Ensiling wild sunflower (Tithonia diversifolia) leaves with sugar cane molasses

Wild sunflower (Tithonia diversifolia) was ensiled with 0, 2, 4 and 6% molasses (dry matter basis). Wild sunflower leaves were harvested within the University of Ado Ekiti community, chopped into small pieces of about 2-3cm before ensiling in 2kg plastic bags for periods of 0, 7, 14, 21 and 28 days. Each ensiling date had the 4 molasses levels in 3 replicates and samples were taken from all the bags for physical examination (smell, colour and odour). Phytochemical determinations and pH, NH₃-N, water extractable DM and N and total N were also carried out.

All samples except samples at 0% molasses level turned yellow-brown on the 7^th day and the colour deepened with increased molasses addition. All silages had attractive smell. pH dropped from an average of 6 at the start of ensiling and continued to decrease with increasing length of duration and increasing levels of molasses. The NH₃-N and water soluble N increased with increasing length of ensiling. Major anti-nutrients in Tithonia diversifolia leaves (phytin, tannin, oxalate, alkaloid and flavonoid) gradually decreased with lengthening duration of ensiling. .

A level of 4% molasses and 14 to 21 days ensiling period appeared optimum and most suitable for effective ensiling of Tithonia diversifolia leaves.

Key words: Fermentation, nitrogen, phytochemical constituents, water extractable dry matter, water extractable nitrogen

Introduction

Tithonia diversifolia (Photos 1-3) commonly known as Mexican sunflower was probably introduced into West Africa as an ornamental plant (Akobundu and Agyakwa 1987). A member of the family Asteraceae, Tithonia is an annual aggressive weed growing to a height of about 2.5m and adaptable to most soils. It had been observed to be widely spread in Nigeria where it is found growing on abandoned/waste lands, along major roads and waterways and on cultivated farmlands. Tithonia has been a subject of research interest because of the relatively high nutrient concentrations found in its biomass and because of its ability to extract relatively high amounts of nutrients from the soil (Jama et al 2000). The reported uses of Tithonia include fodder (Anette 1996; Roothaert and Paterson 1997), poultry feed (Odunsi et al 1996), pig feed (Olayeni et al 2006), fuel-wood (Ng’inja et al 1998), compost (Drechsel and Reck 1998; Ng’inja et al 1998; Jama et al 2000), land demarcation (Ng’inja et al 1998), soil erosion control (Ng’inja et al 1998), building materials and shelter for poultry (Otuma et al 1998). It has also been reported that extracts from Tithonia plant parts protect crops from termites (Adoyo et al 1997) and contain chemicals that inhibit plant growth (Baruah et al 1994; Tongma et al 1997) and control insects (Dutta et al 1993). Extracts of Tithonia also have medicinal value for treatment of hepatitis (Lin et al 1993; Kuo and Chen 1997) and control of amoebic dysentery (Tona et al 1998).

This study aimed at investigating the qualitative and quantitative composition of ensiled Tithonia diversifolia leaves treated with molasses as a prelude to incorporation into diets of monogastric animals.

Materials and methods

Location and procedure

The experiment was carried out in the Nutrition Laboratory of the Department of Animal Production and Health Sciences, School of Agriculture and Agricultural Technology, Federal University of Technology, Akure.

Leaves of Tithonia diversifolia (wild sunflower) were harvested fresh from maturing Tithonia diversifolia plants with sharp knives from the top to the middle of the plant when the first inflorescence had opened in 60-80% of Tithonia plants. Harvesting of the leaves was done in the University of Ado-Ekiti community within Ado-Ekiti, a town in the Southwest Nigeria in the rain forest zone on latitude 7^o 40^’ North of the equator and longitude 5^o 15^’ East of the Greenwich Meridian with ambient temperature, 25-37⁰C; relative humidity, 70%; wind, SSW at 11mph (18km/h); barometric pressure, 29.68’ Hg(F). They were chopped with a sharp kitchen knife into small pieces of about 2-3cm and ensiled in plastic bags of 2kg capacity. Sugar cane molasses were added as silage additive at 0, 2, 4 and 6% (DM basis) of the Tithonia leaves. The plastic bags containing the ensiled Tithonia diversifolia leaves were properly sealed to prevent air contamination and put into large plastic buckets to prevent physical damage to the plastic bags and also to prevent the attack of mice. The plastic buckets were stored at room temperature of 20-30⁰C. The mean total sugar content of the molasses was 77.0 Brix. The four molasses levels (0, 2, 4 and 6%) were repeated to correspond with the ensiling periods of 0, 7, 14, 21 and 28 days.

