https://doi.org/10.17533/udea.rccp.v37n1a5
Fermentative profile, chemical composition, in vitro gas production,
and ruminal degradation kinetics of sugarcane silages associated
with increasing levels of butterfly pea hay
Perfil fermentativo, composición química, producción de gas y cinética de la degradabilidad ruminal
in vitro de ensilajes de caña de azúcar asociados a niveles incrementales de heno de guisante mariposa
Perfil fermentativo, composição química, produção de gases in vitro e cinética da degradação ruminal de
silagens de cana-de-açúcar associadas a níveis incrementais de feno de cunhã
Ery J Nascimento-Ramos1* ; Bruno A Souza-Almeida2 ; Glayciane Costa-Gois1 ; Daniel Ribeiro-Menezes1 ;
Ana P Ribeiro-Silva1 ; Timóteo S Santos-Nunes1 ; Mário A Ávila- Queiroz1* .
1Universidade Federal do Vale do São Francisco, Petrolina, PE, Brasil.
2Universidade Estadual do Sudoeste da Bahia, Itapetinga, BA, Brasil.
To cite this article:
Nascimento-Ramos EJ, Souza-Almeida BA, Costa-Gois G, Ribeiro-Menezes D, Ribeiro-Silva AP, Santos-Nunes TS,
Ávila-Queiroz MA. Fermentative profile, chemical composition, in vitro gas production and ruminal degradation kinetics
of sugarcane silages associated with increasing levels of butterfly pea hay. Rev Colomb Cienc Pecu 2024; 37(1):27–41.
https://doi.org/10.17533/udea.rccp.v37n1a5
Abstract
Background: The ensiling process of sugarcane promotes yeast proliferation during fermentation, requiring the
use of additives. Clitorea ternatea can be used as a natural additive in sugarcane silages to reduce dry matter losses and
modifying the fermentation profile of the silage. Objective: To evaluate the fermentative profile, chemical composition,
in vitro gas production and ruminal degradation kinetics of sugarcane silages associated with different levels of butterfly
pea hay. Methods: Increasing levels of butterfly pea hay (0, 10, 20, and 30% on dry matter basis) were added to sugarcane
silages. A completely randomized design was adopted, with four treatments and four repetitions, totaling 16 experimental
silos that were opened after 60 days of ensiling. Results: Positive changes were observed in terms of fermentative
losses, fermentative profile, chemical composition, in vitro gas production, and ruminal degradation kinetics with the
addition of butterfly pea hay to sugarcane silage (p<0.05). Conclusion: The inclusion of up to 20% butterfly pea hay in
sugarcane silage reduces fermentation losses and improves silage quality, such as increase in protein and energy content
and reduction of the fibrous fractions of the silage, making silage an excellent ingredient to be included in ruminant diets.
Received: March 31, 2022. Accepted: Jun 9, 2023
Corresponding author. Department of Animal Sciences, Universidade Federal do Vale do São Francisco, Petrolina, 56300-990, PE,
Brasil. Phone No. +55 87 2101 4861. E-mail: mario.queiroz@univasf.edu.br
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, which permits unrestricted reuse,
distribution, and reproduction in any medium, provided the original work is properly cited.
eISSN: 2256-2958 Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
© 2024 Universidad de Antioquia. Publicado por Universidad de Antioquia, Colombia.
Fermentative profile, chemical composition, in vitro gas production,
and ruminal degradation kinetics of sugarcane silages associated
with increasing levels of butterfly pea hay
Perfil fermentativo, composición química, producción de gas y cinética de la degradabilidad ruminal
in vitro de ensilajes de caña de azúcar asociados a niveles incrementales de heno de guisante mariposa
Perfil fermentativo, composição química, produção de gases in vitro e cinética da degradação ruminal de
silagens de cana-de-açúcar associadas a níveis incrementais de feno de cunhã
Ery J Nascimento-Ramos1* ; Bruno A Souza-Almeida2 ; Glayciane Costa-Gois1 ; Daniel Ribeiro-Menezes1 ;
Ana P Ribeiro-Silva1 ; Timóteo S Santos-Nunes1 ; Mário A Ávila- Queiroz1* .
1Universidade Federal do Vale do São Francisco, Petrolina, PE, Brasil.
2Universidade Estadual do Sudoeste da Bahia, Itapetinga, BA, Brasil.
To cite this article:
Nascimento-Ramos EJ, Souza-Almeida BA, Costa-Gois G, Ribeiro-Menezes D, Ribeiro-Silva AP, Santos-Nunes TS,
Ávila-Queiroz MA. Fermentative profile, chemical composition, in vitro gas production and ruminal degradation kinetics
of sugarcane silages associated with increasing levels of butterfly pea hay. Rev Colomb Cienc Pecu 2024; 37(1):27–41.
https://doi.org/10.17533/udea.rccp.v37n1a5
Abstract
Background: The ensiling process of sugarcane promotes yeast proliferation during fermentation, requiring the
use of additives. Clitorea ternatea can be used as a natural additive in sugarcane silages to reduce dry matter losses and
modifying the fermentation profile of the silage. Objective: To evaluate the fermentative profile, chemical composition,
in vitro gas production and ruminal degradation kinetics of sugarcane silages associated with different levels of butterfly
pea hay. Methods: Increasing levels of butterfly pea hay (0, 10, 20, and 30% on dry matter basis) were added to sugarcane
silages. A completely randomized design was adopted, with four treatments and four repetitions, totaling 16 experimental
silos that were opened after 60 days of ensiling. Results: Positive changes were observed in terms of fermentative
losses, fermentative profile, chemical composition, in vitro gas production, and ruminal degradation kinetics with the
addition of butterfly pea hay to sugarcane silage (p<0.05). Conclusion: The inclusion of up to 20% butterfly pea hay in
sugarcane silage reduces fermentation losses and improves silage quality, such as increase in protein and energy content
and reduction of the fibrous fractions of the silage, making silage an excellent ingredient to be included in ruminant diets.
Received: March 31, 2022. Accepted: Jun 9, 2023
Corresponding author. Department of Animal Sciences, Universidade Federal do Vale do São Francisco, Petrolina, 56300-990, PE,
Brasil. Phone No. +55 87 2101 4861. E-mail: mario.queiroz@univasf.edu.br
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, which permits unrestricted reuse,
distribution, and reproduction in any medium, provided the original work is properly cited.
eISSN: 2256-2958 Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
© 2024 Universidad de Antioquia. Publicado por Universidad de Antioquia, Colombia.
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4128
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Keywords: acetic acid; additives; chemical composition; diet; dry matter; ensilage; fermentative profile; forage; forage
conservation; gas production; ruminants.
Resumen
Antecedentes: El proceso de ensilaje de caña de azúcar promueve la proliferación de levaduras durante la fermentación, por
lo que se requiere usar aditivos. La Clitorea ternatea se puede utilizar como aditivo natural en ensilajes de caña de azúcar para
reducir la pérdida de materia seca y modificar el perfil de fermentación del ensilaje. Objetivo: Evaluar el perfil fermentativo,
la composición química, la producción de gas in vitro y la cinética de degradación ruminal de ensilajes de caña de azúcar
asociados con varios niveles de heno de guisante mariposa. Métodos: Se agregaron niveles incrementales de heno de guisante
mariposa (0, 10, 20 y 30% con base a materia seca) a los ensilajes de caña de azúcar. Se adoptó un diseño completamente al
azar, con cuatro tratamientos y cuatro repeticiones, totalizando 16 silos experimentales que se abrieron después de 60 días
de ensilado. Resultados: Se observaron cambios positivos en las pérdidas fermentativas, perfil fermentativo, composición
química, producción de gas in vitro y cinética de degradación ruminal con la adición de heno de guisante mariposa al ensilaje
de caña de azúcar (p<0,05). Conclusión: La inclusión de hasta 20% de heno de guisante mariposa en el ensilaje de caña de
azúcar reduce las pérdidas por fermentación y mejora la calidad del ensilaje, aumentando el contenido proteico y energético y
reduciendo la fracción fibrosa del ensilaje de caña de azúcar, haciendo del ensilaje un excelente ingrediente a incluir en la dieta
de rumiantes.
Palabras clave: ácido acético; aditivos; composición química; conservación de forrajes; dieta; ensilado; forraje; materia
seca; perfil fermentativo; producción de gas; rumiantes.
Resumo
Antecedentes: A ensilagem da cana-de-açúcar promove a proliferação da levedura durante a fermentação, sendo necessário
o uso de aditivos. Clitorea ternatea pode ser utilizado como aditivo natural em silagens de cana-de-açúcar atuando na redução
da perda de matéria seca e modificando o perfil fermentativo da silagem. Objetivo: Avaliar o perfil fermentativo, composição
química, produção de gases in vitro e cinética da degradação ruminal de silagens de cana-de-açúcar associadas a diferentes
níveis de feno de cunhã. Métodos: Níveis incrementais de feno de cunhã (0, 10, 20 e 30% na matéria seca) foram adicionados
às silagens de cana-de-açúcar. Adotou-se o delineamento inteiramente casualizado, com quatro tratamentos e quatro repetições,
totalizando 16 silos experimentais que foram abertos após 60 dias de ensilagem. Resultados: Foram observadas alterações
positivas nas perdas fermentativas, perfil fermentativo, composição química, produção de gases in vitro e cinética da degradação
ruminal com a adição de feno de cunhã em silagens de cana-de-açúcar (p<0,05). Conclusão: A utilização de até 20% de cunhã
em silagens de cana-de-açúcar reduz as perdas fermentativas e melhora a qualidade da silagem, com aumento do teor de
proteína e energia e redução da fração fibrosa das silagens de cana-de-açúcar, tornando as silagens um excelente ingrediente a
ser incluído nas dietas para ruminantes.
Palavras-chave: ácido acético; aditivos; composição química; conservação de forragem; ensilagem; dieta; forragem;
matéria seca; perfil fermentativo; produção de gás; ruminantes.
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Keywords: acetic acid; additives; chemical composition; diet; dry matter; ensilage; fermentative profile; forage; forage
conservation; gas production; ruminants.
Resumen
Antecedentes: El proceso de ensilaje de caña de azúcar promueve la proliferación de levaduras durante la fermentación, por
lo que se requiere usar aditivos. La Clitorea ternatea se puede utilizar como aditivo natural en ensilajes de caña de azúcar para
reducir la pérdida de materia seca y modificar el perfil de fermentación del ensilaje. Objetivo: Evaluar el perfil fermentativo,
la composición química, la producción de gas in vitro y la cinética de degradación ruminal de ensilajes de caña de azúcar
asociados con varios niveles de heno de guisante mariposa. Métodos: Se agregaron niveles incrementales de heno de guisante
mariposa (0, 10, 20 y 30% con base a materia seca) a los ensilajes de caña de azúcar. Se adoptó un diseño completamente al
azar, con cuatro tratamientos y cuatro repeticiones, totalizando 16 silos experimentales que se abrieron después de 60 días
de ensilado. Resultados: Se observaron cambios positivos en las pérdidas fermentativas, perfil fermentativo, composición
química, producción de gas in vitro y cinética de degradación ruminal con la adición de heno de guisante mariposa al ensilaje
de caña de azúcar (p<0,05). Conclusión: La inclusión de hasta 20% de heno de guisante mariposa en el ensilaje de caña de
azúcar reduce las pérdidas por fermentación y mejora la calidad del ensilaje, aumentando el contenido proteico y energético y
reduciendo la fracción fibrosa del ensilaje de caña de azúcar, haciendo del ensilaje un excelente ingrediente a incluir en la dieta
de rumiantes.
