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DAIANE CAROLINE DE MOURA
FARELO DE BRASSICA NA ALIMENTAÇÃO DE VACAS LEITEIRAS
Cuiabá-MT
2018
DAIANE CAROLINE DE MOURA
FARELO DE BRASSICA NA ALIMENTAÇÃO DE VACAS LEITEIRAS
Cuiabá-MT 2018
Tese apresentada ao Programa de Pós-Graduação em Ciência Animal da Universidade Federal de Mato Grosso para a obtenção do título de Doutora em Ciência Animal Área de Concentração: Nutrição de Ruminantes Orientador: Prof. Dr. André Soares de Oliveira Co-Orientadores: Prof. Dr. André Fonseca de Brito e Prof. Dr. Nelcino Francisco de Paula
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Aos meus amigos e família. Em especial: A minha mãe Maria Inês de Moura e ao meu esposo
por todo carinho e paciência Flavio Junior Gonçalves Vieira que sempre me apoiaram e
incentivaram em todos os momentos.
Dedico!
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É saber se sentir infinito
Num universo tão vasto e bonito, é saber sonhar
Então fazer valer a pena
Cada verso daquele poema sobre acreditar
Não é sobre chegar
No topo do mundo e saber que venceu
É sobre escalar e sentir que o caminho te fortaleceu
É sobre ser abrigo
E também ter morada em outros corações
E assim ter amigos contigo em todas as situações
(Ana Vilela)
FORÇA, FOCO E FÉ
Trabalhe para seu EU e não para seu EGO. (Fabio de Melo)
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AGRADECIMENTOS A Deus:
“Meu refugio, minha fortaleza,
Meu Deus eu confio em ti”
A minha família:
Á minha amada mãe. EU TE AMO
Ao meu amado esposo Flavio Junior Gonçalves Vieira.
Agradeço a todos, que sempre se preocupam com meus estudos, com meu futuro, sempre
me incentivam, oram por mim que de alguma forma direta ou indiretamente, me ajudam ao longo
desses anos.
Deus abençoe e recompense a todos com suas graças.
Ao meu orientador, pela oportunidade de sua orientada, Professor D.Sc. André Soares de
Oliveira. Obrigada pela compreensão, paciência, pela confiança e amizade, pelos valiosos
ensinamentos.
Agradeço a Capes pela concessão de bolsa de doutorado, o CNPq pela concessão de bolsa
sanduiche e a Fundação de Amparo à Pesquisa do Estado de Mato Grosso (FAPEMAT), projeto
número 483724/2011 PRONEM 006/2011) pelo recurso para execução dos experimentos.
Agradeço a empresa Caramuru Alimentos S.A (Itumbiara, GO, Brasil). pelo fornecimento
do farelo de crambe.
A Universidade Federal de Mato Grosso Campus Sinop e Cuiabá pela minha formação,
pela oportunidade e contribuição científica neste trabalho. Obrigada a todos os funcionários de
uma forma geral.
Aos membros da banca de defesa de tese Dr. André Fonseca de Brito, Dr. Nelcino
Francisco de Paula, Eduardo Henrique Bevitori Kling de Moraes e Erick Darlisson Batista.
A família Peron (Braz, Eloide, Eloir, Josimar, Bia minha amada e Josi), por todo carinho,
atenção e conhecimentos. OBRIGADA POR FAZEREM PARTE DA MINHA FAMÍLIA.
As minhas vaquinhas amadas: Preta, Suja, Miúda, Coração, Mandala, Vermelha, Fumaça,
Tana, Dala, Rapoza, Preta e Capitu por colaborem durante todo o experimento.
A família que conquistei nos Estados Unidos: Adelia, Berquinha e todos de sua família que
me acolherem com muito carinho. Minha querida amada Lia e Rafael por todo carinho cuidados e
passeios. Arlete e família por todo carinho e risadas. A lindíssima Ellen Quirino e suas amigas por
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me receber tão bem. Aos amigos de batalhas do frio Ronan, Simone, Vicent e Geraldo, por todo
carinho, conversas e risadas. A Dover Adult Learning Center por todos os ensinamentos de inglês.
As minhas amadas eternas amigas e irmãs que encontrei em momentos muitos difíceis
Mariane Moreno Ferro, Caren Paludo Ghedini, Luiza Ghedini e Juliane Freire. EU AMO VOCÊS
POR DEMAIS.
Aos amigos e irmãos do Núcleo de Pesquisa em Pecuária de leite: Karine Claudia Alessi,
Suziane Rodrigues Soares, Rodrigo Torres, Henrique Melo, Leticia, Mari Lucia, Indiara, Andrea
Donadia, Juliana Carla, Dhulyeli Silveira e Janaine Poli
Aos amigos que ajudaram para execução dos experimentos Robson Miranda, Suziane
Rodrigues Soares, Fabio Woork, Fernanda Norberto, Silvia, Douglas, Daniely da Silva Sousa,
Poliana Oliveira Cordeiro, Luana Molossi, Viviane, Vitor T. Padilha, Maycon Barbosa, Gabriel
Tschope, Fabricio Marquez, Taynara, Hugo Assis, Rafael Moreno Ferro, Cadu Teles, Natan
Cecconello, Laísa Marangoni, Camila Piovezan, Rodrigo Torres, Silvana, Dâmiris.
Quem tem um amigo tem um grande tesouro
viii
BIOGRAFIA
Daiane Caroline de Moura, filha de Dirceu Figueiredo e Maria Inês de Moura, nasceu em
Vilhena, Rondônia, no dia 20 de janeiro de 1988.
Em agosto de 2006 ingressou no Curso de Graduação em Zootecnia pela Universidade do
Estado de Mato Grosso, no Campus Universitário de Pontes e Lacerda tendo concluído o curso
em fevereiro de 2012.
Em março de 2012, iniciou o curso de Mestrado em Ciência Animal pela Universidade Federal
de Mato Grosso, desenvolvendo estudos na área de Nutrição e Produção de Ruminantes,
submetendo-se à defesa de tese em 26 de fevereiro de 2014.
Em fevereiro de 2014, iniciou o curso de Doutorado em Ciência Animal pela Universidade
Federal de Mato Grosso e em novembro de 2016 realizou doutorado sanduiche na University of
New Hampshire nos Estados Unidos, em ambas Universidade desenvolvendo estudos na área de
Nutrição e Produção de Ruminantes, submetendo-se à defesa de tese em 05 de Março de 2018.
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RESUMO GERAL
MOURA, D. C. Farelo de brassica na alimentação de vacas de leiteiras. 2018. 65f. Tese
(Doutorado em Ciência Animal), Faculdade de Agronomia e Medicina Veterinária, Universidade
Federal de Mato Grosso, Cuiabá, 2018.
Este trabalho está dividido em 2 capítulos. Primeiro capítulo: objetivou-se investigar os efeitos de
inclusão de farelo de crambe (FC, 382,4 g de proteína bruta (PB) / kg de matéria seca (MS), 450
mg de glicosinolatos / kg de MS) na dieta total parcial (DTP; 0, 45, 90 e 135 g / kg DM), sobre o
desempenho produtivo, eficiência de utilização de nutrientes em vacas leiteiras e preferência de
queijo para consumidores não treinados. Foram utilizadas doze vacas leiteiras sendo oito vacas
mestiças de Holandês × Zebu (456 ± 91 kg de peso corporal) e quatros vacas
Jersey (384 ± 32.29 kg de peso corporal) distribuídas em três quadrado latino 4 × 4 simultâneos
com quatro períodos experimentais de 21 dias cada. As DTP foram isonitrogenadas (130 g PB/kg
MS) e oferecido ad libitum entre a ordenha das 7:00 da manhã e 18:30 tarde. As vacas entre o
horário 19:00 da tarde até 6:30 da manhã permaneceram em pastagens Panicum Maximum cv.
Mombaça (564 g de fibra detergente neutro/kg MS e 90,4 g PB/kg MS). A produção de leite e o
consumo de DTP foram registrados do dia 15º ao dia 21º de cada período experimental. As
amostras de leite foram coletadas nos dias 17º e 18º de cada período experimental. As amostras de
fezes de cada animal foram coletadas entre 17º a 21º para estimativa de excreção fecal (usando
dióxido de titânio como indicador externo) e para o consumo de pastagem (utilizando fibra
insolúvel detergente neutro indigestível após 288 incubação ruminal in situ). A inclusão de FC não
afetou o consumo de DTP (P = 0,173; 11,47 ± 0,20 kg de MS/dia), consumo de forragem (P =
0,185; 0,90 ± 0,07 kg MS/dia), consumo de PB (P = 0,481; 1,49 ± 0,01 kg de PB/dia),
digestibilidade da matéria orgânica (P = 0,254; 0,749 ± 0,01 g/g), digestibilidade da PB(P = 0,545;
0,747 ± 0,02 g/g), síntese de proteína microbiana (P = 0,348; 0,83 ± 0,08 kg/d), produção de leite
(P = 0,462; 13,29 ± 0,24 kg/dia), nitrogênio (N) no leite (P = 0,566; 64,2 ± 1,3 g/dia), ureia no
leite (P = 0,178; 10,6 ± 0,94 mg/dL), excreção urinária de nitrogênio (P = 0,717; 90,9 ± 1,9 g/dia),
eficiência de nitrogênio no leite (P = 0,622; 0,268 ± 0,01 g consumo nitrogênio/nitrogênio do
leite), função hepática IU/mL (GGT 32,05 ± 2,94; ALT 15,98 ± 0,44 e AST 48,02 ± 5,71),
rendimento de queijos (0,21 ± 0,01 kg/kg de leite) e análise sensorial (gostei 59,78, 80,00, 76,00,
77,67 % dos provadores). O farelo de crambe pode ser incluso até 135 g/kg de MS na DTP sem
afetar o desempenho produtivo, eficiência na utilização de nutrientes em vacas leiteiras e
preferência de queijo por consumidores não treinados. Segundo capítulo: Utilizamos a abordagem
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meta-analítica para avaliar os efeitos da substituição de diferentes fontes de proteína por dietas de
brassicas sobre a produção de leite e na utilização de nutrientes de vacas leiteiras, de 37 artigos
revisados por pares. O farelo de canola (FC) foi a principal fonte de brassica. Os efeitos foram
comparados pelas diferenças médias brutas (DMB) entre as dietas com FC e fontes de proteínas
(controle) e ponderados pela variância inversa usando modelos de efeito aleatório. O nível de
heterogeneidade foi analisado por estatística I2 (baixa ≤ 25%, moderada = 26 a 50% e alta> 50%).
Em geral, o uso de FC como fonte de proteína aumentou o consumo de MS (DMR = 0,22 ± 0,12
kg de MS/dia, P <0,01; n = 79; I2 = 9,1%) e ingestão de proteína bruta (PB) = (DMR, 14 ± 0,07
kg PB/dia, P <0,01; n = 33; I2 = 21,1%), ambos com baixa heterogeneidade, mas não afetaram a
digestibilidade do trato total da matéria orgânica (P = 0,50; n = 12; I2 = 29,2 %). No geral, o uso
de FC aumentou a produção de leite (DMR = 0,69 ± 0,35 kg/dia, P <0,01; n = 88; I2 = 74,9%),
mas seu efeito depende da comparação entre as fontes proteínas: o FC versus o FS não houve efeito
a produção de leite (DMR = 0,23 ± 0,66 kg/dia, P = 0,50; n = 33), mas a produção de leite aumentou
com a substituição de DDG por FC (DMR = 2,03 ± 1,67 kg/dia; P <0,01; n = 13) e de outras fontes
de proteína por FC (DMR = 0,82 ± 0,43 kg/dia; P <0,01; n = 42). No geral, o uso de FC não afetou
o teor de proteína do leite (P = 0,08; n = 60; I2 = 19,5%) e teor de gordura do leite
(P = 0,20; n = 60; I2 = 16,9%), mas FC aumentou a produção de proteína do leite
(DMR = 0,02 ± 0,01 kg/dia, P <0,01; n = 60; I2 = 0%). O uso de FC reduziu o teor nitrogênio
ureico no leite (N) (DMR = - 0,98 ± 0,31 mg/dL; P <0,01; n = 22; I2 = 32,2%) e aumentou a
eficiência de consumo N no leite (DMR = 0,22% N do leite/consumo de N ± 0,07 mg/dL; P ≤
0,05; n = 34; I2 = 0%), ambos com baixa heterogeneidade. Concluímos que FC é fonte de proteína
semelhante ao FS e é mais eficaz do que o DDG e outras fontes (farelo de algodão, farelo de milho
e farinha de girassol) para vacas leiteiras em lactação. De modo geral os farelos de brassicas (farelo
de crambe e farelo de canola pode ser incluídos em dietas de vacas leiteiras em substituição á
fontes proteicas sem afetar no desempenho produtivo, eficiência de utilização de nutrientes.
