WO2008120940A1

WO2008120940A1 - Assorted feedstuff composition containing chia seed or its extract, method of farming livestock using the same, and meat acquired thereby

Info

Publication number: WO2008120940A1
Application number: PCT/KR2008/001805
Authority: WO
Inventors: Youn Jin Kim; Ho Jun Kim
Original assignee: Youn Jin Kim; Ho Jun Kim
Priority date: 2007-03-29
Filing date: 2008-03-31
Publication date: 2008-10-09

Abstract

The present invention relates to an assorted feedstuff containing a natural antibiotic that can be mass-produced to increase antibacterial activity and immunity in livestock but not to reduce meat flavor and taste, and a method for farming livestock using the natural antibiotic. The feedstuff composition of the present invention contains an ingredient selected from the group consisting of chia seed, chia meal and a mixture thereof. In a preferred embodiment of the present invention, the feedstuff composition contains chia seed by 1 to 10.8 weight% for the total weight of the composition. According to the method for farming livestock of the present invention, the feedstuff containing one of chia seed, chia meal and a mixture thereof as a natural antibiotic is provided to livestock and separately drinking water is provided considering the water absorptiveness of chia seed dietary fiber. The natural antibiotic of the present invention has greater antibacterial activity than the conventional artificial antibiotics, and cause no problem related with the antibiotic residues in the inside of body and antibiotic resistance, unlike the conventional artificial antibiotics. In addition, the present invention is effective in improving quality of meat without reducing flavor or taste.

Description

[DESCRIPTION] [Title of Invention]

ASSORTED FEEDSTUFF COMPOSITION CONTAINING CHIA SEED OR ITS EXTRACT, METHOD OF FARMING LIVESTOCK USING THE SAME, AND MEAT ACQUIRED THEREBY

[Technical Field]

The present invention relates to a feedstuff composition and a method of farming livestock, more particularly a feedstuff composition containing a natural substance as an antibiotic instead of artificial antibiotics and a method of farming livestock such as chickens or pigs by using the same. Further the present invention relates to meat obtained thereby.

[Background Art]

According to the increase of national income and advance of living standard, dietary habit has been changed to consume more meat. Among meat, chicken and pork which are less expensive but contain plenty of high-quality proteins, have been in increasing demand. The livestock such as chickens and pigs might be in high risk of infection because of an insanitary stall environment. Particularly, in a large scaled factory farm that is very crowded with livestock, once some of livestock are infected, it spreads throughout the farm even to the next farm so fast that the whole livestock might be in danger of death. In general, a feedstuff contains grain such as barley, corn, sorghum and wheat; by-products of flour milling; soybean cake; and minerals as a major component. To prevent disease, feedstuff companies have added a significant amount of antibiotics to feed to inhibit or kill pathogens in the body of livestock. In fact, in the present times, at least half of the total antibiotic consumption for animal was consumed as feed additives, suggesting that antibiotics have been used more as a preventive additive than for the treatment of disease. However, although the antibiotics added to the assorted feed prevent disease and improve digestion by killing intestinal pathogens and accelerate growth, they increase resistance against drugs to reduce disease treatment effect. Moreover, the antibiotics are accumulated in the body of livestock and still remain when consumed by human, causing a critical problem that antibiotic- resistance of human is increased.

To prevent the accumulation of the antibiotics in meat, it has been prohibited by the government to add some kind of antibiotics to feedstuff. Even the allowed antibiotics cannot be used for a certain period of time before going to market (so-called withdrawal time). To control the over-use of antibiotics, limit of antibiotic residue in livestock has been determined. Feedstuff companies have also produced a feedstuff for late stage of fattening without antibiotics. However, not so many farmers are keeping the regulations and keep using antibiotic-mixed assorted feed or adding antibiotics separately to feed on their own. Therefore, it is an urgent request to develop a novel additive that can replace the antibiotics in assorted feed.

Prior to the present invention, various methods to reduce the content of antibiotics as an additive to assorted feed or not to use antibiotics have been proposed. For example, clay minerals such as kaolinite, zeolite, bentonite and vermiculite have long been used to improve growth and digestion of livestock, to improve feed efficiency, to control moisture level and to eliminate bad smell of livestock excreta, etc (Pond et al., 1988, Hagedorn et al., 1990, Kovar et al., 1990). Son and Park (1997) proposed the method of adding 0.3% elvan to broiler diets. Kwon (1999) reported the improvement of weight gain rates by adding a material radiating far infrared ray to growing pigs. Yang (1999) reported the example of adding porous scoria to feedstuff for growing pigs and finishing pigs. It is known that germanium has immune enhancing activity and anticancer effect by stimulating the generation of interferons, in addition to be functioning to protect a living body from toxic materials or virus.

Korean Patent Publication No. 2002-34342 (Title of Invention : Method for preparing livestock feed using Rhus verniciflua and method for breeding livestock) describes that Rhus verniciflua sawdust is fermented and heat-distilled and then active ingredients extracted from the above are added to feedstuff for livestock, to reduce the problem caused by antibiotic residue in stock farm products. Korean Patent Publication No. 2004-104075 (Title of Invention: Assorted feedstuff composition and method of making it) describes a assorted feedstuff composition which does not include antibiotics but contains the additive comprising a herb mixture composed of loess powder, gallnut, red wood, tumerio, clove, amud cork tree and pine leaf powder in order to increase immunity of raised chicken and a method for preparing the same. Korean Patent No. 572524 (Title of Invention : Antibacterial additives for feed) describes the use of antibiotic and antiviral sophorolipid produced from bacteria as an alternative feed additive replacing the conventional antibiotics. Korean Patent No. 643596 (Title of Invention: Additive for feed and feed comprising the same) describes a feed additive comprising molokhia and Lonicera japonica having various physiological activities such as anti-oxidant, anti-cancer, anti-bacterial, anti-viral and detoxification activities and distillers dried grains with solubles, the by-product of ethanol production process, to increase immunity of livestock. In addition, feed containing ammonium molybdate (Korean Patent No. 5762), animal feeder containing oligo saccharide (Korean Patent No. 93674), low-cholesterol fat-feed additive (Korean Patent No. 58297), the production method of broiler meat containing broiler raised by the feed containing feather meal and pyridoxine and broiler meat therefrom (Korean Patent No. 647082), feed composition of growing livestock for replacing antibiotics (Korean Patent No. 343367), feed additive for broiler chicks and manufacturing method of that (Korean Patent No. 495605), and animal feed composition comprising an extract of Diospyrous kaki (Korean Patent No. 485057) have been reported. However, according to the methods proposed above, the production of such feed additives requires a complicated procedure or the materials used as additives are not possible to obtain massively, suggesting that mass-production of assorted feed with such feed additives and feed compositions is difficult. Meanwhile, in general, in the case of adding a medicinal herb or aromatic compounds to the feed, the content of such additive has to be carefully determined not to make livestock refuse to eat or to prevent the smell of the material from staying in meat, which makes human feel distaste when eat the meat.