Palabras clave: ácido acético; aditivos; composición química; conservación de forrajes; dieta; ensilado; forraje; materia
seca; perfil fermentativo; producción de gas; rumiantes.
Resumo
Antecedentes: A ensilagem da cana-de-açúcar promove a proliferação da levedura durante a fermentação, sendo necessário
o uso de aditivos. Clitorea ternatea pode ser utilizado como aditivo natural em silagens de cana-de-açúcar atuando na redução
da perda de matéria seca e modificando o perfil fermentativo da silagem. Objetivo: Avaliar o perfil fermentativo, composição
química, produção de gases in vitro e cinética da degradação ruminal de silagens de cana-de-açúcar associadas a diferentes
níveis de feno de cunhã. Métodos: Níveis incrementais de feno de cunhã (0, 10, 20 e 30% na matéria seca) foram adicionados
às silagens de cana-de-açúcar. Adotou-se o delineamento inteiramente casualizado, com quatro tratamentos e quatro repetições,
totalizando 16 silos experimentais que foram abertos após 60 dias de ensilagem. Resultados: Foram observadas alterações
positivas nas perdas fermentativas, perfil fermentativo, composição química, produção de gases in vitro e cinética da degradação
ruminal com a adição de feno de cunhã em silagens de cana-de-açúcar (p<0,05). Conclusão: A utilização de até 20% de cunhã
em silagens de cana-de-açúcar reduz as perdas fermentativas e melhora a qualidade da silagem, com aumento do teor de
proteína e energia e redução da fração fibrosa das silagens de cana-de-açúcar, tornando as silagens um excelente ingrediente a
ser incluído nas dietas para ruminantes.
Palavras-chave: ácido acético; aditivos; composição química; conservação de forragem; ensilagem; dieta; forragem;
matéria seca; perfil fermentativo; produção de gás; ruminantes.
29Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Introduction
Due to seasonality of forage plants during
the dry season, tropical forages do not provide
sufficient nutrients for maintain productive
response of small ruminants in arid and semiarid
regions. Therefore, alternatives to meet the demand
for roughage during this period, such as silage
production, are necessary. Thus, storage of surplus
forage produced during the rainy season for use in
the dry season is a viable strategy (Amorim et al.,
2020).
Sugarcane (Saccharum officinarum L.) is
widely used in tropical regions due to its high
production of dry matter (DM; 25–40 t/ha) (Del
Valle et al., 2019), climate adaptability, and
resistance to pests and diseases. Its high content
of water-soluble carbohydrates (396 g/Kg DM)
and buffering power (25.81%) allow the pH of the
silage made with this forage plant to drop to values
close to 3.5 due to generation of organic acids
such as lactic acid. In addition, the high content of
soluble sugars can result in proliferation of yeasts,
which generate ethanol, and high loss of gases
and effluents, resulting in low DM recovery (Reis
et al., 2022).
To reduce DM losses during the ensiling
process, chemical and microbial additives are
traditionally used in sugarcane ensiling (Rabelo
et al., 2019). Besides additives, mixed silages are
capable to modify the fermentation profile and
reduce DM losses. Compared to other additives,
mix silages can result in even greater increase
in chemical composition and degradability (Del
Valle et al., 2020). The effects depend on the
forages chosen for the association with sugarcane
in the ensiling process, as observed in the report
by Carvalho et al. (2018), with the addition of
Manihot pseudoglaziovii; Silva et al. (2014), with
addition of Atriplex numularia; and Queiroz et al.
(2015) with addition of Typha domingensis.
Butterfly pea (Clitorea ternatea Linn) is an
excellent option as an absorbent additive for
sugarcane ensilage. Butterfly pea belongs to the
Fabacea tribe and originates from tropical Asia.
It can be found in India, China, Africa, Central
America, and Brazil. Its common names are
telang flower, samsamping, dậu biếc, kordofan
pea, guisante mariposa and cunhã. It is a perennial
plant that can grow from 1 to 1,800 m.a.s.l., with
rainfall between 650–1,250 mm, and temperatures
above 27 °C (Pratiwi, 2022; Surya et al., 2022).
Butterfly pea is a legume with root nodules that
play a role in fixing nitrogen in the soil, acting as
a natural fertilizer in agricultural land (Suarna and
Wijaya 2021). Its flowers can be white, lilac, light
blue and dark blue, with corolla in normal and
multilayered variations. The seeds are brown or
green, olive green, 4-7 mm long and 3-4 mm wide
(Pratiwi, 2022).
Due to its high dry matter content (286 to
291 g/Kg in fresh matter) (Araújo et al., 2022)
and crude protein (254.8 g/Kg DM) (Jusoh and
Nur Hafifah, 2018), butterfly pea is widely used
in diets for ruminants (Araújo et al., 2022). In
addition, it contains phenolic compounds, terpenes
and alkaloids with antioxidant and bactericidal
potential (Jaafar et al., 2020; Hariad et al., 2023)
that can modify silage fermentation and improve
the quality of the ensiled mass. Thus, butterfly pea
hay can contribute to improve the nutritional and
fermentative quality of sugarcane silages.
The association of butterfly pea with elephant
grass provides a good fermentative profile of
the silage, with reduced levels of butyric acid,
ammoniacal nitrogen, total losses, and increased
DM recovery, thus improving the nutritional
value of silage (Costa et al., 2022); however, it
can reduce the in vitro digestibility of dry matter
(Lemos et al., 2021). Butterfly pea hay has been
also associated with cactus pear meal in diets
for goats. A mix of 67% butterfly pea hay and
33% cactus pear meal increased digestibility and
weight gain of animals (Araújo et al., 2022),
increased carcass yield, and improved the fatty
acid profile of goat meat (Pereira et al., 2020)
when compared to feeding a diet of 70% elephant
grass. However, to the best of our knowledge,
there is a scarcity of studies on the effects of
including butterfly pea hay in sugarcane silage.
Thus, our hypothesis is that butterfly pea
hay promotes protein increase and improves
fermentation kinetics in sugarcane silage.
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Introduction
Due to seasonality of forage plants during
the dry season, tropical forages do not provide
sufficient nutrients for maintain productive
response of small ruminants in arid and semiarid
regions. Therefore, alternatives to meet the demand
for roughage during this period, such as silage
production, are necessary. Thus, storage of surplus
forage produced during the rainy season for use in
the dry season is a viable strategy (Amorim et al.,
2020).
Sugarcane (Saccharum officinarum L.) is
widely used in tropical regions due to its high
production of dry matter (DM; 25–40 t/ha) (Del
Valle et al., 2019), climate adaptability, and
resistance to pests and diseases. Its high content
of water-soluble carbohydrates (396 g/Kg DM)
and buffering power (25.81%) allow the pH of the
silage made with this forage plant to drop to values
close to 3.5 due to generation of organic acids
such as lactic acid. In addition, the high content of
soluble sugars can result in proliferation of yeasts,
which generate ethanol, and high loss of gases
and effluents, resulting in low DM recovery (Reis
et al., 2022).
To reduce DM losses during the ensiling
process, chemical and microbial additives are
traditionally used in sugarcane ensiling (Rabelo
et al., 2019). Besides additives, mixed silages are
capable to modify the fermentation profile and
reduce DM losses. Compared to other additives,
mix silages can result in even greater increase
in chemical composition and degradability (Del
Valle et al., 2020). The effects depend on the
forages chosen for the association with sugarcane
in the ensiling process, as observed in the report
by Carvalho et al. (2018), with the addition of
Manihot pseudoglaziovii; Silva et al. (2014), with
addition of Atriplex numularia; and Queiroz et al.
(2015) with addition of Typha domingensis.
Butterfly pea (Clitorea ternatea Linn) is an
excellent option as an absorbent additive for
sugarcane ensilage. Butterfly pea belongs to the
Fabacea tribe and originates from tropical Asia.
It can be found in India, China, Africa, Central
America, and Brazil. Its common names are
telang flower, samsamping, dậu biếc, kordofan
pea, guisante mariposa and cunhã. It is a perennial
plant that can grow from 1 to 1,800 m.a.s.l., with
rainfall between 650–1,250 mm, and temperatures
above 27 °C (Pratiwi, 2022; Surya et al., 2022).
Butterfly pea is a legume with root nodules that
play a role in fixing nitrogen in the soil, acting as
a natural fertilizer in agricultural land (Suarna and
Wijaya 2021). Its flowers can be white, lilac, light
blue and dark blue, with corolla in normal and
multilayered variations. The seeds are brown or
green, olive green, 4-7 mm long and 3-4 mm wide
(Pratiwi, 2022).
Due to its high dry matter content (286 to
291 g/Kg in fresh matter) (Araújo et al., 2022)
and crude protein (254.8 g/Kg DM) (Jusoh and
Nur Hafifah, 2018), butterfly pea is widely used
in diets for ruminants (Araújo et al., 2022). In
addition, it contains phenolic compounds, terpenes
and alkaloids with antioxidant and bactericidal
potential (Jaafar et al., 2020; Hariad et al., 2023)
that can modify silage fermentation and improve
the quality of the ensiled mass. Thus, butterfly pea
hay can contribute to improve the nutritional and
fermentative quality of sugarcane silages.
The association of butterfly pea with elephant
grass provides a good fermentative profile of
the silage, with reduced levels of butyric acid,
ammoniacal nitrogen, total losses, and increased
DM recovery, thus improving the nutritional
value of silage (Costa et al., 2022); however, it
can reduce the in vitro digestibility of dry matter
(Lemos et al., 2021). Butterfly pea hay has been
also associated with cactus pear meal in diets
for goats. A mix of 67% butterfly pea hay and
33% cactus pear meal increased digestibility and
weight gain of animals (Araújo et al., 2022),
increased carcass yield, and improved the fatty
acid profile of goat meat (Pereira et al., 2020)
when compared to feeding a diet of 70% elephant
grass. However, to the best of our knowledge,
there is a scarcity of studies on the effects of
including butterfly pea hay in sugarcane silage.
Thus, our hypothesis is that butterfly pea
hay promotes protein increase and improves
fermentation kinetics in sugarcane silage.
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4130
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Therefore, the aim of this study was to evaluate
the fermentative profile, chemical composition,
in vitro gas production, and ruminal degradation
kinetics of sugarcane silage associated with
different levels of butterfly pea hay.
Materials and Methods
Ethical considerations
This study was approved and certified by the
Ethics Committee on Animal Use of the Federal
University of Vale do São Francisco Univasf
(protocol 0010/18042018), Brazil.
Experimental site
The experiment was conducted at the Federal
University of Vale do São Francisco (Univasf),
in Petrolina-PE, Brazil (9º 19 '28” South latitude,
40º 33' 34” West longitude, with an altitude of
393 m.a.s.l.). The climate is hot semi-arid with
rainy season (BSh), with 376 mm average
annual precipitation. Maximum and minimum
temperatures during the experimental period were
33.83 and 24.56°C, respectively, with relative
humidity between 50.50 and 73.56%.
Experimental design and elaboration of
silages
Levels of butterfly pea hay inclusion (0, 10,
20 and 30% on dry matter basis) were evaluated
in sugarcane silage, in a completely randomized
experimental design, with four treatments and
four repetitions, totaling 16 experimental silos.
