milk yield by 23.6% and
kg0.75, respectively). When supplying castor bean cake, the milk yield corrected for 3.5% fat by 36.7% when compared to
intake of DM and NDFap in % BW and the intake of DM in a cottonseed cake.
g kg0.75 reduced (2.7 and 1.0% BW and 131.7 g kg0.75, re- The contents of milk components were altered when using
spectively), on average 15.6, 14.2, and 15, 2% respectively. the byproducts from biodiesel industry in the diet for lactating
The use of byproducts from the biodiesel industry in the cows (Table 6). An increase (P < 0.05) was observed in the
diet for lactating cows did not change the intake of EE (P = protein contents for the standard concentrate in relation to
0.439), and NDFi (P = 0.318) when compared to the standard castor bean cake; however, the milk protein contents were
concentrate (Table 4). similar between the cottonseed cake and sunflower meal diets
Table 5 lists the DMD, CPD, EED, NDFapD, NFCD, and and the standard concentrate. Milk protein content decreased
TDN of diets of lactating cows fed byproducts derived from by 8.2% when lactating cows received castor bean cake com-
the biodiesel production. DMD was higher (P < 0.05) in diets pared to the standard concentrate.
containing standard concentrate (75.7%). This reduction may The contents of milk urea nitrogen were higher (P < 0.05)
have been caused by the high content of NDFap, ADF, NDFi, when the cows were fed with a diet containing cottonseed
and lignin observed in diets containing cottonseed cake, sun- cake (15.4 mg dL−1) compared to those fed standard concen-
flower meal, and castor bean cake (Table 3). trate (13.3 mg dL−1); this corresponds to an increase in milk
The CPD was higher (P < 0.05) in diets containing cotton- urea nitrogen by 13.6%. Values of milk urea nitrogen were
seed cake (79.8%). Diets containing cottonseed cake were similar when the cows fed cottonseed cake, sunflower meal,
superior by 11.0% to diets containing castor bean cake and castor bean cake.
(71.0%); however, the NDFapD of diets containing cotton- The contents of casein and percentage of casein in milk
seed cake (49.6%) was lower (P < 0.05) than those containing protein were higher (P < 0.05) when the cows were fed stan-
standard concentrate (59.0%), but higher than those contain- dard concentrate (29.3 and 79.7%) compared to those fed
ing castor bean cake (39.9%). The TDN of diets containing cottonseed cake and castor bean cake. Diets containing cot-
standard concentrate was 5.0% higher (P < 0.05) compared to tonseed cake and castor bean cake reduced these vari-
the TDN of diets containing sunflower meal and 13% higher ables when compared to the standard concentrate.
than the TDN of diets containing castor bean cake. Casein contents and percentage of casein in milk protein
The inclusion of cottonseed cake, sunflower meal, were similar when the cows received standard concen-
and castor bean cake from the biodiesel industry in trate and sunflower meal (Table 6).
the diet for lactating cows did not change the EED (P Inclusion of cottonseed cake, sunflower meal, and castor
= 0.289) and NFCapD (P = 0.918) when compared to bean cake from the biodiesel industry in the diet for lactating
the standard concentrate (Table 5). cows did not alter the contents of fat (P = 0.123), lactose (P =

Table 5 Apparent digestibility of

dry matter (DMD), crude protein Variable Experimental diet
(CPD), ether extract (EED),
neutral detergent fiber corrected Standard concentrate Cotton Sunflower meal Castor bean P value
for ash and protein (NDFapD), cake cake
and non-fibrous carbohydrates
(NFCD), and total digestible nu- DMD 75.7a ± 3.9 70.9b ± 1.7 69.3b ± 4.9 63.2c ± 4.5 0.000
trients (TDN) in lactating dairy
CPD 75.9ab ± 4.4 79.8a ± 4.9 73.5ab ± 7.1 71.0b ± 3.3 0.009
cows fed diets containing
byproducts derived from biodie- EED 63.5 ± 9.0 61.8 ± 10.4 61.4 ± 8.9 54.0 ± 15.7 0.289
sel production NDFapD 59.0a ± 7.2 49.6b ± 3.7 52.5ab ± 4.4 39.9c ± 5.1 0.000
NFCD 90.4 ± 5.9 91.1 ± 2.1 90.4 ± 3.9 89.8 ± 3.9 0.918
TDN 74.6a ± 4.3 71.3ab ± 2.5 70.7b ± 3.9 64.9c ± 3.3 0.000

