Archivos Latinoamericanos de Producción Animal. 2025 (OctubreDiciembre). 33 (4)

Byproducts replacing ground corn for dairy cows rotationally grazing

elephant grass: milk yield and composition

Recibido: 20250723. Revisado: 20251014. Aceptado: 20251114

1Department of Animal Science, Mato Grosso State University, Pontes e Lacerda 78250000, MT, Brazil.

2Institute for Future Farming Systems, Central Queensland University, Rockhampton, QLD 4701, Australia

3Embrapa SemiArid Region, Brazilian Agricultural Research Corporation, Petrolina 56302970, PE, Brazil.

4Correspondence author: fapsantos@usp.br

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Junio Cesar Martinez 1

Abstract. This study investigated the effects of different byproducts used as feedstuffs to replace ground corn for

supplementing Holstein dairy cows rotationally grazing intensively managed elephant grass. Five experiments were

conducted to evaluate various levels of pelleted citrus pulp (PCP), corn gluten feed (CGF), whole cottonseed (WCS),

wheat middlings (WHM), and soybean hulls (SBH) as alternatives to ground corn in isoproteic and isoenergetic

supplements. The experiments followed a Latin square design and assessed blood and performance parameters such

as milk yield (MY) and milk composition. Blood parameters were not affected (p > 0.05) by any byproducts. The

inclusion of PCP, CGF and SBH up to 75 %, and WHM up to 50 % in place of corn did not affect

(p > 0.05) either MY or milk composition. However, replacing the inclusion of ground corn at the level of 75 % of WCS

resulted in a significant reduction (p < 0.05) in yields of milk, 3.5 % fatcorrected milk, protein, and milk solids, along

with an increase (p < 0.05) in milk urea nitrogen. Results obtained suggest that the replacement of ground corn by

PCP, CGF and SBH up to 75 % and WHM and WCS up to 50 % in the supplement have no adverse shortterm effects

on MY and composition in Holstein cows grazing intensively managed elephant grass.

Keywords: blood parameters, coproducts, forage quality, Holstein, Cenchrus purpureus, tropical pastures.

https://doi.org/10.53588/alpa.330401

Department of Animal Science, Luiz de Queiroz College of Agriculture,

University of São Paulo, Piracicaba 13418900, SP, Brazil

Sustitución del maíz molido por subproductos en vacas lecheras en pastoreo

rotacional de pasto elefante: producción y composición de la leche

Resumen. Este estudio investigó los efectos de reemplazar el maíz molido por diferentes subproductos utilizados

como suplementos para vacas lecheras Holstein en pasturas de pasto elefante manejadas intensivamente bajo

pastoreo rotacional. Se realizaron cinco experimentos para evaluar diferentes niveles de pulpa de cítricos

peletizada (PCP), gluten de maíz (CGF), semilla entera de algodón (WCS), afrecho de trigo (WHM) y cascarilla de

soya (SBH) como alternativas al maíz molido. Los experimentos siguieron un diseño de Cuadrado Latino y

evaluaron parámetros sanguíneos y productivos, tales como la producción y composición de la leche. Los

parámetros sanguíneos no se vieron afectados (p > 0,05) por ninguno de los subproductos. La inclusión de PCP,

CGF y SBH hasta un 75 % y de WHM hasta un 50 % en sustitución del maíz no afectó (p > 0,05) la producción ni

la composición de la leche. Sin embargo, la substitución del maíz molido por 75 % de WCS resultó en una

reducción significativa (p < 0,05) en la producción de leche, leche corregida al 3,5 % de grasa, proteína y sólidos

lácteos, y un incremento(p < 0,05) en el nitrógeno ureico de la leche. Los resultados obtenidos sugieren que la

sustitución del maíz como suplemento por PCP, CGF y SBH hasta un 75 %, y por WHM y WCS hasta un 50%, no

tiene efectos adversos a corto plazo sobre la producción y la composición de la leche de las vacas Holstein

pastoreando pasto elefante manejado intensivamente.

Palabras clave: parámetros sanguíneos, coproductos, calidad del forraje, Holstein, Cenchrus purpureus, pasturas tropicales.

Guilhermo Francklin de Souza Congio

Pedro Guerreiro2

Diogo Fleury Azevedo Costa

Tadeu Vinhas Voltolini3 Carla Maris Machado Bittar Flávio Augusto Portela Santos4

186

Introduction

The bulk of dry matter (DM) consumed by grazing

dairy cows comes from the grazed forage; however, a

significant portion of the energy utilized for

maintenance and lactation may derive from non

structural carbohydrates and lipids supplied by

supplements (Vazquez and Smith, 2000; NRC, 2001).

Corn is the main feed used as supplement for grazing

dairy cows in Brazil (Carmo et al., 2015) and

approximately 72 % of the corn DM is starch (NRC,

2001). However, depending on corn type and

processing methods, the starch content can be as high

or above 90 % DM (Ferraretto et al., 2013). Therefore,

the use of high levels of corn as supplement can cause

nutritional disorders, such as acidosis (Bramley et al.,

2008), if nutrients coming from grazed forages and

supplements are not well balanced.

More recently, the inclusion of humanedible grains

in ruminant diets has been intensely debated in the

light of food security concerns (Wilkinson and Lee,

2018). Contextually, feeding byproducts can supply

valuable nutrients to livestock and allow the use of

feedstuffs that would otherwise be treated as waste

and sent to landfills (Eastridge, 2006). Several by

products have then been tested as alternatives to

replace protein or energy sources in ruminant diets,

especially for dairy cows (Harvatine et al., 2002;

Kononoff et al., 2006; Bales et al., 2024). However, those

studies have been mostly developed for confined cows

fed total mixed rations, and there are considerably

fewer studies conducted under grazing conditions,

particularly in tropical climates, highlighting the need

for research that addresses existing gaps for grazing

dairy systems.