Determination of physical characteristics

Samples were taken from each ensiled plastic bag at different ensiling durations (0, 7, 14, 21 and 28 days) with different molasses levels (0, 2, 4 and 6%) making a total of 20 samples. Physical observation was made of the physical characteristics such as smell and colour and records were taken.

Phytochemical screening for phytin, oxalate, tannin, alkaloid and flavonoid

The extraction and precipitation of phytin in the fresh and ensiled Tithonia leaves were done by the method of Wheeler and Ferrel (1971) while iron in the precipitate was determined as described by Makower (1970). Phytin was determined by using a 4:6 Fe/P ratio to calculate phytin phosphorous and multiplying the phytin phosphorous by 3.55 factor (Young and Greaves 1940). Oxalate content was determined by the titrimetric method of Moir (1953) as modified by Ranjhan and Krishna (1980). Where extracts were intensely coloured, they were decolourised with activated charcoal (Balogun and Fetuga 1980). The polyphenols (tannic acid) were determined by extracting the dried and finely blended fresh and ensiled Tithonia leaves (250mg in 10ml of 70% aqueous acetone) for 2hrs at 30⁰C using Gallenkamp orbital shaker (Survey, UK). Pigments and fats were first removed from the leaves by extracting with diethyl ether containing 1% acetic acid. Thereafter, the total polyphenols (as tannic equivalent) were determined in 0.05, 0.2 or 0.5ml aliquot using Folin Cocalteu (Sigma) and standard tannic acid (0.5mg/ml) as described by Makkar and Goodchild (1996). The HCN (cyanide) was determined after an initial extraction for 2 – 3 min of 5 – 8g material in 0.1M H₃PO₄ by a 2M H₂SO₄ (100⁰C for 50 min) hydrolysis followed by reaction with chloramines-T pyridine barbituric acid (Konig Reaction). KCN dried over concentrated H₂SO₄ was used to calibrate the standard curve from a stock solution containing 75mg KCN/100ml. Alkaloid determination was done using Harbone (1973) method while flavonoid determination was through the method described by Boham and Kocipai-Abyazan (1974).

Other measurements

Similar samples were analysed for total and water extractable dry matter (WV-DM) and nitrogen (N), and fermentation characteristics such as pH and NH₃-N. Water soluble DM, N, crude fibre, NH₃-N were determined according to AOAC (1995).

Statistical analysis

Data were expressed as means + standard deviation of three measurements. One way ANOVA (SPSS 11.0 for Windows, SPSS Inc., Chicago IL, USA) was used to analyse the mean differences of the same parameter. Significant differences were considered where necessary at a level of p < 0.05.

Results

Physical characteristics

All the Tithonia leaf silages were green in colour at the beginning of the ensiling.There was a rapid colour change from green to yellow-brown within the first 7 days except for the silage with 0% molasses that remained green until 21 days when it turned to yellow-brown. The yellow-brown colour was darker at higher concentration of molasses.The smell of all silages including the molasses-free silage was sweet and not offensive but similar to that of vinegar.

Proximate/Chemical characteristics of the Tithonia and molasses

Table 1. Proximate/Chemical characteristics of the Tithonia leaves and molasses
	Tithonia leaves	Molasses
Dry matter, %	11.0	77.1
% of DM
Crude protein	20.6	0.3
Crude fibre	18.9
Ether extracts	4.0
Carbohydrate	42.5	77.0
Ash	14.0	3.4
Antinutrients, mg/100g
Phytin	79.1
Tannin	0.39
Oxalate	1.76
Saponin	2.36
Alkaloid	1.23
Flavonoid	0.87

Chemical composition and pH of Tithonia diversifolia as affected by ensiling duration and molasses levels

At the beginning of the ensiling process (day 0), all treatments had a uniform pH of 6. Except for treatment 1 which had 0% molasses, pH in all treatments later fell below 5 after 14 days (Figure 1; Table 2). A molasses level of at least 4% (fresh basis) appeared to be necessary to ensure pH values of less than 4.