Means followed by the same letter in the row do not differ by the Tukey test (5% probability)
Trop Anim Health Prod (2021) 53:169 Page 7 of 11 169

Table 6 Milk yield (MY), 3.5%

fat-corrected MY (FCMY), and Variable Experimental diet
milk composition of lactating
dairy cows fed diets containing Standard concentrate Cotton Sunflower meal Castor bean P value
byproducts derived from biodie- cake cake
sel production
MY, kg day–1 10.2a ± 2.3 10.6a ± 2.5 9.6ab ± 1.7 8.1b ± 2.7 0.002
FCMY, kg day–1 9.9ab ± 3.1 12.0a ± 3.8 10.1ab ± 2.3 7.6b ± 2.3 0.004
Protein, g kg–1 36.7a ± 3.7 34.6ab ± 4.0 35.4ab ± 3.6 33.6b ± 3.6 0.004
Fat, g kg–1 32.6 ± 11.5 43.1 ± 14.4 37.8 ± 9.8 32.8 ± 11.9 0.123
Lactose, g kg–1 43.5 ± 1.7 43.5 ± 2.2 43.9 ± 1.8 43.6 ± 2.1 0.917
TS, g kg–1 122.6 ± 15.2 131.1 ± 14.2 127.1 ± 10.8 120.0 ± 14.5 0.130
SNF, g kg–1 90.0 ± 3.9 88.0 ± 4.5 89.3 ± 3.7 87.2 ± 4.0 0.071
UN, mg dL–1 13.3b ± 2.5 15.4a ± 3.3 13.6ab ± 2.1 13.4ab ± 2.1 0.027
CAS (%) 29.3a ± 3.5 27.2b ± 4.0 28.2ab ± 3.5 26.4b ± 3.6 0.000
PCAS (%) 79.7a ± 1.6 78.4b ± 2.7 79.3ab ± 2.1 78.4b ± 2.3 0.010
SCC, × 1000 mM–1 75.6 ± 46.1 91.0 ± 86.9 107.0 ± 82.9 82.7 ± 48.1 0.675

TS, total solids; SNF, solids-not-fat; UN, urea nitrogen; CAS, casein; PCAS, percentage of casein in protein; SCC,
somatic cell count
Means followed by the same letter in the row do not differ the Tukey test (5% probability)