[Disclosure] [Technical Problem]

It is an object of the present invention, to overcome the above problems, to provide an assorted feed improving immunity against disease and antibacterial effect by adding natural antibiotics obtainable from the nature massively without reducing flavor and taste of the meat raised with the same.

It is another object of the present invention to provide a method of farming livestock using the natural antibiotics. It is further an object of the present invention to provide meat which is obtained by the method of farming and is free-from side effects such as residual antibiotics caused by artificial antibiotics and has good taste and flavor.

[Technical Solution] The present inventors tested various natural substances. As a result, the inventors found out that chia seed has a strong antibacterial activity against Enterobacteria. The present inventors added chia seed to the feed for chicken and pig. As a result, Enterobacteria were significantly reduced and the function of their livers was improved. In addition, the tolerance against bacteria has been improved and the meat was able to be preserved for longer period. Meat from those chickens and pigs raised with the feed still had good taste and flavor or rather improved taste and flavor, which raises quality of the meat. The characteristics and effects of chia seed used as an additive for feed in the present invention are as follows. Chia (Salvia hispanica or Salvia columbariae) is a member of salvia belonging to Mint genus, Perilla Frutescens family, which is an annual subtropical plant (Hickman, James C. The Jepson Manual Higher Plants of California. University of California Press, Berkeley Los Angeles London. (1993), USDA Natural resources Conservation Service, [web application]. 2006). Chia is native to Mexico, but is commercially cultivated in Argentina, Bolivia, Ecuador and Guatemala. Chia Seed has been used for more than 1000 years as a material to improve endurance, physical strength, health and beauty in South America including Mexico. Traditionally, it has been consumed in Mexico, south western area of USA, and South American countries but has not been known so well in Asia or Europe. Chia Seed is gelatinized and swollen approximately 10 times the volume when it is soaked in water. Since it is swollen even in digestive juice in inside of the body, it makes people feel full long after taking in, so that it has been used as a diet food. Chia seed contains 28% dietary fiber, which is way higher than lettuce, spinach and agar have. Approximately 32% of chia seed is vegetable oil but no cholesterol is contained, making it an excellent natural health food. It also contains plenty of minerals such as calcium, phosphorus, magnesium, potassium, iron, zinc and copper. The molecular weights of polysaccharides composing dietary fiber of chia seed is 74674, 21722, 8066, 2038, and 1050. This kind of low molecular weight or medium molecular weight composition can hold water 5 to 40 times its volume, so that when it is soaked in water for 2 to 30 minutes, it becomes gelatinized (see Fig. 1 and Fig. 2).

The present inventors expected that dietary fiber of chia seed, which is added to feedstuff for livestock as a part of chia seed or chia seed extract, is the key factor for antibacterial activity reducing or killing Enterobacteria.

In a preferred embodiment of the present invention, chia seed is added to feedstuff as it is (as harvested from the nature) or after surface-treated by heat, for example parched, or after steamed and dried. Hereinafter or in the following claims, the term ^λchia seed' includes both original chia seed and treated chia seed by heat including parching or steam and dry treatment.

In another preferred embodiment of the present invention, the residue of chia seed remaining after extracting oil from chia seed, such as chia oil cake, that is chia meal, can be added to the feedstuff instead of chia seed itself as a natural antibiotic. In general, during the oil extraction process, the obtained chia meal is 60 to 65 weight% of chia seed. Hereinafter or in the following claims, the term ^λchia meal' includes the original chia oil cake, the residue of chia seed after oil extraction, chia oil cake powder added with excipients, and chia pellet prepared from the chia oil cake or it's powder.

The feed composition can be mass-produced commercially in a factory as a complete assorted feed to provide to farms and can be added to another conventional assorted feed personally in each farm.

In this description and the claims, the term 'livestock' includes poultry such as chicken, animals such as pig, and cultivated fish.

Particularly, the feedstuff composition of the present invention contains one of chia seed, chia meal and the mixture thereof as a natural antibiotic. In a preferred embodiment of the present invention, the feedstuff composition contains chia seed by 0.84 to 29 weight%, preferably by 1 to 10.8 weight%, and more preferably by 1 to 6.5 weight% for the total weight of the composition. In a modified example, chia meal is added instead of chia seed. In that case, considering the chia meal takes 60 to 65 weight% of chia seed, the content of chia meal is 0.504 to 18.85 weight%, preferably 0.6 to 7.02 weight%, and more preferably 0.6 to 8.45 weight% for the total weight of the composition.

In the method of farming livestock of the present invention, the feedstuff containing one of chia seed, chia meal and the mixture thereof as a natural antibiotic is given to livestock with providing drinking water considering water absorptiveness of dietary fiber of chia seed.

The feedstuff composition is preferably provided to livestock for at least 2 days. The method of farming livestock of the present invention can be effectively used during the withdrawal time of antibiotic administration, but not always limited thereto.

Chia seed can be mixed with various other feed components by a feedstuff company or can be added individually in a farm. At this time, the content of chia seed added to the feedstuff composition is

0.84 to 29 weight%, preferably 1 to 10.8 weight%, and more preferably 1 to 6.5 weight% for the total weight of the composition.

Meat from livestock raised by the method of the present invention can be delivered to human as it is or as being processed.