The food industry is wide with several options of

byproducts to feed ruminants becoming available,

especially in countries devoted to agricultural

production such as Brazil (Silva et al., 2019). Citrus

pulp is a byproduct of the citrus industry, and its

energy content varies from 85 to 90 % of that of corn

(NRC, 2001). It has been shown that citrus pulp

inclusions up to 10 % of diet in DMbasis can

successfully replace ground corn in dairy cows’ diets

improving milk yield (Hartinger et al., 2024). In the

context of tropical agriculture, the effects of increasing

levels of inclusion of citrus pulp replacing flint corn are

likely to be different). In beef cattle, citrus pulp has

been successfully used as a supplement to improve

growth performance of beef cattle (Costa et al., 2019).

Citrus pulp has high contents of soluble carbohydrates

and pectin and low of lignin, which makes its ruminal

fermentation distinct from that of corn. Its acetic

fermentation profile decreases risks of acidosis (García

Rodríguez et al., 2020). Soybean hulls (SBH), a

byproduct from the soybean processing industry, has

little value either as human food and for industrial uses

(Ipharraguerre and Clark, 2003). The nutrient

composition of SBH is variable, but the product is

known for having high pectin content and high

digestible fiber, but low starch content.

Substituição de milho moído por coprodutos para vacas leiteiras em pastejo

rotacionado de capimelefante: produção e composição do leite

Resumo. Este estudo investigou os efeitos de diferentes subprodutos usados como ração para substituir o milho

moído no suplemento de vacas leiteiras holandesas em pastagens de capimelefante manejado intensivamente sob

pastejo rotacionado. Cinco experimentos foram conduzidos para avaliar polpa cítrica peletizada (PCP), farelo de

glúten de milho (CGF), caroço de algodão inteiro (WCS), farelo de trigo (WHM) e casca de soja (SBH) como

alternativas ao milho moído em suplementos isoproteicos e isoenergéticos. Os experimentos seguiram um

delineamento de quadrado latino e avaliaram parâmetros sanguíneos e de desempenho, como produção e

composição do leite. Os parâmetros sanguíneos não foram afetados (p > 0,05) por nenhum subproduto. A inclusão

de PCP, CGF e SBH até 75 % e de WHM até 50 % em substituição ao milho, não afetou (p > 0,05) a produção nem a

composição do leite. Entretanto, a substituição de milho moído por 75 % de WCS resultou em redução significativa

(p < 0,05) na produção de leite, leite corrigido para 3,5% de gordura, proteína e sólidos totais, além de aumento

(p < 0,05) na ureia do leite. Os resultados sugerem que a substituição do milho por PCP, CGF e SBH até 75 % e por

WHM e WCS até 50 % não causa efeitos adversos de curto prazo sobre a produção e composição do leite de vacas

Holandesas em pastejo rotacionado de capimelefante manejado intensivamente.

Palavras chave: parâmetros sanguíneos, coprodutos, qualidade da forragem, Holstein, Cenchrus purpureus,

pastagens tropicais.

Martínez et al.

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187

Byproducts as corn replacements for grazing dairy cows

Materials and Methods

This study comprises a set of five supplementary

feeding experiments carried out from March 2002 to

June 2006 in the same area at the University of Sao

Paulo, Luiz de Queiroz College of Agriculture,

Piracicaba, SP, Brazil (22°42′S, 47°38′W, 546 m.a.s.l.).

The pastures were divided into 40 paddocks (2000 m2

each). The perennial elephant grass (Cenchrus purpureus

(Schumach.) Morrone syn. Pennisetum purpureum

Schumach.) cv. Cameroon and cv. Napier pastures

were established in early 1970, in a high fertility

Eutroferric Red Nitossol soil (NVef) (Batalha et al.,

2022b). According to the Köppen classification, the

climate of the experimental area is humid subtropical

with hot summers and dry winters (Alvares et al.,

2013). The rainy season runs from October to March

and the dry season from April to September. The

historical (19172023) average mean air temperature

and accumulated rainfall during both rainy and dry

seasons are 24.0 °C and 992 mm and 19.6 °C and 282

mm, respectively (Série de Dados Climatológicos do

Campus Luiz de Queiroz de Piracicaba, 2024).

Paddocks were rotationally grazed with fixed

stocking and rest periods. In Experiment 1, the

stocking and rest periods were 3 and 37 days,

respectively, whereas for the remaining experiments

herds were kept only one to two days grazing each

paddock and 23 days resting. Sward surface height

was monitored from ground level to the top leaf

horizon by 20 readings per paddock using an adapted

sward stick (Carnevalli et al., 2021). Ten hand plucked

samples simulating what the animals grazed were

taken per paddock at pregrazing during the grazing

cycles and mixed as a composite sample used to

determine forage nutrient composition (Batalha et al.,

2022a) (Table 1). Pastures were fertilized with 80 kg N

ha1 (as urea, 45 % of N) after each grazing event. In all

experiments replacing ground corn with PCP, CGF,

WCS, WHM, or SBH, supplements were isoproteic

and isoenergetic.

Whole cottonseed (WCS) is a common byproduct

from cotton production (Arieli, 1998). Despite having a

high energy content because of lipids in its

composition, WCS has been classified effective at

stimulating chewing during eating and rumination

because of the linter fraction (Harvatine et al., 2002).