Palavras-Chave: farelo de canola, farelo de crambe, metanálise
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ABSTRAT
MOURA, D. C. Brassica meal for dairy cows. 2018. 65f. Thesis (PhD in Animal Science),
Faculdade de Agronomia e Medicina Veterinária, Universidade Federal de Mato Grosso, Cuiabá,
2018.
This thesis is divided into 2 chapters. The objective of this study was to investigate the effects of
inclusion of crambe meal (CM, 382.4 g crude protein/kg dry matter (DM), 450 mg
glucosinolates/kg DM) in the total partial diet (pTMR, 0, 45, 90 and 135 g/kg DM), on the
productive performance, nutrient utilization efficiency in dairy cows and cheese preference for
untrained consumers. Twelve dairy cows were used: eight crossbred Holstein × Zebu cows (456 ±
91 kg body weight) and four Jersey cows (384 ± 32.29 kg body weight) distributed in three
simultaneous 4 × 4 Latin squares with four experimental periods of 21 days each. The pTMR were
isonitrogenated (130 g CP/kg DM) and offered ad libitum between milking from 7:00 am and 6:30
pm. Cows between the hours of 19:00 in the afternoon until 6:30 in the morning remained in
pastures Panicum Maximum cv. Mombaça (564 g neutral detergent fiber/kg MS and 90.4 g CP/kg
DM). Milk yield and pTMR intake were recorded from day 15th to day 21st of each experimental
period. The milk samples were collected on days 17th and 18th of each experimental period. Fecal
samples from each animal were collected between 17th and 21st to estimate fecal excretion (using
titanium dioxide as an external indicator) and for pasture consumption (using indigestible neutral
detergent insoluble fiber after 288 ruminal in situ incubation). The inclusion of CM did not affect
pTMR intake (P = 0.173, 11.47 ± 0.20 kg DM/day), forage intake (P = 0.185, 0.90 ± 0.07 kg
DM/day), CP intake (P = 0.481, 1.49 ± 0.01 kg CP/day), organic matter digestibility (P = 0.254,
0.749 (P = 0.545, 0.747 ± 0.02 g/g), microbial protein synthesis (P = 0.348, 0.83 ± 0.08 kg/d),
milk yield (P = 0., 64.2 ± 1.3 g/d), milk yield (P = 0.462; 13.29 ± 0.24 kg/d), nitrogen (N) milk
(P = 0.566; 64.2 ± 1.3 g/d), milk urea-N (P = 0.178; 10.6 ± 0.94 mg/dL), N urinary excretion (P =
0.717; 90.9 ± 1.9 g/d), N milk efficiency (P = 0.622; 0.268 ± 0.01 g N milk/g N intake) hepatic
function IU/mL (GGT 32.05 ± 2.94, ALT 15.98 ± 0.44 and AST 48.02 ± 5.71), cheese yield (0.21
± 0.01 kg/kg milk) and sensory analysis (I liked 59.78 , 80.00, 76.00, 77.67% of the tasters).
Crambe meal can be up to 135 g/kg DM in pTMR without affects productive performance,
efficiency of nutrient utilization in dairy cows and cheese preference for untrained consumers. We
used meta-analytical approach to evaluate the effects of replacement of different sources of protein
by brassicas meals on milk production and nutrient utilization of dairy cows, from 37 peer-review
papers. Canola meal (CM) was unique brassica source founded. The effects were compared by
xii
raw mean differences (RMD) between CM diet and control treatment means and weighted by
inverse variance using random-effect models. Heterogeneity level was analyzed by I2 statistic (low
≤ 25%; moderate = 26 to 50%; and high > 50%). In overall, use of CM as protein source increased
DM intake (RMD = 0,22 ± 0.12 kg DM/d; P < 0.01; n = 79; I2 = 9.1%) and crude protein (CP)
intake (RMD = 0,14 ± 0.07 kg CP/d; P < 0.01; n = 33; I2 = 21.1%), both with low heterogeneity,
but it did not affect organic matter total-tract digestibility (P = 0.50; n = 12; I2 = 29.2%). In overall,
use of CM increased milk yield (RMD = 0.69 ± 0.35 kg/d; P < 0.01; n = 88; I2 = 74.9%), but its
effect depends on protein sources comparation: CM versus SBM did not increase milk yield (RMD
= 0.23 ± 0.66 kg/d; P = 0.50; n = 33), but milk yield was increased with replacement of DDG by
CM (RMD = 2.03 ± 1.67 kg/d; P < 0.01; n = 13) and of other protein sources by CM (RMD =
0.82 ± 0.43 kg/d; P < 0.01; n = 42). In overall, CM use did not affect milk protein content (P =
0.08; n = 60; I2 = 19.5%) and milk fat content (P = 0.20; n = 60; I2 = 16.9%), but CM increased
milk protein yield (RMD = 0,02 ± 0.01 kg/d; P < 0.01; n = 60; I2 = 0%). Use of CM reduced milk
urea nitrogen (N) (RMD = - 0,98 ± 0.31 mg/dL; P < 0.01; n = 22; I2 = 32.2%) and increase N
intake milk efficiency (RMD = 0.22% N milk/N intake ± 0.07 mg/dL; P ≤ 0.05; n = 34; I2 = 0%),
both with low heterogeneity. We concluded that CM is similar protein source to SBM and it is
more effective than DDG and other sources (cottonseed meal, corn gluten meal and sunflower
meal) to lactating dairy cows.
Key words: canola meal, crambe meal, meta-analysis
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SÚMARIO
1 - INTRODUÇÃO GERAL ............................................................................................. 1
2- REFERÊNCIA BIBLIOGRAFICA .............................................................................. 6
Capitulo 1- Crambe meal in diets for dairy cows ............................................................ 10
INTRODUCTION ........................................................................................................... 11
MATERIALS AND METHODS .................................................................................... 12
RESULTS ........................................................................................................................ 19
DISCUSSION ................................................................................................................. 20
CONCLUSIONS ............................................................................................................. 22
REFERENCES ................................................................................................................ 22
Capitulo 2 - Meta-analysis of the use of brassicas meals in diets for dairy cows ........... 33
INTRODUCTION ........................................................................................................... 34
MATERIALS AND METHODS .................................................................................... 35
RESULTS AND DISCUSSION ..................................................................................... 37
CONCLUSIONS ............................................................................................................. 39
REFERENCES ................................................................................................................ 40
1
1 - INTRODUÇÃO GERAL
As Brassicaceae são compostas por 51 gêneros e 218 espécies, algumas das quais são
espécies importantes de culturas cultivadas como vegetais, forragens e oleaginosas
(Sabharwal et al., 2006). Após a extração de oleaginosas são produzidos produtos com potencial
de uso na alimentação dos ruminantes como fonte de proteína. O farelo de canola, farelo de
crambe e farelo de carinata são as principais fontes de brassicas inclusas na alimentação animal,
mas seu uso depende dos níveis de glicosinolatos (Tripathi e Mishra, 2007; Moura et al., 2017).
O crambe (Crambe abyssinica H.) representa uma fonte alternativa e promissora de óleo
vegetal para produção de biodiesel conhecida como couve etíope. É uma oleaginosa da família
Brassicaceae e também conhecidas como da família das crucíferas, da qual fazem parte plantas
como a mostarda (Brassica campestris L.), a canola (Brassica napus L.) e a nabiça
(Raphanus raphanistrum L.). O crambe é nativo da região do Mediterrâneo sendo e também
cultivado na África, Ásia, Europa, Estados Unidos, México e América do Sul
(Fontana et al.,1998; Souza et al., 2009).
As plantas de crambe são herbáceas anuais, com altura entre 70 e 90 cm, com ciclo
anual curto (90 dias), ramifica-se próxima ao solo para formar galhos (trinta ou mais), os quais
se ramificam, formando galhos terciários. Suas folhas são ovais e assimétricas. As flores são
brancas, numerosas pequenas e estão agrupadas em cachos (Opliger et al., 1991; Fontana et
al.,1998; Souza et al., 2009). Os frutos são uma siliquia de forma esférica inicialmente verde,
tornando-se amarelo com a maturidade e distribuído por todos os galhos da planta. Cada siliquia
possui uma semente de cor verde ou marrom esverdeado com diâmetro entre 0,8 a 2,5 mm
(Desai et al., 1997).
A cultura do crambe é tolerante a seca e a geadas em grande parte de seu
desenvolvimento, cultivada entre a safra de verão e inverno, podendo ser implantada e cultivada
de forma mecanizada, utilizando implementos da cultura soja (Pitol et al., 2010). Para Brito
(2009) o ponto importante de crambe é a baixa ocorrência de pragas, o que diminui os custos
de sua produção.,
A cultura de crambe foi utilizada pela primeira vez em 1933, na Estação Botânica
Boronez, na antiga União das Republicas Socialistas Soviéticas (Mastebroek et al., 1994). As
pesquisas com crambe e a sua produção comercial se intensificaram a partir dos anos 80, após
2
sua introdução nos Estados Unidos da América, no Reino Unido e em alguns países da Europa,
como a Itália, França e Portugal. Contudo, as áreas plantadas nesses países não aumentaram
expressivamente, devido a competição por área com as principais culturas de safra, como o
milho, a soja e o trigo, sendo que nessas regiões não foi possível cultivar o crambe em safrinhas
(período de entressafra compreendido entre culturas principais e iniciada após a cultura de
verão). Dessa maneira, o crambe vem crescendo muito em outros países, como a Austrália, a
África do Sul, o Paraguai e o Brasil (Pitol et al., 2010).
A introdução da crambe no Brasil ocorreu na década de 90 por meio da introdução de
cultivares vindo do México, os quais foram selecionados por pesquisadores da Fundação Mato
Grosso do Sul, em 1995, originando o primeiro cultivar de crambe (FMS Brilhante) no país,
entretanto, somente a partir de 2003 com o lançamento do Programa Nacional de Biodiesel
(PNB) despertou interesse ao cultivo (Pitol et al., 2010; Pilau et al., 2011). De acordo com Pitol
et al. (2010), a produtividade de semente de crambe no Brasil é de 1.000 a 1.500 kg/ha, com
potencial de produção de 450 kg de óleo/ha. No Brasil central, nas regiões Centro-oeste e
Sudeste, o crambe pode ser cultivado como cultura de “safrinha”, no período de fevereiro a
abril, após as culturas de verão (Fundação MS, 2009).
Estudos realizados na estação de pesquisa da Fundação do Mato Grosso do Sul (2007),
em Maracajú – MS, destacaram como vantagens do crambe tolerância à seca, à geada e depois
de estabelecida, elevado teor de óleo (34% a 38%), precocidade (florescimento aos 35 dias e a
colheita aos 85/90 dias) (Jaspar et al., 2010).
Segundo Carlsson (1983), o óleo produzido não é comestível, porém possui grande
utilidade para matéria prima industrial (lubrificante, inibidor da corrosão, matéria-prima para a
produção de borracha sintética e plástico), pois apresenta cerca de 60% de ácido erúcico
(C22:2). O ácido erúcico é um ácido graxo monoinsaturado de cadeia longa que pode provocar
lesões cardíacas quando ingeridos (Air, 1997). Assim, este óleo não concorre com óleos
destinados ao setor alimentício.
O processo de extração de óleo de crambe gera produtos (farelos ou tortas) com elevado
teor proteico e com grande potencial de uso na alimentação animal em substituições às fontes
tradicionais, como farelo de soja e de algodão.
Entretanto tornando-se necessário, o estudo mais detalhado do perfil aminoacídico do
farelo de crambe para poder substituir o farelo de soja em momentos de escassez ou preço em
3
alta no mercado, sem que este fornecimento de aminoácidos não seja prejudicado. Em estudos
realizados por Liu et al. (1993) e Anderson et al. (1993), verificaram que tanto o teor de
metionina como o de cistina é maior no farelo de crambe do que no farelo de soja.