0.917), total solids (P = 0.130), defatted dry extract (P = feeding (P = 0.437), by lactating cows. Cows maintained a
0.071), and somatic cell count (P = 0.675) when compared longer time in idleness when they received diets containing
to the standard concentrate (Table 6). The contents of fat standard concentrate; however, the idleness time was similar
(36.6 g kg−1), lactose (43.6 g kg−1), total solids (125.2 g between the diets with sunflower meal, cottonseed cake, and
kg−1), defatted dry extract (88.6 g kg−1), and the somatic cell the standard concentrate.
count (77.9 × 1,000 mM−1); these results are close to the The number of chews per day was higher (P < 0.05) when
range, or above the values recommended by the Regulation the cows fed diets containing cottonseed cake (34283.8 times
of Industrial and Sanitary Inspection of Animal Products. This a day). No difference was detected for the number of chews
suggests that the milk from crossbred Holstein-Zebu cows fed per day (P = 0.239), hours spent in chewing per day (P =
diets containing byproducts from biodiesel production has the 0.061), number of chews per cud (P = 0.355), and time in
same characteristics as the milk of crossbred cows fed stan- seconds per chewed cud (P = 0.445). The times spent in feed-
dard concentrate, being considered a normal milk under the ing, rumination, and idleness are consistent with the pattern of
legislation, and can be used by the dairy industries. feeding behavior of confined ruminants (Table 7).
The intake of DM and NDFap in 24 h was higher (P <
0.05) when the cows fed diets containing cottonseed cake
(17.87 and 6.36 kg day−1, respectively) or sunflower meal Discussion
(17.41 and 6.45 kg day−1, respectively). When these animals
consumed castor bean cake, the intake of DM decreased by NRC (2001) recommends a CP intake of between 1.1 and
16.2% (14.69 kg day−1), and the intake of NDFap was reduced 1.3 kg day−1 and TDN of 6.6 and 7.2 kg day−1 for cows with
by 16.5% (5.35 kg day−1); however, the intake of NDFap was a body weight of 550 kg, producing between 8 and 10 kg of
similar between the diets containing cottonseed cake, sun- milk (corrected). In this trial, the CP intake varied between 1.8
flower mean, and standard concentrate. and 2.5 kg day−1 and TDN from 9.6 to 13.0 kg day−1. The
The longest time spent in rumination (average of 9.5 h) protein and TDN in this trial met the requirements suggested
occurred when these animals fed diets containing 20% cotton- by the NRC (2001) with results above the upper limits.
seed cake, consequently, less time spent in idleness was ob- DMI is a determining factor in animal performance, as it is
tained (average of 8.0 h). When cows fed standard concentrate the starting point for nutrient intake, particularly energy and
or sunflower meal, these animals had shorter rumination protein (Oliveira et al. 2011). Brito et al. (2015) did not ob-
times, averaging 8.0 and 8.2 h, respectively. The rumination serve any differences in DMI using sunflower meal, flaxseed,
time of cows fed cottonseed cake or castor bean cake was and soybean meal as protein sources in diets for cows with a
similar (Table 7). There was no difference between the milk yield of 15 kg; however, this intake was lower (15.0 kg
byproducts tested in relation to the time spent (h day−1) on day−1) than that found in the present trial (16.8 kg day−1). Feed
 169 Page 8 of 11 Trop Anim Health Prod (2021) 53:169

Table 7 Feeding behavior of lactating cows fed diets containing byproducts derived from biodiesel production

Variable Experimental diet

Standard concentrate Cotton Sunflower meal Castor bean P value

cake cake

Intake in 24 h, kg day–1
DM 17.3a ± 2.8 18.1a ± 2.2 17.4a ± 1.8 14.7b ± 2.4 0.001
NDFap 5.9ab ± 1.2 6.4a ± 0.7 6.4a ± 0.7 5.3b ± 0.9 0.001
Activity, h day–1
Feeding 6.5 ± 1.2 6.5 ± 1.0 6.5 ± 0.5 7.0 ± 1.2 0.437
Rumination 8.0b ± 1.4 9.5a ± 0.7 8.2b ± 0.7 8.5ab ± 1.0 0.004
Idleness 9.5a ± 1.3 8.0b ± 1.2 9.3ab ± 1.0 8.5ab ± 1.3 0.003
Chewing
Cuds, n day–1 517.7 ± 112.0 599.8 ± 126.1 560.2 ± 92.5 532.7 ± 132.5 0.239
n day–1 28488.7b ± 6408.0 34283.8a ± 4170.1 28843.9b ± 2991.4 29233.1b ± 3655.6 0.003
h day–1 14.5 ± 1.3 15.9 ± 1.2 14.8 ± 1.0 15.5 ± 1.3 0.061
n cud–1 55.7 ± 8.4 59.4 ± 14.4 52.9 ± 11.6 56.7 ± 9.3 0.355
s cud–1 56.8 ± 9.3 58.8 ± 11.9 54.2 ± 9.7 59.1 ± 8.8 0.445