Figure 1. pH of ensiled Tithonia diversifolia as affected by ensiling duration and molasses levels

The solubility of the nitrogenous fraction (WV-N), and the proportion of the N as ammonia-N, increased with length of ensiling time up to 7 days thereafter remaining fairly constant (Table 2). By contrast, the concentrations of all the anti-nutrients decreased with ensiling time. Molasses level had no marked effect on the concentrations of anti-nutrients other than the diluting effect caused by the addition of the molasses.

Table 2. Chemical composition of ensiled Tithonia diversifolia leaves as affected by ensiling duration
Some chemical parameters	Days					SEM	P-value
Some chemical parameters	0	7	14	21	28	SEM	P-value
pH	6.35^a	4.56^b	3.76^c	3.54^c	3.54^c	0.15	0.001
DM, %	17.0^a	20.1^b	17.4^a	17.7^a	17.9^a	0.52	0.002
NH₃-N, mg/kg DM	294^a	968^b	939^b	966^b	1101^b	35.2	0.001
NH₃-N in total N, %	0.86^a	4.12^b	4.05^b	4.07^b	4.25^b	0.31	0.001
N, %DM	2.42^a	2.64^ab	2.76^b	2.78^b	2.89^b	0.04	0.002
WV-DM, % of DM	40.1^a	42.8^a	42.8^a	50.2^b	43.3^a	1.45	0.072
WV-N, % of total N	32.3^a	45.1^b	42.9^b	45.0^b	46.2^b	2.32	0.012
Antinutrients, mg/100g
Phytin	78.5^a	71.4^b	64.3^c	51.1^d	51.0^d	2.25		0.086
Tannin	0.38^a	0.34^a	0.21^b	0.21^b	0.19^b	0.22		0.001
Oxalate	1.75^a	1.45^b	0.40^b	0.33^c	0.30^c	0.05		0.061
Alkaloid	1.23^a	1.19^ab	1.10^b	0.75^c	0.75^c	0.17		0.001
Flavonoid	0.85^a	0.81^a	0.74^b	0.74^b	0.72^b	0.51		0.002
^{a, b, c} Mean values within rows with different superscripts letters are significantly different (P<0.05)

Increasing the molasses level decreased the concentration of ammonia-N in the DM and of ammonia-N as a proportion of the total N (Table 3). The nitrogen solubility was also increased with molasses level.

Table 3. Effect of molasses level on the chemical composition of ensiled Tithonia diversifolia leaves at 28 days of ensiling
Some chemical parameters	Level of molasses, % (fresh basis)				SEM	P-value
Some chemical parameters	0	2	4	6	SEM	P-value
pH	5.3^a	4.9^a	3.8^b	3.7^b	0.18	0.001
DM, %	17.9^a	16.1^b	17.6^a	18.1^a	0.32	0.041
NH₃-N, mg/kg DM	998^a	795^b	702^b	100^c	32.1	0.001
NH₃-N in total N, %	4.02^a	3.85^a	2.45^b	2.18^b	0.31	0.003
N, %DM	2.82^a	2.80^a	2.72^a	2.73^a	0.50	0.007
WV-DM, % in DM	42.9^a	43.2^a	46.5^b	47.1^b	0.05	0.061
WV-N, % of total N	37.3^a	40.1^a	45.7^b	46.1^b	2.01	0.001
Antinutrients, mg/100g
Phytin	49.1^a	47.4^a	40.2^b	40.2^b	0.05	0.002
Tannin	0.17^a	0.16^a	0.08^b	0.08^b	0.44	0.001
Oxalate	0.31^a	0.27^b	0.25^b	0.25^b	0.05	0.061
Alkaloid	0.86^a	0.83^a	0.73^b	0.72^b	2.01	0.001
Flavonoid	0.76^a	0.71^a	0.71^a	0.70^a	0.05	0.002
^{a, b, c} Mean values within rows with different superscripts letters are significantly different (P<0.05)