The whole sugarcane plant used was cultivar
VAT90212, harvested manually seven months
after the last cut (regrowth). Butterfly pea (leaf,
branches, petioles, and pods) was harvested
manually at 90 days after planting (DAP),
when the plants were at the flowering stage,
cut at 10 cm from the ground. Butterfly pea hay
elaboration was conducted in the field, by natural
sun drying. After harvesting, before processing,
butterfly pea (leaf, branches, petioles and pods)
was spread over a plastic canvas to reduce losses,
and remained under dehydration for 48 h, with
the material being turned over after 24 h to
uniform drying. During the night, to avoid losses,
the material was covered with canvas. When the
hay point (87.5% of dry matter) was obtained, the
material was collected and stored in a dry place.
Sugarcane and butterfly pea hay were crushed
in a stationary forage chopper (Nogueira PN PLUS
2000, São Paulo, Brazil) to an average particle
size of approximately 2.0 cm. The material was
manually mixed and ensiled in experimental silos
(25 L capacity) equipped with a Bunsen valve to
allow for fermentation gas escape. A total of 2 Kg
of sand were deposited at the bottom of the silos
and protected with a cotton fabric to prevent the
ensiled material from meeting sand and allowing
the effluent to drain.
Table 1. Chemical composition of sugarcane and butterfly pea hay.
Variable (g/Kg dry matter) Sugarcane Butterfly pea** Butterfly pea hay
Dry matter* 259 829 875
Mineral matter 44 88.9 73
Ether extract 27 33.7 27
Crude protein 16 198 150
Neutral detergent fiber 622 448 470
Acid detergent fiber 360 254 319
Total carbohydrates 913 - 751
Non-fiber carbohydrates 291 - 281
Total digestible nutrients 574 - 603
*in g/Kg natural matter; **Araújo et al. (2022).
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Therefore, the aim of this study was to evaluate
the fermentative profile, chemical composition,
in vitro gas production, and ruminal degradation
kinetics of sugarcane silage associated with
different levels of butterfly pea hay.
Materials and Methods
Ethical considerations
This study was approved and certified by the
Ethics Committee on Animal Use of the Federal
University of Vale do São Francisco Univasf
(protocol 0010/18042018), Brazil.
Experimental site
The experiment was conducted at the Federal
University of Vale do São Francisco (Univasf),
in Petrolina-PE, Brazil (9º 19 '28” South latitude,
40º 33' 34” West longitude, with an altitude of
393 m.a.s.l.). The climate is hot semi-arid with
rainy season (BSh), with 376 mm average
annual precipitation. Maximum and minimum
temperatures during the experimental period were
33.83 and 24.56°C, respectively, with relative
humidity between 50.50 and 73.56%.
Experimental design and elaboration of
silages
Levels of butterfly pea hay inclusion (0, 10,
20 and 30% on dry matter basis) were evaluated
in sugarcane silage, in a completely randomized
experimental design, with four treatments and
four repetitions, totaling 16 experimental silos.
The whole sugarcane plant used was cultivar
VAT90212, harvested manually seven months
after the last cut (regrowth). Butterfly pea (leaf,
branches, petioles, and pods) was harvested
manually at 90 days after planting (DAP),
when the plants were at the flowering stage,
cut at 10 cm from the ground. Butterfly pea hay
elaboration was conducted in the field, by natural
sun drying. After harvesting, before processing,
butterfly pea (leaf, branches, petioles and pods)
was spread over a plastic canvas to reduce losses,
and remained under dehydration for 48 h, with
the material being turned over after 24 h to
uniform drying. During the night, to avoid losses,
the material was covered with canvas. When the
hay point (87.5% of dry matter) was obtained, the
material was collected and stored in a dry place.
Sugarcane and butterfly pea hay were crushed
in a stationary forage chopper (Nogueira PN PLUS
2000, São Paulo, Brazil) to an average particle
size of approximately 2.0 cm. The material was
manually mixed and ensiled in experimental silos
(25 L capacity) equipped with a Bunsen valve to
allow for fermentation gas escape. A total of 2 Kg
of sand were deposited at the bottom of the silos
and protected with a cotton fabric to prevent the
ensiled material from meeting sand and allowing
the effluent to drain.
Table 1. Chemical composition of sugarcane and butterfly pea hay.
Variable (g/Kg dry matter) Sugarcane Butterfly pea** Butterfly pea hay
Dry matter* 259 829 875
Mineral matter 44 88.9 73
Ether extract 27 33.7 27
Crude protein 16 198 150
Neutral detergent fiber 622 448 470
Acid detergent fiber 360 254 319
Total carbohydrates 913 - 751
Non-fiber carbohydrates 291 - 281
Total digestible nutrients 574 - 603
*in g/Kg natural matter; **Araújo et al. (2022).
31Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
The material was compacted aiming to reach a
minimum density of 600 Kg/m³ of natural matter.
Samples of the non-ensiled material (original
material) were collected for further laboratory
analysis (Table 1). After being sealed, the silos
were kept for 60 days in a covered shed.
Determination of density and fermentative
losses of silages
The silos were weighed empty, after ensiling
and weighed again at 60 days of ensiling, during
opening. The density of ensiled mass was
determined by the equation:
D (Kg/m3) = m/V (Equation 1)
where: D=density; m=weight of the ensiled
material; V=volume of ensiled material.
Total dry matter losses (TDML) were obtained
by adding the production of gases and effluents
(Amorim et al., 2020). Dry matter recovery
(DMR), gas losses (GL), and effluent losses
(EL) in silages were determined according to the
equations proposed by Amorim et al. (2020):
DMR=((FMo*DMo) / (FMc*DMc))*100
(Equation 2)
where: FMo=forage mass at the opening;
DMo=dry matter content at the opening;
FMc=forage weight at closing; DMc=dry matter
content of forage at closing.
GL=((WSc - WSo)/FMs*FDMc)*100
(Equation 3)
where: WSc=weight of the silo at closing,
WSo=weight of the silo at opening, FMs=forage
mass in silage; FDMc=forage dry matter content
at silo closure (Amorim et al., 2020).
EL=((WSSo - WSS)/GMSF)*100
(Equation 4)
where: WSSo=weight of the set (silo + sand
+ screen) at the opening; WSS=weight of the set
(silo + sand + screen) in the silage; GMSF=green
mass of silage forage.
Fermentation profile of silages
Sample pH was measured with a portable
digital pH meter (Marconi® MA-552, Piracicaba,
São Paulo, Brazil) immediately after opening the
silos and collecting the material. Organic acids
concentration: acetic (AA), propionic (PA),
and lactic (LA), was measured according to the
methodology of Kung Jr and Ranjit (2001). In 2
mL of filtrate, 1 mL of metaphosphoric acid 20%
v/v was added, and this sample was centrifuged.
Analyzes of organic acids were performed by high
performance liquid chromatography (HPLC).
Chemical composition of silages
Silage samples were collected during the
opening of silos, with the top layer (10 cm) of
each silo being discarded. Samples were pre-
dried in a forced-ventilation oven at 55 °C for
72-h. Then, they were individually processed in a
knife mill (Wiley, Marconi, MA 580, Piracicaba,
Brazil) at 3 mm mesh sieve to determine gas
production and in vitro degradability, and at 1 mm
mesh sieve to determine dry matter (DM; method
967.03), mineral matter (MM; method 942.05),
crude protein (CP; method 981.10), ether extract
(EE; method 920.29), and acid detergent fiber
(ADF; method 973.18) (AOAC 2016). Neutral
detergent fiber (NDF) was determined according
to Van Soest et al. (1991). Klason lignin (KL)
content was determined according to Theander
and Westerlund (1986), and total digestible
nutrients (TDN) was obtained with the equation
proposed by Harlan et al. (1991):
TDN=82.75 – (0.704*ADF) (Equation 5)
Total carbohydrates (TC) were measured
according to Sniffen et al. (1992):
TC (g/Kg DM)=1000 – (CP + EE + MM)
(Equation 6)
Non-fiber carbohydrates (NFC) were
measured according to Hall (2003):
NFC (g/Kg DM)=1000 – (% CP +% EE +%
MM + NDF) (Equation 7)
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
The material was compacted aiming to reach a
minimum density of 600 Kg/m³ of natural matter.
Samples of the non-ensiled material (original
material) were collected for further laboratory
analysis (Table 1). After being sealed, the silos
were kept for 60 days in a covered shed.
Determination of density and fermentative
losses of silages
The silos were weighed empty, after ensiling
and weighed again at 60 days of ensiling, during
opening. The density of ensiled mass was
determined by the equation:
D (Kg/m3) = m/V (Equation 1)
where: D=density; m=weight of the ensiled
material; V=volume of ensiled material.
Total dry matter losses (TDML) were obtained
by adding the production of gases and effluents
(Amorim et al., 2020). Dry matter recovery
(DMR), gas losses (GL), and effluent losses
(EL) in silages were determined according to the
equations proposed by Amorim et al. (2020):
DMR=((FMo*DMo) / (FMc*DMc))*100
(Equation 2)
where: FMo=forage mass at the opening;
DMo=dry matter content at the opening;
FMc=forage weight at closing; DMc=dry matter
content of forage at closing.
GL=((WSc - WSo)/FMs*FDMc)*100
(Equation 3)
where: WSc=weight of the silo at closing,
WSo=weight of the silo at opening, FMs=forage
mass in silage; FDMc=forage dry matter content
at silo closure (Amorim et al., 2020).
EL=((WSSo - WSS)/GMSF)*100
(Equation 4)
where: WSSo=weight of the set (silo + sand
+ screen) at the opening; WSS=weight of the set
(silo + sand + screen) in the silage; GMSF=green
mass of silage forage.
Fermentation profile of silages
Sample pH was measured with a portable
digital pH meter (Marconi® MA-552, Piracicaba,
São Paulo, Brazil) immediately after opening the
silos and collecting the material. Organic acids
concentration: acetic (AA), propionic (PA),
and lactic (LA), was measured according to the
methodology of Kung Jr and Ranjit (2001). In 2
mL of filtrate, 1 mL of metaphosphoric acid 20%
v/v was added, and this sample was centrifuged.
Analyzes of organic acids were performed by high
performance liquid chromatography (HPLC).
Chemical composition of silages
Silage samples were collected during the
opening of silos, with the top layer (10 cm) of
each silo being discarded. Samples were pre-
dried in a forced-ventilation oven at 55 °C for
72-h. Then, they were individually processed in a
knife mill (Wiley, Marconi, MA 580, Piracicaba,
Brazil) at 3 mm mesh sieve to determine gas
production and in vitro degradability, and at 1 mm
mesh sieve to determine dry matter (DM; method
967.03), mineral matter (MM; method 942.05),
crude protein (CP; method 981.10), ether extract
(EE; method 920.29), and acid detergent fiber
(ADF; method 973.18) (AOAC 2016). Neutral
detergent fiber (NDF) was determined according
to Van Soest et al. (1991). Klason lignin (KL)
content was determined according to Theander
and Westerlund (1986), and total digestible
nutrients (TDN) was obtained with the equation
proposed by Harlan et al. (1991):
TDN=82.75 – (0.704*ADF) (Equation 5)
Total carbohydrates (TC) were measured
according to Sniffen et al. (1992):
TC (g/Kg DM)=1000 – (CP + EE + MM)
(Equation 6)
Non-fiber carbohydrates (NFC) were
measured according to Hall (2003):
NFC (g/Kg DM)=1000 – (% CP +% EE +%
MM + NDF) (Equation 7)
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4132
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
In vitro gas production
Gas production and rumen degradation
were carried out according to the methodology
proposed by Menezes et al. (2015). The
inoculum was obtained from rumen fluid jointly
and homogenized from two ruminally fistulated
cattle which were fed a diet with 70% sugarcane
and 30% concentrate based on cottonseed meal
and ground corn and offered water ad libitum.