The present invention provides a pharmaceutical composition for animal having antibacterial activity by containing a component selected from the group consisting of chia seed, chia meal and a mixture thereof.

The pharmaceutical composition contains chia seed by 0.84 to 29 weight%, preferably 1 to 10.8 weight% and more preferably 1 to 6.5 weight% for the total weight of the composition. The pharmaceutical composition herein contains chia meal by

0.504 to 18.85 weight%, preferably 0.6 to 7.02 weight% and more preferably 0.6 to 4.225 weight% for the total weight of the composition.

[Advantageous Effect] As explained hereinbefore, according to the present invention, chia seed that is the natural material obtainable massively from the nature and can be cultivated massively or its extract chia meal is added to feedstuff as a natural antibiotic and livestock is raised using the same. It is also possible to added chia seed or chia meal along with a small amount of conventional antibiotics, instead of complete replacement.

The present inventors presumed that among components of chia seed, it would be the dietary fiber that gave antibacterial effect and immunity enhancing effect. Particularly, specific dietary fiber of chia seed adsorbs decayed proteins or lipids in the intestines or old acidic stercoral adhered on the intestines and decomposed gas to eliminate them from the body, so that it cleans the intestines and eliminates stercoral to make the intestines be functioning normally. Besides, it is understood that the antibacterial component included in the dietary fiber of chia seed can directly inhibit the growth of intestinal pathogens. Meanwhile omega-3 fatty acid plays a subsidiary role in enhancing immunity against disease by strengthening the functions of circulatory system including blood vessels. The antibacterial component of chia seed confirmed by the present inventors not only inhibits enterobacteria but also significantly increases immunity of chicken as it is absorbed and stays in the inside of livestock without addition of artificial antibiotics.

According to the present invention, the addition of chia seed as a natural antibiotic results in the increase of antibacterial effect, which is more powerful than that of the conventional antibiotic, and causes neither problems by antibiotic residue accumulated in the inside of livestock nor antibiotic-resistance. The present invention also brings the additional effect of improving quality of meat without changing taste and flavor. That is, chicken or pork produced by the method of the present invention has advantages of less distasteful flavor unique to meat, soft meat, and high preservability. In particular, the preservability is higher than when artificial antibiotics are used.

[Description of Drawings]

Fig. 1 is a graph of molecular weight pattern of dietary fiber contained in chia seed, verified from GPC using chia seed powder.

Fig. 2 is a table showing molecular weights of various dietary fibers contained in chia seed, verified from GPC using chia seed powder. Fig. 3 is a flow chart for separating effective ingredients from chia seed.

Fig. 4 is a photograph illustrating the test result with MRSA staphylococcus for verifying that the effective ingredients of chia seed inhibit the enterobacteria. Fig. 5 is a graph representing the result of counting the number of enterobacteria in the group administered with chia seed and the group non-administered with chia seed.

Fig. 6 to Fig. 17 are photographs illustrating the changes of enterobacteria in the culture medium. Fig. 6 is a photograph showing the culture media of enterobacteria extracted from the control ll chickens raised for 4 days without being administered with chia seed. Fig. 7 to Fig. 11 are photographs showing the culture mediums of enterobacteria extracted from the experimental chickens raised for 4 days with the administration of chia seed at different doses of 2 g, 4 g, 6 g, 8 g, and 10 g, respectively. Fig. 12 is a photograph showing the culture medium of enterobacteria extracted from the control chickens raised for 6 days without being administered with chia seed. Fig. 13 to Fig. 17 are photographs showing the culture media of enterobacteria extracted from the experimental chickens raised for 6 days with the administration of chia seed at different doses of 2 g, 4 g, 6 g, 8 g, and 10 g, respectively.

Fig. 18 to Fig. 20 are photographs illustrating the livers extracted from the control livestock non-administered with chia seed.

Fig. 21 and Fig. 22 are photographs illustrating the sections of the liver of the control livestock.

Fig. 23 to Fig. 27 are photographs illustrating the livers extracted from the livestock administered with chia seed by 8 g/day for 6 days.

Fig. 28 to Fig. 32 are photographs illustrating the sections of the livers extracted from the livestock administered with chia seed by 8 g/day for 6 days.

Fig. 33 is a photograph illustrating the comparison of livers extracted from other control livestock non-administered with chia seed and livers extracted from livestock administered with chia seed by 10 g/day for 6 days. Fig. 34 to Fig. 38 are are photographs showing the culture media of enterobacteria in the fecal samples of the experimental and the control group pigs. Fig. 34 is a photograph showing the culture medium of enterobacteria in the fecal sample of the control group pigs non-administered with chia seed. Fig. 35 to Fig. 38 are photographs showing the culture media of enterobacteria in the fecal samples of the experimental group pigs administered with chia seed by 30 g, 60 g, 100 g, and 15Og, respectively.

[Best Mode]

Practical and presently preferred embodiments of the present invention are illustrative as shown in the following Examples.

However, it will be appreciated that those skilled in the art, on consideration of this disclosure, may make modifications and improvements within the spirit and scope of the present invention.

Experimental Example 1: Investigation of antibacterial effect of chia seed on enterobacteria in vitro

An experiment was performed to investigate antibacterial activity of chia seed against enterobacteria in livestock.

2,500 g of chia seed was pulverized by using a blender, to which

10,000 ml of methanol was added, followed by vacuum-concentration at 60⁰C for 24 hours. Approximately 450 g of the methanol fraction was obtained, followed by shaking with n-hexane (5,000 ml) at room temperature for 24 hours. Extraction was performed twice. The hexane fraction was filtered by 0.4 μm filter paper, followed by vacuum-concentration at 35°C . As a result, 25 g of hexane fraction was obtained. The fraction proceeded to column chromatography (45 mm x 900 mm) filled with 250 g of silica gel. 175 g of chloroform (CHCb) fraction was obtained by eluting the fraction with chloroform/methanol as a moving phase and 64 g of ethyl acetate (EtOAc) fraction was obtained by eluting the fraction with 2000 ml of ethyl acetate. In addition, butanol (BuOH frc. 27g), water (H₂O frc. 148g) and ether (Et₂O frc. llg) fractions were obtained in that order (see Fig. 3).

