Extrato metanólico de <i>Urochloa humidicola</i> na fermentação ruminal in vitro

O objetivo deste trabalho foi avaliar o efeito da adição de extrato metanólico de Urochloa humidicola em quatro diferentes concentrações (0, 75, 150 e 250 g L^-1) sobre a fermentação ruminal in vitro de Urochloa brizantha. As seguintes variáveis foram avaliadas pela técnica de produção de gás in vitro: parâmetros cinéticos; degradação ruminal da matéria seca; e produção e concentração dos gases metano e dióxido de carbono e dos ácidos graxos de cadeia curta acetato, propionato e butirato. A inclusão do extrato metanólico reduz a produção de gases oriundos da degradação dos carboidratos não fibrosos (fração A) em 9,55, 6,67 e 13,33%, respectivamente, às concentrações de 75, 150 e 250 g L^-1, em comparação ao grupo controle, mas influencia negativamente a degradação da matéria seca de U. brizantha às concentrações de 150 e 250 g L^-1. O extrato apresenta efeito quadrático negativo na produção de gás durante 12 e 24 horas de incubação de U. brizantha. O extrato de U. humidicola reduz a produção de metano e aumenta a produção de ácidos graxos de cadeia curta às concentrações de 75, 150, e 250 g L^-1.

Termos para indexação:
Urochloa humidicola; mitigação de metano; metabolismo ruminal; metabólitos secundários de plantas

Autoria

Rafaela Scalise Xavier de Freitas

Universidade Federal Rural do Rio de Janeiro, BR 465, Km 7, CEP 23890-000 Seropédica, RJ, Brazil. E-mail: rafascalise@hotmail.com, delci_ufrrj@yahoo.com.br, ecmodesto@gmail.com, tpireszootec@gmail.com, joaocarlosbq@gmail.com, leonardofiusa@yahoo.com.brUniversidade Federal Rural do Rio de JaneiroBrazilSeropédica, RJ, BrazilUniversidade Federal Rural do Rio de Janeiro, BR 465, Km 7, CEP 23890-000 Seropédica, RJ, Brazil. E-mail: rafascalise@hotmail.com, delci_ufrrj@yahoo.com.br, ecmodesto@gmail.com, tpireszootec@gmail.com, joaocarlosbq@gmail.com, leonardofiusa@yahoo.com.br

Delci de Deus Nepomuceno

Elisa Cristina Modesto

Tatiana Pires Pereira

João Carlos de Carvalho Almeida

Leonardo Fiusa de Morais

Luiz Gustavo Ribeiro Pereira

Embrapa Gado de Leite, Eugênio do Nascimento, no 610, Dom Bosco, CEP 36038-330 Juiz de Fora, MG, Brazil. E-mail: luiz.gustavo@embrapa.br, fernanda.samarini@embrapa.br, mariana.campos@embrapa.br, thierry.tomich@embrapa.brEmbrapa Gado de LeiteBrazilJuiz de Fora, MG, BrazilEmbrapa Gado de Leite, Eugênio do Nascimento, no 610, Dom Bosco, CEP 36038-330 Juiz de Fora, MG, Brazil. E-mail: luiz.gustavo@embrapa.br, fernanda.samarini@embrapa.br, mariana.campos@embrapa.br, thierry.tomich@embrapa.br

Fernanda Samarini Machado

Mariana Magalhaes Campos

Thierry Ribeiro Tomich

SCIMAGO INSTITUTIONS RANKINGS

Figuras | Tabelas | Fórmulas

Figuras (1)
Tabelas (4)
Fórmulas (3)

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Figure 1.
Cumulative gas production of Urochloa brizantha at different concentrations of the crude methanolic extract of Urochloa humidicola (MEUh).

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Table 1.
Chemical-bromatological composition of Urochloa brizantha, Urochloa humidicola, and U. humidicola methanol extract (MEUh) in percentage of dry matter.

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Table 2.
Estimation of the kinetic parameters⁽¹⁾ of the in vitro gas production of the dry matter of Urochloa brizantha at different concentrations of the methanolic extract of Urochloa humidicola.

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Table 3.
Estimations of in vitro rumen degradation of the dry matter of Urochloa brizantha when crude methanolic extract of Urochloa humidicola was added at different concentrations⁽¹⁾.

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Table 4.
Gas production, dry matter degradation (DMD), pH of ruminal fluid, production of carbon dioxide of the incubated dry matter of Urochloa brizantha (CO₂MLI), production of CO₂ of the degraded dry matter of U. brizantha (CO₂MLD), methane of the incubated dry matter of U. brizantha (CH₄MLI), CH₄ of the degraded dry matter of U. brizantha (CH₄MLD), total production of short-chain fatty acids (SCFAs), acetic acid (HOAc), propionic acid (PPA), and butyric acid (BTA), according to the concentrations of the methanolic extract of Urochloa humidicola at 12 and 24 hours of incubation⁽¹⁾.