Pesquisadores norte-americanos observaram que a substituição integral do farelo de soja
pelo farelo de crambe in natura (10% de inclusão na dieta), com teor de glicosinolatos de
(56 mmol/kg de MS ou 22 g de equivalente sirigin/kg de MS – peso molecular do sirigin de
397) não afetaram o crescimento e características de carcaça de bovinos, bem como o
desempenho reprodutivo e função da tireóide em vacas de corte gestantes e não lactantes
(Anderson et al., 1993; Anderson et al., 2000).
No Brasil, Mizubuti et al. (2011) observaram que a torta de crambe apresenta
degradabilidade ruminal e digestibilidade intestinal da proteína semelhantes ao do farelo de
soja. Conava et al. (2015) verificou que a substituição do farelo de soja pela torta de crambe
(21% de inclusão na dieta, base da matéria seca; teores de glicosinolatos não informados)
reduziu o consumo e digestibilidade da dieta em ovinos, em razão do aumento excessivo do
teor de lipideos na dieta de 7,5% de extrato etéreo (acima do limite padrão de 6%, base da
matéria seca), comprometendo a digestão da fibra.
Diante do exposto, verifica-se ainda há necessidade de maiores investigações dos efeitos
do farelo de crambe e outras brassicas na alimentação de vacas de leite.
A canola (Brassica napus L. var oleífera) é uma oleaginosa e cultura alternativa de
inverno (Zimmermann, 2005). Foi desenvolvida a partir do melhoramento genético da colza,
obtida do cruzamento Brassica napus (colza) e Brassica campestris (mostarda) passando ser
chamada de canola (Canola Council of Canada, 2009). O termo canola significa a abreviação
internacional para Canadian Oil Low Acid, grão com baixos níveis de ácido erúcico no óleo
(menos que 2%) e baixos níveis de glicosinolatos no farelo (menos que 3 mg/g) (Bell, 1984;
Bett et al., 1999; Newkirk, 2009).
No Brasil cultiva-se apenas canola de primavera, da espécie Brassica napus L. var.
oleífera, que início a implantação na década de 80 no estado do Paraná (PR) e em 2003 em
Goiás (GO) sendo esta uma ótima opção nos sistemas de rotação de culturas, auxiliando na
quebra do ciclo de diversas doenças e pragas das gramíneas e leguminosas, isso ocorre pelo
fato de ser uma cultura da família de crucíferas. (Tomm, 2000; Tomm, 2006).
4
A canola é uma planta anual com o hábito de crescimento indeterminado, com sistema
radicular pivotante e ramificação lateral significativa. O caule é herbáceo, ereto, com tamanho
variável de 0,5 a 1,7 m. As folhas inferiores da planta são pecioladas e formam uma roseta.
Após a elongação do caule, as folhas emitidas são lanceoladas e abraçam parcialmente a haste.
As flores, agrupadas em cachos, são pequenas e amarelas, formadas por quatro pétalas dispostas
em cruz, seis estames e pistilo. A duração do período de floração varia conforme com a cultivar.
Os frutos são síliquas com cerca de 6 cm de comprimento. Os números de grãos e o
comprimento das síliquas, varia dependendo a cultivar. (García, 2007).
A semente de canola contém aproximadamente 40,5% de óleo, o que a torna uma das
oleaginosas com maior teor de óleo disponível para a produção de biodiesel (Lardy, 1993). Os
principais ácidos graxos que compõe o óleo são em sua maioria, oléico (C18: 1), linoléico
(C18:2) e linolênico (C18:3) (Sanches 1997).
Após a extração do óleo é gerado o farelo de canola. De acordo com Queiroz (2008), o
grão de canola é moído, esmagado e prensado para extração do óleo. Posteriormente para
retirada do óleo remanescente é adicionado solvente (hexano) e tostado e moído gerando o
farelo de canola.
Hoje, o farelo de canola é a segunda mais consumida no mundo (12% de participação
de mercado), abaixo de apenas a farelo de soja (71% de participação no mercado) (SoyStats,
2018).
O farelo de canola apresenta teores médios de 38,0% de PB, 5,2% de amido; 21,2% de
FDN e 17,2% de FDA e 1,5% de EE (Bell, 1993). O mesmo apresenta aproximadamente 35%
de proteína não degradável no rúmen (NRC, 2001). O farelo de canola apresenta um perfil de
aminoácidos semelhante ao do farelo de soja, porém com teor inferior de lisina e superior de
metionina + cistina (NRC 2001). Piepenbrink and Schingoethe (1998) corroboram afirmando
que o perfil de aminoácidos do farelo de canola e muito semelhante ao encontrado no leite de
bovinos. Assim havendo interesse crescente em balancear as dietas de vacas leiteiras visando
atender as exigências de aminoácidos.
O cultivo de Carinata está sendo implantada na Dakota do Sul, Dakota do Norte,
Minnesota, Montana e na Flórida (EUA) como potencial para cultura de inverno e nova matéria-
prima para a produção de biocombustíveis (Lawrence e Anderson, 2015). Esta oleaginosa
5
pertence a das família Brassica onde estão incluidas as culturas alimentares mais comuns
repolho, couve de Bruxelas, couve-flor, couve, colza / canola e brócolis (Moser, 2010).
Após a extração do óleo, é gerado o farelo de carinata que está havendo um grande
interesse para ser inclusão na alimentação animal, principalmente pelo alto teor proteína 35-
40% na matéria seca, boa fonte de proteína degradável do rúmen (70% de proteína bruta
degradada no rúmen) e digestibilidade total comparável à das farinhas de soja (farelo de soja
96% versus farelo de carinata 94,4%) (Lawrence e Anderson, 2015).
O SDSU Dairy e Food Science Department testou recentemente os efeitos da
alimentação de carinata no desempenho de crescimento de novilhas leiteiras, com nível de
inclusão de 10 % da MS total da dieta em comparação DDGS (grãos secos de destilaria com
solúveis) e verificou que a carinata pode ser inclusa até 10% da dieta, pois apresentou
desempenho de crescimento semelhante em comparação com novilhas alimentadas com dietas
com DDGS (Rodriguez-Hernandez e Jill Anderson, 2016).
Este foi um dos primeiros estudos realizados sobre a alimentação de carinata para
alimentação de bovinos leiteiros. Assim novas pesquisas são necessárias para testar os efeitos
carinata na alimentação de vacas em comparação com outras fontes de proteína
Os capítulos foram formatados de acordo com as normas da Journal of Dairy Science.
6
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BOTINI, L. A. A.; SINHORIN, P.; OGUNADE, I. M.; OLIVEIRA A. S. Crambe meal
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T. M.; DOLL, J. D.; KELLING, K. A.; DURGAN, B. R.; NOETZEL, D. M. Crambe.
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extensão agrícola cooperativa. Universidade de Wisconsin, Madison, 1991.
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(Brassica napus L.) nas variedades CTC-4 e ICIOLA-4 e de oito variedades de canola
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9
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10
Crambe meal in diets for dairy cows 1
Abstract: We evaluated the effects crambe meal (CM; 382.4g crude protein (CP)/kg dry matter 2
(DM); 450 mg glucosinolates/kg DM) inclusion in partial total mixed ration (pTMR; 0, 45, 90 and 3
135 g/kg DM), on productive performance, efficiency of nutrient utilization in dairy cows and 4
cheese preference for untrained consumers. Twelve dairy cows were used: eight crossbred 5
Holstein × Zebu cows (456 ± 91 kg body weight) and four Jersey cows (384 ± 32.29 kg body 6
weight) distributed in three simultaneous 4 × 4 Latin squares with four experimental periods of 21 7
days each. The pTMR were isonitrogenated (130 g CP/kg DM) and offered ad libitum between 8
milking from 7:00 am and 6:30 pm. Cows between the hours of 19:00 in the afternoon until 6:30 9
in the morning remained in pastures Panicum Maximum cv. Mombasa (564 g neutral detergent 10
fiber/kg MS and 90.4 g CP/kg DM). Milk yield and pTMR intake were recorded from day 15th to 11
day 21st of each experimental period. The milk samples were collected on days 17th and 18th of 12
each experimental period. Fecal samples from each animal were collected between 17th and 21st to 13
estimate fecal excretion (using titanium dioxide as an external indicator) and for pasture 14
consumption (using indigestible neutral detergent insoluble fiber after 288 ruminal in situ 15
incubation). Spot urine samples were collected after feeding (3 h) on the 20th day of each period 16
by manual stimulation of the vulva. The inclusion of CM did not affect pTMR intake (P = 0.173, 17
11.47 ± 0.20 kg DM/day), forage intake (P = 0.185, 0.90 ± 0.07 kg DM/day), CP intake (P = 0.481, 18
1.49 ± 0.01 kg CP/day), organic matter digestibility (P = 0.254, 0.749 (P = 0.545, 0.747 ± 0.02 19
g/g), microbial protein synthesis (P = 0.348, 0.83 ± 0.08 kg/d), milk yield (P = 0., 64.2 ± 1.3 g/d), 20
milk yield (P = 0.462; 13.29 ± 0.24 kg/d), nitrogen (N) milk (P = 0.566; 64.2 ± 1.3 g/d), milk 21
urea-N (P = 0.178; 10.6 ± 0.94 mg/dL), N urinary excretion (P = 0.717; 90.9 ± 1.9 g/d), N milk 22
efficiency (P = 0.622; 0.268 ± 0.01 g N milk/g N intake) hepatic function IU/mL (GGT 32.05 ± 23
2.94, ALT 15.98 ± 0.44 and AST 48.02 ± 5.71), cheese yield (0.21 ± 0.01 kg/kg milk) and sensory 24
11
analysis (I liked 59.78 , 80.00, 76.00, 77.67% of the tasters). Crambe meal can be up to 135 g/kg 25
DM in pTMR without affects productive performance, efficiency of nutrient utilization in dairy 26
cows and cheese preference for untrained consumers 27
Key words: brassica; byproducts; intake; glucosinolates 28
29
1 - Introduction 30
Crambe (Crambe abyssinica H) represents alternative source of vegetable oil to produce 31
biodiesel and other industrial use (Souza et al., 2009). Crambe meal (CM) is a byproduct obtained 32
after oilseed extraction and contains 370 to 430 g crude protein (CP)/ kg dry matter (DM) 33
depending on the cultivar, method of oil extraction (Mizubuti et al., 2011; Moura et al., 2017). 34
However, glucosinolates is a potential limiting for use in animal diet (Tripathi e Mishra, 2007). 35
Moura et al. (2017) observed that CM provided for Animal Nutrition Industry in Brazil had low 36
glucosinolates level, and that its use did not affect intake, total-tract digestibility, ruminal microbial 37
protein synthesis and nitrogen (N) balance in ruminant. 38
Crambe meal and soybean meal (SBM) exhibit similar intestinal digestibility the protein 39
(Lambert et al., 1970; Mendonça et al., 2015a; Mendonça et al., 2015b; Moura et al., 2017). The 40
methionine in CM contents are higher than SBM, but CM has low lysine level 41
(Anderson et al., 1993, Liu et al., 1993). 42
Therefore, there is evidence that CM could be an alternative protein source to SBM to 43
lactating dairy cows. However, we did not found studies about use of CM on diet to dairy cows, 44
and so it is necessary to elucidate its effects on nutrient utilization efficiency and dairy cow 45
performance. We hypothesized that inclusion SBM with CM did not affect the efficiency of 46
nutrient utilization and performance the dairy cows. Therefore, we evaluated the effects crambe 47
meal (CM; 382.4g crude protein (CP)/kg dry matter (DM); 450 mg glucosinolates/kg DM) 48
inclusion in partial total mixed ration (pTMR; 0, 45, 90 and 135 g/kg DM), on productive 49
12
performance, efficiency of nutrient utilization in dairy cows and cheese preference for untrained 50
consumers. 51
2 - Materials and methods 52
Care and handling of the animals used in the current study were conducted as outlined in 53
the guidelines of the Universidade Federal de Mato Grosso Institutional Animal Care and Use 54
Committee (IACUC# 23108718743/2016-11). 55
This experiment was conducted at the commercial dairy farm (Dona Hermínia Fazenda, 56
Sinop, MT, Brazil; 55o20`19.802`` O and 11o49`50.988`` S; altitude 344 m above sea level) and 57