Furthermore, the low density of the oil in WCS likely

assists with this ingredient floating in and above the

rumen mat, therefore touching the rumen walls and

stimulating motility. Similarly to WCS, corn gluten

feed (CGF) and wheat middlings (WHM) are by

products noted by their high energy and protein

contents (Armentano and Dentine, 1988; Duncan et al.,

2024). Yet, CGF contains low lignin content being a

source of highly digestible fiber with relatively low

concentration of rapidly fermentable carbohydrates,

decreasing the risk of ruminal acidosis (Kononoff et al.,

2006). Therefore, our objective was to assess milk yield

and composition of dairy cows rotationally grazing

elephant grass pastures and fed increasing levels of

pelletized citrus pulp (PCP), corn gluten feed (CGF),

whole cottonseed (WCS), wheat middlings (WHM),

and soybean hulls (SBH), replacing ground corn as

concentrate ingredients.

Table 1. Pre and postgrazing sward heights and nutrient composition of pregrazing hand pluck forage samples for all five experiments.

Item Exp. 1 Exp. 2 Exp. 3 Exp. 4 Exp. 5 Mean

Pregrazing sward height, cm 154.0 130.0 100.0 142.7 149.0 135.1

Postgrazing sward height, cm 129.5 66.0 60.0 72.0 67.3 79.0

Nutrient composition, %DM

Dry matter 20.3 18.3 18.6 18.5 18.2 18.8

Ash 10.5 10.6 10.9 12.7 11.3 11,2

Crude protein 13.7 18.5 14.7 17.6 17.1 16.3

Neutral detergent fiber 62.9 61.4 63.9 61.5 60.8 62.1

Acid detergent fiber 33.3 32.0 33.5 33.4 33.3 33.1

Ether extract 2.5 2.7 2.5 3.0 2.7 2.7

Lignin 3.3 2.8 3.2 3.2 3.3 3.2

In vitro DM digestibility 68.8 72.9 67.4 70.2 70.3 69.9

Supplements formulation

The supplements of all experiments were formulated

using the Nutrient Requirements of Dairy Cattle model

(NRC, 2001), considering a forage containing 18 % of

crude protein (DM basis) for dairy cows yielding 20 kg

d1 of milk, similar to other experiments carried out in

the same set of paddocks (Danes et al., 2013; Chagas et

ISSNL 10221301. Archivos Latinoamericanos de Producción Animal. 2025. 33 (4): 185  197

188

al., 2021). The formulation of the diets was determined

based on the total diet, which accounted for both the

estimated forage intake and the known daily

supplement consumption. The treatments'

nomenclature refers to the percentage that ground corn

was substituted by each byproduct in each experiment.

Experiment 1 – Citrus pulp

Between March and June 2002, 8 multiparous

Holstein dairy cows averaging (±SD) 523 ± 82 kg body

weight (BW), 82 ± 27 days in milk (DIM), and 20.0 ± 1.5

kg d1 milk yield (MY) at the beginning of the

experiment, were used in a duplicated (n = 2) 4 × 4

Latin square design. Cows were grouped into 2 squares

based on their DIM, MY and BW characteristics. The

experiment consisted of four 20day periods, with 15

days for adaptation and 5 for data collection. Cows

within each square were randomly assigned to the

treatments based on the inclusion levels of finely ground

and pelletized citrus pulp (PCP): 0, 25, 50 and 75 % of

PCP (PCP0, PCP25, PCP50 and PCP75, respectively).

The ingredients and nutrient composition of

experimental concentrates are presented in Table A1.

Cows were offered the supplements individually (6.3 kg

DM d1), in two equal portions after the morning and

before the afternoon milking.

Milk yield was recorded daily during the evaluation

period with samples collected in vials containing a

bronopol preservative pill. Blood samples were taken

from the coccygeal vessels of each cow four hours after

the morning concentrate feeding on the last day of the

sampling period and stored in vacutainers containing

potassium oxalate as an anticoagulant. The body

condition score (BCS) was assessed on the first and last

day of each period based on a 5point scale, as

proposed by Wildman et al. (1982).

Experiment 2 – Corn gluten feed

Sixteen multiparous Holstein dairy cows averaging

468 ± 52 kg BW, 141 ± 52 DIM, and 13.9 ± 2.2 kg d1 MY

at the beginning of the experiment, were used in a

quadruplicated (n = 4) 4 × 4 Latin square design with

four periods of 20 days each (15 + 5 days for adaptation

and evaluation, respectively) from November 2004 to

February 2005. Cows were grouped into 4 squares

based on their DIM, MY, and BW attributes. The

treatments consisted of increasing levels of dried corn

gluten feed (CGF) replacing ground corn in concentrate

supplements as follows: 0, 25, 50 and 75 % of CGF

(CGF0, CGF25, CGF50 and CGF75, respectively). The

ingredients and nutrient composition of experimental

concentrates are presented in Table A2. Each cow

received 4.9 kg DM d1 of the supplementary feed, and

the provision of those, as well as the measurement of

animal performance and metabolic parameters, was

similar to that in Experiment 1.

Experiment 3 – Whole cottonseed

Twelve multiparous Holstein dairy cows averaging

544 ± 73 kg BW, 143 ± 14 DIM, and 20.1 ± 2.5 kg d1 MY

at the beginning of the experiment, were used in a

triplicated (n = 3) 4 × 4 Latin square design with four

periods of 20 days each (15 + 5 days for adaptation and

evaluation, respectively) from March to June 2005.

Cows were grouped into 3 squares based on DIM, MY,

and BW. The treatments consisted of increasing levels

of whole cottonseed (WCS) replacing ground corn in

the concentrate supplement as follows: 0, 25, 50 and 75

% of WCS (WCS0, WCS25, WCS50 and WCS75,

respectively) to correspond to inclusion levels of

approximately 0, 7, 14 and 21 % of WCS in the total

diet. The ingredients and nutrient composition of the

experimental concentrates are shown in Table A3. Each

cow received 5.9 kg DM d1 of the supplementary

feeds, and the provision of those, as well as the

measurement of animal performance and metabolic

parameters was similar to that in Experiment 1.

