KR20060027075A - Feed additive using citrus peel and method for manufacturing thereof - Google Patents

Abstract

The present invention relates to a feed additive using a citrus gourd and a method for preparing the same, and more particularly, to a feed additive containing a culture solution inoculated with bacteria of Bacillus in a citrus gourd or citrus gourd extract and a method for producing the same. The feed additive of the present invention retains the effective substance and flavor of citrus gourds, and has a high antibacterial effect and exhibits a formal effect of inhibiting the growth of harmful bacteria in the intestines of livestock, and also for feed utilization, weight gain and disease of livestock. It shows the effect of improving the resistance.

Description

Feed additive using citrus gourd and its manufacturing method             

1 is a mixture of citrus gourd 0.5% calcium hydroxide (calcium hydroxide, Ca (OH) ₂ ) and then dehydrated through a screw extruder to separate the solid and liquid parts, the solid part is dried to produce a citrus foil pellets After preparing a feed, and cultured by adding 20 parts by weight of Bacillus strain culture medium, 80 parts by weight of water and 0.1% tryptone to each of the citrus gourd extract and citrus gourd (100 parts by weight) of the liquid portion, containing the strain culture solution It is a process of manufacturing a feed additive to

Figure 2 is a feed additive, ampicillin, kanamycin, kanamycin, streptomycin, tetracycline prepared from Bacillus subtilis cultures for the pathogenic bacteria Staphylococcus aureus, Salmonella gallinarum and E. coli. It is a graph comparing the antimicrobial activity of tracycline, vancomycin, vancomycin, spectinomycin, and novobiocin (wherein "Bacillus super." Is a feed additive prepared from Bacillus subtilis culture) ).

The present invention relates to a feed additive using a citrus gourd and a method for preparing the same, and more particularly, to a feed additive containing a culture solution inoculated with bacteria of Bacillus in a citrus gourd or citrus gourd extract and a method for producing the same.

Recently, with the development of agricultural technology, the production of fruits such as citrus fruits is also increasing. This increase in production has led to industrial necessity of increasing the availability of citrus fruits due to price imbalances and supply and demand imbalances, thereby increasing the production of processed products such as citrus juice and citrus liquor. However, along with increased production, the generation of citrus peels and debris as by-products or wastes has also increased, and the remaining citrus gourds after these citrus juices do not rot well and thus become a serious environmental factor.

On the other hand, citrus gourds are known to contain essential oils and various flavonoids, alkaloids and vitamins. In addition, the fragrant smell of citrus fruits is due to the essential oil component is the main ingredient is limonene (d-limonene), the fragrant odor induces the animal to eat more feed. Therefore, in recent years, methods for effectively treating citrus gourds have been studied in various ways, and methods for making citrus gourds by pulverizing and drying them to feed, fertilizers, or compost, and citrus gourd fermented feed production apparatuses have been studied.

Among them, citrus peels are juiced and separated into juices and juices. Candida utilis , Kluyveromyces lactis , and Saccharomyces cerevisiae (Candida utilis ) are known from the juices. Saccharomyces cerevisiae ) to obtain a yeast culture by culturing any of the yeasts selected from, and then the yeast culture is added to the isolated juice gourd as it is, and the mixture is dried, the method for producing a feed using citrus peel Korean Patent No. 0055364). However, the method requires a high cost as a huge facility, requires a lot of auxiliary devices, has a problem that the utilization value is very low because the process is complicated, and does not exhibit a special antibacterial effect.

On the other hand, there is a patent (Korean Patent Registration No. 0381726) regarding a citrus fermentation broth obtained by fermenting citrus fruits, its juice or juice concentrate with anaerobic microorganisms such as lactic acid bacteria, yeast, photosynthetic bacteria, and feed for fish farming containing the citrus fermentation broth. have. However, the patent is a method for increasing the weight of fish by using the citrus juice itself as a feed additive, it does not recycle citrus gourd and does not exhibit a special antibacterial effect.