Ruminal inoculum was collected through the
cannula and stored in an anaerobic environment
in a thermal bottle. The solid part was collected
from the rumen through the cannula and manually
pressed to separate the solid from the liquid part.
Ruminal inoculum was filtered through four
layers of gauze, constantly injecting CO2 to
maintain the anaerobic environment, and kept in
a water bath at 39 °C.
One gram of sample was added to glass vials
(160 mL), to which 90 mL nutrient medium
(buffer solution, pH indicator solution, macro,
and micro mineral solution, 1 molar sodium
hydroxide solution, and reducing solution) was
added. Subsequently, 10 mL of ruminal fluid
was added to each flask, which was kept under
a CO2 atmosphere. Then they were sealed with
rubber stoppers and aluminum seals. The same
procedure was applied to the blanks (flask
containing inoculum and medium, without
samples). Four flasks were used as a blank.
The pressure (P; in psi) originated by the gases
accumulated in the upper part of the vials was
measured with a portable pressure transducer (GE
Druck Series DPI 705) connected at its end to a
needle (0.6 mm). Pressure readings were taken
more frequently during the initial fermentation
period and subsequently reduced (2, 4, 6, 8, 10,
12, 14, 17, 20, 24, 28, 34, 48, 72, and 96 h of
incubation).
Pressure data were converted to gas volume
(1 psi=4.859 mL gas). From each pressure
reading, the total produced by the bottles without
substrate (blank) was subtracted from each
sample. Cumulative gas production data were
analyzed by the Gompertz two-compartment
model (Schofield et al., 1994):
V=Vf1 / (1 + exp (2 - 4*m1*(T - L))) + Vf2 /
(1 + exp (2 - 4*m2*(T - L))) (Equation 8)
where: V=total gas volume; Vf1=maximum
volume of gas production from non-fibrous
carbohydrates; Vf2=maximum volume of
gas production from fibrous carbohydrates;
m1=degradation rate (%/h) of the fraction of non-
fibrous carbohydrates; m2=rate of degradation (h)
of the fibrous carbohydrate fraction; T=incubation
time (h); L=time of colonization (h).
The Gompertz two-compartment model
was chosen assuming that gas production
rate is proportional to microbial activity, but
proportionality decreases with incubation time,
which can be attributed to the loss of efficiency
of fermentation rate in time (Cunha et al., 2022).
In vitro ruminal degradation kinetics
In vitro DM degradability was estimated by
inserting nylon bags (20 mg cm-2 weight and 50
microns of porosity) containing 600 mg sample
in flasks with 60 mL buffer solution (combination
of solutions A + B with pH 6.8) and 15 mL
ruminal inoculum. Samples were incubated for
0, 2, 6, 12, 24, 48, 96 and 120 h. After in vitro
fermentation, bags were washed, and oven dried
at 105 ºC for 4-h and weighed. Samples at time
0 were just washed with distilled water at 39 °C
for 5 minutes and then dried and weighed (Tilley
and Terry 1963).
To determine potential degradability (PD),
effective degradability (ED), and the non-
degradable fraction C, the Ørskov and McDonald
(1979) models were used:
PD=A + B (1 -exp-ct) (Equation 9)
where: PD=is potential degradability; A=is the
water-soluble fraction; B=is the water-insoluble
fraction, but potentially degradable; c=is the
degradation rate of fraction B and t=is incubation
time (h).
ED=a + (b*c) / (c + kp) (Equation 10)
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
In vitro gas production
Gas production and rumen degradation
were carried out according to the methodology
proposed by Menezes et al. (2015). The
inoculum was obtained from rumen fluid jointly
and homogenized from two ruminally fistulated
cattle which were fed a diet with 70% sugarcane
and 30% concentrate based on cottonseed meal
and ground corn and offered water ad libitum.
Ruminal inoculum was collected through the
cannula and stored in an anaerobic environment
in a thermal bottle. The solid part was collected
from the rumen through the cannula and manually
pressed to separate the solid from the liquid part.
Ruminal inoculum was filtered through four
layers of gauze, constantly injecting CO2 to
maintain the anaerobic environment, and kept in
a water bath at 39 °C.
One gram of sample was added to glass vials
(160 mL), to which 90 mL nutrient medium
(buffer solution, pH indicator solution, macro,
and micro mineral solution, 1 molar sodium
hydroxide solution, and reducing solution) was
added. Subsequently, 10 mL of ruminal fluid
was added to each flask, which was kept under
a CO2 atmosphere. Then they were sealed with
rubber stoppers and aluminum seals. The same
procedure was applied to the blanks (flask
containing inoculum and medium, without
samples). Four flasks were used as a blank.
The pressure (P; in psi) originated by the gases
accumulated in the upper part of the vials was
measured with a portable pressure transducer (GE
Druck Series DPI 705) connected at its end to a
needle (0.6 mm). Pressure readings were taken
more frequently during the initial fermentation
period and subsequently reduced (2, 4, 6, 8, 10,
12, 14, 17, 20, 24, 28, 34, 48, 72, and 96 h of
incubation).
Pressure data were converted to gas volume
(1 psi=4.859 mL gas). From each pressure
reading, the total produced by the bottles without
substrate (blank) was subtracted from each
sample. Cumulative gas production data were
analyzed by the Gompertz two-compartment
model (Schofield et al., 1994):
V=Vf1 / (1 + exp (2 - 4*m1*(T - L))) + Vf2 /
(1 + exp (2 - 4*m2*(T - L))) (Equation 8)
where: V=total gas volume; Vf1=maximum
volume of gas production from non-fibrous
carbohydrates; Vf2=maximum volume of
gas production from fibrous carbohydrates;
m1=degradation rate (%/h) of the fraction of non-
fibrous carbohydrates; m2=rate of degradation (h)
of the fibrous carbohydrate fraction; T=incubation
time (h); L=time of colonization (h).
The Gompertz two-compartment model
was chosen assuming that gas production
rate is proportional to microbial activity, but
proportionality decreases with incubation time,
which can be attributed to the loss of efficiency
of fermentation rate in time (Cunha et al., 2022).
In vitro ruminal degradation kinetics
In vitro DM degradability was estimated by
inserting nylon bags (20 mg cm-2 weight and 50
microns of porosity) containing 600 mg sample
in flasks with 60 mL buffer solution (combination
of solutions A + B with pH 6.8) and 15 mL
ruminal inoculum. Samples were incubated for
0, 2, 6, 12, 24, 48, 96 and 120 h. After in vitro
fermentation, bags were washed, and oven dried
at 105 ºC for 4-h and weighed. Samples at time
0 were just washed with distilled water at 39 °C
for 5 minutes and then dried and weighed (Tilley
and Terry 1963).
To determine potential degradability (PD),
effective degradability (ED), and the non-
degradable fraction C, the Ørskov and McDonald
(1979) models were used:
PD=A + B (1 -exp-ct) (Equation 9)
where: PD=is potential degradability; A=is the
water-soluble fraction; B=is the water-insoluble
fraction, but potentially degradable; c=is the
degradation rate of fraction B and t=is incubation
time (h).
ED=a + (b*c) / (c + kp) (Equation 10)
33Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
where: kp=is the rate of passage (a 5%/h pass
rate was admitted).
The undegradable fraction (C) was calculated
with the following equation:
C=100 - (A + B) (Equation 11)
Gas production rate obtained by the semi-
automated gas production technique (m1+m2)
was used to estimate the rate of passage used in
the degradability test.
Statistical analysis
Data were subjected to the normality test
and analysis of variance. When significant, the
parameters of the regression equations were
determined by GLM and REG procedures,
respectively, with 5% significance. The results
were analyzed with the SAS program, version
9.4 (2013) software (Statistical Analysis System;
SAS Institute Inc., Cary, NC, USA; 2013).
Results
Increased levels of butterfly pea hay in
sugarcane silage provided a linear reduction
in density (p<0.001) and EL (p<0.001) silages
(Table 2). Quadratic effect was observed for
GL (p<0.001), TDML (p<0.001), and DMR
(p<0.001), with the increase in the levels of
butterfly pea hay in the sugarcane silage (Table 2).
The pH (p<0.001) and PA (p<0.001) of the
sugarcane silages increased linearly according to
increased inclusion of butterfly pea hay (Table 2).
A quadratic effect was observed for AA
(p<0.001) and TFA (p<0.001) content in silages
of sugarcane associated with increasing levels
of butterfly pea hay (Table 2). There was no
effect of the inclusion of butterfly pea hay in
sugarcane silage on LA (p>0.05) (Table 2).
In relation to chemical composition, the DM
(p<0.001), MM (p<0.001), CP (p<0.001), KL
(p<0.001) and TDN (p<0.001) content increased
linearly as the proportions of butterfly pea
hay increased in the composition of sugarcane
silages (Table 3). The opposite effect was
observed NDF (p<0.001) and ADF (p<0.001),
whose content decreased linearly according
to increased levels of butterfly pea hay in the
composition of the sugarcane silages (Table 3).
Table 2. Density, losses, and fermentative profile of sugarcane silages with increasing levels of butterfly pea hay.
Variable Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Density (Kg/m3) 566.35 434.90 405.75 398.55 17.57 <0.0011 <0.511
Gas losses (g/Kg DM) 89.0 131.8 103.0 74.0 0.56 <0.341 <0.0012
Effluent losses (Kg/ton NM) 25.54 6.07 3.63 2.45 2.43 <0.0013 <0.3201
Total dry matter losses (g/Kg DM) 112.8 137.3 106.5 76.5 0.57 <0.201 <0.0014
Dry matter recovery (g/Kg DM) 887.1 862.8 893.7 923.5 0.57 <0.425 <0.0015
pH 3.27 3.64 3.74 3.86 0.06 <0.0016 <0.451
Lactic acid (g/Kg DM) 4.54 4.57 4.62 4.73 0.05 0.257 0.729
Acetic acid (g/Kg DM) 5.11 10.30 11.60 10.15 6.69 <0.471 <0.0017
Propionic acid (g/Kg DM) 0.47 0.64 1.08 1.40 0.10 <0.0018 0.175
Total fatty acids (g/Kg DM) 5.58 10.94 12.68 11.55 6.75 <0.121 <0.0019
SEM: Standard error of the mean; DM: Dry matter; NM: Natural matter; L: Linear effect; Q: Quadratic effect. Significant at the
5% probability level. Equations: ¹ŷ=531.27-5.325x, R2=0.77; 2ŷ=9.256+0.465x-0.018x², R2=0.83; ŷ3=20.181-0.717x, R2=0.72;
ŷ4=11.555+0.269x-0.014x², R2=0.88; ŷ5=88.423-0.265x+0.013x², R2=0.88; ŷ6=3.345+0.019x, R2=0.89; ŷ7=51.675+6.611x-
0.166x2, R2=0.91; ŷ8=0.415+0.033x; R2=0.92; ŷ9=52.127+6.634x-0.165x2, R2=0.91.
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
where: kp=is the rate of passage (a 5%/h pass
rate was admitted).