Experiment 4 – Wheat middlings

Twelve multiparous Holstein dairy cows averaging

532 ± 34 kg BW, 84 ± 13 DIM, and 20.1 ± 1.8 kg d1 MY

at the beginning of the experiment, were used in a

triplicated (n = 3) 4 × 4 Latin square design with four

periods of 20 days each (15 + 5 days for adaptation and

evaluation, respectively) from November 2005 to

February 2006. Cows were grouped into 3 squares

based on DIM, MY, and BW. The treatments consisted

of increasing levels of wheat middlings (WHM)

replacing ground corn in the concentrate supplement

as follows: 0, 25, 50 and 75 % of WM (WHM0, WHM25,

WHM50 and WHM75, "respectively". Each cow

received 5.6 kg DM d1 of supplementary feeds, and the

provision of those, as well as the measurement of

animal performance and metabolic parameters was

similar to that in Experiment 1. The ingredients and

nutrient composition of experimental concentrates are

presented in Table A4.

Experiment 5 – Soybean hulls

Twelve multiparous Holstein dairy cows averaging

509 ± 57 kg BW, 91 ± 11 DIM, and 20.6 ± 1.4 kg d1 MY

at the beginning of the experiment, were used in a

triplicated (n = 3) 4 × 4 Latin square design with four

periods of 20 days each (15 + 5 days for adaptation and

Martínez et al.

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189

periods of 20 days each (15 + 5 days for adaptation and

evaluation, respectively) from March to June 2006.

Cows were grouped into 3 squares based on DIM, MY,

and BW. The treatments consisted of increasing levels

of soybean hulls (SBH) replacing ground corn in

concentrate supplements as follows: 0, 25, 50 and 75 %

of SBH (SBH0, SBH25, SBH50 and SBH75,

respectively). Each cow received 5.8 kg DM d1 of the

supplementary feed, and the provision of those, as well

as the measurement of animal performance and

metabolic parameters was similar that in Experiment 1.

Ingredients and nutrient composition of experimental

concentrates are shown in Table A5.

Laboratory analysis

Forage and concentrate samples were ground

through a 1mm screen (Wiley Mill, Thomas Scientific,

Philadelphia, PA) before analysis of chemical

composition. Dry matter, ash, and ether extract (EE)

were determined according to Helrich (1990). Neutral

detergent fiber (NDF) and acid detergent fiber (ADF)

were determined by an automatic fiber analyzer

(ANKOM Technology Corp., Macedon, NY, USA)

adding sodium sulfite and heatstable αamylase as

proposed by Van Soest et al. (1991). Lignin was also

determined according to Van Soest et al. (1991). Total

nitrogen (N) content was determined by the Dumas

combustion method using a N analyzer (Leco FP2000;

Leco Instruments Inc., St. Joseph, MI, USA), and crude

protein (CP) was calculated as N × 6.25. Forage in vitro

DM digestibility (IVDMD) was determined by the two

stage procedure of Tilley and Terry (1963) modified by

Goering and Van Soest (1970).

Milk samples were analyzed for fat, protein, lactose,

total solids, and urea nitrogen (MUN) using infrared

procedures (MilkoScan FT+; Foss North America Inc.,

Eden Prairie, MN). Blood samples were centrifuged

(3,000 × g, 4°C, 20 min) and plasma analyzed for

glucose (YSI 2700 Select, Biochemistry analyzer, Yellow

Spring, OH, USA), ureaN (PUN; Kit N 535, Sigma

Chemical Co., St. Louis, MO, USA), and nonesterified

fatty acids (NEFA; NEFAC kit, Wako Chemicals

GmbH, Richmond, VA, USA) modified by Johnson and

Peters (1993).

Statistical analysis

All data were analyzed with SAS (release 8.0, SAS

Institute Inc.). Data were examined for outliers using

the REG procedure and outliers were removed based on

an absolute studentized residue value > 3. Then all

datasets were tested for model additivity, independence

of errors, normality, and homoscedasticity before

analysis of variance. Analysis of variance for all five

experiments was performed using the GLM procedure.

Means were obtained using the least squares method

and compared using the Tukey’s test. Differences were

declared significant at p ≤ 0.05, and trends were

declared at 0.05 ≤ p ≤ 0.10.

Results and Discussion

Experiment 1 – Citrus pulp (PCP)

Increasing levels of PCP up to 75 % replacing ground

corn in the supplement did not affect (p > 0.05)

performance, milk composition and blood parameters

in midlactation Holstein cows grazing intensively

managed elephant grass (Table 2). These results agree

with those from Assis et al. (2004) who reported no

differences in nutrient intake, BW change as well as

MY and composition of Holstein cows fed total mixed

rations with increasing levels of PCP replacing up to

100 % of corn. In cows maintained in pens, some

authors have reported an increase in milk fat content

due to the higher inclusion levels of dried citrus pulp

in the supplementary feed, at the expense of corn

(Drude et al., 1971; Van Horn et al., 1975). This increase

in milk fat content, usually observed when the lipids

content in the basal diet was less than 3.5 %, is likely

due to a more efficient rumen fermentation because of

a greater proportion of highly degradable NDF and

readily fermentable soluble sugars found in PCP

(GarcíaRodríguez et al., 2020), which is the case of the

diets in this study (Table 1). Conversely, other studies

have shown no increase in milk fat content when the

lipid content in the basal diet was higher than 3.5 %

(Harvatine et al., 2002).

Experiment 2 – Corn gluten feed (CGF)

Increasing levels of CGF in supplements did not

influence (p > 0.05) the production performance, milk

composition and blood parameters of the dairy cows

rotationally grazing elephant grass pastures (Table 3).

