KR102769334B1 - Composition for weight gain in livestock comprising fecal microbiota - Google Patents

Abstract

The present invention relates to a livestock weight gain composition comprising fecal microbiota. The livestock weight gain composition comprising fecal microbiota according to the present invention was confirmed to have a remarkable weight gain effect in the growing period (12 months) and fattening period (24 months). In addition, it was confirmed that the carcass weight, loin area, and intramuscular fat grade after slaughter were excellent, and thus, it can be utilized in various ways in the livestock industry.

Description

{Composition for weight gain in livestock comprising fecal microbiota}

The present invention relates to a composition for livestock weight gain comprising fecal microflora.

Calf diarrhea is known to be a multifactorial disease with a variety of causes. In addition to various environmental factors such as the cleanliness of the barn, milk quality of the mother cow, temperature, and humidity, infections by viruses (e.g., bovine viral diarrhea virus, bovine coronavirus, and group A rotavirus), bacteria (e.g., Salmonella sp., and Clostridium perfringens), and protozoa (e.g., Eimeria zuernii) have been identified as causes of calf diarrhea. However, recent studies on neonatal diarrhea in mammals including ruminants have shown that infections with the aforementioned pathogenic microorganisms are frequently observed in healthy individuals regardless of diarrheal symptoms (Case-control study of microbiological etiology associated with calf diarrhea (2013), Vet microbiol). Conversely, infections with pathogenic microorganisms causing diarrhea were not found at all in individuals with severe diarrheal symptoms.

When examining the formation process of the intestinal microbial community of newborn calves, aerobic microbes such as Proteobacteria are dominant among the intestinal microbial community derived from the mother cow in the early stage of birth, and these consume oxygen in the intestinal environment and create an environment in which anaerobic microbes can settle (Proteobacteria: microbial signature of dysbiosis in gut microbiota (2015), Trends Biotechnol). This early intestinal microbial community formation process is known to provide colonization resistance to the intestinal environment, thereby reducing infection by allochthonous enteric pathogens. Therefore, the incomplete formation of the initial intestinal microbial community of newborn calves and the resulting gut microbial dysbiosis can cause various intestinal diseases including diarrhea.

Diarrhea is a common disease in young livestock during cattle breeding, and it causes enormous economic damage due to its high mortality rate. However, there is no alternative to the use of antibiotics or microbial agents that can survive under aerobic conditions. When antibiotics are used, it affects the growth of livestock that depend on microorganisms, and it also causes various problems such as the problem of residual antibiotics and the emergence of super bacteria due to antibiotic resistance. Microbial agents developed so far are very easy to isolate, do not have a clear biological basis for their mechanism of action, and are microorganisms that easily grow under aerobic conditions, but in the intestines of actual livestock, anaerobic strains have a much greater effect on immunity and metabolism.

Meanwhile, Fecal Microbiota Transplantation (FMT) is a method developed in the United States to treat patients with severe diarrhea caused by Clostridium difficile infection by transplanting the feces of healthy people selected according to strict criteria. The importance of maintaining a balanced microbiota is being highlighted rather than indiscriminate use of antibiotics.

Accordingly, the inventors of the present invention developed a livestock feed composition containing fecal microbiota and confirmed the composition's effect on increasing body weight and improving slaughter grade, thereby completing the present invention.

Therefore, the purpose of the present invention is to provide a feed composition for livestock weight gain comprising a homogeneous fecal microflora.

Another object of the present invention is to provide a feed additive composition for livestock weight gain comprising a homogeneous fecal microflora.

Another object of the present invention is to provide a livestock mass increase method comprising the steps of: (a) obtaining feces from livestock; (b) mixing the feces of step (a) with an electrolyte and glycerol to prepare a fecal mixture containing fecal microbiota of the same species; and (c) administering the fecal mixture of step (b) to a livestock in need thereof.

Another object of the present invention is to provide a feed composition for improving meat quantity and meat quality comprising a homologous fecal microflora.

To achieve the above purpose, the present invention provides a feed composition for livestock weight gain containing a homogeneous fecal microbiota.

The present invention also provides a feed additive composition for livestock weight gain comprising a homogeneous fecal microflora.

In addition, the present invention provides a livestock mass increase method comprising: (a) obtaining feces from livestock; (b) mixing the feces of step (a) with an electrolyte and glycerol to prepare a fecal mixture containing fecal microbiota of the same species; and (c) administering the fecal mixture of step (b) to a livestock in need thereof.

The present invention also provides a feed composition for improving meat quantity and quality, comprising a homogeneous fecal microbiota.

The livestock weight gain composition containing the fecal microbiota according to the present invention was confirmed to have a remarkable weight gain effect in the growing period (12 months) and fattening period (24 months). In addition, it was confirmed that the carcass weight, loin area, and intramuscular fat grade after slaughter were excellent, so it can be utilized in various ways in the livestock industry.