Dairy Cattle Research Lab at the Universidade Federal de Mato Grosso - Sinop (55o28`51.532`` 58
O and 11o51`49.517`` S; altitude 378 m above sea level), from 11th of August to 7th of november 59
2014 (dry to rain transition), totaling 84 experimental days. CM was provided by Camaruru Ltda 60
(Itubiaria, Goiás, Brazil). 61
Animals, experimental design and pasture management 62
Twelve cows eight Holstein × Zebu dairy cows (456 ± 91 kg body weight (BW) and 73 ± 63
64 days in milk) and four Jersey (384 ± 32.29 kg body weight (BW) and 73 ± 64 days in milk) 64
cows were blocked by days in milk and randomly assigned with squares to treatment sequences in 65
tree replicated 4 x 4 Latin squares (1 Latin square Jersey and 2 Latin square Holstein × Zebu dairy 66
cows). Treatment sequences within Latin squares were balanced for carryover effects in 67
subsequent four periods (Williams, 1949). Each period lasted 21 d and consisted of 14 d for diet 68
adaption and 7 d for data and samples collection. At the end of the experiment all, all cows less 69
than 150 d of gestation. 70
Cows were fed on pasture plus pTMR system. The cows received four isonitrogenous 71
pTMR (130 ± 0.10 g crude protein/kg DM) containing four levels inclusion of CM 72
(0, 45, 90 and 135 g/kg DM). The chemical composition of the feeds is presented in Table 1. The 73
13
proportion and chemical composition of the pTMR are described in Table 2. Each cow individually 74
received pTMR ad libitum between the morning and afternoon milking in a tie-stall barn, totality 75
16 h of feeding. The pTMR were fed four daily at 0700, 0900, 1200 am and 0300 pm Amounts of 76
pTMR offered to the cows were adjusted daily to allow refusals equal to 5 to 10% of intake. The 77
DM content from weekly composites of the silages and concentrated mixture were used to adjust 78
as-fed pTMR composition to maintain constant dietary ingredient over the trial. After the last 79
milking (6:00 pm), the cows went to 1.3 acre of pasture of Panicum maximum cv. Mombaça, with 80
access to water. 81
Sampling and Experimental procedures 82
The average sward was determined by measuring the height at pre-grazing and post-grazing 83
by 15 measurements in each paddock. The forage mass in pre-grazing was determined using a 84
square of 0.25 m2, with two measurements per paddock. After collection, the components were 85
separated by leaf, green stem and dead material, and were weighed and stored (-15oC) for 86
subsequent pre-drying, grinding and chemical analysis. During the collection period (7 d) the 87
pasture intake was sampled daily by simulated grazing. Immediately, the samples were stored (-88
15oC). At the end of collection period (d 7) pasture sample by simulated grazing were removed 89
from the freezer, thawed at room temperature, and blended manually to obtain a composite sample 90
per each period. The composite sample of pasture (simulated grazing) was predried in a forced-air 91
oven at 55°C for 72 h. 92
Were sampled individual ingredients of concentrate mixture in each mixture preparation 93
(21 d) and kept in a freezer (−15°C) for subsequent pre-drying, ground and chemical analysis. 94
Daily intake of pTMR was determined by the difference between the weight of DM offered and 95
refused. Twice daily were weighed corn silage, concentrate mixture offered and pTMR refusals 96
(removed) for each cow. Twice daily were sampled (approximately 100 g) of the offered diet and 97
refusals and stored in plastic bags, labeled, and kept in a freezer (−15°C). At the end of collection 98
14
period (d 7), corn silage and refusal samples of each animal were removed from the freezer, thawed 99
at room temperature, and blended manually to obtain a composite sample per animal for each 100
period. The composite sample of corn silage and refusal were predried in a forced-air oven at 55°C 101
for 72 h. 102
Titanium dioxide (TiO2) was used as external marker to estimate the fecal excretion 103
(Titgemeyer et al., 2001) and indigestible NDF (iNDF (indigestible neutral detergent fiber)); after 104
288 hours of in situ rumen) was used as internal marker to estimate intake. After morning milking, 105
each cow received 15 g of TiO2 during the 16th to 21st d of each period. Feces were directly 106
collected from the rectum once daily at 0800, 1000, 1200, 1400 and 1600 h, from 16th to 21st day 107
of each period, and stored (-15oC) for subsequent predrying, grinding and chemical analysis. After 108
predrying and grinding, a single composite fecal sample was obtained per cow per period. 109
The dry matter intake (DMI) from pasture (DMIpasture) per cow was estimated as: 110
DMIpasture (kg/d) = [(FE × iNDFf) – iNDFc intake]/iNDFp 111
where: FE, the fecal excretion (kg/d); iNDFf, iNDF in feces (kg/kg DM); iNDFc intake, iNDF 112
concentrate intake (kg/d) = supplement DM intake (kg/d) × iNDF in suppplement (kg/kg DM); 113
iNDFp, iNDF in pasture by simulated grazing (kg/kg DM). 114
Feces, feed and pasture samples were dried in a forced-ventilation oven at 55 ºC for 72 115
hours, ground in a Wiley mill (Tecnal Equipamentos para Laboratório, Piracicaba, São Paulo, 116
Brazil) with 1-mm screen for chemical analysis and 2-mm screen for ruminal incubation in situ 117
(iNDF). 118
Cows were mechanically milked twice daily (0600 a.m and 0500 p.m.) and milk yields 119
were recorded daily from the 15th to the 21st of each period. Milk samples from a.m. and p.m. 120
milking were collected on 18th and 19th d of each period. Composite samples were prepared daily 121
according to milk production and two aliquots ware taken from each composite sample: the first 122
aliquot (50 mL) forwarded to analyze fat, protein and lactose in milk using Mast Classic milk 123
15
analyzer (Milkotester LTD, Model: LM2, Serial: 11624); the second aliquot was deproteinized 124
with trichloroacetic acid (20 mL of milk mixed with 10 mL of trichloroacetic acid 250 g/L), filtered 125
(Whatman #1 filter paper) and stored (-15 ºC) for subsequent analysis of allantoin and urea. Milk 126
yield was adjusted to 40 g fat/kg as NRC (2001). Body weights and body condition score (1 to 5; 127
Ferguson et al. 1994) were measured 21th d of each period. 128
Blood samples were collected in EDTA Vacutainer® tubes from coccygeal vein, 3 h after 129
a.m. supplement feeding on 19th d of each period. Blood samples were immediately centrifuged 130
(2,300 x g, for 15 minutes, room temperature) and plasma was stored (-15oC) for subsequent 131
analysis of urea nitrogen and enzymes gamma glutamyl transpeptidase, alanine 132
aminotransaminase and aspartate aminotransaminase. 133
Spot urine samples were obtained at approximately 3 h supplement post feeding on 20th d 134
of each period by mechanical stimulation of the vulva. The urine samples were filtered and aliquots 135
of 10 mL were immediately diluted into 40 mL of 0.072 N H2SO4 and stored (-15°C) for 136
subsequent analysis of nitrogen, urea, allantoin, uric acid and creatinine. Immediately before the 137
analysis, the urine samples for each cow/period were thawed and centrifuged (2,300 x g, for 15 138
minutes, room temperature). The balance of nitrogenous compounds (N) was obtained by 139
calculating the difference of intake N, feces N, urine N and milk N. 140
Estimation of rumen microbial protein synthesis 141
The microbial protein synthesis in the rumen (MPS, kg/d) was calculated as a function of 142
purine absorbed (PA; mmol/d): 143
MPS (g/d) = [(70 × PA) / (0.83 × 0.116 × 1000] × 6.25 144
where: 70 is the N content in purines (mg N/mmol); 0.83 is the intestinal digestibility of microbial 145
purines microbial purine digestibility; and 0.116 is the purine-N:total bacterial-N 146
(Chen & Gomes, 1992). 147
16
The PA was calculated from total excretion of purine derivatives (EPD), where EPD is sum 148
of urinary allantoin and uric acid excreted, and allantoin secreted in milk, as: 149
EPD (mmol/d) = 0.85 × PA + 0.442 × BW0.75 150
where: 0.85 is efficiency of intestinal absortion of purines (Verbic et al., 1980) and 0.442 × BW0.75 151
the contribution to endogenous excretion of purines obtained from dairy cows (Oliveira, 2014). 152
Urine volume was estimated using creatinine concentration as a marker and assuming a daily 153
creatinine excretion of 24.05 mg/kg of BW (Chizzotti et al., 2008). 154
Chemical analysis 155
The GIs (glucosinolates) were extracted of the CM with methanol and quantified by high-156
performance liquid chromatography (C18 5 µm 250 mm x 4.6 mm I.D. column, from Vertical 157
Chromatography Co, Nonthaburi, Thailand). The GIs were detected at 233 nm and quantified 158
using sinigrin (Sinigrin hydrate, FLUKA Sigma, St. Louis MO) as an external standard (standard 159
curve: 1000, 500, 400, 300, 200, 100, 50 and 5 mg / L). The results were expressed as sinigrin 160
equivalents (mg / kg DM). The sum of the peak areas observed between 5 and 18 minutes were 161
considered as GIs total (Lee et al., 2006) 162
Samples of feeds concentrate, pasture, refusals, and feces were analyzed for the 163
concentrations of dry matter (DM method no. 934.01), organic matter (OM method no. 942.05), 164
crude protein (CP method no. 954.01), ether extract (EE, method no. 920.39) according to the 165
AOAC (1990). Neutral detergent fiber (NDF) was determined using heat stable amylase without 166
sodium sulfite and corrected for residual ash (Mertens, 2002) and N (Licitra et al., 1996) 167
(aNDFom), with an Ankom® fiber analyzer (Ankom technology, Fairport, NY). Neutral detergent 168
insoluble N (NDIN) was measured according to Licitra et al. (1996). The levels of non-fibrous 169
carbohydrates corrected for ash and protein (NFC) were calculated as: NFC = 100 - [(CP - CP 170
from urea + urea in the supplement) + aNDFom + EEt + ashes] (Hall 2000). Fecal titanium 171
concentration was obtained as Myers et al. (2004). Indigestible NDF of feeds, refusals and feces 172
17
was obtained after in situ ruminal incubation in polyester bag (Ankon®, Ankon®, filter bag 57, 173
Ankom technology, Fairport, NY, USA) for 264 h (Casali et al., 2008) 174
Analyses of blood plasma GGT, ALT and AST were performed by optimized ultraviolet 175
kinetic method (Schumann et al., 2002 a, b) using commercial kits (Labtest Diagnostica S.A., 176
Lagoa da Santa, Minas Gerais, Brazil). Urea in plasma and milk samples was measured by 177
enzymatic colorimetric assay, with commercial kits (Urea CE Ref. 27, Labtest Diagnostica S.A., 178
Lagoa da Santa, Minas Gerais, Brazil). Urinary uric in urine samples was determined using the 179
enzymatic-Trinder method with commercial kits (Ácido úrico Liquiform Ref. 73, Labtest 180
Diagnostica SA, Lagoa da Santa, Minas Gerais, Brazil). Allantoin in milk and urine samples was 181
determined colorimetrically as Young and Conway (1942). 182
In situ ruminal degradability 183
For in situ evaluation we used a half-breed cattle, cannulated in the rumen, weighing 500 184
kg and an average age of 36 months. The cattle fed on forage grass Panicum Maximum cv Massai 185
and concentrated feed containing corn, soybean meal and 300 g/kg crambe meal, which was given 186
2 kg of feed once a day in the morning (0800 am) following recommendations (Nocek, 1988). 187
Water and commercial mineral mixture (Matsuda ® 805 FOS) were administered ad libitum. To 188
determine the degradability were used in nylon bags measuring 10x20 cm, with porosity of 50 189
micrometers, which was added 4 g of dried sample (pasture, corn silage, corn, soy meal and crambe 190
meal) in air respecting 20 mg/cm2 (Nocek, 1988). The samples were incubated in the rumen, 191