Table 4. Chemical composition of ensiled Tithonia diversifolia leaves as affected by ensiling duration
	pH		DM, %		NH₃-N, g/kg DM		WV-DM, %DM		WV-N, % of total N
Molasses levels→	0%	4%	0%	4%	0%	4%	0%	4%	0%	4%
Day↓
0	6.5^a	6.3^a	17.0^a	18.2^c	0.22^d	0.21^d	31.0^a	35.0^b	22.0^d	32.0^a
7	6.2^a	5.5^b	20.1^b	20.1^b	1.33^c	0.78^ab	22.0^e	44.5^c	38.0^a	42.0^b
14	5.5^b	4.2^c	17.1^a	18.1^c	1.22^bc	0.72^a	30.5^a	37.0^b	25.0^d	48.0^bc
21	5.3^b	3.8^c	17.1^a	18.3^c	1.02^b	0.83^b	32.0^ab	54.0^d	38.0^ab	41.0^b
28	5.3^b	3.8^c	17.2^a	18.1^c	1.38^c	0.92^b	28.5^a	47.0^c	31.0^a	47.0^bc
SEM	0.19		0.32		32.1		0.31		2.01
P-value	0.032		0.041		0.001		0.003		0.001
^{a, b, c….} Mean values within columns with different superscripts letters are significantly different (P<0.05)

The increase in water-soluble DM with addition of 4 or 6% molasses cannot be explained by the addition of the soluble carbohydrates in molasses (sucrose and reducing sugars).

Discussion

The increase in the concentration of NH₃-N particularly within the first 7 days of ensiling agreed with the report of McDonald et al (2002) that proteolytic organisms mainly Clostridia are active while the pH is still relatively high resulting in the breakdown of protein to amino acids, amines and NH₃. The significant increase in water-extractable DM particularly with increased addition of molasses is in conformity with the report from a similar study with Taro leaves (Colocasia esculenta L. Shott) revealing about 100% increase in water-extractable DM at 4% and 6% molasses addition (Chittavong et al 2008).

It is noteworthy that all the major anti-nutrients identified in fresh Tithonia leaves were significantly reduced during the ensiling period. This is in agreement with several previous studies. Pham Sy Tiep et al (2006) reported a reduction of about 79% in calcium oxalate content of ensiled Alocasia macrorrhiza leaves with rice bran and molasses as additives. Savage and Dubois (2006) and Chittavong et al (2008) reported a significant reduction of about 86% in the oxalate content of the raw leaves of Taro (Colocasia esculenta (var.) Schott).

Conclusions

Acknowledgements

We gratefully acknowledge the technical and laboratory assistance of Mr. Mike Oguntokun of the Animal Nutrition Laboratory, Federal University of Technology, Akure.

References

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Ensiling wild sunflower (Tithonia diversifolia) leaves with sugar cane molasses

A O Fasuyi, F A S Dairo and F J Ibitayo

Department of Animal Production and Health Sciences, Faculty of Agricultural Sciences, University of Ado-Ekiti,
P.M.B. 5363, Ado-Ekiti, Ekiti State, Nigeria
dejifasuyi@yahoo.com

Abstract

Introduction

Materials and methods

Location and procedure

Determination of physical characteristics

Phytochemical screening for phytin, oxalate, tannin, alkaloid and flavonoid

Other measurements

Statistical analysis

Results

Physical characteristics

Proximate/Chemical characteristics of the Tithonia and molasses

**Chemical composition and pH of Tithonia diversifolia as affected by ensiling duration and molasses levels**

Discussion

Conclusions

Acknowledgements

References


Photo 1. Tithonia diversifolia leaves	Photo 2. Tithonia diversifolia at flowering

Photo 3. Tithonia diversifolia growing wild and predominant as major vegetation in a recently cleared plot in Ado-Ekiti, South Western Part of Nigeria

Ensiling wild sunflower (Tithonia diversifolia) leaves with sugar cane molasses

A O Fasuyi, F A S Dairo and F J Ibitayo

Department of Animal Production and Health Sciences, Faculty of Agricultural Sciences, University of Ado-Ekiti, P.M.B. 5363, Ado-Ekiti, Ekiti State, Nigeria dejifasuyi@yahoo.com

Abstract

Introduction

Materials and methods

Location and procedure

Determination of physical characteristics

Phytochemical screening for phytin, oxalate, tannin, alkaloid and flavonoid

Other measurements

Statistical analysis

Results

Physical characteristics

Proximate/Chemical characteristics of the Tithonia and molasses

Chemical composition and pH of Tithonia diversifolia as affected by ensiling duration and molasses levels

Discussion

Conclusions

Acknowledgements

References

Department of Animal Production and Health Sciences, Faculty of Agricultural Sciences, University of Ado-Ekiti,
P.M.B. 5363, Ado-Ekiti, Ekiti State, Nigeria
dejifasuyi@yahoo.com

**Chemical composition and pH of Tithonia diversifolia as affected by ensiling duration and molasses levels**