addition in maize stover-based substrate fermented in vitro
| Livestock Research for Rural Development 32 (6) 2020 | LRRD Search | LRRD Misssion | Guide for preparation of papers | LRRD Newsletter | Citation of this paper |
The objective of this study was to evaluate the effect of cinnamon powder on methane emission and degradation of maize stover-based substrate incubated in an in vitro fermentation medium. The substrate incubated was maize stover and concentrate mixture in ratio 6: 4. The cinnamon powder was added to the substrate at levels of 0, 5, 10, 15 and 30 mg/g dry matter (DM) to make five treatments. The experiment was laid out in a completely randomized design with six replicates per treatment. Phytochemical composition of cinnamon powder was determined. Approximately 200 mg of the substrate was weighed into 100 ml glass syringes and 30 ml of rumen fluid and a buffer solution (1:2 v/v) was added. Total gas volume, methane production, dry matter and fibre degradation were measured after 48 h incubation period.
Cinnamon powder had high flavonoids, saponin and tannin contents. Methane production in total gas reduced with the addition of cinnamon powder in the substrate. Percentage of methane reduction was between 7 and 14 %. Addition of 5 mg/g of cinnamon power reduced methane production without decreasing dry matter and fibre degradation in maize stover-based substrate.
Keywords: crop residue, feed degradation, phytogenic additive, gas production
Crops residues are important feed resources for ruminants to alleviate feed deficits, particularly in the tropics when natural pastures are not available (Tesfaye and Chairatanayuth 2007). Maize stover is one of the main crop straws widely distributed, highly abundant, of low cost and not competitive in its usage (Li et al 2014). It is considered the most abundant in the world for ruminant feeding (Tang et al 2011). However, degradation in the rumen due to their high fibre content is associated with high methane emissions (Yanti and Yayota 2017; Berhanu et al 2019).
Methane emission poses a great challenge to environmental sustainability being a major contributor to greenhouse gas (Chuntrakort et al 2014). Besides its environmental hazard, it represents a loss of carbon sources culminating to unproductive dietary energy use, with considerable loss of up to 12% of dietary energy intake (Kim et al 2012). Therefore, regulating methane losses from these feed resources becomes essential.
Current dietary strategies for mitigating enteric methane targets modifying rumen fermentation with the use of plant secondary metabolites (Kumar et al 2014; Bhatta 2015; Patra and Saxena 2010). Plant secondary metabolites are bioactive compounds in plants such as saponins, tannins, essential oils, organosulphur compounds and flavonoids (Patra and Saxena 2010). The methane suppressing effect of plant secondary metabolites is mainly due to their antimicrobial properties (Hague et al 2018). Herbs and spices have been studied as sources of plants secondary metabolites to alter rumen fermentation towards methane reduction (Garcia-Gonzalez et al 2008; Chaudhry and Khan 2012; Pawar et al 2014; Medjekal et al 2017). They are considered as natural and safe alternatives to chemical feed antibiotics (Yang et al 2015).
Cinnamon species is one of the most popular spices used worldwide, known also for its antimicrobial effect (Hajimonfarednejad et al 2019). The main bioactive compounds in cinnamon are polyphenols and cinnamaldehyde (Ribeiro-Santos et al 2017). Studies of Shihabudeen et al (2011) also confirms the presence of flavonoids, tannins, saponins, steroid, glycosides, coumarins, anthraquinones and alkaloids in cinnamon bark extract. Methane inhibitory effects of cinnamon extracts have been reported (Chaudhry and Khan 2012; Pawar et al 2014). However, the effectiveness of these plant additives in the rumen is inconsistent and dependent on several factors, one of which is the diet composition (Patra and Saxena 2009). Levels of phytogenic additives in the diet at which substantial reduction in enteric methane can be attained without adverse effect on rumen fermentation of fibrous feeds are also scarce in the literature. The objective of this study was to evaluate the effect of cinnamon powder on methane emission and degradation of maize stover-based substrates in an in vitro medium.
This study was conducted at the laboratory of the Animal Nutrition Department, College of Animal Science and Livestock Production, Federal University of Agriculture, Abeokuta, Ogun State, Nigeria.