sequentially, to be taken together at the end of the incubation period, being used times 0, 2, 4, 8, 192
16, 24, 36, 48 and 72 h (NRC, 2001). 193
Immediately after incubation in the rumen, the bags were immersed in cold water 194
(± 0 ° C) and then washed manually with tap water at room temperature until it was clear. The 195
soluble fractions (time zero of incubation) were determined by the same procedures, but without 196
ruminal incubation, washed only under running water. After washing the bags were taken to 197
18
dryness in forced ventilation oven at a temperature of 65 ° C for 72 h, then placed in a desiccator 198
for 30 minutes and weighed to determine the concentrations of DM 199
(method no. 934.01) and CP (method no. 954.01), according to the AOAC (1990). 200
The effective degradability (ED) of DM and CP were estimated using the asymptotic 201
growth model of the first order as Orskov and McDonald (1979) described by the function: Yt = a 202
+ b * (1-e -kd * t); wherein: Yt = fraction degraded at time "t" (%); a = soluble fraction (%); b = 203
potentially degradable insoluble fraction (%); b = kd degradation rate (h-1); and independent 204
variable t = time (h). 205
Cheese production and sensory analysis 206
Milk samples (1 L cow/period) were pasteurized by heating at 65 ° C for 30 minutes. After 207
cooling the milk to 37 ° C was added 0.4 ml of calcium chloride and 0.9 mL of rennet per 1 L of 208
milk. After 40 minutes standing, yielded the 'mass' of cheese was cut into cubes, remaining at rest 209
for 3 minutes to whey removal, the "mass" was moved slowly for 20 minutes. Was added 3.5% by 210
weight of milk by cheese sodium chloride (NaCl), which was added as 500 g and 24 h after turning 211
made. Thereafter the cheeses were unpacked, heavy, cut into small squares. After three days, about 212
¼ of the cheese was used to determine the crude protein content (N x 6.38) modified by the 213
Kjeldahl method (AOAC, 1995), moisture content by direct drying in an oven at 105 ° C, the rest 214
was used for sensory analysis. 215
The cheeses of each cow/period were subjected to sensory test with untrained consumers. 216
with an average of 10/cheese/period, totaling 30 reviews/level CM of inclusion. Each person 217
received a paper containing radio liked and disliked and guidance on how to perform the test. 218
Statistical analysis 219
Data were analyzed as three simultaneous 4 x 4 Latin squares, using a mixed model 220
(Littell et al., 1997) through the PROC MIXED procedure of SAS (SAS Institute, 1999-2000): 221
Yijkl = µ + Si + Pj + Ck(i) + Tl + STil + eijkl, 222
19
where: Yijkl is the dependent variable; µ is the overall mean; Si is the random effect of square i; Pj 223
is the random effect of period; Ck(i) is the random effect of cow k (within Latin square i); Tl is the 224
fixed effect of treatment l (four CM level on supplement); STil is random effect of interaction 225
between square I and treatment l; and eijkl = random error associated with each observation, 226
assumed NID (0, σ2). Linear, quadratic and cubic effect of CM level on pTMR were tested by 227
partitioning freedom degrees for diet into single freedom degrees variable corresponding to linear, 228
quadratic and cubic effects. Significance was declared at P ≤ 0.05. All reported values were as 229
least square means. 230
Cubic effects were not significant. Thus, the P-values for those components are not 231
presented in Tables 4–6 232
3 - Results 233
Crambe meal inclusion did not affect pTMR intake (P = 0.173), pasture intake (P = 0.185), 234
total DM intake (P = 0.481), OM intake (P = 0.420), EE intake (P = 0.216), CP intake (P = 0.481), 235
aNDFrom intake (P = 0.078) and TDN intake ((P = 0.262) (Table 4). In addition, CM inclusion 236
did not affect total-tract digestibility of the DM (P = 0.229), OM (P = 0.254), aNDFrom (P = 237
0.451), NFC (P = 0.255), TDN (P = 0.120), ruminal microbial protein (P = 0.338) and serum 238
concentrations of GGT (P = 0.529), ALT (P = 0.654) and AST (P = 0.432). Increased levels of 239
CM in pTMR linearly reduced NFC intake (P = 0.002) and it linearly increased EE digestibility 240
(P = 0.052) (Table 4). 241
Crambe meal inclusion in pTMR did not affect body weight (P = 0.379), body condition 242
score (P = 0.693), milk yield (P = 0.462) and milk lactose (P = 0.139), protein (P = 0.626) and fat 243
(P = 0.191) contents, and feed efficiency (P = 0.800) (Table 5). Crambe meal inclusion inclusion 244
did not affect cheese DM (P = 0.484) and CP (P = 0.847), cheese yield (P = 0.314) and cheese 245
preference for untrained consumers (P = 0.299) (Table 5). 246
20
Crambe meal inclusion did not affect Plasma urea-N (P = 0.258), milk urea-N (P = 0.178), 247
urinary N excretion (P = 0.717), N balance (P = 0.598) and N-efficiency to lactation (P = 0.10) 248
(Table 6). 249
4 - Discussion 250
The average stocking rate was 12.24 AU/ha and the total available forage mass was 251
8380.66 ± 669.50 kg DM/ha, composed of 2421.54 ± 595.24 kg of foliar blade/kg of DM, 4904.22 252
± 1011.82 kg of stalk/kg of MS (Figure 1). 253
Low-glucosinolates CM inclusion in pTMR (up to 135 g/kg DM) does not affect diet 254
intake, diet total-tract digestibility, ruminal microbial protein synthesis, hepatic function, milk 255
yield and composition, N dietary efficiency of crossbreed Holstein × Zebu cows, and cheese 256
preference for untrained consumers. Therefore, our hypothesis that CM is similar protein source 257
to SBM for lactating dairy cows was confirmed. 258
The absence of effects on intake and digestibility probably occurred due to low 259
glucosinolates content of CM used (450 mg/kg DM), and confirm our previous studies (Moura et 260
al., 2017). Brassicas meal are classified with low glucosinolates contents when they present values 261
between 2800 and 13000 mg/kg DM (Tripathi and Mishara, 2007, adapted). However, the 262
reduction in NFC intake can be explained by the increase in the inclusion of crambe meal and 263
reduction of corn and soybean meal in the diet, which resulted in lower starch content in diets with 264
a higher inclusion level of crambe meal. The causes of increase in EE digestibility with CM is 265
unclear, but as CM had high content of erucic acid (22:1 ω-9), its may have improve fatty acid 266
intestinal digestibility in relation SBM (rich in unsaturated C:18). 267
The absence of effects on ruminal microbial protein synthesis was due no CM effect no 268
intake and digestibility, that did not affect N and energy availability for ruminal microbial growth. 269
Therefore, absence of CM effects on milk production and milk composition is a response to intake, 270
21
digestibility and ruminal microbial protein synthesis, because energy and metabolizable protein 271
are main nutritional drivers of lactation (NRC, 2001). Absence of effects on the milk composition 272
and milk N- efficiency can be explained by the composition of the diets (isoproteic and energetic), 273
as well as the similarity of the amino acid profile of crambe meal, to soybean meal (Anderson et 274
al., 1993). 275
With rupture and contact of the enzyme myrosinase with glucosinolates, a hydrolysis 276
occurs through the cleavage of the glucose in its connection with the sulfur atom, releasing the 277
isothiocyanates, thiocyanates and nitriles, potentially causing liver damage and other organs 278
(Halkier and Gershenzan, 2006). The major enzymes for liver function indicators are gamma 279
glutamyltranspeptidase (GGT), alanine aminotransaminase (ALT), aspartate aminotransaminase 280
(AST). Although the observed values of ALT and AST were not influenced by diets, they were 281
outside the recommended reference range for species 22 to 28 IU/L for ALT, 60 to 280 IU/L for 282
AST (Radostites et al., 2002). These results were observed for all inclusion levels even for a diet 283
without inclusion of CM. In addition to levels, we must consider factors such as an animal species, 284
race, age, sex, breeding systems can interfere with levels, hepatic enzymes, without being 285
associated with pathological factors (Gregory, 1995). However, the GGT activity the mean values 286
of all treatments remained within the physiological standard (25-50 IU/L) reference values 287
observed by Gregory et al. (1999). 288
The effects found for composition and yield of cheeses can have occurred due to the 289
absence of effects on the milk composition of cows fed with CM inclusion. The absence of effects 290
on cheese preference indicate that CM with low glucosinolates level does not have chemical 291
composts that affect cheese sensorial characteristic in relation to typical diets with SBM. 292
293
22
5 - Conclusions 294
Crambe meal with low glucosinolates levels can be included in pTMR (up 135 g/kg DM) 295
without affects productive performance, efficiency of nutrient utilization in Jersey and Holstein × 296
Zebu crossbreed dairy cows with 132 kg of milk and cheese preference for untrained consumers. 297
298
Acknowledgments 299
Family Peron (Dairy Farm Dona Herminia) to provide the structure, Fundação de Amparo à 300
Pesquisa do Estado de Mato Grosso for financial support (FAPEMAT, project number 301
483724/2011 PRONEM 006/2011; Chair: André Soares de Oliveira), Caramuru Alimentos S.A 302
(Itumbiara, Goiás, Brazil) for providing crambe meal, Conselho Nacional de Desenvolvimento 303
Científico e Tecnológico (CNPq, Brazil) and Coordenação de Aperfeçoamento de Pessoal de Nível 304
Superior (CAPES; Brazil) for scholarships provided author D. C. Moura. 305
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410
411
412
413
414
27
415
416
Figure 1 - Available pasture mass of Panicum maximum cv grass Mombaça in the experimental 417
periods 418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
0.00
1000.00
2000.00
3000.00
4000.00
5000.00
6000.00
7000.00
8000.00
1st period 2nd period 3rd period 4th period
Pas
ture
Mas
s (K
g D
M/a
cre)
Stem
Foliar blade
28
433
434
435
436
Table 1. Chemical composition of Panicum maximum cv. Mombaça obtained by simulated grazing 437
and ingredients used in the partial total mixed ration supplements 438
Itensa Feedsa
Pasture CS CG SM CM
Dry matter (DM), g/kg 403.3 287.0 897.5 915.2 915.7
Organic matter, g/kg DM 924.1 956.8 984.1 930.9 928.0
Ether extract, g/kg DM 14.1 15.4 26.7 18.1 30.1
Crude protein, g/kg DM 90.4 66.1 92.2 510.0 381.8
NDIPb, g/kg N 276.2 212.9 75.6 30.5 159.6
ADIPc, g/kg N 54.5 80.2 19.2 26.2 30.1
aNDFomd , g/kg DM 563.5 532.4 96.1 114.6 333.5
Non-fibrous carbohydrates, g/kg DM 256.0 344.3 769.1 288.3 182.7
Lignin, g/kg DM 38.6 35.6 7.5 4.6 101.5
NDF indigestiblee, g/kg DM 128.2 153.0 17.2 7.7 262.6
Glucosinolatesf, mg/kg DM - - 450.0
aPasture; CS, corn silage; CM, corn grain ground; SM, soybean meal; CM, and crambe meal. 439 bNDIP, neutral detergent insoluble protein. 440 cADIP, acid detergent insoluble protein. 441 daNDFom, neutral detergent insoluble fiber corrected for ash and N. 442 eObtained after 264 hours in situ rumen incubation. 443 f sirigin equivalents 444 445
446
447
448
449
450
451
452
29
453
454
455
456
Table 2. Ingredients and chemical composition of the partial total mixed ration (pTMR) with 457
different inclusions of crambe meal. 458
Ingredients CM in pTMR (g/kg DM)
0 45 90 135
Ingredient composition, g/kg DM
Corn silage 700 700 700 700
Corn grain ground 183.0 166.5 150.0 133.5
Soybean meal 102.0 73.5 45.0 16.5
Crambe meal (CM) 0 45.0 90.0 135.0
Urea/ammonium sulfate 9:1 4.5 4.5 4.5 4.5
Mineral mixturea 10.5 10.5 10.5 10.5