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Figure 1. Cumulative gas production of Urochloa brizantha at different concentrations of the crude methanolic extract of Urochloa humidicola (MEUh).

Table 1. Chemical-bromatological composition of Urochloa brizantha, Urochloa humidicola, and U. humidicola methanol extract (MEUh) in percentage of dry matter.

Variable⁽¹⁾	Urochloa brizantha ⁽²⁾	Urochloa humidicola ⁽³⁾	MEUh⁽³⁾
Dry matter (DM)	91.77	89.36	81.42
Crude protein (CP)	12.56	5.17	10.20
Ether extract (EE)	1.59	1.57	35.00
Mineral matter (MM)	11.58	8.14	16.14
NFC	6.24	9.53	38.52
NDF	68.03	75.59	0.14
ADF	39.86	40.77	0.18
Lignin	-⁽⁴⁾	7.19	0.17

⁽¹⁾
NFC, non-fibrous carbohydrates; NDF, neutral detergent fiber; and ADF, acid detergent fiber. ⁽²⁾Results of the chemical-bromatological analysis of U. brizantha (sample 2012613 of Embrapa Gado de Leite, protocol number 20,120,089). ⁽³⁾Results of the chemical-bromatological analysis of U. humidicola and U. humidicola methanol extract obtained at the laboratory of animal nutrition of Universidade Federal Rural do Rio de Janeiro. ⁽⁴⁾Not analyzed.

Table 2. Estimation of the kinetic parameters⁽¹⁾ of the in vitro gas production of the dry matter of Urochloa brizantha at different concentrations of the methanolic extract of Urochloa humidicola.

Concentration (g L^-1)	A (mL)	L (min)	T/2 (hours)	µ (mL h^-1)
0	203	97	21.23	0.036
75	283	0.008	12.91	0.048
150	273	0.003	11.81	0.073
250	275	0.004	13.40	0.050

⁽¹⁾
Obtained by the model of France et al. (1993)FRANCE, J.; DHANOA, M.S.; THEODOROU, M.K.; LISTER, S.J.; DAVIES, D.R.; ISAC, D. A model to interpret gas accumulation profiles with “in vitro” degradation of ruminant feeds. Journal of Theoretical Biology, v.163, p.99-111, 1993. DOI: 10.1006/jtbi.1993.1109.
https://doi.org/10.1006/jtbi.1993.1109... : Y = A {1 -exp [(- b(t-L) - c) x (√t-√L)]}, where A is the maximum potential gas production, L is the lag phase, T/2 is the time to reach half of the asymptote, and µ is the fractional rate of gas production.

Table 3. Estimations of in vitro rumen degradation of the dry matter of Urochloa brizantha when crude methanolic extract of Urochloa humidicola was added at different concentrations⁽¹⁾.

Concentration (g L^-1)	Parameter			DE			R²
Concentration (g L^-1)	a (%)	b (%)	c (%)	2	5	8	R²
0	4.24^ns	69.80	0.03	47.52	31.80	24.46	0.99
75	13.79	49.79	0.02	38.55	27.91	23.66	0.95
150	10.91	25.27	0.04	27.75	22.14	19.33	0.98
250	17.57	4.18	0.04^ns	20.30	19.37	18.91	0.63

⁽¹⁾
a, soluble fraction of dry matter; b, potentially degradable insoluble fraction of dry matter; c, degradation rate by fermentative action of b; DE, effective rumen degradability; K, estimated passage rates of 2, 5, and 8% per hour, representing animals with low, medium, and high consumption, respectively; and R², coefficient of determination. The results were obtained through the mean degradation times (12, 24, 48, and 96 hours). *Significant by the analysis of variance for nonlinear regression, at 5% probability. ^nsNonsignificant.

Table 4. Gas production, dry matter degradation (DMD), pH of ruminal fluid, production of carbon dioxide of the incubated dry matter of Urochloa brizantha (CO₂MLI), production of CO₂ of the degraded dry matter of U. brizantha (CO₂MLD), methane of the incubated dry matter of U. brizantha (CH₄MLI), CH₄ of the degraded dry matter of U. brizantha (CH₄MLD), total production of short-chain fatty acids (SCFAs), acetic acid (HOAc), propionic acid (PPA), and butyric acid (BTA), according to the concentrations of the methanolic extract of Urochloa humidicola at 12 and 24 hours of incubation⁽¹⁾.