Cinnamon sticks were sourced from a local market. The sticks were free of foreign materials and were ground to pass through a 1 mm sieve. The ground cinnamon sticks were subsequently added as a non-nutritive phytogenic additive to a maize stover-based feed substrate for in vitro studies.
The substrate (Table 1) was a complete feed of maize stover and concentrate mixture in ratio 6: 4 (dry matter basis). Cinnamon powder was added at levels of 0, 5, 10, 15 and 30 mg/g of substrate. The experiment was laid out in a completely randomized design with five treatments and each treatment was replicated six times.
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Table 1. Nutrient composition (%) of maize stover and concentrate used as a substrate |
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|
|
Maize stover |
Concentrate# |
|
Crude protein |
5.27 |
15.0 |
|
Organic matter |
92.2 |
93.6 |
|
Nitrogen free extract |
50.6 |
56.4 |
|
Neutral detergent fibre |
68.8 |
51.3 |
|
#concentrate contained 44% wheat offal, 38% rice bran, 15% brewers’ dried grain, 2% bone meal and 1% salt |
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The in vitro gas production technique of Menke and Steingass (1988) was used to measure total gas production from microbial fermentation of substrates. Approximately 200 mg dried and milled sample from each substrate was weighed into 100 ml calibrated glass syringes in six replicates. Each of the syringes was filled with 30 ml inoculum which was a mixture of a medium solution and rumen liquor (2: 1 v/v). The medium contained macro and micro mineral solutions, resazurin and bicarbonate buffer solution as outlined by Menke and Steingass (1988). The rumen fluid was collected from three goats through suction tubes and strained through four-layered sieve cloth. The goats were previously fed grass and concentrate diet to ensure high microbial harvest in rumen fluid. The tip of the syringes was fitted with a rubber tube and sealed to prevent the escape of gas. Syringes were incubated at 39℃ for 48 h and gas readings were recorded at three hours interval from 0 to 48 h. Incubation procedure was handled under continuous anaerobic conditions. Three blanks were included in the run to correct for gas production not arising from substrate fermentation. Methane was determined from the total gas produced at 48 h according to the procedure of Fievez et al (2005) and the percentage of methane from total gas was estimated.
Substrate degraded (mg/200 mg) was determined by subtracting the un-degraded residue for each sample from the initial quantity incubated. The residue was recovered by filtering and drying at 105℃ for 12 h. Neutral detergent fibre degradation was also determined by subtracting the residue neutral detergent fibre from that of the substrate incubated. The difference was then related as a percentage of the substrate incubated.
Oven-dried samples (n=3) of maize stover and concentrate mixture used in this experiment were analyzed for the proximate composition according to AOAC (2000). Fibre fractions in maize stover, concentrate and residues were determined following the procedure of Van Soest et al (1991). The tannin content in cinnamon powder was determined using the Folin Ciocalteu method according to Makkar (2003). Total phenol was determined using Folin Ciocalteu reagent method following Do et al (2014). Flavonoids were determined using aluminium chloride colourimetric method as described by Nasseri et al (2019). Oxalate was determined by permanganate titration method as described by Mishra et al (2017). Saponin was determined as described by Mir et al (2016) while alkaloid was determined by a gravimetric method as described by Adeniyi et al (2009).
Data were analyzed using one-way analysis of variance procedure of SAS (2002). Significant differences between means were separated using Duncan multiple range test (SAS 2002).
Cinnamon powder contained a high proportion of flavonoids in addition to saponin, tannin, total phenol, alkaloids and oxalate (Table 2). Total gas production declined with the addition of cinnamon powder from 10 mg/g up to 30 mg/g in substrate degraded in vitro (Table 3; Figure 1).