Byproducts as corn replacements for grazing dairy cows

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Table 2. Performance, milk, and blood parameters of dairy cows rotationally grazing elephant grass and supplemented with

increasing levels of pelletized citrus pulp (PCP) replacing ground corn.

Item(1) PCP0(2) PCP25 PCP50 PCP75 SEM(3) pvalue

BCS 2.50 2.53 2.53 2.47 0.04 0.61

Milk yield, kg d1 18.3 18.9 18.7 18.8 0.44 0.78

3.5% FCM, kg d1 18.2 18.8 18.7 19.0 0.43 0.62

Fat, kg d1 0.630 0.660 0.650 0.670 0.01 0.57

Protein, kg d1 0.550 0.580 0.560 0.560 0.01 0.57

Milk solids, kg d1 2.30 2.46 2.51 2.42 0.12 0.83

Fat, % 3.53 3.50 3.54 3.55 0.06 0.95

Protein, % 3.06 3.12 3.06 3.02 0.03 0.27

Lactose, % 4.38 4.43 4.37 4.35 0.02 0.12

Total solids, % 12.9 12.9 13.3 12.9 0.43 0.83

MUN, mg dL1 15.1 15.1 14.8 14.9 0.35 0.90

Plasma glucose, mg dL1 76.5 76.1 73.4 77.6 1.38 0.20

Plasma urea, mg dL1 36.3 36.7 36.0 36.4 0.81 0.94

PUN, mg dL1 16.9 17.1 16.8 16.9 0.72 0.99

NEFA, µEq L1 492 429 443 461 38.3 0.68

(1)BCS, body condition score; 3.5 % FCM (fatcorrected milk) = [(0.4324 × milk yield) + (16.216 × fat yield)]; MUN, milk urea nitrogen; PUN, plasma urea nitrogen;

NEFA, nonesterified fatty acids. (2)PCP0, 0 % of PCP; PCP25, 25 % of PCP; PCP50, 50 % of PCP; PCP75, 75 % of PCP. (3)SEM, standard error of the mean.

Table 3. Performance, milk, and blood parameters of dairy cows rotationally grazing elephant grass and supplemented with

increasing levels of corn gluten feed (CGF) replacing ground corn.

Item(1) CGF0(2) CGF25 CGF50 CGF75 SEM(3) pvalue

BCS 2.45 2.44 2.42 2.36 0.03 0.18

Milk yield, kg d1 12.4 12.6 12.4 12.2 0.23 0.58

3.5% FCM, kg d1 12.3 12.5 12.2 12.3 0.23 0.73

Fat, kg d1 0.425 0.434 0.420 0.430 0.09 0.72

Protein, kg d1 0.396 0.407 0.395 0.398 0.08 0.70

Milk solids, kg d1 1.46 1.49 1.45 1.45 0.27 0.68

Fat, % 3.50 3.50 3.47 3.60 0.07 0.67

Protein, % 3.30 3.31 3.31 3.35 0.08 0.55

Lactose, % 4.26 4.22 4.25 4.20 0.02 0.13

Total solids, % 11.9 11.9 11.9 11.9 0.07 0.78

MUN, mg dL1 14.3 13.9 14.2 14.0 0.22 0.52

Plasma glucose, mg dL1 66.7 66.7 66.1 65.0 1.69 0.84

Plasma urea, mg dL1 41.5 42.3 41.0 39.0 1.88 0.61

PUN, mg dL1 19.4 19.8 19.1 18.2 0.90 0.61

NEFA, µEq L1 385 373 353 357 16.6 0.51

(1)BCS, body condition score; 3.5 % FCM (fatcorrected milk) = [(0.4324 × milk yield) + (16.216 × fat yield)]; MUN, milk urea nitrogen; PUN, plasma urea nitrogen;

NEFA, nonesterified fatty acids. (2)CGF0, 0 % of CGF; CGF25, 25 % of CGF; CGF50, 50 % of CGF; CGF75, 75 % of CGF. (3)SEM, standard error of the mean.

The results obtained showed that CGF can replace

corn up to 75 % in the supplement with no side effects

in the case of the late lactating dairy cows grazing

intensively managed elephant grass. These results are

similar to others obtained in with highproducing

Holstein cows fully maintained in corrals (Armentano

and Dentine, 1988; Pedroso et al., 2009). Other authors

have reported increased MY when CGF was included

in the diet (Kononoff et al., 2006; Firkins et al., 1991);

however, in those studies, CGF replaced not only the

ground corn but also other roughage and concentrate

ingredients, resulting in higher DM intake, which

explained greater MY. The results obtained in other

studies for milk composition parameters, especially the

protein, fat, and ureaN contents, were variable, with

some studies showing changes (Pedroso et al., 2009)

whilst others did not (Firkins et al., 1991). Such

variability is likely due to which ingredients were

replaced by CGF, was only corn or corn plus other

forage and concentrate ingredients.

Martínez et al.

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191

Table 4. Performance, milk, and blood parameters of dairy cows rotationally grazing elephant grass and supplemented with

increasing levels of whole cottonseed (WCS) replacing ground corn.

