Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 54 issue 7 (july 2020) : 851-855

Chemical properties, antioxidant activities and in vitro fermentation profiles of some shrubs of North Western Tunisia

H. Selmi1, M. Hasnaoui2, A. Bahri2, C. Abbes1, S. Dallali1, S. Jedidi1, H. Rouissi2
1Sylvo-Pastoral Institute, University of Jandouba, Tabarka, Tunisia.
2Department of Animal Productions, Higher Institute of Agriculture, University of Carthage, Mateur, Tunisia.
Cite article:- Selmi H., Hasnaoui M., Bahri A., Abbes C., Dallali S., Jedidi S., Rouissi H. (2018). Chemical properties, antioxidant activities and in vitro fermentation profiles of some shrubs of North Western Tunisia . Indian Journal of Animal Research. 54(7): 851-855. doi: 10.18805/ijar.B-953.

ABSTRACT

The main aims of this study were to analyze the chemical composition and total phenolic, and to evaluate the in vitro fermentation and antioxidant activities from some shrubs (Erica arborea, Myrtus communis, Arbutus unedo and Phillyrea latifolia). The determination of the chemical composition, total phenolic, flavonoid contents and the antioxidant activity were investigated by using in vitro methods. The in vitro gas production and ruminal parameters were determined in the presence of goat rumen juice. Total nitrogen content was significantly high with Myrtus communis (p <0.01). Total phenolic and flavonoid content vary by species and season (p <0.01). It can be concluded that all parameters were significantly affected by the seasonal period and the shrubs nature. Expect the OM that was similar regardless the seasonal period.

INTRODUCTION

The inhabitants of the mountainous regions of the Mediterranean practice goats breeding in view of the production and management characteristics of this specie. Indeed, spontaneous vegetation, especially bushy and shrubby strata could contribute to meeting the needs of ruminants raised in this area (Selmi et al., 2015). This could reduce production costs and excessive use of concentrated foods. The north-western region of Tunisia is characterized by rugged mountainous terrain and unsuitable soils, which are occupied by natural forests. The climate is typically Mediterranean with mild and wet winters and hot dry summers. Average annual rainfall is 940 mm, with 77% occurring between October and April. The average temperature is 12.8°C in winter and 32.3°C in summer (Mebirouk-Boudechiche et al., 2015). These climatic conditions make goat farming the main activity of small-scale breeders in this region. Research on the nutritional characterization of this vegetation underlines its richness in secondary compounds, particularly tannins which form complexes with proteins and other macro-molecules, thus preventing their digestion and affecting their nutritional value (Selmi et al., 2010). However, gas production by these shrubs and in particular methane which is the main source of energy loss in ruminants depends on the chemical composition and digestibility of the plant wall. Thus, the aim of this study was to analyze the chemical composition and total phenolic and to evaluate the in vitro fermentation and antioxidant activities from some shrubs (Erica arborea, Myrtus communis, Arbutus unedo and Phillyrea latifolia).

MATERIALS AND METHODS

Four shrubs (Erica arborea, Myrtus communis, Arbutus unedo and Phillyrea latifolia), were sampled in November, January and March. Half of each sample is placed in an oven at 105°C for 48 hours. The other half of the samples were set at 60°C for 24 h and then were ground to pass through a 1mm sieve in a Willey mill. Samples were analyzed for organic matter (OM), by ashing in a muffle furnace at 550°C (AOAC 1990). Crude protein (CP) and FAT were measured according to the combustion method (AOAC 1990). The NDF and ADF contents were determined using an ANKOM200/220 Fibre Analyzer based on the methods described by Van Soest et al., (1991).
       
Determination of the phenolic compounds is carried out using the method of Singleton et al., (1999) using the Folin-Ciocalteu. The principle of this assay consists in: Preparing in the test tubes the following mixture: 500 μl of the methanol extract, 500 μl of the Folin-Ciocalteu reagent (10 times) and 1 mL of Na2 CO3 (20%).
       
The Flavonoids contained in the methanol extracts of plants are estimated by aluminum chloride spectrophotometric assay (Yi et al., 2007). The mixture is vigorously stirred. After 30 minutes of incubation, the absorbance is read at 430 nm, a calibration curve established by quercetin, carried out under the same operating conditions as the samples. While, the evaluation of the total content of condensed tannin was determined using a method described by Sun et al., (1998). The mixture was left in the dark for 15 minutes and the absorbance was measured at 500 nm. The tannin content is determined from a standard Catechin range. The results were expressed as µg of catechin equivalent per gram of dry matter.
       
Plant extracts were tested for the scavenging effect on DPPH radical according to the method of Braca et al., (2001). After 30 min incubation at room temperature, the absorbance was read against a blank at 517 nm in a UV/Vis Jenway ­6300 (Jenway Ltd., United Kingdom).
 
       
Where: I was DPPH inhibition (%), A0 was the absorbance of the control and A1 was the absorbance of the extract/standard.
       
The rumen content is then homogenized and filtered to remove the solid phase. The contents of the flask were emptied into an industrial mixer and purged simultaneously with CO2 to maintain anaerobic conditions (Grant and Mertens 1992). After mixing, the rumen fluid was transferred to a 100 ml glass syringe.  In each syringe we mixed: 10 ml of filtered rumen juice, 20 ml of artificial saliva and 300 mg of crushed substrate (3 replicates per sample). Then the syringe were stored in a water bath at 39°C, purged with CO2 and continued as recommended by Goering and Van Soest (1970).
       