Accordingly, the present inventors prepare a feed additive containing a culture medium cultured by inoculating Bacillus sp. Bacteria in citrus gourd or citrus gourd extract, and the feed additive not only increases feed utilization and weight gain of livestock, but also has a unique antibacterial effect. The present invention was completed by confirming the improvement of resistance to disease.

It is an object of the present invention to provide a feed additive having a high antimicrobial activity from a citrus gourd or a citrus gourd extract and a method for producing the same.

In order to achieve the above object, the present invention provides a feed additive containing a culture solution cultured by inoculating Bacillus sp.

In addition, the present invention comprises the steps of 1) obtaining a citrus gourd extract from citrus gourd; 2) provides a method of preparing a feed additive comprising the step of obtaining a culture solution by inoculating the citrus gourd or the citrus gourd extract obtained in step 1 with Bacillus bacteria.

Hereinafter, the present invention will be described in detail.

The present invention provides a feed additive containing a culture solution cultured by inoculating Bacillus sp. Citrus gourd in the present invention refers to the remaining residue after citrus juice, in the present invention can be usefully recycled not only the liquid extract of citrus gourd but also citrus gourd.

At this time, Bacillus genus bacteria used for inoculation may be applied to all kinds of Bacillus genus bacteria, but preferably Bacillus licheniformis ( Bacillus licheniformis ), Bacillus subtilis ( Bacillus subtilis ) and Bacillus coagulance ( Bacillus coagulance) It is preferably selected from the group consisting of), and most preferably Bacillus rickeniformis. Bacillus liqueniformis is a strain that forms resistant spores and produces a large amount of cellulose-degrading enzymes and grows them, and is a strain that can be used as a probiotics (effective probiotic) and has the characteristics of producing polysaccharides and crude proteins. It can be used as a probiotic for feed.

In order to prepare the feed additive, it is preferable to add 1 to 20 parts by weight, and more preferably 10 to 20 parts by weight, of the seed culture of Bacillus bacteria in 100 parts by weight of citrus fruit or citrus fruit extract. As a result of confirming the antimicrobial effect of the feed additive of the present invention by the agar disk diffusion test method, it shows significant antimicrobial activity against pathogenic bacteria (see Table 1 ). In particular, the feed additive prepared from the Bacillus subtilis culture shows excellent antimicrobial activity compared to the rest of the conventional antibiotics except ampicillin (ampicillin) compared to conventional antibiotics (see Figure 2 and Table 2 ).

In addition, the present invention is more preferably added in 10 to 20% of the feed additive to the general formula feed. It provides an antimicrobial feed, and there is no limitation on the type of compound feed, and means a compound feed commonly used for cattle or pigs (see Table 3 ). When the feed additive of the present invention is added to the blended feed, when fed to beef cattle, the daily gain (unit: kg) and the carcass rate are improved compared to the feed requirement (see Table 4 ).

In addition, the feed additive of the present invention is added to a general blended feed (Total mixed ration (TMR)) when fed to milking cows, milk production and maintenance rate is increased, milk protein and solid solid content is also increased in milk components (See Table 5 ). On the other hand, as a result of examining the somatic cell count of the milking cows, a significant decrease in the somatic cell count was found, which may be effective in preventing breast cancer.

In addition, when the feed additive of the present invention is added to the general formula feed, when fed to pigs, the carcass performance is excellent, the meat quality is improved, such as intramuscular fat deposition and lean meat increase. On the other hand, the digestibility of nutrients is improved, in particular, protein digestibility in the growing season, protein and crude fiber and soluble nitrogen (NFE) digestibility in the growing season.

In addition, the present invention comprises the steps of 1) obtaining a citrus gourd extract from citrus gourd; 2) provides a method of preparing a feed additive comprising the step of obtaining a culture solution by inoculating the citrus gourd or the citrus gourd extract obtained in step 1 with Bacillus bacteria.

Step 1 of the present invention is a step of obtaining a citrus gourd extract from citrus gourd. In a preferred embodiment of the present invention, the citrus gourd was mixed with calcium hydroxide and then dehydrated to obtain a citrus gourd extract.