Item(1) WCS0(2) WCS25 WCS50 WCS75 SEM(3) pvalue(4)

BCS 2.29 2.25 2.33 2.28 0.03 0.37

Milk yield, kg d1 17.7a 17.4a 16.9a 15.3b 0.31 <0.01

3.5% FCM, kg d1 17.8a 17.8a 17.8a 16.2b 0.38 0.01

Fat, kg d1 0.627 0.635 0.648 0.593 0.01 0.18

Protein, kg d1 0.512a 0.494a 0.470a 0.426b 0.01 < 0.01

Milk solids, kg d1 2.06a 2.03a 2.00a 1.80b 0.04 < 0.01

Fat, % 3.55 3.65 3.80 3.80 0.11 0.27

Protein, % 2.90 2.84 2.80 2.83 0.03 0.06

Lactose, % 4.23 4.20 4.17 4.00 0.07 0.06

Total solids, % 11.6 11.6 11.7 11.5 0.16 0.81

MUN, mg dL1 13.4b 15.0a 15.9a 15.8a 0.46 < 0.01

Plasma glucose, mg dL1 49.4 51.2 52.0 53.8 1.71 0.10

Plasma urea, mg dL1 49.8 48.3 49.0 51.8 1.48 0.16

PUN, mg dL1 23.2 22.6 22.9 24.2 0.69 0.16

NEFA, µEq L1 368.3 371.5 365.3 362.3 18.2 0.60

(1)BCS, body condition score; 3.5 % FCM (fatcorrected milk) = [(0.4324 × milk yield) + (16.216 × fat yield)]; MUN, milk urea nitrogen; PUN, plasma urea nitrogen;

NEFA, nonesterified fatty acids. (2)WCS0, 0 % of WCS; WCS25, 25 % of WCS; WCS50, 50 % of WCS; WCS75, 75 % of WCS. (3)SEM, standard error of the mean.

(4)Means followed with the same lowercase letter in the rows do not differ by Tukey’s test, at 5 % probability.

The substitution of ground corn at the level of 75 % of

WCS in the supplement decreased yields of milk, 3.5 %

FCM, protein, and milk solids of dairy cows in the

current study. This is likely due to a decrease in the

total DM intake and its consequences on microbial

synthesis and the supply of metabolizable protein to

the duodenum (Clark et al., 1992). Milk protein content

tended to be lower with increasing levels of WCS,

which suggests either a lower supply of microbial

protein to the duodenum or a poorer amino acid

profile of this protein source. Previous studies with

housed dairy cows have shown that inclusion of WCS

between 1518 % in the diet did not have negative

effects on MY and composition (Fernandes et al., 2002).

For dairy cows yielding 4050 kg d1 of milk, WCS

levels up to 16 % have increased milk and fat yields

compared with 0 and 24 % WCS (Bales et al., 2024). We

also observed an increase in MUN when WCS was fed.

It corroborates previous studies, and it is likely due to

the high degradability of the WCS protein (Arieli,

1998). Even though diets had the same crude protein

and net energy for lactation contents, the increase in

MUN when WCS was added to the diets might be

explained due to poorer synchronism of protein and

energy as the energy provided by WCS comes from lipids

and not carbohydrates.

Experiment 4 – Wheat Middlings (WHM)

Increasing levels of WHM in the supplement of dairy

cows rotationally grazing elephant grass had no influence

(p > 0.05) on animal performance and blood parameters

(Table 5). The substitution of ground corn for 75 % of

WHM resulted in a slight decline in MY and 3.5 % FCM

(p < 0.06), and the MUN tended to increase along with

the level of substitution of maize with WHM (p < 0.05)

in dairy cows.

Overall, dairy cows were not affected by increasing

levels of WHM in the supplement replacing corn, which

is consistent with previous research with housed animals

(Soares et al., 2004; Tufarelli and Laudadio, 2010).

Conversely, Acedo et al. (1987) reported a decrease in MY

of dairy cows when WHM was fed at 60 %. The latter

aligns with this study with a high level of WCS inclusion

reached with the 75 % substitution. Further, cows fed

WHM at 75 % of corn substitution had higher MUN

compared to other levels. This is likely due to the greater

ruminal degradability of WHM protein and the lower

availability of fermentable carbohydrates in the rumen

(Soares et al., 2004). Other studies have shown that this

greater protein degradability linked to lower energy

availability in the rumen provided by higher levels of

WHM inclusion also resulted in an increased PUN

(Acedo et al., 1987); however, the results obtained in this

study did not corroborate such findings.

Experiment 3 – Whole cottonseeds (WCS)

Dairy cows fed supplements including 75 % of WCS

had lower (p < 0.05) yields of milk, 3.5 % FCM, protein,

and milk solids compared to other levels of inclusion

(Table 4). The inclusion of WCS in supplements increased

MUN (p < 0.05) of dairy cows regardless of the level.

Byproducts as corn replacements for grazing dairy cows

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192

Table 5. Performance, milk, and blood parameters of dairy cows rotationally grazing elephant grass and supplemented with

increasing levels of wheat middlings (WHM) replacing ground corn.

Item(1) WHM0(2) WHM25 WHM50 WHM75 SEM(3) pvalue(4)

BCS 2.25 2.31 2.23 2.27 0.02 0.11

Milk yield, kg d1 19.6 19.7 19.6 18.6 0.30 0.06

3.5% FCM, kg d1 19.1 19.5 19.4 18.5 0.38 0.29

Fat, kg d1 0.653 0.676 0.673 0.646 0.02 0.55

Protein, kg d1 0.576 0.594 0.586 0.558 0.01 0.10

Milk solids, kg d1 2.05 2.09 2.10 1.98 0.05 0.28

Fat, % 3.35 3.46 3.45 3.52 0.06 0.34

Protein, % 2.96 3.04 3.00 3.02 0.04 0.51

Lactose, % 4.19 4.24 4.20 4.23 0.03 0.53

Total solids, % 10.5 10.6 10.7 10.7 0.17 0.73

MUN, mg dL1 13.8b 14.3b 14.6b 15.9a 0.30 < 0.01

Plasma glucose, mg dL1 69.8 68.8 73.5 68.3 3.52 0.72

Plasma urea, mg dL1 39.3 41.3 46.0 44.3 1.75 0.11

PUN, mg dL1 18.3 19.3 21.5 20.7 0.82 0.11

NEFA, µEq L1 262 268 280 238 12.4 0.12

(1)BCS, body condition score; 3.5 % FCM (fatcorrected milk) = [(0.4324 × milk yield) + (16.216 × fat yield)]; MUN, milk urea nitrogen; PUN, plasma urea nitrogen;

NEFA, nonesterified fatty acids. (2)WHM0, 0% of WHM; WHM25, 25 % of WHM; WHM50, 50 % of WHM; WHM75, 7 5% of WHM. (3)SEM, standard error of the

mean. (4)Means followed with the same lowercase letter in the rows do not differ by Tukey’s test, at 5 % probability.