The recovered chia seed fractions were tested for their antibacterial activities by solid medium DISC diffusion method (Division of Mcrobiology and Immuology, General Experimental Microbiology, Taehak-sa, pi 16, 1998). First, MRSA staphylococcus and salmonella were purchased from Korean Culture Center of Microorganisms (KCCM). The bacteria were taken from them by platinum loop and then suspended in distilled water, which was smeared on a solid blood agar plate, followed by culture for 24 hours at 25°C . Each fraction (20 μl) was loaded on Whatman paper disc (8 mm in diameter) at the concentration of 25 μg/ml, followed by culture for 24 hours at 25^°C . Ampicillin (25 μg/ml) was used as the control antibiotic.

Figure 4 is a photograph illustrating the test result with MRSA staphylococcus. As shown in this figure, butanol fraction was confirmed to have the strongest inhibiting effect on MRSA staphylococcus and ethyl acetate fraction and ether fraction follows.

In the meantime, Table 1 illustrates the sizes of inhibition zones (clear zones) indicating different antibacterial activities of each fraction against salmonella.

Table 1

Fraction Size of inhibition zone

Methanol fraction 24 ± 0.2

Water fraction 12 ± 1.0

Chloroform fraction 15 ± 0.5

Ethyl acetate fraction 16 ± 1.2

Hexane fraction 10 ± 2.5

Butanol fraction 28 ± 0.0

Ether fraction 16 ± 0.5

As shown in this Table, chia seed butanol fraction and methanol fraction had the strongest antibacterial activity against salmonella.

The clear zones caused by the death of the bacteria were approximately 28±0.0mm and 24±0.2mm in diameter, respectively.

Other fractions were confirmed to have medium level antibacterial activities.

Considering solvent specificity of chia seed antibacterial activity, the natural antibacterial component included in chia seed seems to have the medium polarity between non-polar and polar. And the dietary fiber included largely in outer envelope of chia seed is presumably the key material of such natural antibacterial component.

Experimental Example 2: Investigation of antibacterial activity and improvement of liver functions with chicken farming Chicken farming

To investigate antibacterial activity and improvement of liver functions by chia seed, broiler chickens were raised in a large scale and clinical test was performed. 150 improved Korean native chickens at 25 days from hatching were taken from a farm in Goesan, Chungcheongbuk-do, Korea.

These chickens were raised separated from other chickens in a territory divided by net. After hatching, these chickens were administered with enrofloxacin diluted in drinking water five times, on the 2^nd day, on the 8^th day, on the 11^th day, on the 15^th day and on the

21^st day from hatching. No additional antibiotic administration was performed during this experiment.

The feedstuff added with chia seed was provided to 125 out of

150 chickens separated on the 26^th day from hatching. In the meantime, general feed not containing chia seed was provided to the control chickens (25 chickens) by the conventional method.

Particularly, 125 chickens were divided into 25 groups (5 chickens per group). To each experimental group consisting of 5 chickens, assorted feed each containing chia seed by 2 g, 4 g, 6 g, 8 g, and 10 g was given. And the duration of feeding was arranged differently as follows: 2 days, 4 days, 6 days, 8 days and 10 days for each group.

So, the 125 chickens were raised until the 28^th to 36^th day from hatching.

Examination and identification of enterobacteria Intestines were extracted from the raised chickens and enterobacteria were investigated. First, 0.5 g of sample was suspended in 10 ml of triply distilled water. 90 ml of triply distilled water was added to the suspension to dilute the solution to 1/10. 0.1 ml of the solution was taken by a sterilized pipette, which was distributed on a blood agar plate. The sample was spread evenly over the medium using a glass rod. The medium was face-downed, followed by culture for 24 hours in a 25⁰C incubator. After culture, colony formation was investigated and if there was, the number or cells was counted.

Table 2 summarizes the result of cell counting and Figure 5 illustrated the result.

Table 2

As shown in Table 2 and Figure 5, the number of enterobacteria was significantly reduced in the group administered with chia seed, compared with the group non-administered with chia seed. In the meantime, the number of bacteria was significantly reduced until 4^th day of experiment in both chia seed administered group and non- administered group, but since then the number of bacteria began to increase. This result is presumably attributed to the artificial antibiotic administered diluted in drinking water until 5 days before chia seed administration to the experimental group treated with chia seed which still had effect therein until 4 days after the experiment started. The number of enterobacteria began to increase after 4 to 6 days in both chia seed administered group and non-administered group, which seemed to be because the artificial antibiotic was no more functioning. However, after losing the functions of the conventional antibiotic, chia seed still acts as a powerful antibiotic to kill enterobacteria.

Changes of enterobacteria were detected in the chia seed administered group after around 4 to 6 days from the experiment start. As confirmed in the following bacteria identification test, the number of pathogens was decreased until 4 to 6 days after the initiation of the experiment which was believed to be the effect of antibiotics, but from then on, pseudomonas sp. degrading high-molecular dietary fiber of chia seed took the position of a dominant species. From 6^th day of the experiment, the number of bacteria began to decrease slightly in both chia seed administered group and non-administered group, which seemed partially because of competition between bacterias that had the effect of inhibiting bacteria growth. Figure 6 to Figure 17 are photographs illustrating the changes of enterobacteria in the culture medium. Figure 6 is a photograph showing the culture media of enterobacteria extracted from the control chickens raised for 4 days without being administered with chia seed. Figure 7 to Figure 11 are photographs showing the culture mediums of enterobacteria extracted from the experimental chickens raised for 4 days with the administration of chia seed at different doses of 2 g, 4 g, 6 g, 8 g, and 10 g, respectively. Figure 12 is a photograph showing the culture medium of enterobacteria extracted from the control chickens raised for 6 days without being administered with chia seed. Figure 13 to Figure 17 are photographs showing the culture media of enterobacteria extracted from the experimental chickens raised for 6 days with the administration of chia seed at different doses of 2 g, 4 g, 6 g, 8 g, and 10 g, respectively.