Step 2 of the present invention is a step of obtaining a culture cultured by inoculating Bacillus bacteria in citrus gourd or citrus gourd extract obtained in step 1 above. In this case, Bacillus genus bacteria can be applied to all kinds of Bacillus genus bacteria, preferably Bacillus licheniformis ( Bacillus licheniformis ), Bacillus subtilis ( Bacillus subtilis ) and Bacillus coagulance ( Bacillus coagulance ) group It is preferably selected from, and most preferably Bacillus rickeniformis. Bacillus liqueniformis is a strain that forms resistant spores and produces a large amount of cellulose-degrading enzymes and grows them, and is a strain that can be used as a probiotics (ie, an effective probiotic) and produces polysaccharides and crude proteins. It can be used as a probiotic for feed. The bacterium of the Bacillus is to grow in the citrus gourd or citrus gourd extract because the bacterium decomposes pectin in citrus gourd or citrus gourd extract using nutrients produced by the bacterium Can be.

Thereafter, in order to inoculate Bacillus bacteria, the seed culture of Bacillus bacteria is preferably added in an amount of 1 to 20 parts by weight, more preferably 10 to 20 parts by weight based on 100 parts by weight of the citrus fruit or citrus fruit extract. Do.

In addition, it may further comprise the step of preparing a powder by drying the culture solution obtained after step 2 of the present invention. In a preferred embodiment of the present invention by spray-drying the culture solution was prepared a feed additive (see Figure 1 ).

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

< Example  1> Citrus gourd  Preparation of Feed Additives

The strains used in the present invention are Bacillus licheniformis , B. subtilis , and B. coagulans , respectively, isolated from Cheonggukjang or Doenjang. For the practice of the present invention, each strain was incubated in LB suspension medium (tryptone 1.0%, yeast extract 0.5%, sodium chloride 0.5%) for one day, and then added 20% glycerol to -70 ° C in a 1.5 ml vial. Stored. Then, each strain was incubated in a 25 ℃ fermentor (Korea Fermentor) for 12 hours was used as a seed culture.

Corresponds to the remaining residue after the juice of citrus foil citrus, citrus foil with 0.5% calcium (calcium hydroxide, Ca (OH) 2) as a mixture after, citrus foil extract of dehydrated through a screw extruder a solid phase portion and liquid phase portion Separated.

Thereafter, 1 kg of the seed culture medium was inoculated and mixed in a culture tank containing 10 g / L tryptone and 10.0 kg of citrus fruit or citrus fruit extract adjusted to pH 3.5-8.0, followed by incubation at 35 ° C. for 24 hours. Obtained. The culture solution was directly used as a feed additive, or the culture solution was spray-dried and used as a feed additive (3.0 kg) ( FIG. 1 ).

< Example  2> Citrus gourd  Confirmation of Antimicrobial Effect of Feed Additives

Agar disk diffusion test (Kirby-Bauer) was carried out to confirm the antimicrobial effect of the feed additive using the citrus gourd of the present invention (hereinafter referred to as "citrus gourd feed additive") (Bauer, AW, .. kirby, WMM, Sherris, JC, Turck, M., Am J. Clin Pathol, 45:. 943, 1966).

Specifically, the pathogenic bacteria Staphylococcus aureus , Salmonella Galinarum ( Salmonella) 0.5 ml (10 ⁶ -10 ⁷ CFU / ml) of gallinarum ) and E. coli were diluted in soft agar (0.8%, 4.5 ml) and flattened on tryptic soy broth agar (1.5%) plate. Smeared. After drying at room temperature for about 30 minutes, four to five steel wells were placed on the TSB agar plate.

Thereafter, the feed additive prepared from the culture medium of the Bacillus bacteria (Bacillus rikenimoform, Bacillus subtilis and Bacillus coagulans) of Example 1 was removed by sterile filtering, and then the steel well. 100 μl each. After incubation for 12 hours at 37 ℃, and observed the antimicrobial active ring to compare the antimicrobial activity by the size (unit: mm) ( Table 1 ).

