The invention generally relates to the field of animal nutrition. More specifically, it relates to a herbal extract-based composition for replacing synthetic methionine in livestock diet and a method thereof.
BACKGROUND OF THE INVENTION
Methionine is a sulfur-containing amino acid that is essential for healthy and productive poultry, and is important for many different functions in the body. It is also essential for cell proliferation and development. There are two methionine requirements in livestock: one for maximal performance (i.e., maximum body weight gain or egg production), and a lower level needed for adequate but reduced growth or egg production while still raising a healthy, productive animal.
Why methionine is important in livestock?
Methionine is considered to be the first limiting amino acid in poultry birds and cattle. In poultry birds, adequate dietary level of this amino acid is needed to support optimum growth and carcass yield of fast-growing commercial broilers. Methionine plays an important role in energy production, protein synthesis, enhancing overall growth performance, feed efficiency utilization and livability in broilers. Methionine provides methyl groups, which are needed for several metabolic reactions such as synthesis of epinephrine, choline, carnitine and creatine. Methionine also provides the building blocks for immune cells and tissues. This includes the nonspecific mechanisms such as the skin and mucosa, and the specific mechanisms that include T and B lymphocytes. Thereby, methionine is particularly important for newly hatched poultry chicks that are highly susceptible to infection during the first two weeks of life. Deficiency in methionine can have serious results as below:
- Poor feed conversion
- Retarded growth in meat birds
- Reduced egg production
- Poor feather growth

Increased feather pecking. A methionine-deficient bird will tend to eat feathers in an attempt to obtain enough methionine. Feather pecking can quickly turn into cannibalistic behavior in a flock. In cattle, methionine is a limiting, essential amino acid not synthesized within the animal's tissues. Therefore, it must be present in the diet or provided by the rumen microbes. However, it is difficult for growing cattle to get enough methionine in their diets. Supplemental methionine is especially important for late-gestation cows when they consume poor-quality, dormant native-range forages and hay. Adding a high-quality methionine supplement has been shown to support oxidative balance and improve feed efficiency, proper metabolic activity and lean growth, which all lead to improved performance, better reproduction and increased profits {Reference: www.progressivecattlexom/topics/feed-nutrition/5670-maximize-performance-with-methionine). Natural sources of methionine in poultry diets are typically corn and soybean meal. Grains are typically low in lysine, and legumes (e.g., soybeans) are low in methionine. With this combination of feed ingredients, methionine is typically the first limiting amino acid. One way of meeting the methionine requirement is to feed excessive protein so that the daily intake of methionine is met. However, this results in an excess of nitrogen excretion and is not environment friendly.
Commercial Importance of Methionine
Most of the methionine produced in the world is used for animal feed in livestock production. In 2013, the world market amounted to over 6,00,000 tons/year. However in organic farming there is a ban or strong limitation of using synthetic methionine. In 2005, India allowed a maximum of 15 % nonorganic feed (dry matter) for ruminants and 20 % for non-ruminants, with a reduction of 5 % each by 2010. However, synthetic appetizer, synthetic growth promoters, pure amino acids or abattoir waste as well as genetically modified organisms (GMOs) are prohibited. (Reference: www.ncbi.nlm.nih.gov/pubmed/25381187). For this reason, supplements which can substitute L-methionine are the need of the hour. Existing Sources of Methionine
Existing sources of methionine in feeds are of two types- natural and synthetic. Both have their benefits and limitations as discussed below:

Natural sources of methionine and their limitations Plant sources
i. Oilseed meals such as soybean meal are common high protein poultry feeds which are
industrial bio-waste of vegetable oil industries. However, the extraction process uses
chemical solvents and the remaining meal is not permitted in organic production.
ii. Sesame meal has a high methionine content (1.06%); however, the methionine is not
well digested and is also low in lysine.
iii. Sunflower meal has a methionine content similar to soybean meal but same is present
in hull. The chicken while feeding cannot remove the hull of whole sunflower seeds,
due to which intake of the methionine rich portion gets drastically reduced.
iv. Flaxseeds have a methionine content of 0.62% but when added at high concentration,
lead to an offensive fishy flavor. Hence, their addition in feed is limited to no more than 30%. Animal Sources
i. Animal slaughter by-products (i.e., meat and bone meal) are important ingredients in
conventional poultry diets, but they are banned in organic livestock feed production.
ii. Fishmeal is a good source of methionine (1.68%) in organic livestock production.
However, fish meal also suffers from limitations such as
- Most of the fishmeal contains synthetic preservative ethoxyquine, which is not allowed in organic production.
- Fishmeal can be used only in small amounts because it taints the flavor of meat and eggs.
- Eggs or chicken marketed as "Vegetarian-Fed" will not be able to use fishmeal as a source of methionine.
iii. Earthworms and insects are high-quality proteins, similar to fishmeal, and high in methionine. Worms and insects can convert wastes such as food scraps or animal manure to high-quality protein. However, since worm and insect meal is considered an animal by-product, it cannot be fed in certified organic poultry rations. (Reference :http://articles.extension.org/pages/69042/synthetic-methionine-and-organic poultry-diets

Synthetic sources of methionine
Synthetic DL-Methionine is mainly produced by chemical synthesis from methyl mercaptan, acrolein and hydrogen cyanide. Methionine in tissues typically occurs in the L-form. Although the D-form is not biologically active, poultry have the ability to utilize both D- and L- forms. Common forms of synthetic methionine are
Dry DL methionine: in the form of dry D, L-methionine which is 99% pure. - Liquid DL methionine hydroxy analog-free acid: is equivalent to 88% methionine after the conversion of the analog to the biologically active form. (Reference: http://dx.doi.org/10.15242/IICBE.C0215133) Limitations of synthetic methionine
i. Expensive: Raw materials for DL- methionine synthesis include oil, natural gas, air and water. As the cost of oil goes up, the cost of synthetic methionine also goes up. ii. Harmful and Toxic: Synthetic methionine is metabolized into highly toxic compounds such as methyl-propionate, thereby, adversely altering the performance of livestock. iii. Prohibited in Organic farming: Use of synthetic methionine is prohibited/banned in organic farming practices.
PRIOR ART
Kumari et al. 2012 found Herbal Amino Acid (Methiorep) supplement equally effective as
Synthetic DL Methionine on Broiler Growth Performance and Carcass Quality Traits.
(International Journal of Scientific and Research Publications. 2; 8. 1-6.).
Methiorep and all commercially available herbal methionine supplements are in solid powder
form, which makes it difficult to achieve a homogeneous dispersion of methionine in the food and
to be able to measure methionine with precision. The use of methionine in solid form has
drawbacks that make handling and dosing difficult. The use of liquid products makes it possible
to achieve homogeneous dispersion at easily controllable concentrations.
No patent was found in the Indian Patent Database.