Chemical composition
Dry matter (DM), g/kg as fed 459.0 459.0 459.0 459.0
Organic matter, g/kg DM 945.0 944.0 943.0 942.0
Ether extract, g/kg DM 17.0 18.0 18.0 19.0
Crude protein, g/kg DM 127 128 129 131
NDIPb, g/kg CP 100 116 132.0 148.0
ADIPc, g/kg CP 42.0 43.0 44.0 44.0
aNDFomd, g/kg DM 402.0 412.0 422.0 432.0
Non-fibrous carbohydrates, g/kg DM 406.0 393.0 380.0 368.0
NDF indigestiblee, g/kg DM 111.1 120.0 129.0 138.0
Lignin, g/kg DM 27.0 31.0 35.0 40.0
Rumen degradable protein, g/kg DM 118.0 121.0 123.0 126.0
Glucosinolatesf, mg/kg DM nd 20.25 40.50 60.75`
aCommercial mixture mineral (guarantee levels per kg of product: 160g of calcium; 80g of 459
phosphorus; 12g of sulfur; 114 g of sodium; 15g of magnesium; 100mg of cobalt; 700mg of 460
copper; 100 mg of iodine; 15g of manganese; 30 mg of selenium; 2500mg of zinc; 3000mg (max) 461
fluorine). 462 bNDIP, neutral detergent insoluble protein CP. 463
30
cADIP, acid detergent insoluble protein CP. 464 daNDFom, neutral detergent insoluble fiber corrected for ash and N. 465 eObtained after 288 hours in situ rumen incubation. 466 f equivalent sirigin 467
Table 3. Effect crambe meal (CM) inclusion in partial total mixed ration (pTMR) on intake, total-468
tract digestibility and ruminal microbial protein synthesis (MPS) of grazing dairy cows 469
Item CM in pTMR (g/kg DM)
SEDb P-valuec
0 45 90 135 L Q
Intake, kg/day
pTMR dry matter (DM) 11.68 11.60 11.30 11.31 0.679 0.173 0.848
Pasture 0.835 0.856 0.914 0.992 0.272 0.185 0.726
Total DM 12.51 12.46 12.22 12.33 0.8062 0.481 0.729
Organic matter 11.91 11.85 11.61 11.71 0.765 0.420 0.734
Ether extract 0.203 0.210 0.216 0.212 0.012 0.216 0.325
Crude protein 1.48 1.49 1.50 1.50 0.056 0.481 0.627
aNDFoma 5.35 5.41 5.42 5.71 0.414 0.078 0.353
Non-fibrous carbohydrates 4.83 4.69 4.43 4.29 0.278 0.002 0.951
Total digestible nutrient 9.14 9.01 8.80 9.34 0.617 0.754 0.262
pTMR intake:total DM, g/g 0.933 0.934 0.927 0.920 0.020 0.1349 0.547
Digestibility, g/g
Dry matter 0.715 0.717 0.715 0.746 0.016 0.229 0.389
Organic matter 0.740 0.745 0.746 0.767 0.015 0.254 0.609
Ether extract 0.756 0.733 0.782 0.818 0.023 0.052 0.224
Crude protein 0.735 0.740 0.750 0.773 0.017 0.094 0.545
aNDFoma 0.726 0.720 0.726 0.744 0.018 0.451 0.483
Non-fibrous carbohydrates (NFC) 0.756 0.766 0.760 0.789 0.017 0.255 0.578
Total digestible nutrient, g/g DM 0.782 0.768 0.780 0.815 0.020 0.120 0.121
MPS, kg / dia 0.880 0.771 0.735 0.922 0.217 0.887 0.338
g MPS / kg TDN intake 99.82 90.22 90.14 99.13 20.76 0.978 0.621
Plasma, IU/mL
Gamma glutamyl transpeptidase 32.47 30.52 35.13 31.18 5.967 0.930 0.592
Alanine aminotransaminase 16.40 15.52 16.40 15.34 2.125 0.654 0.937
31
Aspartate aminotransaminase 50.33 46.58 47.41 44.64 9.458 0.432 0.913 aaNDFom, neutral detergent insoluble fibre corrected for ash and N. 470 bSED = standard error of the least squares means. 471 cProbability of linear (L) or quadratic (Q) effect of CM level in pTMR. 472 473 Table 5. Effect of the crambe meal (CM) inclusion in partial total mixed ration (pTMR) on 474
productive performance of grazing dairy cows 475
Item
CM in pTMR
(g/kg DM) SEDa P-valueb
0 45 90 135 L Q
Body weigth, kg 483.52 466.52 475.91 471.40 49.64 0.399 0.379
Body condition score (1 a 5) 2.78 2.77 2.76 2.80 0.253 0.833 0.693
Milk yield, kg/d 13.16 13.63 13.10 13.25 0.849 0.793 0.462
4.0% fat correted milk, kg/d 14.25 14.65 13.70 14.24 0.887 0.563 0.852
Milk lactose, g/kg 48.16 47.78 47.44 47.49 0.637 0.139 0.516
Milk crude protein, g/kg 30.97 30.86 30.77 30.90 0.410 0.778 0.626
Milk fat, g/kg 45.81 45.13 43.67 45.07 3.096 0.302 0.191
Milk lactose, kg/d 0.634 0.651 0.617 0.628 0.038 0.874 0.113
Milk crude protein, kg/d 0.407 0.420 0.402 0.406 0.024 0.550 0.573
Milk fat, kg/d 0.600 0.614 0.565 0.594 0.040 0.531 0.770
Feed efficiency, kg/kg 1.15 1.18 1.14 1.16 0.078 0.853 0.800
Yield and sensory analysis the cheese
Dry matter (DM), g/kg 35.96 36.34 36.92 35.94 1.668 0.903 0.484
Crude protein kg/d 5.70 5.56 5.74 5.72 0.733 0.874 0.847
Yield kg / kg the milk 0.227 0.222 0.216 0.208 0.021 0.314 0.928
Like, % consumers 59.78 80.00 76.73 77.68 9.113 0.239 0.299
Disliked, % consumers 40.22 20.00 23.27 22.32 9.113 0.239 0.299 aSED = standard error of the least squares means. 476 bProbability of linear (L) or quadratic (Q) effect of CM level in pTMR. 477
478
479
480
481
482
32
483
484
485
Table 6. Effect of the crambe meal (CM) inclusion in partial total mixed ration (pTMR) on nitrogen 486
(N) efficiency of grazing dairy cows 487
Item
CM in Ptmr
(g/kg DM) SEDa P-valueb
0 45 90 135 L Q
Plasm urea-N, mg/dL 10.98 10.80 11.68 10.75 0.370 0.889 0.258
Milk urea-N, mg/dL 11.86 10.18 10.86 9.70 2.103 0.178 0.767
Urinary urea-N, g/d 20.08 24.68 25.50 28.58 11.15 0.587 0.943
Urinary urea-N:total N, g/g 0.519 0.466 0.474 0.351 0.082 0.169 0.646
Intake N, g/d 236.60 239.19 240.64 240.01 8.952 0.481 0.627
Feces N, g/d 62.30 62.50 60.13 54.37 5.083 0.195 0.481
Urinary N, g/d 91.70 89.27 89.40 93.30 11.53 0.899 0.717
Milk N, g/d 63.94 65.95 62.91 63.91 3.914 0.566 0.673
Balance N, g/d 20.09 24.68 25.50 28.58 11.16 0.587 0.943
N efficiency, g/g intake N
Feces N 0.264 0.259 0.254 0.222 0.018 0.100 0.405
Urine N 0.383 0.366 0.368 0.389 0.041 0.914 0.622
Milk N 0.271 0.274 0.261 0.264 0.013 0.100 0.920
Balance N 0.084 0.106 0.112 0.117 0.046 0.598 0.856
aSED = standard error of the least squares means. 488 bProbability of linear (L) or quadratic (Q) effect of CM level in pTMR. 489
490
491
492
493
494
495 496
497 498 499
33
Meta-analysis of the use of brassicas meals in diets for dairy cows 500
501
ABSTRACT: We used meta-analytical approach to evaluate the effects of replacement of 502
different sources of protein by brassicas meals on milk production and nutrient utilization of 503
dairy cows, from 37 peer-review papers. Canola meal (CM) was unique brassica source 504
founded. The effects were compared by raw mean differences (RMD) between CM diet and 505
control treatment means and weighted by inverse variance using random-effect models. Control 506
protein sources (28.6 ± 6.9 kg milk/d) were soybean meal (SBM; 57%), dried destillers grains 507
(DDG; 21%), corn gluten meal (7%), cottonseed meal (5%), sunflower meal (2%) and other 508
protein sources (9%). Heterogeneity level was analyzed by I2 statistic (low ≤ 25%; moderate = 509
26 to 50%; and high > 50%). In overall, use of CM as protein source increased DM intake 510
(RMD = 0,22 ± 0.12 kg DM/d; P < 0.01; n = 79; I2 = 9.1%) and crude protein (CP) intake 511
(RMD = 0,14 ± 0.07 kg CP/d; P < 0.01; n = 33; I2 = 21.1%), both with low heterogeneity, but 512
it did not affect organic matter total-tract digestibility (P = 0.50; n = 12; I2 = 29.2%). In overall, 513
use of CM increased milk yield (RMD = 0.69 ± 0.35 kg/d; P < 0.01; n = 88; I2 = 74.9%), but 514
its effect depends on protein sources comparation: CM versus SBM did not increase milk yield 515
(RMD = 0.23 ± 0.66 kg/d; P = 0.50; n = 33), but milk yield was increased with replacement of 516
DDG by CM (RMD = 2.03 ± 1.67 kg/d; P < 0.01; n = 13) and of other protein sources by CM 517
(RMD = 0.82 ± 0.43 kg/d; P < 0.01; n = 42). In overall, CM use did not affect milk protein 518
content (P = 0.08; n = 60; I2 = 19.5%) and milk fat content (P = 0.20; n = 60; I2 = 16.9%), but 519
CM increased milk protein yield (RMD = 0,02 ± 0.01 kg/d; P < 0.01; n = 60; I2 = 0%). Use of 520
CM reduced milk urea nitrogen (N) (RMD = - 0,98 ± 0.31 mg/dL; P < 0.01; n = 22; I2 = 32.2%) 521
and increase N intake milk efficiency (RMD = 0.22% N milk/N intake ± 0.07 mg/dL; P ≤ 0.05; 522
n = 34; I2 = 0%), both with low heterogeneity. We concluded that CM is similar protein source 523
34
to SBM and it is more effective than DDG and other sources (cottonseed meal, corn gluten meal 524
and sunflower meal) to lactating dairy cows. 525
Key words: canola meal, effect sizes, heterogeneity 526
INTRODUCTION 527
528 Brassicaceae is composed of 51 genera and 218 species, which some are important 529
species of crops cultivated as vegetables, forages and oilseeds (Sabharwal et al., 2006). After 530
oilseed extraction are produced products with potential of use in the animal as protein source. 531
Colza, canola, crambe and carinata are main brassicas meals used in animal nutrition, but its 532
use depends of glucosinolates levels (Tripathi and Mishra, 2007; Moura et al., 2017). 533
Glucosinolates are goitrogenic compounds that reduce the availability of iodine to the 534
animal and diminish the synthesis of thyroxine (Grenet and Journet, 1971). A lower intake and 535
animal performance were also reported in diets containing brassica meal with high 536
glucosinolates levels (Papas et al., 1978; Thomke, 1981; Tripathi et al., 2001). Therefore, only 537
lower glucosinolates brassicas meals (< 30 μmol/g oil-free meal) are considered a suitable 538
feedstuff for livestock (Tripathi and Mishra, 2007). 539
Canola is a term specifically to identify colza varieties containing less than 2% erucic 540
acid in the oil portion and lower glucosinolates level on meals (Bell, 1984; Newkirk, 2009). 541
Canadian plant breeders were the first to develop canola cultivars in 1970’s year 542
(Stefansson and Kondra, 1975). Today, canola meal is the second oilseed meal consumed in the 543
world (12% market share), below only of the soybean meal (71% market share) 544
(SoyStats, 2018). 545
Have been reported that canola meal (CM) was similar to soybean meal (SBM; Brito 546
and Broderick, 2007; Huhtanen et al., 2011; Matineau et al., 2013), but more effective protein 547
source to lactating dairy cows that cottonseed meal (Brito and Broderick, 2007) and dry 548
35
distillated grain (DDG; Swanepoel et al., 2014), due differences on protein intestinal 549
digestibility and first-limiting AA profile to milk production (i.e. Lys, Met and His). In 550
previous meta-analysis were confirmed that replacement of SBM by CM did not affect milk 551
yield (Huhtanen et al., 2011; Matineau et al., 2013), but replacement of other sources 552
(not specified) improved milk production (Matineau et al., 2013). However, is not clear yet 553
which protein source the CM may be more effective to milk production. 554
We hypothesis is that CM use in replacement to SBM does not affect dairy cows 555
performance, but its use in replacement to DDG, cottonseed meal and other oilseeds meal 556
reduces milk production of dairy cows. Therefore, we used a meta-analytical approach to 557
evaluate the effects of replacing of different sources of protein by brassicas meals on milk 558
production and nutrient utilization by dairy cows. 559
560
MATERIALS AND METHODS 561
Literature Search 562
Papers were identified by searching for peer-reviewed manuscripts that were published 563
in English using online manuscript retrieval databases [Web of Science 564
(https://login.webofknowledge.com), PubMed (https://www.ncbi.nlm.nih.gov/pubmed), 565
Google Scholar (http://www.scholar.google.com/), ScienceDirect 566
(http://www.sciencedirect.com/)]. Up to 274 publications were retrieved using search terms 567
including “brassica,” “brassica aleracea,” “brassica rapa” “canola meal,” “rapessed meal,” 568
“ crambe meal,” “canerita meal,” “dairy cows. Of the papers that were retrieved, only those that 569
satisfied the predetermined inclusion criteria were included in the analysis. 570
Inclusion Criteria 571
A flowchart explaining the process of study identification and selection for analyzing 572
the effects of replacement of different protein sources by brassica meal is shown in Figure 1. 573
36