DM, dry matter; NDFap, neutral detergent fiber corrected for ash and protein; Cuds (n day–1 ), number of cuds chewed per day; n day–1 , number of
chews per day; h day–1 , hours expended on chewing per day; n cud–1 , number of chews per cud; s cud–1 , time in seconds per chewed cud
Means followed by the same letter in the row do not differ by the Tukey test (5% probability)

intake predictions are essential to optimize the use of dietary Digestibility is closely related to the NDF content and its
nutrients, which can increase the productive, economic, and consumption (Gonzaga Neto et al. 2015). The higher protein
environmental performance of dairy systems. However, these content, improve the NDF viability by microorganisms in the
predictions are restricted to specific geographic and environ- rumen and, consequently, improve the digestibility (Polizel
mental conditions (Souza et al. 2014). Neto et al. 2014). In contrast, a reduction in feed digestibility
The NDFap intake (1.2% BW) for diets containing cotton- is observed in some situations when there is a higher fat content
seed cake and sunflower meal was similar to the value (1.2% in the diet, which may compromise the energy value resulting
BW) reported by Mertens (1992) as the intake-limiting level. in a decreased total tract digestibility (Oliveira et al. 2013).
However, several studies on dairy cows in tropical countries The decrease in CP digestibility of castor bean cake was
have already shown that animals with lower milk yield, com- probably due to a decrease in CP intake. Reduction in the
pared to purebred animals from temperate climates, have a digestibility of NDFap occurred due to the enlargement of
greater fiber-intake capacity without having their milk yield the lignin content when castor bean cake is included
decreased, as observed by Cavalcanti et al. (2008). (Oliveira et al. 2010).
The decrease in DMI with castor bean cake may be asso- Sunflower meal as a source of protein, increases production
ciated with higher lignin content. The increase in the indigest- and microbial efficiency, increasing the contribution of micro-
ible fraction (lignin) promoted the filling effect of the rumen; bial protein to the duodenum, in addition to increasing the feed
once the disappearance of this fraction is through the passage passage rate (Mendes et al. 2006). According to Hashimoto
rate, while the disappearance of the potentially digestible frac- et al. (2007), higher fiber content reduces the DM digestibility
tion of NDF (NDF-iNDF) is affected by the passage rate and due to a reduction in non-structural carbohydrates, which are
digestion (Allen and Mertens 1988; Oliveira et al. 2011). rapidly degraded in the rumen. However, an increase in fiber
The lower DM digestibility observed in castor bean intake tends to stimulate rumination and, consequently, sali-
cake diets is related to the higher content of NDF and vation, improving the ruminal environment, maintaining the
ADF castor bean cake, consequently also increasing the ruminal pH at adequate levels, bettering the development and
lignin concentration (24.5%) (Table 2). These results maintenance of the cellulolytic bacteria and increasing the
indicate that the DM digestibility was affected in ani- NDF degradation.
mals receiving this diet. The highest coefficients of DM Diets with alternative ingredients must be efficient, safe,
apparent digestibility were found in animals that re- economical, and allow similar productive performance that
ceived supplementation with standard concentrate. the animals fed traditional ingredients (Pina et al. 2006).
Trop Anim Health Prod (2021) 53:169 Page 9 of 11 169