Regardless of the changes shown above, it was clearly understood that the lines indicating the populations of bacteria in the experimental groups administered with chia seed were located down from the lines indicating the populations of the control chickens non- administered with chia seed, in Figure 5. Based on the founding that the population of enterobacteria was decreased in the chia seed administered groups, it is believed that chia seed has an antibacterial activity so that it can act as a natural antibiotic inhibiting bacteria growth without affecting the normal growth of chicken. In fact, the population of enterobacteria was decreased in every experimental group. As for the effect of antibacterial activity of chia seed, the effective dose of chia seed for livestock is 2 to 10 g or more and the preferable administration period is 2 to 10 days or more. As shown in Table 2 and Figure 5, it is more preferred to treat livestock with chia seed at the dose of 4 g or more for 4 or more days. The culture media for the bacteria were observed by the naked eye. As a result, 1 to 4 kinds of bacteria were detected in each medium. The bacteria were then identified. To do so, some of each detected bacteria were isolated and suspended in distilled water. Then, the strains were identified by using Easy 24E plus identification kit (Hanil Komed Co., Ltd., Korea) and Easy View program.

From the identification, pathogenic bacteria such as Acinetobacter baumannii (MDR) resistant to imipenem; Shigella spp. causing dysentery; Yersinia aldorae, the bacillus causing diarrhea and high fever; Klebsiella ozaenae causing pneumonia and decreased immune response; Morganella morganii; Enterobacter cloaceae; and Pseudomonas aeruginosa were detected all the time during the experiment in the chia seed non-administered group. In addition, hemolysis bacteria was detected intermittently. On the other hand, such pathogenic bacteria were significantly reduced in the chia seed administered groups. Particularly, Pseudomonas putida, the food spoilage microorganism capable of degrading high molecular substances, was detected in the experimental group administered with chia seed by 2 g for at least 4 days. In some of the experimental groups administered with high dose for a long period, Pantoea (Enterobacter) agglomerans degrading polysaccharides and E. coli inactive form were detected.

As explained hereinbefore, the number of enterobacteria was reduced and the kind of enterobacteria in the chia seed administered group was changed. And, a stinking smell from intestinal extract and feces was gone in the chia seed administered group, unlike in the chia seed non-administered group. Therefore, it is presumed that chia seed includes a component that inhibits the growth of a pathogen containing sulfide. Plenty of polysaccharide dietary fiber in the intestines changes the intestinal environment to let probiotics and E. coli degrading the polysaccharide dietary fiber be dominant, so that the development of a disease caused by pathogenic microorganisms can be significantly reduced in livestock.

Liver test

To investigate the effect of chia seed on the improvement of liver functions in livestock raised with chia seed, the liver was extracted. The inside and outside color of the liver was observed by the naked eye and photographed. Figure 18 to Figure 20 are photographs illustrating the livers extracted from the control livestock non-administered with chia seed. Figure 21 and Figure 22 are photographs illustrating the sections of the liver of the control livestock. In the meantime, Figure 23 to Figure 27 are photographs illustrating the livers extracted from the livestock administered with chia seed by 8 g/day for 6 days. Figure 28 to Figure 32 are photographs illustrating the sections of the livers.

Figure 33 is a photograph illustrating the comparison of livers extracted from other control livestock non-administered with chia seed and livers extracted from livestock administered with chia seed by 10 g/day for 6 days. In this photograph, livers on the left were extracted from the control livestock. And livers on the center and the right were extracted from the livestock administered with chia seed.

As shown in those photographs, in the control livestock non- administered with chia seed, liver color was not vivid, bile secretion was not normal and its color was dim yellow, elasticity was comparatively weak, and yellow fat layer was observed. On the contrary, in the livestock administered with chia seed, fat layer was not observed (photographs in Figures 19, 20, 23 and 27; white parts indicate external intestines not completely eliminated), liver color was clear red, and outward shape was elastic so that even not a professional could judge the liver as healthy and active.

Liver is involved in almost every metabolism in vivo, and particularly plays an important role in detoxification, antibody production, elimination of unnecessary materials, and keeping lives healthy. So, as long as this important organ liver is healthy by the method of the present invention, immunity of the livestock raised by the method of the present invention can improve significantly.

Experimental Example 3: Investigation of antibacterial activity and improvement of liver functions with pig farming Pig farming

To investigate antibacterial activity and improvement of liver functions by chia seed, pigs were raised and clinical test was performed. 8 cross-bred black pigs at approximately 180 days old were taken from a farm in Goesan, Chungcheongbuk-do, Korea. The 8 pigs were divided into 4 groups, 2 pigs per group. One of the two was female and the other was male in each group. Chia seed assorted feed containing 30 g, 60 g, 100 g and 150 g of Australian chia seed, respectively was given to the 4 experimental groups for 10 days. Two pigs were selected for the control group which was raised without being administered with antibiotics. Both the experimental groups and the control group were not treated with antibiotics before experiment started.

Examination and identification of enterobacteha Feces were taken from the pigs raised for 10 days and enterobacteria were investigated. First, 0.5 g of sample was suspended in 10 ml of triply distilled water. 90 ml of triply distilled water was added to the suspension to dilute the solution to 1/10. 0.1 ml of the solution was taken by a sterilized pipette, which was distributed on a blood agar plate. The sample was spread evenly over the medium using a glass rod. The medium was face-downed, followed by culture for 24 hours in a 25⁰C incubator. After culture, colony formation was investigated and if there was, the number was counted

Figure 34 to Figure 38 are photographs showing the culture media of enterobacteria in the fecal samples of the experimental and the control group pigs. Figure 34 is a photograph showing the culture medium of enterobacteria in the fecal sample of the control group pigs non-administered with chia seed. Figure 35 to Figure 38 are photographs showing the culture media of enterobacteria in the fecal samples of the experimental group pigs administered with chia seed by 30 g, 60 g, 100 g, and 15Og, respectively. Table 3 shows the summary of the result of counting bacteria.