In view of foregoing, there exists a need for natural source of methionine for replacement of synthetic methionine which can be added in drinking water or can be mixed properly in feed e.g. by spraying etc., thus making the administration as well as dose monitoring easier.
Innovative solution provided by Inventors
5 The product of the present invention is a herbal composition which is not just a mere admixture of
herbs but a well-defined extract in liquid form which can be easily and uniformly dispensed in
water leading to proper intake as a dietary additive and achieving desired biological effects. The
composition is prepared by mixing and grinding specific herbs in optimized concentrations and
then preparing an extract in liquid form under acidic and alkaline conditions. The 11 herbs are
10 Allium cepa L., Allium sativum L., Azadirachta indica L., Boerhaavia diffusa L., Cicer arietinum
L., Eclipta prostrata L., Glycine max L., (Full fat soya), Mucuna pruriens L., Phaseolus mungo L., Trigonella foenumgraecum L. and Triticum sativum L.
Advantages
i. Replacement of synthetic methionine effectively-The composition can replace synthetic
15 methionine completely or partially in livestock feeds.
ii. Ease of administration- Being in liquid form, the composition can be added in drinking
water or mixed properly in feed, making the administration as well as dose monitoring
quite easy which is not possible with solid powdered form of methionine.
iii. Dosage required is very low- Dosage of the composition on daily basis is very less, which
20 makes its use easy and economical e.g. 5 ml per 100 birds in chicks, 10 ml per 100 birds
in growers, and 20 ml per 100 birds in broiler finishers.
OBJECTS OF THE PRESENT INVENTION
The object of the present invention is to disclose a herbal extract-based composition for replacement of synthetic methionine in livestock feed.
25 Another object is to disclose a method of preparation of the said herbal extract composition in
liquid form.
A last object is to disclose the comparative results of the composition with synthetic methionine.
6

SUMMARY OF THE INVENTION
The present invention discloses a liquid herbal extract-based composition for replacement of
synthetic methionine used in livestock feed. The same is an extract of 11 herbs viz. Allium cepa
L., Allium sativum L., Azadirachta indica L., Boerhaavia diffusa L., Cicer arietinum L., Eclipta
5 prostrata L., Glycine max L., (Full fat soya), Mucuna pruriens L., Phaseolus mungo L., Trigonella
foenumgraecum L. and Triticum sativum L which are mixed and powdered in specific
concentrations and extracted using a specific method of extraction involving acidic and alkaline
conditions. The composition can replace synthetic methionine completely or partially in livestock
feeds. Being in liquid form, the composition can be added in drinking water or mixed properly in
10 feed, making the administration as well as dose monitoring quite easy which is not possible with
solid powdered form of methionine. Dosage of the composition on daily basis is very less, which makes its use easy and economical e.g. 5ml per 100 birds in chicks, 10ml per 100 birds in growers, and 20ml per 100 birds in broiler finishers.
DETAILED DESCRIPTION OF THE DIAGRAMS
15 -NIL-
DETAILED DESCRIPTION OF THE INVENTION
The present invention discloses a liquid herbal extract-based composition for replacement of
synthetic methionine used in livestock feed. The same is an extract of 11 herbs viz. Allium cepa
L., Allium sativum L., Azadirachta indica L., Boerhaavia diffusa L., Cicer arietinum L., Eclipta
20 prostrata L., Glycine max L., Mucuna pruriens L., Phaseolus mungo L., Trigonella
foenumgraecum L. and Triticum sativum L which are mixed and powdered in specific concentrations and extracted using a specific method of extraction involving acidic and alkaline conditions.
All the herbs used in the composition are available commercially and can be procured online
25 sources also.
The method is described in detail below and consists of the following steps:
7

Step 1: Blending and grinding of specific herbs in optimized ratios: 11 specific herbs at specific concentrations are taken, blended and ground to obtain a coarse powder of mesh size 10 as per Table 1 below:
Table 1: Composition of the Herbal Methionine Replacer
5
S. No. Scientific name of the herb Common name of the herb/Part used Quantity %
1 Allium cepa Linn. Onion (Bulb) 0.20-0.30 , optimally 0.25
2 Allium sativum Garlic (Bulb) 0.20-0.30 , optimally 0.25
3 Azadirachta indica Neem (Leaves) 0.30-0.40 , optimally 0.35
4 Boerhaavia diffusa Red hogweed (Punarnava) (whole plant) 0.40-0.60 , optimally 0.50
5 Cicer arietinum Chickpea (Seeds) 0.50-0.70, optimally 0.60
6 Eclipta prostrata False daisy (whole plant) 0.30-0.40 , optimally 0.35
7 Glycine max (Full fat soya) Soybean (Seeds) 0.30-0.40 , optimally 0.35
8 Mucuna pruriens Velvet bean (Seeds) 0.70-0.80 , optimally 0.75
9 Phaseolus mungo Black gram (Seeds) 0.55-0.65 , optimally 0.60
10 Trigonella foenumgraecum Fenugreek (Seeds) 0.40-0.50 , optimally 0.45
11 Triticum sativum Bread Wheat (Seeds) 0.50-0.70, optimally 0.60
Step 2: Preparation of acid soluble extract- Concentrated Hydrochloric Acid and 30% Hydrogen Peroxide are mixed in the ratio 4:1 and added to herbal powder in the ratio 10:1 (one part herbal powder and 10 parts of acidic solution). The above mixture is stirred at 10-20 rpm for 3 hours and filtered to get acid-soluble extract in liquid form.
10 Step 3: Preparation of alkali soluble extract- The herbal powder residue left after filtration is
used for preparation of alkali soluble extract. To the herbal powder residue, 0.4% to 0.6%,
8