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Table 2. Plant secondary metabolites in cinnamon powder |
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|
Concentration % DM |
|
|
Tannin |
5.64 |
|
Flavonoids |
7.21 |
|
Total phenol |
4.41 |
|
Saponin |
6.02 |
|
Oxalate |
1.45 |
|
Alkaloids |
3.23 |
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Table 3. Effect of cinnamon powder on gas production of maize stover-based substrate in an in vitro medium |
|
|
Cinnamon powder |
Gas |
|
0 |
32.5a |
|
5 |
31.2ab |
|
10 |
28.5bc |
|
15 |
27.3c |
|
30 |
24.8 |
|
SEM |
0.75 |
|
P |
0.003 |
|
|
| Figure 1. Curvilinear trend between gas production andcinnamon powder
addition in maize stover-based substrate fermented in vitro |
Methane as a percentage of total gas volume reduced with cinnamon powder added to the substrate at 5 to 30 mg/g (Table 4; Figure 2). The percentage reduction relative to the control was between 7 to 14%. Addition of cinnamon powder reduced substrate dry matter (Table 4; Figure 3) and fibre degradation (Table 4) in vitro at 10 to 30 mg/g.
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Table 4. Effect of cinnamon powder on methane production and degradation of maize stover-based substrate |
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|
CTRL |
CN5 |
CN10 |
CN15 |
CN30 |
SEM |
p |
|
|
Methane production, % |
35.5a |
28.6b |
25.2bc |
21.9c |
22.5c |
1.10 |
<0.001 |
|
Substrate degraded, % |
69.0a |
56.5ab |
51.1b |
49.8b |
47.3b |
2.30 |
0.013 |
|
NDF degradation, % |
61.7a |
58.8ab |
54.2bc |
50.3c |
43.7d |
1.47 |
<0.001 |
|
abcd Means in the same row without common letters are different at P<0.05; CTRL: substrate without cinnamon powder; CN5: substrate with 5 mg/g cinnamon powder; CN10: substrate with 10 mg/g cinnamon powder; CN15: substrate with 15 mg/g cinnamon powder CN30: substrate with 30 mg/g cinnamon powder; SEM: standard error of mean |
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 |
| Figure 2. Curvilinear trend between methane production and cinnamon powder addition in maize stover-based substrate fermented in vitro |
 |
| Figure 3. Curvilinear trend between substrate degradation and cinnamon powder addition in maize stover-based sub strate fermented in vitro |
Cinnamon powder contained plant secondary metabolites which have been confirmed to exert antimicrobial properties (Wallace 2004; Compean and Ynalvez 2014; Yang et al 2015.). The presence of flavonoids, tannin, saponin and alkaloids have similarly been reported in cinnamon bark extracts (Shihabudeen et al 2011). The reduction effect of cinnamon powder on methane production and substrate degradation from our study was attributed to the phytochemicals in cinnamon. Methane inhibitory effect of flavonoids (Oskoueian et al 2013; Kim et al 2015), saponin (Wang et al 2009) and tannin (Anantasook et al 2014) have been well documented. Studies of Chaudhry and Khan (2012) similarly reported a reduction in methane production with cinnamon addition in wheat and rye grass-based diets fermented in vitro. Several mechanisms by which phytogenic feed additives reduce rumen methane have been proposed, part of which includes via direct inhibition of methanogens or indirectly through protozoa inhibition (Patra and Saxena 2010). Methane reduction potentials of plant secondary metabolites have also been attributed to inhibition of fibre degradation (Aderao et al 2018).
Similar to our result, methane reduction with a concomitant decrease in feed degradation had previously been reported with phytogenic feed additives (Hess et al 2006; Bodas et al 2012; Oskoueian et al 2013; Cobellis et al 2016; Joch et al 2019). In crop residue-based diets, this could imply efficient fibre degradation in the rumen. A positive correlation between methane production and Ruminococcaceae, which are fibre degrading microbes have been reported (Joch et al 2018). Reducing methane production without substantially suppressing fibre degradation would be beneficial in high fibre diets to ensure the adequate microbial breakdown of fibre within the rumen. However, this has been considered as challenging (Goel and Makkar 2012). Our study proposed that 5 mg/g of cinnamon powder addition in maize stover-based diets could reduce enteric methane without adverse effect on rumen fibre degradation. To the best of our knowledge, inclusion levels of phytogenic additives in maize stover-based diets for ruminants at which methane can be reduced without negative effect on fibre degradation have not previously been proposed.
The authors acknowledge the laboratory materials support for this research from the Nigeria Tertiary Education Trust Fund.
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Received 19 April 2020; Accepted 25 April 2020; Published 1 June 2020