This can be observed in this study when comparing milk yield (2009) observed this behavior and reported that the longest
and fat-corrected milk yield of cows fed cottonseed cake and rumination times occurred early and late at night, and in the
cotton meal with cows fed standard concentrate. coldest periods of the day.
Corroborating the results found in this study, Bernard According to Van Soest (1994), the time spent by an ani-
(1997) stated that the cottonseed cake is efficient to maintain mal in rumination activities is influenced by the nature of the
milk yield at the same levels as soybean meal when the CP diet, the physical form, and NDF content. The pattern of the
content of the diet is higher than 16%. search for feed by feedlot cattle is very characteristic, with two
The decrease in milk production when castor cake is in- main —early morning and late afternoon—, and the daily time
cluded in the diet can be explained by the lower carbohydrate spent in this activity (4.5 h) by lactating dairy cows
intake (NFC and NDF) that results in a reduction in the flow of (Damasceno et al. 1999) may vary from 1 (high-energy
glucose from the blood to the mammary gland. Furthermore, a feeds) to 6 h or more (low-energy feeds). The 6.6, 8.5, and
reduction in rumen microbial N synthesis might also have 8.8 h day–1 spent in feeding, ruminating, and idle times by the
contributed to a decrease in milk production, considering a cows in this trial are within the range mentioned.
decreased flow of amino acids to the mammary gland. In the process of balancing the feed for dairy cows, the cost
The milk protein content found in this study (33.6 to 36.6 g of ingredients should be observed, since these can be signifi-
kg−1) are close to those reported in the literature (Pedroso et al. cantly reduced by including byproducts. The feed cost pro-
2009; Pereira et al. 2011; Coutinho et al. 2014). Haugen et al. duction with cottonseed cake, sunflower meal, and castor bean
(2006) stated that the protein evaluation system for dairy cattle cake (57.55, 60.06, and 57.71%) is lower compared to diets
recognizes that there are differences between feedstuffs with formulated with commonly ingredients (72.02%); which
respect to the intestinal digestibility of proteins. shows the importance of evaluating and comparing by-
A reduction in the flow of glucose to the mammary gland products with other raw materials.
resulted from the decreasing carbohydrate intake might also Consumption and total cost of food were equal or higher
have contributed to the milk CP depression; since the secret- for diets with standard concentrate, cottonseed cake, and sun-
ing cells of the mammary gland mainly use glucose to provide flower meal and lower for the diet with castor bean cake (R$
energy for the amino acids capture, synthesis, and milk protein 10.25, 9.44, 9.06, and 7.31), demonstrating that these diets
transport (Fonseca 1995; Mackle et al. 2000). (standard concentrate, cottonseed cake, and sunflower meal)
UN concentrations in milk must remain between 12 and showed greater acceptability. The cost of the concentrate was
20 mg dL−1 (Guimarães Junior et al. 2007), and an increase in higher for the control diet (standard concentrate) and lower for
this concentration indicates an excess of protein, low quantity the diet containing castor bean cake.
or quality of carbohydrates, or an imbalance in the ratio A higher total cost of feeding was observed with the stan-
protein:energy in the diet. Therefore, it can be observed that dard diet, while the lowest total cost of feeding was observed
the diets did not influence the increase in the UN concentra- with the castor bean cake diet. The lowest cost of milk pro-
tion in milk above the value determined by Guimarães Junior duction was observed in cows fed with cottonseed cake (R$
et al. (2007), presenting a mean of 13.9 mg dL−1 UN. 0.89), and the highest, for the standard diet (R$ 1.01).
The main advantages of using alternative sources are asso- Consequently, the highest profit was observed with cotton-
ciated with cost reduction and higher availability of ingredi- seed cake diet, with a gross margin of R$ 2.75, higher than
ents to balance the diet, considering that these ingredients the gains obtained with standard, sunflower meal, and castor
contain desired nutrients and are easily processed (Santos bean cake (R$ 1.48, 1.98, and 2.00) diets.
et al. 2020). It is assumed that diets used did not differ in terms The use of cottonseed cake, sunflower meal, and castor
of ruminal production of volatile fatty acids, which resulted in bean cake decreased the total feed costs by 7.9, 11.6, and
a similar percentage of milk fat between treatments. 28.7% compared to the standard diet. A higher gross margin
The times observed for feeding, rumination, and idle activ- is highlighted for the cottonseed cake diet.
ities are consistent with the feeding behavior pattern of feedlot Our conclusion suggests that the inclusion of cotton-
ruminants. Our results—276 ± 55.8, 482.4 ± 43.8, and 606 ± seed cake and sunflower meal in the diet changes the
126 min day−1 (feeding, rumination, and idleness)—were sim- intake, digestibility of nutritional compounds, milk yield
ilar to those reported by researchers who worked with cattle in and composition, and feeding behavior of lactating
a feedlot system. (Oliveira et al. 2007; Correia et al. 2012). cows. Castor bean cake is not recommended for dairy
However, those values were higher than those reported by cows because it reduces the intake of DM, CP, NDFap,
Carlotto et al. (2010), where the mean rumination times NFCap, and TDN, as well as negatively affects the ap-
ranged from 1.5 to 2.1 h during the daytime period, with no parent digestibility of nutrient compounds, milk yield,
significant difference being observed between the diets tested. and composition, and feeding behavior. Therefore, cot-
It is supported that these values were low because most of the tonseed cake and sunflower meal can be included in up
rumination activity takes place during the night. Zanine et al. to 20% in the diet of lactating cows.
 169 Page 10 of 11 Trop Anim Health Prod (2021) 53:169