optimally 0.5% w/v solution of sodium hydroxide is added in the ratio 1: 10 [One part of herbal powder and 10 parts of the sodium hydroxide (0.5%)]. The mixture is stirred at 10- 20 rpm for 3 hours at 60-70°C and filtered to get the alkali soluble extract in liquid form.
Step 4: Mixing of the extracts: The acid extract and alkali extracts are mixed to get clear liquid.
5 Step 5: Addition of preservatives and pH adjustment: Preservatives namely Methyl Paraben
Sodium (0.2%w/v), Propyl Paraben Sodium (0.02%w/v), and Sodium Benzoate (0.2% w/v) are
added to ensure product stability for sufficient period of about 36 months to ensure commercial
distribution and stocking. pH of the liquid composition is adjusted to acidic range (pH 4-6) by
addition of sodium hydroxide or hydrochloric acid. The final product obtained is clear brown
10 colored liquid and can be stored at room temperature for about 36 months without any deterioration
in quality.
Recommended Dosage for replacement of synthetic methionine in livestock diet
The composition in a very low concentration, henceforth referred to as the recommended dosage,
added to water daily i.e. 5ml per 100 birds in chicks, 10ml per 100 birds in growers, and 20ml per
15 100 birds in broiler finishers can partially replace up to 50% of the dietary requirements for
synthetic methionine .
Evaluation and scientific validation of the composition
The performances of herbal extract composition of the present invention and synthetic methionine were evaluated on commercial broilers (Vencobb). 20
Experimental design
Three hundred unsexed day-old commercial broiler chicks (Vencobb) of the same hatch and of
similar live body weights were used in the experiment. The chicks were randomly divided into
three equal groups as below:
25 Group I supplemented with the herbal extract composition at the recommended dosage and
synthetic methionine at 0.05% of feed
Group II supplemented with the herbal extract composition at twice the recommended
dosage
9

Group III supplemented with synthetic methionine at 0.1% of feed for 6 weeks.
Results- The chicks fed with herbal extract composition were found to be better than the synthetic
methionine fed chicks in terms of growth, dressing percentage and edible meat yield. Results are
5 given in Table 2 below:
Table 2: Comparison of Herbal Composition of present invention with Synthetic Methionine

Parameters Group -I
(Supplemented with herbal composition at recommended dosage and synthetic methionine at 0.05% of feed) Group-II
(Supplemented with herbal composition at twice the recommended dosage) Group-III
(Fed synthetic methionine at 0.1% of feed for 6 weeks)
Body weight Day -0 Day-42 42.05 g 2522.32 g 41.84 g 2446.26 g 42.12 g 2410.56 g
Dressing percentage 71.20% 70.86% 70.26%
Edible meat yield 1672.67 g 1612.34 g 1574.67 g
The extract can help to significantly reduce feed cost, leading to promotion of organic natural alternatives to synthetic methionine.
Novelty, Inventive Step and Industrial Application
10 Novelty- The herbal replacer for synthetic methionine in livestock diet, consisting of acid and
alkali soluble extracts of 11 specific herbs in specific ratios, has not been disclosed in prior art.
Inventive step- The technical advancement of knowledge lies in disclosing that synthetic methionine being used in livestock diet can be replaced by an herbal composition, in form of a well-defined liquid composition which can be easily added to water. Further, the invention offers
10