Table 3

As shown in those Figures and the Table, the population of enterobacteria was significantly reduced in the chia seed administered group, compared with the chia seed non-administered group. Both the experimental and the control group pigs were raised without treatment of antibiotics before experiment. So, the number of bacteria in the control pigs was very high. On the contrary, only a few bacteria were detected in the fecal samples from the pigs administered with chia seed. Considering the difference of the number of bacteria in the fecal samples between the control and the experimental group, it was suggested that the number of bacteria active in the intestines would also be very different between the control and the experimental group.

Therefore, it was confirmed that enterobacteria were significantly reduced in the chia seed administered groups. And from the result, it was suggested that chia seed contains a component that is capable of inhibiting the growth of enterobacteria.

Liver test

On the 7^th day of 10 day raising, blood sampling was performed from both experimental and control pigs by a veterinarian. Component analysis of the blood sample was performed by Eone Reference Laboratory, Dongjak-gu, Seoul, Korea. During the blood sampling, one of the two control pigs missed the blood sampling by the veterinarian's mistake. And the results of the analysis are as follows.

Table 4

As an index for blood sugar, glucose level was measured. As a result, blood glucose level was 72 in the control pig, whereas the levels of the chia seed administered group pigs were half the level in the control. The decrease of blood glucose in the chia seed administered pigs indicates that hormone secretion, one of the liver functions, was normal and much healthier in the chia seed administered pigs, compared with the control pigs. There was not much difference in GPT between the control and the experimental group pigs. But GOT in the chia seed administered group pigs was much reduced to 1/10, compared with the control group pigs. No significant difference in cholesterol levels between the control and the experimental group pigs were observed. In the case of HDL (high density lipoprotein), the factor eliminating cholesterol from vessel wall, a slight increase was detected in the chia seed administered groups but not significant. Free fatty acid was also slightly increased in the chia seed administered groups, compared with the control group. Among the experimental groups, there was not much difference according to the dose of chia seed.

From the results of examination of blood glucose, GOT, HDL cholesterol, and free fatty acid, it was confirmed that chia seed enhanced liver functions and health conditions significantly in pigs. Since the liver playing an important role in detoxification, antibody production, elimination of unnecessary materials, and thus keeping livestock healthier is treated with chia seed, the immunity of the livestock will be significantly improved owing to the healthier liver. Experimental Example 4: Additional experiment-weight gaining bv chia

The present inventors noticed during the experiment of Experimental Example 2 that the weight gain rates of livestock administered with chia seed might be affected as the dose increased and the treatment period was longer. So, the present inventors weighed pigs of the experimental group administered with chia seed for 10 days at different doses of 2 g, 4 g, 6 g, 8 g, and 10 g. The control group pigs were raised for 10 days without administration of chia seed. The results are shown in Table 5.

Table 5

As shown in the above Table, in the pigs administered with chia seed for 10 days at the dose of 8 g, the weight gain inhibition was significant, compared with the control pigs. But, in the pigs administered with 10 g of chia seed every day, the weight gain inhibition was not observed.

Therefore, considering the weight gain rate, preferable dose of chia seed per day was approximately 6 g or less. However, 10 g administration had no much problem since it reduces the weight gain inhibition.

When the pigs were administered with chia seed at the daily dose of 30 g or 60 g, weight gaining inhibition effect was not so great, but the pigs were administered with chia seed at the daily dose of 100 g or 150 g, weight was reduced by 3 to 5 kg.

Usability of chia seed as a natural antibiotic and preferable dosage thereof

The present inventors confirmed the antibacterial activity against enterobacteria and improvement effect on liver functions of chia seed by in vitro experiments with livestock including chicken and pig and clinical tests with them, etc. If considering the antibacterial activity and liver function improvement effect alone, chia seed is effective enough at the dosage of 2 to 10 g per day and the administration period is 2 to 10 days or more, and more preferably at the dose of 4 to 10 g for at least 4 days. But, to prevent weight gain inhibition, it is not recommended to treat chia seed more than 6 g per day. However, more than 10 g of administration is not a big problem in weight gain rate.

In conclusion, it is preferred to treat chia seed to chicken for farming by 2 to 10 g per day for at least 2 days. It is more preferred to treat chia seed by 4 to 6 g per day. If chia seed is administered to chickens from 27^th day (required feed: 83.9 g: according to standard chicken raising program provided by Cargill, Inc., USA) from hatching to 54^th day (required feed: 149.Ig) throughout the period or intermittently, daily dose of chia seed is preferably 1% to 10.8 weight% for the daily requirement of feed. And 2.5% to 6.5% is more preferred.

If chia meal is provided to broiler chickens instead of chia seed (2 g/day), the preferable dose of chia meal is 1.2 g per day, considering that chia meal takes 60 to 65% of chia seed.

In the pigs administered with 30 g to 150 g of chia seed each day as described in Experimental Example 3, significant antibacterial activity and liver function improvement effect were observed. If chia meal is provided to pigs instead of chia seed (more than 30 g/day), the preferable dose of chia meal is more than 18 g per day.

Example 1: Chicken farming and examination of meat characteristics Chicken farming with feeding chia seed

Separately from the above Experimental Example 2, approximately 1,000 to 10,000 baby chicks per week were raised with feeding chia seed by 5 g per day for 8 months in a farm in Goesan, Chungcheongbuk-do, Korea. The feeding of chia seed started from 27 to 28 days after hatching. Chickens at 32 to 35 days were firstly shipped for Samgyetang (chicken broth with ginseng). And the rest of the chickens were additionally treated with chia seed for 1 week to 10 days and then shipped for the roast chicken. Storage of chicken

Even if thiobarbituric acid reactive substance (TBARS) or protein denaturation (VBN) in the raised chicken meat was not quantified, the chickens obtained from the control and the group administered with chia seed by 4g per day for 10 days were left at room temperature, not in a refrigerator, for 7 hours and then sensory evaluation was performed. As a result, the chicken obtained from the control group was degraded so that it smelled badly. In the meantime, the chicken obtained from the group administered with chia seed by 4 g per day was smelled like the meat in a refrigerator.

The resistance against bacteria or the improvement of storage property of the chicken was presumably because that the component having antibacterial activity included in chia seed not only successfully works in the intestines but also remains in the tissues to be functioning via digestion and absorption by the chicks.