The study selection criteria were (1) publication in English in a peer-reviewed journal, (2) use 574
of concurrent negative control and treatment groups, (3) use of lactating dairy cows, (4) use of 575
at least one Brassica meal as a dietary treatment, (5) presentation of least squares means and 576
standard errors of the means for DMI our milk yield. The study exclusion criteria were: (1) did 577
not involve feeding a TMR, (2) involved a study that was not randomized. We only included 578
peer-reviewed publications in the study because the peer review process is a proxy for assessing 579
the quality of studies (Weisz et al., 1995). 580
Data Extraction 581
Based on the inclusion criteria, 37 articles were reviewed by selected pairs and ranked 582
by the first author, publication reference, genetic type, forage in diet (% DM), brassica meal 583
type, glucosinolates mg/kg DM, replaced culture, days in milk (begin experiment), number 584
cows, experimental design, type of diet (pasture, TMR, partial TMR) , days experiment, diet 585
composition, milk yield of cows, number of replicates, significance, and variances (SE, SD) 586
were extracted for the following response variables for control and brassicas treatments: 587
nutrient intake, diet total-tract digestibility, feed efficiency, milk yield, milk composition and 588
nitrogen metabolism. 589
Statistical Analysis 590
Meta-analysis was conducted using the metafor package of R Software 591
(Viechtbauer, 2010) version 3.2.3. The effects of different sources of protein by brassica meal 592
were evaluated by examining the raw mean differences (RMD) between brassica meal and 593
inoculated treatment means (effect size), which were weighted by the inverse of the variance in 594
the respective studies using the method proposed by DerSimonian and Laird (1986) for a 595
random effect model. When were not reported sufficient information to use variance 596
(i.e SEM or SD), we weighted the observations by replication number per treatment. 597
37
Between-study variability (i.e., heterogeneity of effect size) was evaluated using the chi-598
squared (Q) test and the I2 statistic, which measures the percentage of variation due to 599
heterogeneity (Higgins et al., 2003). An I2 value less than 25% indicated low heterogeneity, 600
whereas values between 25 to 50% denoted moderate heterogeneity and those above 50 high 601
heterogeneity (Higgins et al., 2003). Publication bias was assessed using funnel plots 602
(Light and Pillemer, 1984) and was tested for funnel plot asymmetry 603
(indicative of publication bias) by Egger’s regression method between RMD and SE (Egger et 604
al., 1997). We adopted a level of P = 0.05 of probability for significance and P = 0.10 of 605
probability for trend. 606
607
RESULTS AND DISCUSSION 608
We founded only CM as brassica source used in diets to lactating dairy cows. Therefore, 609
we evaluated only the effects of CM on milk production and nutrient utilization of dairy cows. 610
Raw mean differences to feed efficiency and N-milk efficiency were weight by replication 611
number per treatment and the others response variables by inverse of the variance. Protein 612
source of the control treatments were soybean meal (57%), dried destillers grains (21%), corn 613
gluten meal (7%), cottonseed meal (5%), sunflower meal (2%) and other protein sources (9%). 614
Experimental design use in the studies were Latin square (53.3%), randomized block (27.8%), 615
changeover (14.4%) and not reported experimental design (4.4%). Total mixed ration was uses 616
in 96.2% of studies. 617
Overall, the replacement of several protein sources for CM did not, organic matter (OM) 618
total-tract digestibility (P = 0.50), ruminal pH (P = 0.50) and propionate concentration 619
(P = 0.36) (Table 1). However, CM use in comparation with all protein sources evaluated 620
reduced CP total-tract digestibility (P < 0.01) and it increased (P < 0.01) DM, OM, CP and 621
NDF intakes, and ruminal acetate concentration (P = 0.03) (Table 1). Overall, the replacement 622
38
of several protein sources for CM increased milk yield (P < 0.01), milk protein yield (P < 0.01), 623
tended to increase protein milk content (P = 0.08), but it did not affect milk fat content 624
(P = 0.20) and yield (P = 0.24), and feed efficiency (P > 0.05). As result of the increase in milk 625
yield and N dietary efficiency, CM use in comparation with all protein sources evaluated 626
reduced milk urea-N (P < 0.01) and increased milk N efficiency (P < 0.05) (Table 1). 627
Low and medium heterogeneity (I2 statistic < 25 and 26-50%) were observed for all 628
response variables, except for milk yield (I2 = 74.5%) (Table 1). There was no evidence of 629
funnel plot asymmetry (P > 0.10) for any response (Figure 2), indicating that publication bias 630
was not evident. We identified that protein source used to compare with CM was a main 631
heterogeneity source to milk yield response. Replacement of SBM by CM did not increase milk 632
yield (RMD = 0.23 ± 0.66 kg/d; P = 0.50; n = 33; Figure 3), but milk yield was increased with 633
replacement of DDG by CM (RMD = 2.03 ± 1.67 kg/d; P < 0.01; n = 13; Figure 4) and of other 634
protein sources (cottonseed meal, corn gluten meal and sunflower meal) by CM 635
(RMD = 0.82 ± 0.43 kg/d; P < 0.01; n = 42; Figure 5). 636
We confirmed our hypothesis that CM use in replacement to SBM does not affect dairy 637
cows performance, but its use in replacement to DDG, cottonseed meal and other oilseeds meal 638
reduces milk production of dairy cows. Similar milk production response of CM with SBM 639
may be attributed to similar ruminal protein degradation, intestinal digestibility and/or better 640
first limiting essential amino acids (EAA) profile to lactating (i.e. Lys, Met and His) (NRC, 641
2001). There is evidence that DGG in diet reduced intestinal digestibility and N retention in 642
ruminant, due higher ADIN (acid detergent insoluble nitrogen) and lower Lys than SBM 643
(Mjoun et al., 2010; Stotzer, 2017). As CM has similar protein value that DDG, its suggest that 644
CM has higher intestinal digestibility or Lys content than DDG. Similarly, higher milk response 645
of the CM versus other sources (cottonseed meal, corn gluten meal and sunflower meal) may 646
39
be attributed high ruminal protein degradation, intestinal digestibility and/or better first limiting 647
EAA profile to lactation (i.e. Lys, Met and His). 648
Although we identified reduction on CP total-tract digestibility with CM versus all 649
sources, the reduction on MUN and increase on N-milk efficiency suggest that CM increase 650
anabolic use of AA to lactating and reduce AA losses by catabolism process. In the meta-651
analysis of Huhtanen et al. (2011) and Martineau et al. (2013) also reported positive responses 652
to DMI when CM replaced other dietary proteins (mainly SBM). The increase in DMI can be 653
attributed to the increase nutrient demand, where the more balanced supply of AA increases 654
lactation performance and, consequently, increases energy demand, which stimulates feed 655
intake (NRC, 2001). The increase in rumen acetate concentration when CM replaced other 656
protein sources can be due increase of NDF intake. 657
CONCLUSIONS 658
Canola meal is a similar protein source compared to SBM to lactating dairy cows. 659
However, CM is more effective than DDG and other sources (cottonseed meal, corn gluten 660
meal and sunflower meal) to lactating dairy cows because it increases milk yield and N 661
efficiency to lactation. Possibly this increase in milk production is more associated with an 662
improvement in the balance between AA and energy, but is necessary to elucidate the causes 663
of higher N efficiency to lactating of the CM to compare DDG and other protein sources. 664
665
ACKNOWLEDGMENTS 666
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil) and 667
Coordenação de Aperfeçoamento de Pessoal de Nível Superior (CAPES; Brazil) for 668
scholarships provided author D. C. Moura. Núcleo de Pesquisa em Pecuária de Leite 669
40
responsibility A. S. Oliveira (Universidade Federal de Mato Grosso, Campus Universitário de 670
Sinop) 671
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675 Brito, A. F., G. A. Broderick, and S. M. Reynal. 2007. Effects of different protein supplements 676
on omasal nutrient flow and microbial protein synthesis in lactating dairy cows. J. Dairy 677 Sci. 90:1828–1841. 678
679 DerSimonian, R., and N. Laird. 1986. Meta-analysis in clinical trials. Control. Clin. Trials 680
7:177–188. 681 682 Egger, M., G.D. Smith, M. Schneider, and C. Minder. 1997. Bias in meta-analysis detected by 683
a simple, graphical test. BMJ 315:629– 634. 684 685 Grenet, N., and M. Journet. 1971. Rapeseed oil meal in animal feeding. III. Influence of the 686
processing method and the proportion of oil meal in concentrate fed to dairy cows on feed 687 intake, milk production and composition. Ann. Zootech. 20:437–449. 688
689 Higgins, J. P. T., S. G. Thompsom, J. J. Deeks, and D. G. Altman. 2003. Measuring 690
inconsistency in meta-analysis. BMJ 327:557–560. 691 692 Huhtanen, P., M. Hetta, and C. Swensson. 2011. Evaluation of canola meal as a protein 693
supplement for dairy cows: A review and a metaanalysis. Can. J. Anim. Sci. 91:529–543. 694 695 Light, R. J., and D. B. Pillemer. 1984. Summing Up: The Science of Reviewing Research. 696
Harvard University Press, Cambridge, MA. 697 698 Martineau, R., D. R. Ouellet, and H. Lapierre. 2013. Feeding canola meal to dairy cows: A 699
meta-analysis on lactational responses. J. Dairy Sci. 96:1701–1714 700 701 Mjoun, K., K. F. Kalscheur ,1 A. R. Hippen, and D. J. Schingoethe. 2010. Ruminal 702
degradability and intestinal digestibility of protein and amino acids in soybean and corn 703 distillers grains products. J Dairy Sci. 93:4144-4154. 704
705 Moura, D. C., T. S. Fonseca, S. R. Soares, H. M. Silva, F. J. G. Vieira, L. A. Botini, A. P. 706
Sinhorin, I. M. Ogunaded and A. S. Oliveira. 2017. Crambe meal subjected to chemical and 707 physical treatments in sheep feeding. Livest Sci. 203:136-140. 708
709 Newkirk, R. 2009. Canola Meal: Feed Industry Guide. 4th ed. Canadian International Grains 710
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756 757 758 759 760 761 762 763 764 765
42
Table 1. Effect replacement of several protein sources for canola meal on the performance and 766 metabolism of dairy cows 767
Item Control1
mean (SD) N2
RMD3 (95% CI) Heterogeneity4
Random effect P-value P-value I2 (%)
DMI, kg/d 20.94 (3.38) 79 0.22 (0.09, 0.34) <0.01 0.25 9.06
OM intake, kg/d 18.66 (2.61) 12 0.56 (0.23, 0.90) <0.01 0.23 21.74
CP intake, kg/d 3.36 (0.46) 10 0.14 (0.08, 0.21) <0.01 0.08 41.02
NDF intake, kg/d 7.52 (0.82) 14 0.40 (0.28, 0.52) <0.01 0.13 30.90
DM digestibility, % 62.04 (19.54) 20 -0.45 (-0.96, 0.05) 0.08 0.06 35.00
OM digestibility, % 72.27 (3.82) 12 0.20 (0.38, 0.78) 0.50 0.16 29.23
CP digestibility, % 62.17 (21.87) 12 -1.44 (-2.03, -0.86) <0.01 0.33 11.35
NDF digestibility, % 49.57 (12.52) 12 3.20 (2.34, 4.07) <0.01 0.81 0.0
Acetate, mmol/L 59.30 (6.29) 19 0.53 (0.05, 1.01) 0.03 0.86 0.0
Propionate, mmol/L 18.58 (1.98) 19 0.18 (-0.20, 0.57) 0.36 0.85 0.0
pH ruminal 6.38 (0.27) 18 0.02 (-0.06, 0.01) 0.23 0.19 21.96
Milk yield, kg/d 28.61 (6.95) 88 0.69 (0.32, 1.06) <0.01 <0.01 74.95
Milk yield 4%, kg/d 26.09 (8.13) 60 0.82(0.58, 1.05) <0.01 0.54 0.0
Feed efficiency 1.38 (0.19) 79 0.001 (0.00, 0.002) >0.05 0.80 0.0
Milk fat, % 3.78 (0.61) 67 0.03 (-0.07, 0.01) 0.20 0.12 16.86