The undegradable fraction (C) was calculated
with the following equation:
C=100 - (A + B) (Equation 11)
Gas production rate obtained by the semi-
automated gas production technique (m1+m2)
was used to estimate the rate of passage used in
the degradability test.
Statistical analysis
Data were subjected to the normality test
and analysis of variance. When significant, the
parameters of the regression equations were
determined by GLM and REG procedures,
respectively, with 5% significance. The results
were analyzed with the SAS program, version
9.4 (2013) software (Statistical Analysis System;
SAS Institute Inc., Cary, NC, USA; 2013).
Results
Increased levels of butterfly pea hay in
sugarcane silage provided a linear reduction
in density (p<0.001) and EL (p<0.001) silages
(Table 2). Quadratic effect was observed for
GL (p<0.001), TDML (p<0.001), and DMR
(p<0.001), with the increase in the levels of
butterfly pea hay in the sugarcane silage (Table 2).
The pH (p<0.001) and PA (p<0.001) of the
sugarcane silages increased linearly according to
increased inclusion of butterfly pea hay (Table 2).
A quadratic effect was observed for AA
(p<0.001) and TFA (p<0.001) content in silages
of sugarcane associated with increasing levels
of butterfly pea hay (Table 2). There was no
effect of the inclusion of butterfly pea hay in
sugarcane silage on LA (p>0.05) (Table 2).
In relation to chemical composition, the DM
(p<0.001), MM (p<0.001), CP (p<0.001), KL
(p<0.001) and TDN (p<0.001) content increased
linearly as the proportions of butterfly pea
hay increased in the composition of sugarcane
silages (Table 3). The opposite effect was
observed NDF (p<0.001) and ADF (p<0.001),
whose content decreased linearly according
to increased levels of butterfly pea hay in the
composition of the sugarcane silages (Table 3).
Table 2. Density, losses, and fermentative profile of sugarcane silages with increasing levels of butterfly pea hay.
Variable Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Density (Kg/m3) 566.35 434.90 405.75 398.55 17.57 <0.0011 <0.511
Gas losses (g/Kg DM) 89.0 131.8 103.0 74.0 0.56 <0.341 <0.0012
Effluent losses (Kg/ton NM) 25.54 6.07 3.63 2.45 2.43 <0.0013 <0.3201
Total dry matter losses (g/Kg DM) 112.8 137.3 106.5 76.5 0.57 <0.201 <0.0014
Dry matter recovery (g/Kg DM) 887.1 862.8 893.7 923.5 0.57 <0.425 <0.0015
pH 3.27 3.64 3.74 3.86 0.06 <0.0016 <0.451
Lactic acid (g/Kg DM) 4.54 4.57 4.62 4.73 0.05 0.257 0.729
Acetic acid (g/Kg DM) 5.11 10.30 11.60 10.15 6.69 <0.471 <0.0017
Propionic acid (g/Kg DM) 0.47 0.64 1.08 1.40 0.10 <0.0018 0.175
Total fatty acids (g/Kg DM) 5.58 10.94 12.68 11.55 6.75 <0.121 <0.0019
SEM: Standard error of the mean; DM: Dry matter; NM: Natural matter; L: Linear effect; Q: Quadratic effect. Significant at the
5% probability level. Equations: ¹ŷ=531.27-5.325x, R2=0.77; 2ŷ=9.256+0.465x-0.018x², R2=0.83; ŷ3=20.181-0.717x, R2=0.72;
ŷ4=11.555+0.269x-0.014x², R2=0.88; ŷ5=88.423-0.265x+0.013x², R2=0.88; ŷ6=3.345+0.019x, R2=0.89; ŷ7=51.675+6.611x-
0.166x2, R2=0.91; ŷ8=0.415+0.033x; R2=0.92; ŷ9=52.127+6.634x-0.165x2, R2=0.91.
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4134
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
For the gas production kinetics and ruminal
degradability in vitro, the increase in the
levels of butterfly pea hay in the composition
of the sugarcane silages increased the Vf1
(p<0.001), A (p<0.001), Kd (p<0.001), and
IVDMD (p<0.001), reduced L (p<0.001)
and B (p<0.001) (Table 4) and promoted
a quadratic effect for Vf2 (p<0.001), V
(p<0.001), C (p<0.001), PD (p<0.001), ED
(p<0.001), and pH (p=0.002) (Table 4).
Higher cumulative rates of in vitro gas
production were observed in the initial incubation
times. In 48 h of incubation, the average values
of cumulative gas production were close
between the control treatment and the sugarcane
silages containing 10 and 20% butterfly pea hay.
Sugarcane silages containing 10, 20 and 30%
butterfly pea hay stabilized the cumulative gas
production at 72 and 96 h of incubation, while
the control treatment (0% butterfly pea hay)
continued to increase gas production (Figure 1).
Discussion
The association of butterfly pea hay with
sugarcane in mixed silages increased the
dry matter content (Table 1). This increase
difficulted the process of compaction, reducing
silage density. A similar result was reported by
Reis et al. (2022) who, by including Moringa
oleifera hay in sugarcane silage, increased DM
content (from 253.1 to 441.1 g/Kg NM) and
reduced silage density (from 503.9 to 408.3
Kg/m3) with increasing levels of Moringa
oleifera. The increased DM content -reduced
moisture- allowed for proper fermentation of
the ensiled material, which possibly reduced the
development of yeasts (which consume water-
soluble carbohydrates and produce ethanol, CO2,
water, and ATP, generating losses) (Auerbach et
al., 2020; Kim et al., 2021), thus improving DMR
at increasing levels of butterfly pea. In addition,
butterfly pea hay acted as a hygroscopic barrier
absorbing the effluent generated by sugarcane
fermentation, preventing it from leaching, thus
reducing effluent losses.
Gas losses derive from carbohydrate
and protein fermentation, which results in
the production of CO2, N2O and N-NH3,
representing most of the TDML. The
fermentative profile of ensiled material
influences gas losses (Zanine et al., 2020). The
highest GL were obtained with 12.94% butterfly
pea hay inclusion, representing a loss of 122.6
g/Kg DM. The lowest GL was obtained with
30% inclusion of butterfly pea hay in the silage.
Table 3. Chemical composition of sugarcane silages associated with increasing levels of butterfly pea hay.
Variable (g/Kg dry matter) Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Dry matter* 205.7 282.8 347.6 413.0 1.99 <0.0011 0.026
Mineral matter 52.8 61.6 62.6 63.9 0.12 <0.0012 0.003
Crude protein 18.2 47.2 67.2 82.2 0.62 <0.0013 <0.001
Neutral detergent fiber 740.6 679.8 626.9 599.7 1.40 <0.0014 0.0003
Acid detergent fiber 461.3 431.6 425.7 386.3 0.72 <0.0015 0.316
Klason lignin 74.2 81.8 85.4 88.0 0.14 <0.0016 0.061
Total digestible nutrients 497.7 524.1 530.3 553.5 0.52 <0.0017 0.526
Acetic acid (g/Kg DM) 5.11 10.30 11.60 10.15 6.69 <0.471 <0.0017
Propionic acid (g/Kg DM) 0.47 0.64 1.08 1.40 0.10 <0.0018 0.175
Total fatty acids (g/Kg DM) 5.58 10.94 12.68 11.55 6.75 <0.121 <0.0019
*g/Kg natural matter; SEM: Standard error of the mean; L: Linear effect; Q: Quadratic effect. Significant at the 5% probability
level. Equations: ŷ1=20.927+0.686x, R2=0.99; ŷ2=5.509+0.034x, R2=0.67; ŷ3=2.199+0.212x, R2=0.97; ŷ4=73.306-0.476x,
R2=0.96; ŷ5=46.086-0.231x, R2=0.86; ŷ6=7.561+0.045x, R2=0.82; ŷ7=50.038+0.174x, R2=0.91.
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
For the gas production kinetics and ruminal
degradability in vitro, the increase in the
levels of butterfly pea hay in the composition
of the sugarcane silages increased the Vf1
(p<0.001), A (p<0.001), Kd (p<0.001), and
IVDMD (p<0.001), reduced L (p<0.001)
and B (p<0.001) (Table 4) and promoted
a quadratic effect for Vf2 (p<0.001), V
(p<0.001), C (p<0.001), PD (p<0.001), ED
(p<0.001), and pH (p=0.002) (Table 4).
Higher cumulative rates of in vitro gas
production were observed in the initial incubation
times. In 48 h of incubation, the average values
of cumulative gas production were close
between the control treatment and the sugarcane
silages containing 10 and 20% butterfly pea hay.
Sugarcane silages containing 10, 20 and 30%
butterfly pea hay stabilized the cumulative gas
production at 72 and 96 h of incubation, while
the control treatment (0% butterfly pea hay)
continued to increase gas production (Figure 1).
Discussion
The association of butterfly pea hay with
sugarcane in mixed silages increased the
dry matter content (Table 1). This increase
difficulted the process of compaction, reducing
silage density. A similar result was reported by
Reis et al. (2022) who, by including Moringa
oleifera hay in sugarcane silage, increased DM
content (from 253.1 to 441.1 g/Kg NM) and
reduced silage density (from 503.9 to 408.3
Kg/m3) with increasing levels of Moringa
oleifera. The increased DM content -reduced
moisture- allowed for proper fermentation of
the ensiled material, which possibly reduced the
development of yeasts (which consume water-
soluble carbohydrates and produce ethanol, CO2,
water, and ATP, generating losses) (Auerbach et
al., 2020; Kim et al., 2021), thus improving DMR
at increasing levels of butterfly pea. In addition,
butterfly pea hay acted as a hygroscopic barrier
absorbing the effluent generated by sugarcane
fermentation, preventing it from leaching, thus
reducing effluent losses.
Gas losses derive from carbohydrate
and protein fermentation, which results in
the production of CO2, N2O and N-NH3,
representing most of the TDML. The
fermentative profile of ensiled material
influences gas losses (Zanine et al., 2020). The
highest GL were obtained with 12.94% butterfly
pea hay inclusion, representing a loss of 122.6
g/Kg DM. The lowest GL was obtained with
30% inclusion of butterfly pea hay in the silage.
Table 3. Chemical composition of sugarcane silages associated with increasing levels of butterfly pea hay.
Variable (g/Kg dry matter) Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Dry matter* 205.7 282.8 347.6 413.0 1.99 <0.0011 0.026
Mineral matter 52.8 61.6 62.6 63.9 0.12 <0.0012 0.003
Crude protein 18.2 47.2 67.2 82.2 0.62 <0.0013 <0.001
Neutral detergent fiber 740.6 679.8 626.9 599.7 1.40 <0.0014 0.0003
Acid detergent fiber 461.3 431.6 425.7 386.3 0.72 <0.0015 0.316
Klason lignin 74.2 81.8 85.4 88.0 0.14 <0.0016 0.061
Total digestible nutrients 497.7 524.1 530.3 553.5 0.52 <0.0017 0.526
Acetic acid (g/Kg DM) 5.11 10.30 11.60 10.15 6.69 <0.471 <0.0017
Propionic acid (g/Kg DM) 0.47 0.64 1.08 1.40 0.10 <0.0018 0.175
Total fatty acids (g/Kg DM) 5.58 10.94 12.68 11.55 6.75 <0.121 <0.0019
*g/Kg natural matter; SEM: Standard error of the mean; L: Linear effect; Q: Quadratic effect. Significant at the 5% probability
level. Equations: ŷ1=20.927+0.686x, R2=0.99; ŷ2=5.509+0.034x, R2=0.67; ŷ3=2.199+0.212x, R2=0.97; ŷ4=73.306-0.476x,
R2=0.96; ŷ5=46.086-0.231x, R2=0.86; ŷ6=7.561+0.045x, R2=0.82; ŷ7=50.038+0.174x, R2=0.91.
35Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Table 4. In vitro gas production and ruminal degradation kinetics of sugarcane silages with increasing levels of
butterfly pea hay inclusion.
Variable Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Vf1 (mL/g DM) 56.32 60.47 60.31 64.67 3.34 <0.0011 0.892
Vf2 (mL/g DM) 37.99 20.90 19.55 21.45 7.91 <0.371 <0.0012
V (ml/g DM) 94.31 81.37 79.86 86.12 5.92 <0.561 <0.0013
L (h) 17.55 15.69 13.45 10.47 2.81 <0.0014 0.157
A (g/Kg DM) 152.0 191.8 194.9 192.2 1.87 <0.0015 <0.521
B (g/Kg DM) 457.0 469.5 367.9 355.5 5.32 <0.0016 0.183
C (g/Kg DM) 391.0 338.8 437.2 452.3 4.64 <0.001 <0.0017
Kd (%/h) 0.048 0.070 0.075 0.088 0.02 <0.0018 0.199
PD (g/Kg DM) 609.0 661.3 562.8 547.7 4.64 <0.421 <0.0019
ED (g/Kg DM) 375.8 462.5 414.3 418.7 3.33 0.278 <0.00110
pH 6.64 6.96 6.92 6.92 0.04 0.129 0.00211
IVDMD (g/Kg) 358.3 364.4 381.0 452.7 0.88 <0.00112 <0.291
Vf1: Maximum volume of gas production from non-fibrous carbohydrates; Vf2: Maximum volume of gas production of fibrous
carbohydrates; V: Total gas volume; L: Time of colonization (h); A: Water-soluble fraction; B: Water-insoluble fraction, but
potentially degradable; C: Non-degradable fraction; PD: Potential degradability; ED: Effective degradability; IVDMD: In vitro
dry matter degradability; DM: dry matter; SEM: Standard error of the mean; L: Linear effect; Q: Quadratic effect; Significant
at the 5% probability level. Equations: ŷ1=56.708+0.249x, R²=0.74; ŷ2=37.3647-1.934x+0.048x2, R²=0.93; ŷ3=94.122-
1.701x+0.048x2, R²=0.96; ŷ4=17.812-0.235x, R²=0.93; ŷ5=16.419+0.127x, R²=0.58; ŷ6=47.335-0.406x, R²=0.79; ŷ7=37.929-
0.223x+0.017x2, R²=0.63; ŷ8=0.052+0.0013x, R²=0.78; ŷ9=62.071+0.223x-0.017x2, R²=0.63; ŷ10=38.518+0.697x-0.021x2,
R²=0.48; ŷ11=6.663+0.032x-0.00081x2, R²=0.67; ŷ12=34.413+0.299x, R²=0.76.
Figure 1. Cumulative gas production (mL/g DM) at different incubation times of sugarcane silages associated
with butterfly pea hay levels (0, 10, 20 and 30% on dry matter basis).
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Table 4. In vitro gas production and ruminal degradation kinetics of sugarcane silages with increasing levels of
butterfly pea hay inclusion.
Variable Butterfly pea hay levels (%DM) SEM P-value
0 10 20 30 L Q
Vf1 (mL/g DM) 56.32 60.47 60.31 64.67 3.34 <0.0011 0.892
Vf2 (mL/g DM) 37.99 20.90 19.55 21.45 7.91 <0.371 <0.0012
V (ml/g DM) 94.31 81.37 79.86 86.12 5.92 <0.561 <0.0013
L (h) 17.55 15.69 13.45 10.47 2.81 <0.0014 0.157
A (g/Kg DM) 152.0 191.8 194.9 192.2 1.87 <0.0015 <0.521
B (g/Kg DM) 457.0 469.5 367.9 355.5 5.32 <0.0016 0.183
C (g/Kg DM) 391.0 338.8 437.2 452.3 4.64 <0.001 <0.0017
Kd (%/h) 0.048 0.070 0.075 0.088 0.02 <0.0018 0.199
PD (g/Kg DM) 609.0 661.3 562.8 547.7 4.64 <0.421 <0.0019
ED (g/Kg DM) 375.8 462.5 414.3 418.7 3.33 0.278 <0.00110
pH 6.64 6.96 6.92 6.92 0.04 0.129 0.00211
IVDMD (g/Kg) 358.3 364.4 381.0 452.7 0.88 <0.00112 <0.291
Vf1: Maximum volume of gas production from non-fibrous carbohydrates; Vf2: Maximum volume of gas production of fibrous
carbohydrates; V: Total gas volume; L: Time of colonization (h); A: Water-soluble fraction; B: Water-insoluble fraction, but
potentially degradable; C: Non-degradable fraction; PD: Potential degradability; ED: Effective degradability; IVDMD: In vitro
dry matter degradability; DM: dry matter; SEM: Standard error of the mean; L: Linear effect; Q: Quadratic effect; Significant
at the 5% probability level. Equations: ŷ1=56.708+0.249x, R²=0.74; ŷ2=37.3647-1.934x+0.048x2, R²=0.93; ŷ3=94.122-
1.701x+0.048x2, R²=0.96; ŷ4=17.812-0.235x, R²=0.93; ŷ5=16.419+0.127x, R²=0.58; ŷ6=47.335-0.406x, R²=0.79; ŷ7=37.929-
0.223x+0.017x2, R²=0.63; ŷ8=0.052+0.0013x, R²=0.78; ŷ9=62.071+0.223x-0.017x2, R²=0.63; ŷ10=38.518+0.697x-0.021x2,
R²=0.48; ŷ11=6.663+0.032x-0.00081x2, R²=0.67; ŷ12=34.413+0.299x, R²=0.76.
Figure 1. Cumulative gas production (mL/g DM) at different incubation times of sugarcane silages associated
with butterfly pea hay levels (0, 10, 20 and 30% on dry matter basis).
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4136
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
The efficiency in reducing GL with the greatest
inclusion of butterfly pea hay may be related
to greater production of acetic acid, which,
according to Ávila and Carvalho (2020), is
negatively correlated with CO2 production inside
the silo, which may indicate low occurrence of
gas-producing microorganisms such as clostridia
and possible absence of secondary fermentations
(Costa et al., 2022). Corroborating our findings,
when Costa et al. (2022) included butterfly pea
in elephant grass silage also observed reduced
GL compared to exclusive elephant grass silage.
Addition of butterfly pea hay increased the
pH of sugarcane silage. However, the pH value
considered adequate for properly fermented
silages (3.8 to 4.2) (Pereira et al., 2019) was only
observed for sugarcane silages containing 30%
butterfly pea hay. This increase could possibly be
related to reduce levels of soluble carbohydrates
in sugarcane silage with increasing butterfly
pea hay proportions. In addition, by increasing
pH and osmotic pressure of the environment
(Figueiredo et al., 2022), a greater inclusion
of butterfly pea hay in the sugarcane silages
changed the environment -previously favorable
to the development of yeasts- becoming
inappropriate, thus reducing GL. As butterfly
pea has antioxidant and bactericidal substances
(Jaafar et al., 2020) such as anthocyanins,
anthocyanidin, and flavanol (Multisona et al.,
2023), these compounds may have inhibited
growth of undesirable microorganisms during
fermentation, resulting in decreased ethanol
production and fermentative losses.
Contrarily, Carvalho et al. (2014) observed
a pH reduction (3.7 – 3.4) in sugarcane silages
at increasing levels of Manihot pseudoglaziovii.
They emphasized that pH is not a good indicator
of quality for sugarcane silage when considered
in isolation, since the main concern when
ensiling sugarcane is the occurrence of yeasts
which develop even at low pH, and ethanol
itself can act as microbial inhibitor; therefore,
control of losses and ethanol production should
be the focus in sugarcane silage.
Among the fatty acids evaluated, butyric acid
concentration was below the detection limit.
Acetic acid is a potential inhibitor of undesirable
fungi and yeasts in silage. High levels of AA in
sugarcane silage containing butterfly pea hay
may result from enterobacteria and secondary
fermentations in the silos. Acetic acid increased
with inclusion of up to 20% butterfly pea hay;
however, this concentration decreases as the
fermentation process progresses. Studying mixed
sugarcane silage with forage peanut, Costa et al.
(2022) observed that AA increased up to 50%
forage peanut inclusion in the sugarcane silages,
then declined. They emphasized that high AA
content (above 2%) is desirable for silages rich
in water-soluble carbohydrates, as they decrease
yeast activity, reducing gas losses and improving
aerobic stability after opening the silos. Thus,
addition of up to 20% butterfly pea hay in
sugarcane silage could be an alternative to reduce
yeast activity during fermentation in sugarcane
silage since microorganisms represent one of the
main problems when ensiling this crop. However,
with mean values between 10.15 and 11.60 g/Kg
DM, AA concentrations in all silages containing
butterfly pea hay are considered acceptable,
according to Santos et al. (2020).
Propionic acid is associated with conversion
of lactic acid to acetic acid and 1,2-propanediol,
which in turn is converted to PA and 1-propanol
by naturally occurring microorganisms
in silages (Gonzalez-García et al., 2017).
Increased butterfly pea hay levels in sugarcane
silage increased the PA content in silages from
0.47 (control) to 1.40 g/Kg DM, staying below
5 g/Kg DM, without affecting silage quality
(Borreani et al., 2018). As butterfly pea has
high buffering capacity (57.25 e.mg/100 g
DM) (Lemos et al., 2021) heterofermentative
species possibly increased -corroborated by
PA increase- which have antifungal properties
and inhibit yeast growth and, consequently,
decrease ethanol production (Costa et al., 2022).
However, as ethanol was not quantified in this
study, additional research is needed.
The increase in DM, MM, CP, and TDN
contents in silage is directly related to the
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
The efficiency in reducing GL with the greatest
inclusion of butterfly pea hay may be related
to greater production of acetic acid, which,
according to Ávila and Carvalho (2020), is
negatively correlated with CO2 production inside
the silo, which may indicate low occurrence of
gas-producing microorganisms such as clostridia
and possible absence of secondary fermentations
(Costa et al., 2022). Corroborating our findings,
when Costa et al. (2022) included butterfly pea
in elephant grass silage also observed reduced
GL compared to exclusive elephant grass silage.
Addition of butterfly pea hay increased the
pH of sugarcane silage. However, the pH value
considered adequate for properly fermented
silages (3.8 to 4.2) (Pereira et al., 2019) was only
observed for sugarcane silages containing 30%
butterfly pea hay. This increase could possibly be
related to reduce levels of soluble carbohydrates
in sugarcane silage with increasing butterfly
pea hay proportions. In addition, by increasing
pH and osmotic pressure of the environment
(Figueiredo et al., 2022), a greater inclusion
of butterfly pea hay in the sugarcane silages
changed the environment -previously favorable
to the development of yeasts- becoming
inappropriate, thus reducing GL. As butterfly
pea has antioxidant and bactericidal substances
(Jaafar et al., 2020) such as anthocyanins,
anthocyanidin, and flavanol (Multisona et al.,
2023), these compounds may have inhibited
growth of undesirable microorganisms during
fermentation, resulting in decreased ethanol
production and fermentative losses.