Size of Antimicrobial Activity Pills from Citrus Foil Feed Additives Microorganisms Used in the Preparation of Citrus Foil Feed Additives Pathogenic fungi Staphylococcus aureus Salmonella Galinarum Escherichia coli Bacillus subtilis 20 16 23 Bacillus rickeniformis 18 14 18 Bacillus coagulant 17 0 18

As a result, the feed additive prepared from the Bacillus subtilis culture had the highest antimicrobial activity against all three pathogenic bacteria, and the cultures inoculated with Bacillus coagulans had the low antibacterial activity, but all showed significant antimicrobial activity. Indicated.

In addition, in the pathogenic bacterium, the antimicrobial activity of the feed additive prepared from Bacillus subtilis culture and the conventional antibiotic ampicillin against Gram-positive bacteria, Staphylococcus aureus Gram-negative bacteria, Salmonella gallinarum and Escherichia coli antibacterial activity of (ampicillin), kanamycin, kanamycin, streptomycin, tetracycline, vancomycin, spectinomycin, novobiocin (all purchased from Sigma) Comparison was made ( FIG. 2 and Table 2 ).

Comparison of Antimicrobial Activity between Feed Additives and Various Antibiotics S. Aureus (Gram +) S. Galina Room (Gram-) Escherichia coli (gram-) density Antibacterial ring (mm) density Antibacterial ring (mm) density Antibacterial ring (mm) B. Feed Additives Prepared from Subtilis (Non-treated) 10% 28 10% 15-16 10% 17-20 [Feed additives Pretreatment ] 65 ℃, 30 minutes heat treatment 10% ND 10% ND 10% ND 100 ℃, 10 minutes heat treatment 10% ND 10% ND 10% ND Wet Sterilization (121 ℃, 20 minutes) 10% 0 10% 7 10% 7 Protease K, 37 ° C. 2 hours treatment 1 mg / ml ND 1 mg / ml ND 1 mg / ml ND Lipase, 37 ° C. 2 hours treatment 1 mg / ml 12 1 mg / ml 7 1 mg / ml 7 Amylase, 2 hours treatment at 37 ° C 1 mg / ml 17 1 mg / ml 7 1 mg / ml 8 Catalase, 37 ° C. 2 hours treatment 1 mg / ml 15 1 mg / ml 7 1 mg / ml 9 SSF (Solid Culture) 10% 0 10% 7 10% 7 50% methanol, treated at 37 ° C for 1 hour 10 20-22 10% 13-14 10% 8-13 [Other antibiotics] Ampicillin 1 mg / ml 40 1 mg / ml 26-27 1 mg / ml 18-19 Kanamycin 3 mg / ml 20 3 mg / ml 18-22 3 mg / ml 19-20 Streptomycin 1 mg / ml 18-19 1 mg / ml 18 1 mg / ml 7 Tetracycline 3 mg / ml 25-28 3 mg / ml 22 3 mg / ml 19-22 Banokomycin 1 mg / ml 19 1 mg / ml 0 1 mg / ml 0 Spectinomycin 1 mg / ml 13 1 mg / ml 14 1 mg / ml 12-15 Novobiocin 1 mg / ml 27-30 1 mg / ml 0 1 mg / ml 0

As a result, it was confirmed that the feed additive prepared from the Bacillus subtilis culture showed excellent antimicrobial activity compared to the conventional antibiotics except for ampicillin (ampicillin) when compared with the existing antibiotics.

< Example  3> beef cattle, Milking cow , Upbringing On fattening pigs  Pay effect

We tested the feeding effect on the beef cattle, milking cows, and finishing pigs of the citrus gourd feed additive prepared in Example 1 .

Specifically, the citrus gourd feed additive prepared by using the Bacillus rikenipomis according to the method of Example 1 is a general compound feed (crude protein 17%, crude fat 2.0%, crude fiber 5.5%, crude ash 8.0%, calcium 0.7%, 0.7% phosphorus and lysine).