Component analysis of chicken meat

Chickens fed with chia seed by 4 g per day for 15 days were sacrificed. Cholesterol and fatty acid contents in the chicken breast were measured by the general component test method of Korean Food Code (2006). Component analysis was performed by Eone Reference Laboratory, Dongjak-gu, Seoul, Korea. Cholesterol levels are shown in Table 6 and fatty acid levels are shown in Table 7. Table 6

Table 7

As shown in the tables, cholesterol content in chicken was 21% reduced, but omega-3 fatty acid was 2 fold increased. The cholesterol level in chicken was higher than that in pig, which seems because chicken is lighter than pig and the length of chicken intestines is shorter than those of pig so that the effect of chia seed is brought faster in chicken. Sensory evaluation of chicken

The raised chickens were sacrificed and cooked to make Samgyetang, Baksuk (boiled chicken), and oven baked chicken. 25 sensory evaluating agents were participated in the evaluation including the test of smell and quality of meat.

As a result, higher grade was given to chicken meat obtained from the chickens raised with chia seed added feed, compared with chicken meat obtained from the chickens raised with the conventional feed. Particularly, chicken meat treated with chia seed had good taste and flavor of meat. And, fat floating on Samgyetang was reduced, the soup was clearer and meat was softer. The reduction of distasteful smell was resulted from the decrease of enterobacteria by chia seed, meaning intestinal environment was improved and the liver functions were improved so that the health condition of chicken was enhanced.

Further investigation on characteristics of the raised chickens and their meat

The chickens raised by the method of the present invention had clear red combs, had good color and gloss on feather, compared with chickens raised by the conventional method. This difference was so peculiar that even the farmer could distinguish the chia seed treated chickens from the chickens non-treated with the naked eye. During the sacrificing, the movement and resistance of the chia seed treated chickens were tougher than chickens raised by the conventional method. After sacrificing, it was more difficult to pluck feathers from the chia seed treated chicken. Besides, bad smell of feces was significantly reduced. The chickens, in particular intestines, raised by the method of the present invention were healthier and had enhanced strength and accordingly had increased immunity. After sacrificing, chicken was stored in a refrigerator. Even after then, chicken smelled not bad and rather good. Even chicken foot or intestines smelled not distasteful. As mentioned hereinbefore, even after storing chicken not in a refrigerator but at room temperature for many hours, it hardly smelled, suggesting excellent preservability. The chickens raised by the method of the present invention were additionally confirmed that they had increased resistance against bacteria and enhanced immunity even without using the conventional antibiotics. The chicken was left at room temperature and then stored in a refrigerator for 2 days. Then, the meat was taken out for cook. In that case, the meat obtained from the chicks raised with the conventional feed was spoiled with bad smell, whereas the meat obtained from the chicks raised with chia seed was maintained without bad smell and was similar to the meat stored in a refrigerator right after sacrificing. So, since the meat obtained from the chicks raised by the method of the present invention has no distasteful smell, it is not necessary to added spices to eliminate bad smell.

In this invention, the meat obtained from the chicks raised with chia seed was so soft that the breast could be torn apart easily by wooden chopsticks, whereas the chicken breast obtained from the chicks raised with the conventional feed was so hard that wooden chopsticks were almost broken in the attempt to draw the flesh apart.

The chickens raised by the method of the present invention had largely thin skin and thin subcutaneous fat in between skin and flesh. The low fat property was peculiar in neck, perianal area, and intestines, which are representative regions generally having thick fat layer. The skin color was whiter and clearer in those chickens raised with chia seed containing feed, compared with the chickens raised with the conventional feed. When Samgyetang was cooked with the chicks raised with the feed of the present invention, fat floating in the soup was white, compared with the yellow fat in the conventional Samgyetang, and the amount of fat floating was also reduced, so that soup itself was clearer. According to the comparatively high unsaturated fatty acid level, even after cooled down, the fat in the soup was hardly solidified.

Example 2: Sensory evaluation of pork Pig farming with feeding chia seed

Separately from the above Experimental Example 3, 3 cross- bred white male pigs at 6 month having around 100 kg of weight were raised with chia seed. Along with approximately 3 kg (daily dose) of assorted feed, 200 g/day of chia seed was fed to those pigs for 12 days. At this time, chia seed was pre-mixed with the assorted feed before provided. One of them was sacrificed. Determination of 200 g per day dosage of chia seed was compared with the result of Experimental Example 3 that when at least 150 g/day of chia seed was provided, antibiotic like effect and weight gain effect were observed. So, 200 g/day of chia seed was fed to the above white pigs, followed by sensory evaluation.

Component analysis of pork

One of those pigs raised by the above method was sacrificed. 100 g of sirloin was taken and tested for measuring fatty acid content by the general component test method of Korean Food Code (2006). Component analysis was performed by National Institute of Animal Science, Suwon-si, Gyeonggi-do, Korea. As a control, a pig was raised under the same condition in the same farm except chia seed was not fed. The results of the analysis are shown in Table 8.

Table 8

There was no big difference between the chia seed administered group and the control, but unsaturated fatty acid was slightly increased and EPA, which was not detected in the control, was detected in the chia seed administered group.

Weight changes in pigs according to the feeding of chia seed

Despite the amount of feed taking in the experimental group fed with chia seed was more than in the control without chia seed, the pigs of the experimental group were 4 to 5 kg lighter than the control pig, after 12 days of farming. The weights of those pigs were reduced by 1 to 2 kg, compared with the black pigs fed with 150 g/day of chia seed for 15 days. However, no significant deviations by chia seed were observed.

Sensory evaluation of meat taste

Each part of the pig sacrificed was processed. Some were grilled and some were boiled and some were added to Kimchi pot-stew. 20 sensory evaluating agents were participated in the test investigating smell and taste of the meat according to different dishes.