Parameter	T (hours)	Concentration (g L^-1)				MSE	CV (%)	Effect	R²	p-value
Parameter	T (hours)	0	75	150	250	MSE	CV (%)	Effect	R²	C	DT	CxDT
Gas production (mL)	12	51.33	127.67	144.00	132.67	40.81	50.21	Q	0.87	*	**	ns
Gas production (mL)	24	102.67	185.67	210.00	172.33	43.79	45.23	Q	0.85	*	**	ns
DMD (%)	12	27.33	24.13	20.73	19.19	1.10	7.23	L	0.88	*	ns	*
DMD (%)	24	41.02	32.73	25.19	25.72	1.45	8.46	L	0.95	*	ns	*
pH	12	6.88	6.57	6.28	5.73	0.02	0.49	L	0.88	*	**	ns
pH	24	6.69	6.42	6.15	5.43	0.02	0.51	L	0.89	*	**	ns
CO₂MLI (mL g^-1)	12	4.39	12.17	12.57	12.15	0.74	8.13	Q	0.84	*	ns	***
CO₂MLI (mL g^-1)	24	9.56	24.77	25.19	25.67	0.94	7.63	Q	0.80	*	ns	***
CO₂MLD (mL g^-1)	12	16.37	50.55	61.14	63.68	14.09	37.40	Q	0.92	*	ns	*
CO₂MLD (mL g^-1)	24	23.34	75.87	100.36	131.92	20.53	43.03	Q	0.91	*	ns	*
CH₄MLI (mL g^-1)	12	0.52	1.28	0.21	0.00	0.01	5.20	Q	0.32	*	**	**
CH₄MLI (mL g^-1)	24	2.00	4.69	0.92	0.02	0.02	2.05	Q	0.41	*	**	**
CH₄MLD (mL g^-1)	12	1.93	5.50	1.02	0.06	0.14	6.41	Q	0.29	*	***	***
CH₄MLD (mL g^-1)	24	4.90	14.52	3.70	0.12	0.23	6.37	Q	0.41	*	***	***
SCFA (µmol mL^-1)	12	12.55	32.68	23.11	23.22	1.49	10.07	Q	0.39	*	***	ns
SCFA (µmol mL^-1)	24	18.45	42.55	26.79	25.44	1.77	10.85	Q	0.32	*	***	ns
HOAc (µmol mL^-1)	12	7.69	20.28	19.61	17.55	0.66	6.41	Q	0.81	*	ns	ns
HOAc (µmol mL^-1)	24	11.70	24.93	22.21	18.31	0.79	7.12	Q	0.45	*	ns	ns
PPA (µmol mL^-1)	12	3.40	10.43	-	4.07	0.16	4.04	Q	0.89	*	***	ns
PPA (µmol mL^-1)	24	4.49	12.72	-	9.04	0.21	4.21	Q	0.82	*	***	ns
BTA (µmol mL^-1)	12	1.46	5.44	3.50	1.60	0.04	2.33	Q	0.52	*	ns	ns
BTA (µmol mL^-1)	24	2.25	7.36	4.58	1.10	0.05	2.69	Q	0.63	*	ns	ns

⁽¹⁾
T, incubation time (hours); MSE, mean standard error; CV, coefficient of variation; Effect, effect of linear (L) and quadratic (Q) regressions; R², coefficient of determination; C, concentration (g L^-1); DT, degradation time; and CxDT, interaction between concentration and degradation time. *, **, and ***Significant at 0.01, 1, and 5% probability. ^nsNonsignificant at 5% probability. -, error due to co-eluted peak that did not allow quantification.

Y = A {1 - \exp [(- b (t - L) - c) \times (\sqrt{t} - \sqrt{L})]}

Y_{i j k} = μ + α_{j} + ε_{(i) j} + β_{j} + {(α β)}_{i j} + ω_{k} + e_{i j k},

Y_{i j k} = μ + α_{j} + ε_{(i) j} + δ_{j k} + β_{j} + {(α β)}_{i j} + ω_{k} + e_{i j k},

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Extrato metanólico de <i>Urochloa humidicola</i> na fermentação ruminal in vitro

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[TFN1] ⁽¹⁾
NFC, non-fibrous carbohydrates; NDF, neutral detergent fiber; and ADF, acid detergent fiber. ⁽²⁾Results of the chemical-bromatological analysis of U. brizantha (sample 2012613 of Embrapa Gado de Leite, protocol number 20,120,089). ⁽³⁾Results of the chemical-bromatological analysis of U. humidicola and U. humidicola methanol extract obtained at the laboratory of animal nutrition of Universidade Federal Rural do Rio de Janeiro. ⁽⁴⁾Not analyzed.