Experiment 5 – Soybean hulls (SBH)

The animal performance and blood parameters in

dairy cows rotationally grazing elephant grass were

not affected (p > 0.05) by increasing levels of SBH

replacing ground corn in the supplement up to 75 %

(Table 6).

Table 6. Performance, milk, and blood parameters of dairy cows rotationally grazing elephant grass and supplemented with

increasing levels of soybean hulls (SBH) replacing ground corn.

Item(1) SBH0(2) SBH25 SBH50 SBH75 SEM(3) pvalue

BCS 2.17 2.25 2.23 2.19 0.05 0.63

Milk yield, kg d1 17.8 17.8 17.4 17.3 0.31 0.50

3.5% FCM, kg d1 16.5 16.8 16.6 16.4 0.48 0.92

Fat, kg d1 0.540 0.560 0.557 0.547 0.03 0.93

Protein, kg d1 0.500 0.517 0.485 0.479 0.02 0.54

Milk solids, kg d1 1.73 1.78 1.73 1.68 0.05 0.59

Fat, % 3.06 3.21 3.22 3.20 0.13 0.80

Protein, % 2.81 2.92 2.78 2.78 0.09 0.70

Lactose, % 3.95 4.02 3.97 3.82 0.13 0.74

Total solids, % 9.71 10.0 9.90 9.70 0.23 0.65

MUN, mg dL1 15.7 15.9 15.0 14.5 0.54 0.24

Plasma glucose, mg dL1 75.3 81.0 75.5 76.8 1.29 0.06

Plasma urea, mg dL1 29.8 30.8 34.3 32.8 1.28 0.16

PUN, mg dL1 13.9 14.4 16.0 15.3 0.60 0.16

NEFA, µEq L1 358 371 361 342 18.8 0.75

(1)BCS, body condition score; 3.5 % FCM (fatcorrected milk) = [(0.4324 × milk yield) + (16.216 × fat yield)]; MUN, milk urea nitrogen; PUN, plasma urea nitrogen;

NEFA, nonesterified fatty acids. (2)SBH0, 0 % of SBH; SBH25, 25 % of SBH; SBH50, 50 % of SBH; SBH75, 75 % of SBH. (3)SEM, standard error of the mean.

Previous research has tested SBH replacing corn in

the concentrate to supply 0, 10, 20, 30 or 40 % of the

dietary DM of housed Holstein cows (Ipharraguerre et

al., 2002), and found that the yields of 3.5 % FCM,

protein, and total solids as well as protein content and

MUN were not affected by such levels of replacement.

On the other hand, milk fat and total solids contents as

well as fat yield increased linearly with increasing

levels of SBH. Ipharraguerre and Clark (2003) did not

report any significant effects of replacing corn with

SBH in dairy cow diets using a metaanalysis

approach. The authors have attributed the results to

the average dietary inclusion level of SBH in the

dataset. However, based on a smaller group of studies,

the latter authors speculated that levels greater than 30

% of dietary DM as SBH in high grain diets (≥ 50 %),

would limit the physically effective fiber. It is

important to emphasise that the latter may increase the

concentration of acids in the rumen resulting in

reduced DM intake (Ipharraguerre and Clark, 2003).

Martínez et al.

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193

The use of PCP, CGF and SBH replacing corn with

up to 75 % of the supplement and WCS and WHM up

to 50 %, has no adverse shortterm effects on milk yield

and composition of Holstein cows grazing intensively

managed elephant grass.

Conclusions

Acknowledgements

We would like to thank all the staff from CTRH

(Centro de Treinamento em Recursos Humanos) and

interns from CPZ (Centro de Práticas Zootécnicas) at

the Department of Animal Science, USP/ESALQ.

Conflicts of Interest: The authors declare no conflicts of interest.

Approval of Animal Ethics: This study was conducted according to research protocols approved by the Animal

Care and Use Committee at the University of Sao Paulo, Luiz de Queiroz College of Agriculture (USP/ESALQ).

Author Contributions: Conceptualization, J.C.M. and F.A.P.S.; methodology, J.C.M. and F.A.P.S.; software, J.C.M.;

validation, J.C.M. and F.A.P.S.; formal analysis, J.C.M.; investigation, J.C.M. and T.V.V.; resources, J.C.M. and

F.A.P.S.; data curation, J.C.M.; writing—original draft preparation, J.C.M., G.F.S.C., D.F.A.C., P.G., T.V.V.,

C.M.M.B. and F.A.P.S.; writing—review and editing, J.C.M., G.F.S.C., D.F.A.C., P.G., T.V.V., C.M.M.B. and

F.A.P.S.; visualization, J.C.M., G.F.S.C., D.F.A.C., P.G. and F.A.P.S.; supervision, F.A.P.S.; project administration,

F.A.P.S.; funding acquisition, F.A.P.S. All authors have read and agreed to the published version of the manuscript.

Funding: This research was funded by São Paulo Research Foundation (FAPESP), grant numbers 2001/117118,

2001/129541, 2003/091408, and 2006/535674.