Before the sensory evaluation, all agents agreed that meat had clear color and even marbling. The results were as follows. Meat was chewy but soft. Particularly, fat was chewy instead of being crumbly. Even the outer layer was soft. Despite the sacrificed pig was a male, it did not smell bad regardless of the kind of dish. Even after the meat was cold, it was not stiff and rather still soft, unlike the general pig raised with general feed. Compared with black pig however, it was less tasteful and had more fat.

In the meantime, one of black pigs used in Experimental Example 3 was sacrificed and examined for sensory evaluation. As a result, the taste of meat was more excellent than other pigs or the above white pigs. Waste fat resulted from the grilling was dropped in cold water. As a result, unlike the general pig fat (which is supposed to be hardened rightly), the pig fat came out of the pig raised with the feed of the present invention was not hardened and only sticky.

Example 3: Modified example

As described hereinbefore, the feedstuff of the present invention is prepared by adding chia seed to the conventional feedstuff. Herein, chia seed can be mixed with feed ingredients such as grains, brans, expeller cakes, and other processed food residues by a feedstuff company. Or a farmer can mix chia seed directly with assorted feed containing the above feed ingredients purchased.

As explained above, oil can be extracted from chia seed. And the remaining chia cake which is called chia meal can be added instead of chia seed as a natural antibiotic. It is also possible to add chia seed together with chia meal. The administration of chia seed as a natural antibiotic to livestock is particularly effective during the withdrawal time of antibiotic administration.

Those skilled in the art will appreciate that the conceptions and specific embodiments disclosed in the foregoing description may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present invention. Those skilled in the art will also appreciate that such equivalent embodiments do not depart from the spirit and scope of the invention as set forth in the appended claims.

Claims

[CLAIMS]

[Claim 1]

A feedstuff composition comprising an ingredient selected from the group consisting of chia seed, chia meal and a mixture thereof.

[Claim 2]

The feedstuff composition according to claim 1, wherein the chia seed is present in an amount ranging from 0.84 to 29 weight% in the total weight of the composition.

[Claim 3]

The feedstuff composition according to claim 2, wherein the chia seed is present in an amount ranging from 1 to 10.8 weight% in the total weight of the composition.

[Claim 4]

The feedstuff composition according to claim 3, wherein the chia seed is present in an amount ranging from 1 to 6.5 weight% in the total weight of the composition.

[Claim 5]

The feedstuff composition according to claim 1, wherein the chia meal is present in an amount ranging from 0.504 to 18.85 weight% in the total weight of the composition. [Claim 6]

The feedstuff composition according to claim 5, wherein the chia meal is present in an amount ranging from 0.

6 to 7.02 weight% in the total weight of the composition.

[Claim 7]

The feedstuff composition according to claim 6, wherein the chia meal is present in an amount ranging from 0.6 to 4.225 weight% in the total weight of the composition.

[Claim 8]

The feedstuff composition according to claim 1, wherein the chia seed is acting as a natural antibiotic.

[Claim 9]

The feedstuff composition according to claim 1, wherein the livestock is selected from the group consisting of poultry including chicken, animals including pig, and fish.

[Claim 10]

A method for farming livestock comprising the step of feeding the feedstuff containing one of the feedstuff compositions of claim 1 to claim 9 to livestock.

[Claim 11] The method for farming livestock according to claim 10, further comprising the step of providing the livestock with drinking water.

[Claim 12] The method for farming livestock according to claim 10, wherein the livestock is selected from the group consisting of poultry including chicken, animals including pig, and fish.

[Claim 13] The method for farming livestock according to claim 12, wherein the feedstuff is provided at least 2 days.

[Claim 14]

The method for farming livestock according to claim 13, wherein the feedstuff is provided during the withdrawal time of antibiotic administration.

[Claim 15]

The method for farming livestock according to claim 14, wherein the feedstuff composition containing chia seed by 1 to 10.8 weight% for the total weight of the composition is fed to livestock.

[Claim 16]

The method for farming livestock according to claim 15, wherein the livestock are broiler chicks at 27 days after hatching and the chia seed is fed at least 2 g per day.

[Claim 17]

The method for farming livestock according to claim 16, wherein the chia seed is fed at most for 15 days.

[Claim 18]

The method for farming livestock according to claim 15, wherein the livestock are pigs and the chia seed is fed at least 30 g per day.

[Claim 19]

The method for farming livestock according to claim 14, wherein the feedstuff composition containing chia meal by 0.6 to 7.02 weight% for the total weight of the composition is fed to livestock.

[Claim 20]

The method for farming livestock according to claim 19, wherein the livestock are broiler chicks at 27 days after hatching and the chia meal is fed at least 1.2 g per day.

[Claim 21]

The method for farming livestock according to claim 20, wherein the chia meal is fed at most for 15 days.

[Claim 22] The method for farming livestock according to claim 19, wherein the livestock are pigs and the chia meal is fed at least 18 g per day.

[Claim 23] A meat of livestock meat produced by one of the methods for farming livestock of claim 10 to claim 22.

[Claim 24]

A processed meat product produced by the meat obtained from the livestock raised by one of the methods for farming livestock of claim 10 to claim 22.

[Claim 25]

A pharmaceutical composition for animal having antibacterial activity, which comprises an ingredient selected from the group consisting of chia seed, chia meal and a mixture thereof.

[Claim 26]

The pharmaceutical according to claim 25, wherein the chia seed is present in an amount ranging from 0.84 to 29 weight% in the total weight of the composition.

[Claim 27]

The pharmaceutical composition according to claim 26, wherein the chia seed is present in an amount ranging from 1 to 10.8 weight% in the total weight of the composition.

[Claim 28]

The pharmaceutical composition according to claim 27, wherein the chia seed is present in an amount ranging from 1 to 6.5 weight% in the total weight of the composition.

[Claim 29]

The pharmaceutical composition according to claim 25, wherein the chia meal is present in an amount ranging from 0.504 to 18.85 weight% in the total weight of the composition.

[Claim 30]

The pharmaceutical composition according to claim 29, wherein the chia meal is present in an amount ranging from 0.6 to 7.02 weight% in the total weight of the composition.

[Claim 31]

The pharmaceutical composition according to claim 30, wherein the chia meal is present in an amount ranging from 0.6 to 4.225 weight% in the total weight of the composition.