<3-1> Salary Effects on Beef Cattle

Comparing total weight gain and daily weight gain (unit: kg) for beef cattle when 0%, 20% and 40% of the blended feed was supplemented with citrus gourd feed additives for 90 days It was tested ( Table 3 ).

Growth Performance of Beef Cattle When Supplemented with Citrus Foil Feed Additives (Unit: ㎏) division Compound feed salary Combined feed (80%) + citrus gourd feed additive (20%) Combined feed (60%) + citrus gourd feed additive (40%) Test Start Weight 210 197 204 Weight at end 281 264 252 Total weight 71 67 48 Daily weight gain 0.78 0.71 0.54

As a result, there was no difference in the weight gain of beef cattle in mixed feed and mixed feed (80%) + citrus gourd feed additives (20%), but the mixed feed (60%) + citrus gourd feed additive (40%). In the mixed feed group, the weight gain decreased. In other words, the addition ratio of the appropriate citrus gourd feed additive is estimated to be about 20%.

In addition, the daily weight gain (unit: kg), carcass rate (carcass pecent) and feed requirement for the beef cattle when citrus meal feed additives were mixed with the blended feed were compared ( Table 4 ). In this case, the carcass ratio refers to the ratio of carcass weight to the live weight of the livestock. The weight immediately before slaughtering the livestock is called bioweight, and the rest of the carcass after removing the ankle, head, and internal organs after slaughter and peeling is called carcass. . The carcass rate is determined by the breed of the animal, the food left in the digestive organs, the level of fattening and the method of slaughtering.

Effects of Feeding Citrus Gourd Feed Additives on Beef Cattle division Effect of adding citrus gourd feed additive 0% 7% 14% 21% Weight at end of test (kg) 333 339 333 341 Daily weight gain (㎏) 1.18 1.08 1.03 1.11 Conductor Rate (%) 61.6 60.7 60.1 61.2 Daily feed requirement (kg) 8.35 8.72 8.2 8.46 Feed requirement per gain (kg) 7.07 8.15 8.0 7.78

As a result, weight gain and carcass rate were similar regardless of salary level.

<3-2> Milking cows  Pay effect

When the dietary supplement of the citrus gourd feed additive of the present invention was supplemented, the milk yield and maintenance rate of cows were compared, and the increase or decrease of the somatic cell count was analyzed.

Specifically, the separated feed of the blended feed and citrus gourd feed additive, the total mixed ratio (TMR) feed of the citrus gourd feed additive, and the TMR feed of the 30% of the blended feed replaced with the citrus gourd feed additive. We analyzed the milk yield (kg) and milk retention (%) for cows ( Table 5 ). In the present invention, the TMR feeding method is a method of adjusting the feed amount based on the nutrient requirement while feeding the citrus meal feed additive and the blended feed.

The oil production was analyzed by crude oil flow, and the oil production by individual was measured once a month. The maintenance factor and somatic cell count were analyzed as milk quality factors, and the method and timing were the same as in the above acid flow analysis. Retention rate was used for the Multispec-C-M (Infra red milk analyzer), somatic cell count was analyzed using a Fossomatic 300/90 (Fossomatic 300/90).

Effects of Separation or TMR Supplementation division Separate Feeder (Combination Feed + Citrus Foil Feed Additive) TMR Feeders of Citrus Foil Feed Additives 30% TMR Dietary Supplement for Citrus Foil Feed Additives Feed intake (kg / two / day) Compound Feed 9.0 (7.9) 9.6 (8.39) 9.4 (8.22) Citrus gourd feed additive 19.9 26.4 (5.03) 25.8 (4.92) hay 4.7 (4.14) 3.8 (3.35) 3.7 (3.92) Acid flow rate (kg / two / day) 16.39 17.94 16.92 % Retention 3.50 3.62 3.58

As a result, the milk yield was the highest in the TMR feed group of citrus gourd feed additives, followed by the TMR feed group in which 30% of the blended feed was replaced with citrus gourd feed additives, followed by the separated feed group. Retention rates tended to increase slightly with TMR. Therefore, the proper mixing amount of the citrus gourd feed additive for milking cows is when 30% of the blended feed is replaced with the citrus gourd feed additive.