Edited by: Omar AraujoFebres

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APPENDIX

Table A1. Ingredients and nutrient composition of experimental supplements including increasing levels of pelletized citrus

pulp (PCP) – Experiment 1.

Item PCP0(1) PCP25 PCP50 PCP75

Ingredient, % DM

Ground corn 70.0 51.4 33.5 16.4

Pelletized citrus pulp  17.1 33.5 49.1

Soybean meal 24 25.5 27.0 28.5

Urea 1.0 1.0 1.0 1.0

Minerals and vitamins 5.0 5.0 5.0 5.0

Nutrient composition(2)

Crude protein, %DM 21.8 21.8 21.8 21.8

NEL, Mcal kg1 DM 2.0 2.0 2.0 2.0

(1)PCP0, 0 % of PCP; PCP25, 25 % of PCP; PCP50, 50 % of PCP; PCP75, 75 % of PCP indicating the level of substitution of ground corn by PCP. (2)According to NRC

(2001); NEL, net energy for lactation.

Table A2. Ingredients and nutrient composition of experimental supplements including increasing levels of corn gluten feed

(CGF) – Experiment 2.

Item CGF0(1) CGF25 CGF50 CGF75

Ingredient, % DM

Ground corn 80.1 61.3 43.4 23.6

Corn gluten feed  20.4 43.4 70.8

Soybean meal 12.4 11.7 7.2 

Urea 1.9 1.0 0.4 

Minerals and vitamins 5.6 5.6 5.6 5.6

Nutrient composition(2)

Crude protein, %DM 19.0 19.0 19.0 19.0

NEL, Mcal kg1 DM 2.0 2.0 2.0 2.0

(1)CGF0, 0 % of CGF; CGF25, 25 % of CGF; CGF50, 50 % of CGF; CGF75, 75 % of CGF indicating the level of substitution of ground corn by CGF. (2)According to

NRC (2001); NEL, net energy for lactation.

Table A3. Ingredients and nutrient composition of experimental supplements including increasing levels of whole cottonseed

(WCS) – Experiment 3.

Item WCS0(1) WCS25 WCS50 WCS75

Ingredient, % DM

Ground corn 80.1 63.2 43.4 23.6

Whole cottonseed  21.1 43.4 70.7

Soybean meal 12.4 8.7 7.1 

Urea 1.9 1.4 0.5 0.1

Minerals and vitamins 5.6 5.6 5.6 5.6

Nutrient composition(2)

Crude protein, %DM 19.0 19.0 19.0 19.0

NEL, Mcal kg1 DM 2.0 2.0 2.0 2.0

(1)WCS0, 0 % of WCS; WCS25, 25 % of WCS; WCS50, 50 % of WCS; WCS75, 75 % of WCS indicating the level of substitution of ground corn byWCS. (2)According to

NRC (2001); NEL, net energy for lactation

Martínez et al.

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197

ISSNL 10221301. Archivos Latinoamericanos de Producción Animal. 2025. 33 (4): 185  197

Table A4. Ingredients and nutrient composition of experimental supplements including increasing levels of wheat middlings

(WHM) – Experiment 4.

Item WHM0(1) WHM25 WHM50 WHM75

Ingredient, % DM

Ground corn 80.1 60.6 40.7 20.4

Wheat middlings  20.1 40.7 61.1

Soybean meal 12.4 12.4 12.3 12.9

Urea 1.9 1.3 0.7 

Minerals and vitamins 5.6 5.6 5.6 5.6

Nutrient composition(2)

Crude protein, %DM 19.0 19.0 19.0 19.0

NEL, Mcal kg1 DM 2.0 2.0 2.0 2.0

(1)WCS0, 0 % of WCS; WCS25, 25 % of WCS; WCS50, 50 % of WCS; WCS75, 75 % of WCS indicating the level of substitution of ground corn byWCS. (2)According to

NRC (2001); NEL, net energy for lactation

Table A5. Ingredients and nutrient composition of experimental supplements including increasing levels of soybean hulls (SBH)

– Experiment 5.

Item SBH0(1) SBH25 SBH50 SBH75

Ingredient, % DM

Ground corn 80.1 61.1 40.7 19.8

Wheat middlings  20.4 40.7 59.5

Soybean meal 12.4 11.2 11.6 14.5

Urea 1.9 1.7 1.4 0.6

Minerals and vitamins 5.6 5.6 5.6 5.6

Nutrient composition(2)

Crude protein, %DM 19.0 19.2 19.3 19.2

NEL, Mcal kg1 DM 2.0 1.9 1.9 1.8

(1)SBH0, 0 % of SBH; SBH25, 25 % of SBH; SBH50, 50 % of SBH; SBH75, 75 % of SBH indicating the level of substitution of ground corn by SBH. (2)According to NRC

(2001); NEL, net energy for lactation.

Table A6. Nutrient composition of the feed ingredients.

Nutrient composition, %DM GC(1) SBM PCP CGF WCS WHM SBH

Dry matter 89.2 89.8 93.9 89.9 93.7 90.0 90.0

Ash 1.1 6.1 8.5 3.9 3.4 4.3 4.2

Crude protein 10.4 49.0 7.6 23.9 23.7 19.0 10.8

Neutral detergent fiber 9.7 10.4 25.2 35.8 60.5 37.0 60.5

Acid detergent fiber 3.2 6.7 24.4 14.8 38.7 12.8 44.6

Ether extract 4.1 1.2  3.3 19.2 4.4 2.5

(1)GC, ground corn; SBM, soybean meal; PCP, pelletized citrus pulp; CGF, corn gluten feed; WCS, whole cottonseed; WHM, wheat middlings; SBH, soybean hulls.

Byproducts as corn replacements for grazing dairy cows