economic advantage in using easily available herbs and being required in very less quantity, thus making its use on large scale affordable.
Industrial application- The herbal extract composition can be easily prepared at commercial level by industries making veterinary herbal products.
While the invention is described in detail in its preferred embodiments, it will be obvious to those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention. Such changes and modifications shall be regarded as being within the scope of the invention.

claim

1. A herbal extract-based composition for replacement of synthetic methionine in livestock feed WHEREIN the said composition consists of acid and alkali soluble extracts of 11 specific herbs mixed in specific quantities as below:

S.No. Scientific name of the herb Common name of the herb/Part Quantity (in percent)
1 Allium cepa L. Onion/ Bulb 0.20-0.30 , optimally 0.25
2 Allium sativum L. Garlic/ Bulb 0.20-0.30 , optimally 0.25
3 Azadirachta indica L. Neem/ Leaves 0.30-0.40, optimally 0.3 5
4 Boerhaavia diffusa L. Red hogweed (Punarnava)/ whole plant 0.40-0.60 , optimally 0.50
5 Cicer arietinum L. Chickpea/ seeds 0.50-0.70, optimally 0.60
6 Eclipta prostrata L. False daisy/ whole plant 0.30-0.40, optimally 0.3 5
7 Glycine max L. (Full fat soya) Soybean/seeds 0.30-0.40, optimally 0.3 5
8 Mucuna pruriens L. Velvet bean/ seeds 0.70-0.80 , optimally 0.75
9 Phaseolus mungo L. black gram/ seeds 0.55-0.65 , optimally 0.60
10 Trigonella foenumgraecum L. Fenugreek/seeds 0.40-0.50 , optimally 0.45
11 Triticum sativum L. Bread Wheat/ seeds 0.50-0.70, optimally 0.60
2. A herbal extract-based composition for replacement of synthetic methionine in livestock feed as claimed in claim 1 WHEREIN the said composition is in liquid form.
3. A method of making the composition as claimed in claim 1 WHEREIN the method involves the following steps:
Step 1: Blending and grinding of specific herbs in optimized ratios- 11 specific herbs
at specific concentrations as given in Claim 1 are taken, blended and ground to coarse
powder.
Step 2: Preparation of acid soluble extract- Concentrated Hydrochloric Acid and
Hydrogen Peroxide are mixed in the ratio 4:1 and added to herbal powder in the ratio 10:1

(one-part herbal powder and 10 parts of acidic solution). The above mixture is stirred at
10-20 rpm for 3 hours and filtered to get acid soluble extract in liquid form.
Step 3: Preparation of alkali soluble extract- The herbal powder residue left after
filtration is used for preparation of alkali soluble extract. To the herbal powder residue, 0.4
to 0.6%) w/v optimally 0.5% w/v solution of sodium hydroxide is added in the optimal ratio
of 1:10 (One part of herbal powder and 10 parts of the sodium hydroxide (0.5%>). The
mixture is stirred at 10-20 rpm for 3 hours at 60-70°C and filtered to get the alkali soluble
extract in liquid form.
Step 4: Mixing of the extracts in specific ratio- The acid extract and alkali extract are
mixed to get a clear brown liquid.
Step 5: Addition of preservatives and pH adjustment- Preservatives, namely Methyl
paraben sodium (0.2%w/v), Propyl paraben sodium (0.02%w/v), Sodium benzoate
(0.2%w/v), are added to ensure product stability and pH adjusted to acidic range (pH 4-6).