The digestibility of organic matter (DOM) is calculated using the formula proposed by Menke and Steingass (1988). The metabolizable energy (ME) content as well as the volatile fatty acids produced, was calculated according to the method of Makkar (2002).
       
Variance was performed by the software SAS (2000) according to the following model:
 
                                Yijk = μ + Ai + Sj eijk.
 
Where, Yijk, observed variable; μ the overall mean; Ai, effect of species; Sj, effect of season; eijk, random error.

RESULTS AND DISCUSSION

The season affected significantly (p <0.05) the DM content of the shrubs (Table 1 and Table 2). The DM content was significantly affected by the species (p <0.05). Phillyrea latifolia has the highest content compared to other shrubs, while Erica arborea has the lowest value (54.2 in autumn; 43.66 in winter and 40.7% in spring). This result was in agreement with the result found by Grant and Mertens (1992). The FAT content is highly significant (p <0.01) between species and inter-season (p <0.01). In fact, Phillyrea latifolia has the highest values   in Crude Ash, ADF, NDF and Lignin compared to other species. This is similar to the results found by Mebirouk-Bouchiche et al., (2015). Similarly, the incorporation of broken in dosa resulted in a significant increase in crude protein, crude fat, crude fibre, ash, total soluble sugar and total dietary fiber content as compared to control dosa (Subodh et al., 2013).
 

Table 1: Chemical composition (% DM) of maquis shrubs according to the season.


 

Table 2: Parietal composition (% DM) of shrubs studied according to the season.


       
The analysis of total phenols (Table 3) showed that Erica arborea and Arbutus unedo had the highest average concentrations (p <0.01) with 55.52 and 51.74 mg GAE/g, respectively, compared to Phillyrea latifolia and Myrtus communis. In spite of the species, the most prominent total phenol concentrations were observed in spring (p <0.05). This can be explained by the high concentrations of parietal constituents (ADF, NDF and Lignin) with (69.25 ± 5.7; 25.6 ± 2.46; 37.04 ± 1.68 and 56.23 ± 4.92 mg GAE/g) respectively for Arbutus unedo, Phillyrea latifolia, Myrtus communis and Erica arborea. The Erica arborea has the highest value compared to other shrubs. Statistical analysis showed that the season affects the concentration of Flavonoids (p <0.01). Concerning the overall concentration of tannins, the Erica arborea had the highest grade (p <0.01) for the three seasons (6.7g catechin equivalent / kg DM), while the lowest value was displayed for the Phillyrea latifolia (2.35 g EC/ Kg DM). Priyanka and Sudesh (2014) showed that legume and cereal industries by-products had also different concentrations of total phenolic content and antioxidant activity.
 

Table 3: Variation of secondary metabolite concentrations of studied shrubs.


       
Phillyrea latifolia samples collected from winter had significantly the highest inhibition percentages (62.94-59.17%), followed by Arbutus unedo collected from spring (58.25–57.69%). Moreover, this activity was also estimated by evaluating the IC50 values (Table 4), which correspond to the amount of extract required to scavenge the 50% of the DPPH radical’s amount in the reaction mixture. Higher is the IC50 value, lower is the antioxidant activity of the tested sample. The DPPH scavenging activity of the extract is dependent on various bio chemicals besides the polyphenolic contents (Agbor et al., 2005; Rout et al., 2011; Asadujjaman et al., 2013).
 

Table 4: The DPPH radical scavenging activities (%) of methanol extracts.


 
       
The kinetics of gas production of the different species follows an exponential model of 1st degree (Fig 1), whatever the nature of the substrate and the season. In autumn, the gas volume at 24 h of incubation was higher (p <0.05) for Arbutus unedo and Phillyrea latifolia. While Myrtus communis occupies an intermediate position and Erica arborea has the lowest value. This trend continues during the winter season with a dominance of the Phillyrea latifolia which the largest volume accompanied by a significant digestibility has seen the high levels of CP and FAT at this shrub thus promoting a better proliferation microbial rumen. So during the spring season, Arbutus unedo has the highest values (p <0.05) compared to other shrubs. With the exception of Phillyrea latifolia, potential gas production (b) was significantly elevated (p <0.05) for the four species studied during the spring.
 

Fig 1: Gas production (mL per 300mg DM) of shrubs studies.


       
Similarly the in vitro degradability of forage legumes resulted in different volumes of gases produced ranged from 61.38 to 70.37 ml (Katsande et al., 2015).The digestibility of the OM, the metabolizable energy (ME) and the concentration of volatile fatty acids (VFA) are grouped in (Table 5). In fact, the shrubs studied have comparable digestibility values (p> 0.05). The averages are of the order of 50.67; 50.37; 46.9 and 47.27% respectively for Arbutus unedo, Phillyrea latifolia, Myrtus communis and Erica arborea. While the season significantly (p <0.05) affects the digestibility of OM. Mehtap et al., (2016) concluded that the digestible DM and OM were higher when harvested at the mature stage of vegetation. Similarly, the metabolizable energy and total VFA concentration of scrub shrubs were statistically similar (p> 0.05) inter-species, but varied significantly (p <0.01) depending on the season. Spring is the season when ME and VFA levels are highest compared to autumn and winter. This can be explained by the good digestive use of these shrubs by goats, this later translates into intense gas production (Djeridane et al., 2006). The cumulative gas concentration produced by the different shrubs studied shows a significant variability (p <0.05).
 

Table 5: Organic matter digestibility, Metabizable energy and volatile fatty acid (VFA) production according to the season.

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