Milk fat yield, kg/d 1.06 (0.29) 66 0.008 (-0.005, 0.02) 0.24 0.21 11.60
Milk protein, % 3.29 (0.24) 74 -0.01 (-0.03. 0.00) 0.08 0.08 19.52
Milk protein yield, kg/d 0.93 (0.21) 49 0.02 (0.007, 0.02) <0.01 1.00 0.0
Milk lactose, % 4.80 (0.20) 52 0.0006 (-0.01, 0.01) 0.93 0.86 0.0
Milk lactose yield, kg/d 1.45 (0.34) 45 0.02 (0.01, 0.05) <0.01 0.99 0.0
MUN, mg / dL 11.78 (2.75) 22 -0.98 (-1.29, -0.67) <0.01 0.07 32.27
N efficiency5, % 28.12 (4.88) 34 0.22 (0.20, 0.25) <0.05 0.70 0.00 2N = number of comparisons between canola meal and all different protein sources (complete data set is available 768
in Supplementary file S1). 769 3RMD = raw mean difference between brassica meal and different protein sources. 770 4I2 = proportion of total variation of size effect estimates that is due to heterogeneity; P-value to χ2 (Q) test of 771
heterogeneity. 772 5 Milk N intake in proportion of N intake. 773
774
775
776 777 778 779
43
780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804
805 806 807 808 809 810 811 812 Figure 1. Flow diagram showing the inclusion criteria for selection of the studies used to 813 perform the meta-analysis of the effects of substitution of different sources of protein by 814 brassica meal. 815 816 817 818 819 820 821 822 823 824
Records identified by prior search WEB of SCIENCE
“Dairy Cow, “Canola meal”, “Crambe meal”, “Brassica”, “Brassica aleracea”, “Brassica rapessed”, “Rapessed meal” and “Canerita
meal” (n = 274)
Records automatically excluded for “duplicate” and “no article (n = 81)
Records after excluded for “duplicate” and “no article (n = 193)
Records manually excluded (n =156)
Studies include in meta-analysis to performance and efficiency utilization
(n = 37)
44
825
826
827 828
Figure 2. Funnel plots showing the effects of replacing different sources of protein for brassica 829
meal on the performance of dairy cows. The horizontal line indicates the raw mean difference 830
(RMD) estimate and the vertical line indicates their corresponding standard errors (SE). P-value 831
refers to the test for funnel plot asymmetry by Egger`s regression method between RMD and 832
SE. Funnel plot asymmetry is indicative of publication bias. 833
834
835
836
837
Milk Fat (%) Milk Protein (%)
Milk yield (kg/d)
DMI (kg/d)
P = 0.52 n = 79
P = 0.10 n = 88
P = 0.67 n = 67
P = 0.99 n = 74
DM digestibility (%)
P = 0.86 n = 20
45
838
Figure 3. Forest plot showing the effects of replacement soybean meal by canola meal on milk 839
yield (kg/d) of dairy cows. The x-axis shows the raw mean difference (RMD); diamonds to the 840
left of the solid line represent a reduction in the measure, whereas diamonds to the right of the 841
line indicate an increase. Each diamond represents the mean size effect for that study, and the 842
size of the diamond reflects the relative weighting of the study to the overall size effect estimate 843
with larger diamonds representing greater weight. The lines connected to the diamond 844
represents the upper and lower 95% confidence interval for the size effect. The dotted vertical 845
line represents the overall size effect estimate. The diamond at the bottom represents the mean 846
response across the studies, and the solid vertical line represents a mean difference of zero or 847
no effect. 848
849
Overall (I2 =74.9%, P = 0.50)
RMD, kg/d
100% 0.23 [-0.42, 0.87]
46
850
851
Figure 4. Forest plot showing the effects of replacement dry distillated grains by canola meal 852
on milk yield (kg/d) of dairy cows. The x-axis shows the raw mean difference (RMD); 853
diamonds to the left of the solid line represent a reduction in the measure, whereas diamonds to 854
the right of the line indicate an increase. Each diamond represents the mean size effect for that 855
study, and the size of the diamond reflects the relative weighting of the study to the overall size 856
effect estimate with larger diamonds representing greater weight. The lines connected to the 857
diamond represents the upper and lower 95% confidence interval for the size effect. The dotted 858
vertical line represents the overall size effect estimate. The diamond at the bottom represents 859
the mean response across the studies, and the solid vertical line represents a mean difference of 860
zero or no effect. 861
862
863
864
865
Overall (I2 = 18.0%, P < 0.01)
RMD, kg/d
47
866
867
Figure 5. Forest plot showing the effects of replacement other protein sources (cottonseed meal, 868
corn gluten meal and sunflower meal) grain by canola meal on milk yield (kg/d) of dairy cows. 869
The x-axis shows the raw mean difference (RMD); diamonds to the left of the solid line 870
represent a reduction in the measure, whereas diamonds to the right of the line indicate an 871
increase. Each diamond represents the mean size effect for that study, and the size of the 872
diamond reflects the relative weighting of the study to the overall size effect estimate with larger 873
diamonds representing greater weight. The lines connected to the diamond represents the upper 874
and lower 95% confidence interval for the size effect. The dotted vertical line represents the 875
overall size effect estimate. The diamond at the bottom represents the mean response across the 876
studies, and the solid vertical line represents a mean difference of zero or no effect. 877
Overall (I2 = 67.0%, P < 0.01)
RMD, kg/d
100% 0.82 [0.39, 1.24]
48
Appendix. List of references used in meta-analysis 878
879
Acharya, I. P., D. J. Schingoethe, K. F. Kalscheur, and D. P. Casper. 2015. Response of lactating 880 dairy cows to dietary protein from canola meal or distillers' grains on dry matter intake, 881 milk production, milk composition, and amino acid status. Can. J. Anim. 95:267-279. 882
883 Brito, A. F. and G. A. Broderick. 2007. Effects of Different Protein Supplements on Milk 884
Production and Nutrient Utilization in Lactating Dairy Cows1. J. Dairy Sci. 90:1816-1827. 885 886 Broderick, G. A., A. P. Faciola, and L. E. Armentano. 2015. Replacing dietary soybean meal 887
with canola meal improves production and efficiency of lactating dairy cows. J. Dairy Sci. 888 98:5672-5687. 889
890 Choi, C. W., A. Vanhatalo, S. Ahvenjärvi, and P. Huhtanen. 2002. Effects of several protein 891
supplements on flow of soluble non-ammonia nitrogen from the forestomach and milk 892 production in dairy cows. Anim. Feed Sci. Technol. 102:15-33. 893
894 Christen, K. A., D. J. Schingoethe, K. F. Kalscheur, A. R. Hippen, K. K. Karges, and M. L. 895
Gibson. 2010. Response of lactating dairy cows to high protein distillers grains or 3 other 896 protein supplements. J. Dairy Sci. 93:2095-2104. 897
898 DePeters, E. J. and D. L. Bath. 1986. Canola Meal Versus Cottonseed Meal as the Protein 899
Supplement in Dairy Diets. J. Dairy Sci.69:148-154. 900 901 Emanuelson, M., K. Å. Ahlin, and H. Wiktorsson. 1993. Long-term feeding of rapeseed meal 902
and full-fat rapeseed of double low cultivars to dairy cows. Livest Sci. 33:199-214. 903 904 Fisher, L. J. and D. S. Walsh. 1976. Substitution of rapeseed meal for soybean meal as a source 905
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911 Gidlund, H., M. Hetta, S. J. Krizsan, S. Lemosquet, and P. Huhtanen. 2015. Effects of soybean 912
meal or canola meal on milk production and methane emissions in lactating dairy cows fed 913 grass silage-based diets. J. Dairy Sci.98:8093-8106. 914
915 Ingalls, J. R. and H. R. Sharma. 1975. Feeding of Bronowski, Span and commercial rapeseed 916
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923 Kokkonen, T., M. Tuori, V. Leivonen, and L. Syrjälä-Qvist. 2000. Effect of silage dry matter 924
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49
Kokkonen, T. M. Tuori and L. SyrjaÈlaÈ-Qvist. 2000.Effect of silage dry matter content and 927 rapeseed meal supplementation on dairy cows 2. Rumen fermentation and digesta passage rate. 928 Anim. Feed Sci. Technol. 84: 229-242. 929 930 Laarveld, B. and D. A. Christensen. 1976. Rapeseed Meal in Complete Feeds for Dairy Cows. 931
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937 Maiga, H. A., D. M. Harris, M. E. Meyer, C. R. Dahlen, and M. L. Bauer. 2011. Effect of 938
pelleted high-oil canola meal from on-farm biodiesel production on rumen fermentation in 939 lactating Holstein dairy cows. Prof. Anim. Sci. 27:29-34. 940
941 Maxin, G., D. R. Ouellet, and H. Lapierre. 2013. Effect of substitution of soybean meal by 942
canola meal or distillers grains in dairy rations on amino acid and glucose availability. J. 943 Dairy Sci. 96:7806-7817. 944
945 McClean, C. and B. Laarveld. 1991. Effect of somatotropin and protein supplement on thyroid 946
function of dairy cattle. Can. J. Anim. 71:1053-1061. 947 948 McDonnell, R. P. and M. V. Staines. 2017. Replacing wheat with canola meal and maize grain 949
in the diet of lactating dairy cows: Feed intake, milk production and cow condition 950 responses. J. Dairy Sci. 84:240-247. 951
952 Moate, P. J., S. R. O. Williams, C. Grainger, M. C. Hannah, E. N. Ponnampalam, and R. J. 953
Eckard. 2011. Influence of cold-pressed canola, brewers grains and hominy meal as dietary 954 supplements suitable for reducing enteric methane emissions from lactating dairy cows. 955 Anim. Feed Sci. Technol. 166–167:254-264. 956
957 Mulrooney, C. N., D. J. Schingoethe, K. F. Kalscheur, and A. R. Hippen. 2009. Canola meal 958
replacing distillers grains with solubles for lactating dairy cows. J. Dairy Sci. 92:5669-5676. 959 960 Murphy, J. J., J. F. Connolly, and G. P. McNeill. 1995. Effects on cow performance and milk 961
fat composition of feeding full fat soyabeans and rapeseeds to dairy cows at pasture. Livest. 962 Prod. Sci. 44:13-25. 963
964 Oba, M., G. B. Penner, T. D. Whyte, and K. Wierenga. 2010. Effects of feeding triticale dried 965
distillers grains plus solubles as a nitrogen source on productivity of lactating dairy cows. 966 J. Dairy Sci. 93:2044-2052. 967
968 Paula, E. M. G. A. Broderick, M. A. C. Danes, N. E. Lobos, G. I. Zanton, and A. P. Faciola. 969
2018. Effects of replacing soybean meal with canola meal or treated canola meal on ruminal 970 digestion, omasal nutrient flow, and performance in lactating dairy cows. J. Dairy Sci. 971 101:1–12. 972
973 Puhakka, L., S. Jaakkola, I. Simpura, T. Kokkonen, and A. Vanhatalo. 2016. Effects of 974
replacing rapeseed meal with fava bean at 2 concentrate crude protein levels on feed intake, 975
50
nutrient digestion, and milk production in cows fed grass silage–based diets. J. Dairy Sci. 976 99:7993-8006. 977
978 Razzaghi, A., R. Valizadeh, A. A. Naserian, M. D. Mesgaran, A. J. Carpenter, and M. H. 979
Ghaffari. 2016. Effect of dietary sugar concentration and sunflower seed supplementation 980 on lactation performance, ruminal fermentation, milk fatty acid profile, and blood 981 metabolites of dairy cows. J. Dairy Sci. 99:3539-3548. 982
983 Robinson, P. H. 1996. Evaluation of a seal by-product meal as a feedstuff for dairy cows. Anim. 984
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993 Sanchez, J. M. and D. W. Claypool. 1983. Canola Meal as a Protein Supplement in Dairy 994
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1000 Swanepoel, N., P. H. Robinson, and L. J. Erasmus. 2014. Determining the optimal ratio of 1001
canola meal and high protein dried distillers grain protein in diets of high producing 1002 Holstein dairy cows. Anim. Feed Sci. Technol.189:41-53. 1003
1004 Vicente, G. R., J. A. Shelford, R. G. Peterson, and C. R. Krishnamurti. 1984. Effects of feeding 1005
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