In addition, analysis of milk composition showed significant increases in retention and total solids (p <0.05) and increased levels of milk protein and plain solids in citrus meal supplements, especially TMR. In addition, the pH and titratable acidity of milk were generally in the range of the appropriate level.

On the other hand, it was the result of examining the somatic cell, is 66% Class I (2.5x10 ⁵ or less), deungoe (7.5x10 ⁵ or more) are the need for a rather good side or improve the level of 2% was observed. However, in the TMR feed group, a marked decrease in the number of somatic cells was found. On the other hand, the suspension of the citrus gourd feed additives and the somatic cell count increased when switching to the conventional breeding method in the TMR feed, barring the TMR using the citrus gourd feed additives may be effective in preventing breast cancer.

<3-3> upbringing On fattening pigs  Pay effect

Citrus gourd feed additives of the present invention when pigs were fed and analyzed for weight gain, feed intake, digestibility and carcass performance.

Specifically, 36 days of landracex large white hybrids 41 days after delivery were performed for 115 days, the control group without addition of the citrus gourd feed additive (T0), the experimental group with the addition of 2% citrus gourd feed additive (T1) The weight gain, feed intake, digestibility and carcass results were analyzed in the experimental group (T2) supplemented with 8% citrus fruit feed additives.

As a result, the experimental group (T1) in the growing period was slightly lower than the control group (T0), but the feed intake was slightly lower, indicating that the feed demand was improved. However, the gains in the experimental group (T2) were generally reduced (P <0.05).

In addition, the feed intake decreased in the order of control group (T0)> experiment group (T1)> experiment group (T2), whereas the weight gain increased in the experimental group (T1), and the feed demand rate was the best. However, the gains in the experimental group (T2) were lowered as in the growing phase (P <0.05).

On the other hand, in terms of the carcass results, in the experimental group (T2), the weight gain and the carcass rate were slightly increased, and the back fat thickness was getting thinner. It was effective in improving meat quality such as increasing lean meat.

In addition, all of the nutrient digestibility except crude fiber digestibility was higher in the growing season, especially in the experimental group (T2), protein digestibility in the growing season and protein and crude fiber and soluble nitrogen (NFE) digestibility were improved in the growing period. .

As described above, feed additives containing a culture cultured by inoculating Bacillus bacteria into citrus gourd or citrus gourd extract can not only recycle the citrus gourd, but also improve feed utilization, weight gain, and disease resistance. Intestinal effective bacteria in livestock can be increased. In addition, due to the antimicrobial activity of the feed can be expected to inhibit the growth of harmful bacteria inhabiting the intestines of the livestock.

Claims (7)

Antimicrobial feed additive containing a culture solution inoculated with citrus bacterium or citrus gourd extract. The method of claim 1, wherein the Bacillus genus bacteria is Bacillus licheniformis ( Bacillus licheniformis ), Bacillus subtilis ( Bacillus subtilis ) and Bacillus coagulance ( Bacillus coagulance ), characterized in that one selected from the group consisting of additive. The feed additive of claim 2, wherein the Bacillus genus bacteria is Bacillus licheniformis . According to claim 1, wherein the bacteria of the genus Bacillus feed additives, characterized in that 1 to 20 parts by weight based on 100 parts by weight of citrus gourd or citrus gourd extract. Antimicrobial feed comprising 10 to 20% of the feed additive of claim 1 in the general formula feed. (1) obtaining a citrus gourd extract from the citrus gourd; (2) a method of producing a feed additive according to claim 1, comprising the step of inoculating the citrus gourd or the citrus gourd extract obtained in step 1 and inoculating Bacillus bacteria. 7. The method of claim 6, further comprising the step of preparing a powder by drying the culture solution obtained after step 2.

Images