Contrarily, Carvalho et al. (2014) observed
a pH reduction (3.7 – 3.4) in sugarcane silages
at increasing levels of Manihot pseudoglaziovii.
They emphasized that pH is not a good indicator
of quality for sugarcane silage when considered
in isolation, since the main concern when
ensiling sugarcane is the occurrence of yeasts
which develop even at low pH, and ethanol
itself can act as microbial inhibitor; therefore,
control of losses and ethanol production should
be the focus in sugarcane silage.
Among the fatty acids evaluated, butyric acid
concentration was below the detection limit.
Acetic acid is a potential inhibitor of undesirable
fungi and yeasts in silage. High levels of AA in
sugarcane silage containing butterfly pea hay
may result from enterobacteria and secondary
fermentations in the silos. Acetic acid increased
with inclusion of up to 20% butterfly pea hay;
however, this concentration decreases as the
fermentation process progresses. Studying mixed
sugarcane silage with forage peanut, Costa et al.
(2022) observed that AA increased up to 50%
forage peanut inclusion in the sugarcane silages,
then declined. They emphasized that high AA
content (above 2%) is desirable for silages rich
in water-soluble carbohydrates, as they decrease
yeast activity, reducing gas losses and improving
aerobic stability after opening the silos. Thus,
addition of up to 20% butterfly pea hay in
sugarcane silage could be an alternative to reduce
yeast activity during fermentation in sugarcane
silage since microorganisms represent one of the
main problems when ensiling this crop. However,
with mean values between 10.15 and 11.60 g/Kg
DM, AA concentrations in all silages containing
butterfly pea hay are considered acceptable,
according to Santos et al. (2020).
Propionic acid is associated with conversion
of lactic acid to acetic acid and 1,2-propanediol,
which in turn is converted to PA and 1-propanol
by naturally occurring microorganisms
in silages (Gonzalez-García et al., 2017).
Increased butterfly pea hay levels in sugarcane
silage increased the PA content in silages from
0.47 (control) to 1.40 g/Kg DM, staying below
5 g/Kg DM, without affecting silage quality
(Borreani et al., 2018). As butterfly pea has
high buffering capacity (57.25 e.mg/100 g
DM) (Lemos et al., 2021) heterofermentative
species possibly increased -corroborated by
PA increase- which have antifungal properties
and inhibit yeast growth and, consequently,
decrease ethanol production (Costa et al., 2022).
However, as ethanol was not quantified in this
study, additional research is needed.
The increase in DM, MM, CP, and TDN
contents in silage is directly related to the
37Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–41
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
nutritional characteristics of butterfly pea hay
in relation to sugarcane (Table 1). This result
was already expected and corroborates previous
research by Carvalho et al. (2018), Costa et al.
(2022) and Reis et al. (2022) by including
legumes in the composition of sugarcane silage.
The DM value of sugarcane silage associated
with 20% butterfly pea hay is within the limit
of 30 to 35% established by McDonald et al.
(1991) for good forage fermentation. Dry matter
values below 30% make the silage susceptible
to the action of undesirable microorganisms,
promoting effluent losses. Percentages above
35%, make compaction difficult, causing
undesirable phenomena due to air entering the
silo, which was verified for control, 10, and 30%
butterfly pea hay inclusion.
Mixed silages intercropping legumes and
grass forage plants aims to increase CP content
of the silage. Since sugarcane has low CP content
(16 g/Kg DM) (Table 1), protein content of the
silage was favored by butterfly pea hay, which
has 150 g/Kg DM (Table 1). However, only with
30% butterfly pea hay inclusion in the sugarcane
silage it was possible to obtain the necessary CP
content (70 g/Kg DM) (Pereira et al., 2019) for
adequate ruminal fermentation.
Reduction in NDF and ADF content in the
silages was promoted by adding butterfly pea
hay due to reduction in the sugarcane: butterfly
pea hay ratio; the concentration of digestible
components probably increased. Carvalho et
al. (2018) also observed a reduction of NDF
and ADF contents of sugarcane silages by
including Manihot pseudoglaziovii. The highest
concentration of NDF and ADF was found in the
silage not added with butterfly pea hay (Control)
(Table 3), which would be related to sugarcane
composition (Table 1).
Lignin behaved opposite to other cell wall
components. Lignin content is an important
parameter to be considered because it is the
main limiting factor during degradation of the
fibrous fraction of silages (Pereira et al., 2019).
Even with the reduction of NDF, with increased
inclusion of butterfly pea hay, the result is a linear
increase in lignin that influenced the quality
of potentially degradable fraction (fraction
B) and non-degradable fraction (fraction C)
(Table 4). This reduction is explained by the
lower concentration of structural carbohydrates
in the legume cell wall compared to the grass,
contributing to improve silage digestibility
(Silva et al., 2018). Agreeing with our results,
Hawu et al. (2022) also observed that legume
hay addition to corn silage reduced fiber
concentration, probably due to hemicellulose
hydrolysis into monosaccharides, which
provides extra carbohydrates for generating
lactic acid during fermentation. Our results are
like the maximum NDF limit recommended by
Van Soest (1994) for ruminant diets, which is
60% NDF.
Butterfly pea hay increased cumulative gas
production from non-fibrous carbohydrates
in sugarcane silage, as well as fraction A
concentration. Regarding gas production from
the fibrous fraction, inclusion of up to 20%
butterfly pea hay reduced gas production as
well as fraction B concentration; however, gas
production increased at 30% butterfly pea hay
inclusion due to increased degradation of this
fraction, which was reflected in the total volume
of gas produced. This is possibly associated
with the fibrous carbohydrate content, which is
related to the rate of degradation and the time
of colonization by ruminal microorganisms
(Ribeiro et al., 2019).
The latency phase is the time between the
beginning of incubation and the microbial
action on the substrate. Latency depends on the
presence of readily fermentable substrates and
chemical characteristics of the sample, which
can favor microbial fermentation (Hamill et
al., 2020). Addition of butterfly pea hay to
sugarcane silage reduced the latency period,
which is positive, since microorganisms will
adhere to the particles more quickly, promoting
rapid fiber degradation (Yansari et al., 2017).
According to Xue et al. (2020), the energy
used by microorganisms during the first
hours of incubation comes from fermentation
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
nutritional characteristics of butterfly pea hay
in relation to sugarcane (Table 1). This result
was already expected and corroborates previous
research by Carvalho et al. (2018), Costa et al.
(2022) and Reis et al. (2022) by including
legumes in the composition of sugarcane silage.
The DM value of sugarcane silage associated
with 20% butterfly pea hay is within the limit
of 30 to 35% established by McDonald et al.
(1991) for good forage fermentation. Dry matter
values below 30% make the silage susceptible
to the action of undesirable microorganisms,
promoting effluent losses. Percentages above
35%, make compaction difficult, causing
undesirable phenomena due to air entering the
silo, which was verified for control, 10, and 30%
butterfly pea hay inclusion.
Mixed silages intercropping legumes and
grass forage plants aims to increase CP content
of the silage. Since sugarcane has low CP content
(16 g/Kg DM) (Table 1), protein content of the
silage was favored by butterfly pea hay, which
has 150 g/Kg DM (Table 1). However, only with
30% butterfly pea hay inclusion in the sugarcane
silage it was possible to obtain the necessary CP
content (70 g/Kg DM) (Pereira et al., 2019) for
adequate ruminal fermentation.
Reduction in NDF and ADF content in the
silages was promoted by adding butterfly pea
hay due to reduction in the sugarcane: butterfly
pea hay ratio; the concentration of digestible
components probably increased. Carvalho et
al. (2018) also observed a reduction of NDF
and ADF contents of sugarcane silages by
including Manihot pseudoglaziovii. The highest
concentration of NDF and ADF was found in the
silage not added with butterfly pea hay (Control)
(Table 3), which would be related to sugarcane
composition (Table 1).
Lignin behaved opposite to other cell wall
components. Lignin content is an important
parameter to be considered because it is the
main limiting factor during degradation of the
fibrous fraction of silages (Pereira et al., 2019).
Even with the reduction of NDF, with increased
inclusion of butterfly pea hay, the result is a linear
increase in lignin that influenced the quality
of potentially degradable fraction (fraction
B) and non-degradable fraction (fraction C)
(Table 4). This reduction is explained by the
lower concentration of structural carbohydrates
in the legume cell wall compared to the grass,
contributing to improve silage digestibility
(Silva et al., 2018). Agreeing with our results,
Hawu et al. (2022) also observed that legume
hay addition to corn silage reduced fiber
concentration, probably due to hemicellulose
hydrolysis into monosaccharides, which
provides extra carbohydrates for generating
lactic acid during fermentation. Our results are
like the maximum NDF limit recommended by
Van Soest (1994) for ruminant diets, which is
60% NDF.
Butterfly pea hay increased cumulative gas
production from non-fibrous carbohydrates
in sugarcane silage, as well as fraction A
concentration. Regarding gas production from
the fibrous fraction, inclusion of up to 20%
butterfly pea hay reduced gas production as
well as fraction B concentration; however, gas
production increased at 30% butterfly pea hay
inclusion due to increased degradation of this
fraction, which was reflected in the total volume
of gas produced. This is possibly associated
with the fibrous carbohydrate content, which is
related to the rate of degradation and the time
of colonization by ruminal microorganisms
(Ribeiro et al., 2019).
The latency phase is the time between the
beginning of incubation and the microbial
action on the substrate. Latency depends on the
presence of readily fermentable substrates and
chemical characteristics of the sample, which
can favor microbial fermentation (Hamill et
al., 2020). Addition of butterfly pea hay to
sugarcane silage reduced the latency period,
which is positive, since microorganisms will
adhere to the particles more quickly, promoting
rapid fiber degradation (Yansari et al., 2017).
According to Xue et al. (2020), the energy
used by microorganisms during the first
hours of incubation comes from fermentation
Rev Colomb Cienc Pecu 2024; 37(1, Jan-Mar):27–4138
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
of non-fibrous carbohydrates, which are
readily available for degradation, resulting
in shorter fermentation time. After reducing
non-fibrous carbohydrates, fermentation of
fibrous carbohydrates continues, since they are
fermented more slowly.
Although IVDMD increased linearly due
to increased soluble fraction (A), potential
and effective degradability were influenced
by increase in fraction C and remained below
levels considered adequate for satisfactory
degradability (Magalhães et al., 2019).
In conclusion, inclusion of butterfly pea hay
in sugarcane silage improves fermentation, loss
reduction, and nutritional value. Inclusion of
up to 20% butterfly pea hay in sugarcane silage
reduces fermentation losses, improving silage
quality: specifically, by increasing protein and
energy content and reducing the fibrous fractions
of sugarcane silage.
Declarations
Funding
Not applicable.
Conflicts of interest
The authors declare they have no conflicts of
interest with regard to the work presented in this
report.
Author contributions
Conceptualization: MAAQ, DRM; data
acquisition and design of methodology: EJNR,
BASA, APRS, TSSN; data analysis: EJNR,
MAAQ; writing original draft: EJNR, MAAQ,
GCG; writing-review and editing: MAAQ, GCG.
Use of artificial intelligence (AI)
No AI or AI-assisted technologies were used
during the preparation of this work.