Acknowledgements The authors thank the National Council of Scientific from upland cottonseed, Pima cottonseed, or extruded Pima cotton-
and Technological Development (CNPq) and the State University of seed on production of lactating dairy cows. Journal of Dairy
Southwest of Bahia (UESB) for financial support. Science, 96, 2374–2386. doi:https://doi.org/10.3168/jds.2012-5723.
Carlotto, S.B., Medeiros, R.B., Pellegrini, C.B., Garcia, R.P.A., Lisboa,
C.A.V. and Saibro, J.C. 2010. Diurnal ingestive behaviour of preg-
Code availability Not applicable. nant heifers grazing on natural grasslands invaded by Eragrostis
plana Ness as affected by protein and mineral supplements in the
Authors’ contribution Marcus Vinícius Gonçalves Lima is the author different climatic seasons. Revista Brasileira de Zootecnia, 39, 454–
responsible for submission. 461. doi:https://doi.org/10.1590/S1516-35982010000300002.
Casali, A.O., Detmann, E., Valadares Filho, S.C., Pereira, J.C.,
Henriques, L.T., Freitas, S.G. and Paulino, M.F. 2008. Influence
Funding This research was funded by the National Council of Scientific of incubation time and particle size on indigestible compounds con-
and Technological Development (CNPq) and the State University of tents in cattle feeds and feces obtained by in situ procedures. Revista
Southwest of Bahia (UESB) awarded a scholarship to the first author. Brasileira de Zootecnia, 37, 335–342. doi:https://doi.org/10.1590/
S1516-35982008000200021.
Data availability The datasets generated during and/or analyzed during Cavalcanti, M.C.A., Batista, A.M.V., Guim, A., Lira, M.A., Ribeiro, V.L.
the current study are available from the corresponding author on reason- and Ribeiro-Neto, A.C., 2008. Intake and ingestive behavior of
able request. sheep and goats fed with cactus pear (Opuntia ficus-indica Mill)
and prickly pear (Opuntia sp.). Acta Science. Animal Science, 30,
Declarations 173–179. doi:https://doi.org/10.4025/actascianimsci.v30i2.4684.
Correia, B.R., Oliveira, R.L., Jaeger, S.M.P.L., A. R. Bagaldo, Carvalho,
G.G.P., Oliveira, G.J.C., Lima, F.H.S. and Oliveira, P.A. 2012.
Consent to participate Not applicable.
Ingestive behavior and physiological parameters of steers fed with
biodiesel cakes. Archivos de Zootecnia, 61, 79–89. doi:https://doi.
Consent for publication Not applicable. org/10.4321/S0004-05922012000100009.
Coutinho, D.A., Branco, A.F., Santos, G.T., Osmari, M.P., Teodoro, A.L.
Conflict of interest The authors declare no competing interests. and Diaz., T.G. 2014. Intake, digestibility of nutrients, milk produc-
tion and composition in dairy cows fed on diets containing cashew
Animal rights This study was conducted in accordance with the institu- nut shell liquid. Acta Science. Animal Science, 36, 311–316. doi:
tional committee on animal use (Protocol 64/2014). https://doi.org/10.4025/actascianimsci.v36i3.23512.
Damasceno, J.C., Bacari-Júnior, F. and Targa, L.A. 1999. Behavior re-
sponses of holstein dairy cows with constant or limited access to
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