References
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Bezerra LR, Araújo MJ, Silva AL, Mielezrski
F, Nascimento KS. Fermentation profile and
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Rodrigues RTS, Menezes DR. Nutritional value,
feeding behavior, physiological parameters,
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Borreani G, Tabacco E, Schmidt RJ, Holmes
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https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
of non-fibrous carbohydrates, which are
readily available for degradation, resulting
in shorter fermentation time. After reducing
non-fibrous carbohydrates, fermentation of
fibrous carbohydrates continues, since they are
fermented more slowly.
Although IVDMD increased linearly due
to increased soluble fraction (A), potential
and effective degradability were influenced
by increase in fraction C and remained below
levels considered adequate for satisfactory
degradability (Magalhães et al., 2019).
In conclusion, inclusion of butterfly pea hay
in sugarcane silage improves fermentation, loss
reduction, and nutritional value. Inclusion of
up to 20% butterfly pea hay in sugarcane silage
reduces fermentation losses, improving silage
quality: specifically, by increasing protein and
energy content and reducing the fibrous fractions
of sugarcane silage.
Declarations
Funding
Not applicable.
Conflicts of interest
The authors declare they have no conflicts of
interest with regard to the work presented in this
report.
Author contributions
Conceptualization: MAAQ, DRM; data
acquisition and design of methodology: EJNR,
BASA, APRS, TSSN; data analysis: EJNR,
MAAQ; writing original draft: EJNR, MAAQ,
GCG; writing-review and editing: MAAQ, GCG.
Use of artificial intelligence (AI)
No AI or AI-assisted technologies were used
during the preparation of this work.
References
Amorim DS, Edvan RL, Nascimento RR,
Bezerra LR, Araújo MJ, Silva AL, Mielezrski
F, Nascimento KS. Fermentation profile and
nutritional value of sesame silage compared to usual
silages. Italian J Anim Sci 2020; 19(1):230–239.
https://doi.org/10.1080/1828051X.2020.1724523
AOAC. Official methods of analysis. Association
of Analytical Chemists International. 20th ed.
Washington (DC): G.W. Latimer Jr; 2016.
Araújo EJB, Pereira FDS, Nunes TSS, Cordeiro
AE, Silva HC, Queiroz MAA, Gois GC,
Rodrigues RTS, Menezes DR. Nutritional value,
feeding behavior, physiological parameters,
and performance of crossbred Boer goats
kids fed butterfly pea hay and cactus pear
meal. Spanish J Agr Res 2022; 20:e0603.
https://doi.org/10.5424/sjar/2022202-18690
Auerbach H, Theobald P, Kroschewski B, Weiss
K. Effects of various additives on fermentation,
aerobic stability and volatile organic compounds
in whole-crop rye silage. Agron 2020; 10(1):1–17.
https://doi.org/10.3390/agronomy10121873
Ávila CLS, Carvalho BF. Silage fermentation—
updates focusing on the performance of micro-
organisms. J Appl Microbiol 2020; 128(4):
966–984. https://doi.org/10.1111/jam.14450
Borreani G, Tabacco E, Schmidt RJ, Holmes
BJ, Muck RE. Silage review: factors
affecting dry matter and quality losses in
silages. J Dairy Sci 2018; 101(5):3952–3979.
https://doi.org/10.3168/jds.2017-13837
Carvalho FAL, Resende PX, Benício CA, Siqueira
JO, Menezes DR, Queiroz MAA. Chemical-
bromatological composition and in vitro ruminal
kinetics of sugarcane silage with maniçoba.
Acta Sci Anim Sci 2018; 40(e42569):1–7.
https://doi.org/10.4025/actascianimsci.v40i1.42569
Carvalho FAL, Queiroz MAA, Silva JG,
Voltolini TV. Características fermentativas na
ensilagem de cana-de-açúcar com maniçoba.
Ci Rural 2014; 44(1):2078–2083. http://dx.doi.
org/10.1590/0103-8478cr20131471
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https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
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drying temperature towards physicochemical
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su14116743
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journal/v21i2/7-NutritiveShokri.pdf
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FS, Souza JSR, Andrade AP, Lima IE, Oliveira
LP and Nascimento DB. Chemical and mineral
composition, kinetics of degradation and in vitro
gas production of native cactus. J Agric Stud
2019; 7(4):119–137. https://doi.org/10.5296/jas.
v7i4.15315
https://doi.org/10.17533/udea.rccp.v37n1a5Ruminal degradation of sugarcane silage with butterfly pea hay
Costa ER, Mello ACL, Guim A, Costa SBM, Abreu
BS, Silva PHF, Silva VJ, Simões Neto DE. Adding
corn meal into mixed elephant grass–butterfly pea
legume silages improves nutritive value and dry
matter recovery. J Agric Sci 2022; 160:185–193.
https://doi.org/10.1017/S0021859622000284
Costa DR, Ribeiro KG, Cruz GFL, Silva
TC, Cardoso LL, Pereira OG. Mixed silages
of sugarcane and forage peanut treated with
Lactobacillus buchneri. Ci Anim Bras 2022;
23:e–72352E. https://doi.org/10.1590/1809-
6891v23e-72352E
Cunha DS, Rodrigues JMCS, Costa CJP,
Lima RS, Araújo CA, Oliveira GF, Campos
FS, Magalhães ALR, Araújo GGL, Gois GC.
Mineral profile, carbohydrates fractionation,
nitrogen compounds and in vitro gas production
of elephant grass silages associated with cactus
pear. Bulletin Nat Res Centre 2022; 46:e257.
https://doi.org/10.1186/s42269-022-00948-0
Del Valle TA, Antonio G, Zenatti TF, Campana
M, Zilio EMC, Ghizzi LG, Gandra JR, Osório
JAC, Morais JPG. Effects of xylanase on the
fermentation profile and chemical composition of
sugarcane silage. JAgric Sci 2019; 156:1123–1129.
https://doi.org/10.1017/S0021859618001090
Figueiredo MRP, Teixeira ACB, Bittencourt
LL, Moreira GR, Ribeiro AJ, Silva FSG,
Santos ALP, Costa MLL. Elephant grass
silage with addition of regional by-products.
Acta Sci Anim Sci 2022; 44:e56616.
https://doi.org/10.4025/actascianimsci.v44i1.56616
Gonzalez-Garcia RA, McCubbin T, Navone L,
Stowers C, Nielsen LK, Marcellin E. Microbial
propionic acid production. Ferment 2017; 3(1):
1–20. https://doi.org/10.3390/fermentation3020021
Hall MB. Challenges with non-fiber carbohydrate
methods. J Anim Sci 2003; 81(12):3226–3232.
https://doi.org/10.2527/2003.81123226x
Harlan DW, Holter JB, Hayes HH. Detergent
fiber traits to predict productive energy of
forages fed free choice to non lactanting dairy
cattle. J Dairy Sci 1991; 74(4):1337–1353.
https://doi.org/10.3168/jds.S0022-0302(91)78289-1
Hariadi H, Karim MA, Hanifah U, Haryanto
A, Novrinaldi, Surahman DN, Hendarwin,
Astro M, Assalam S, Lubis RFL. Effect of
butterfly-pea powder (Clitoria ternatea L.) and
drying temperature towards physicochemical
characteristics of butterfly-pea milk powder with
vacuum drying method. Food Sci Technol 2023;
43:e109622. https://doi.org/10.1590/fst.109622
Hawu O, Ravhuhali KE, Mokoboki HK,
Lebopa CK, Sipango N. Sustainable use of
legume residues: Effect on nutritive value and
ensiling characteristics of maize straw silage.
Sustain 2022; 14:e6743. https://doi.org/10.3390/
su14116743
Jaafar NF, Ramli ME, Salleh RM. Optimum
extraction condition of Clitorea ternatea
flower on antioxidant activities, total
phenolic, total flavonoid and total anthocyanin
contents. Trop Life Sci Res 2020; 31(2):1–17.
https://doi.org/10.21315/tlsr2020.31.2.1
Jusoh S, Nur Hafifah CS. Nutritive value,
palatability and selectivity of 10 different legume
herbages by rabbits. Malaysian J Anim Sci 2018;
21(2):69–75. http://mjas.my/mjas-v2/rf/pages/
journal/v21i2/7-NutritiveShokri.pdf
Kung Jr L, Ranjit NK. The effect of
Lactobacillus buchneri and other additives on
the fermentation and aerobic stability of barley
silage. J Dairy Sci 2001; 84(5):1149–1155.
https://doi.org/10.3168/jds.S0022-0302(01)74575-4
Lemos MF, Mello ACL, Guim A, Cunha MV,
Silva PHF, Atroch TMA, Simões Neto DE,
Oliveira Neto PM, Medeiros AS, Clemente
JVF. Grass size and butterfly pea inclusion
modify the nutritional value of elephant grass
silage. Pesq Agropec Bras 2021; 56:e02409.
https://doi.org/10.1590/S1678-3921.pab2021.v56.02409
Magalhães ALR, Teodoro AL, Gois GC, Campos
FS, Souza JSR, Andrade AP, Lima IE, Oliveira
LP and Nascimento DB. Chemical and mineral
composition, kinetics of degradation and in vitro
gas production of native cactus. J Agric Stud
2019; 7(4):119–137. https://doi.org/10.5296/jas.
v7i4.15315
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Chalcombe Publications; 1991.
Menezes DR, Costa RG, Araújo GGL,
Pereira LGR, Nunes ACB, Henrique LT,
Rodrigues RTS. Ruminal kinetics of diets
containing detoxicated castor bean meal. Arq
Bras Med Vet Zootec 2015; 67(2):636–641.
https://doi.org/10.1590/1678-7040
Multisona RR, Shirodkar S, Arnold M,
Gramza-Michalowska A. Clitoria ternatea
flower and its bioactive compounds: potential
use as microencapsulated ingredient for
functional foods. Appl Sci 2023; 13:e2134.
https://doi.org/10.3390/app13042134
Ørskov ER, Mcdonald I. The estimation of protein
degradability in the rumen from incubation
measurements of feed in weighted according to
rate passage. J Agric Sci 1979; 92(2):499–503.
https://doi.org/10.1017/S0021859600063048
Paulino AS, Almeida VVS, Oliveira AC, Oliveira
HC, Garcia R, Silva RR, Santos P, Silva YA, Bispo
SB, Lima Júnior DM. Influence of increased
doses of detoxified castor bean meal on chemical
composition and characteristics of sugarcane
silage. Chilean J Agric Res 2018; 78:503–510.
https://doi.org/10.4067/S0718-58392018000400503
Pereira FDS, Menezes DR, Araújo EJB,
Rodrigues RTS, Andreo N, Mattos CW, Quadros
CP, Costa CF, Wagner R, Vendruscolo RG. Diets
containing cunhã (Clitoria ternatea L.) hay and
forage cactus (Opuntia sp.) meal on production
and meat quality of Boer crossbred goat. Trop
Anim Health Prod 2020; 52(5):2707–2713.
https://doi.org/10.1007/s11250-020-02225-6
Pereira DS, Lana RP, Carmo DL, Costa YKS.
Chemical composition and fermentative losses
of mixed sugarcane and pigeon pea silage.
Acta Scient Anim Sci 2019; 41(e43709):1-5.
https://doi.org/10.4025/actascianimsci.v41i1.43709
Pratiwi DY. Potential use of butterfly pea (Clitoria
ternatea) as fish feed ingredient: a minireview.
Asian J Fisheries Aquatic Res 2022; 18(2):1–5.
https://journalajfar.com/index.php/AJFAR/
article/view/412/823
Queiroz MAA, Silva JG, Galati RL,
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