Figure 1 is a diagram showing the schedule for fecal mass transplantation, weight measurement, and slaughter.
Figure 2 shows the results of measuring the body weight of calves according to fecal mass transplantation (FMT: fecal mass transplantation group, CON: electrolyte administration group, ABX: antibiotic administration group).
Figure 3 shows the results of analyzing weight gain based on the body weight measurement results (FMT: fecal mass transplant group, CON: electrolyte administration group, ABX: antibiotic administration group).
Figure 4 shows the results of calculating the body weight, intramuscular fat grade, and loin cross-sectional area after slaughter (FMT: fecal mass transplant group, CON: electrolyte administration group, ABX: antibiotic administration group).

Hereinafter, the present invention will be described in detail.

According to an aspect of the present invention, the present invention provides a feed composition for livestock weight gain comprising a homogeneous fecal microbiota. The present invention also provides a feed additive composition for livestock weight gain comprising a homogeneous fecal microbiota.

In the present invention, microbiota refers to a microscopic organism group in a certain area, and means the combination of microbiota and microfauna of a certain area.

In the present invention, fecal microbiota refers to a set of microorganisms (bacteria, yeast and fungi) found in feces.

In a specific example of the present invention, the fecal microbiota is fecal microbiota derived from healthy livestock, and it is preferable that the healthy livestock is livestock satisfying at least one condition selected from the group consisting of the following conditions.

(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;

(ii) There shall be no record of administration of antibiotics, antifungal agents or antiviral agents.

(iii) When livestock feces are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, feces corresponding to stages 3 and 4 should be excreted.

(iv) The diagnostic multiplex PCR will not be positive for one or more of the following pathogens: group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus , bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli, shigatoxigenic Escherichia coli , enterohemorrhagic Escherichia coli , and Eimeria zuernii.

In a specific embodiment of the present invention, it is preferred that the composition comprises a mixture of an electrolyte and glycerol containing the homogeneous fecal microflora.

In a preferred embodiment of the present invention, the mixture is preferably, but not limited to, a mixture of 5 g of feces and 50 ml of a solution containing 4% electrolyte and 10% glycerol.

In a specific embodiment of the present invention, the livestock is preferably at least one selected from the group consisting of cattle, sheep, chickens, ducks, geese, pheasants, pigs, cows, goats, dogs and cats, but is not limited thereto.

In a preferred embodiment of the present invention, the livestock is preferably a livestock showing diarrhea symptoms. In an embodiment of the present invention, it was confirmed that even in livestock showing diarrhea symptoms, when the composition of the present invention was administered, a remarkable weight gain effect was observed.

In a specific embodiment of the present invention, the composition is preferably for fattening or increasing body weight in the breeding or fattening phase.

The above-mentioned growing period is from 3-4 months of age to 12-13 months for Korean cattle, and is the period for growing and developing young animals that have been weaned, so the specific period may vary depending on the farm and livestock type. The fattening period is the period when livestock gain weight and is a growth stage where fat is mainly accumulated, and refers to the period from the growing period to before shipping.

The feed composition of the present invention may be a feed additive or a compound feed.

In the present invention, a feed additive means a substance added in small amounts to feed for nutritional or specific purposes, and the functional feed additive of the present invention may be the homogeneous fecal microbiota itself or may be mixed with known feed additives at a certain ratio. When mixed, the homogeneous fecal microbiota may be applied in an amount of 0.01 to 99 wt% of the total weight of the feed additive.

The feed composition of the present invention comprises 0.1 to 5 parts by weight of a feed additive containing a homogeneous fecal microflora as an effective ingredient, per 100 parts by weight of compound feed. As an example, the compound feed is composed of 53.71 wt% of corn, 23.2 wt% of soybean meal, 1.0 wt% of rapeseed meal, 2.0 wt% of corn germ meal, 3.0 wt% of distillers' meal, 2.7 wt% of wheat bran, 0.3 wt% of potato starch, 2.8 wt% of fat, 9.9 wt% of calcium carbonate, 0.68 wt% of dibasic calcium phosphate, 0.23 wt% of sodium chloride, 0.03 wt% of phosphorus decomposition enzyme, 0.05 wt% of choline chloride, 0.2 wt% of methionine, 0.1 wt% of mineral mix, and 0.1 wt% of vitamin mix. In addition, it goes without saying that a conventional compound feed for poultry can be used. In addition, it goes without saying that compound feed can be used for cattle, pigs, ducks, etc. depending on the type of livestock being raised.

Livestock can be raised by feeding the above feed composition, and when raising livestock, the same species of fecal microbiota can be diluted in water and provided as drinking water. At this time, 100 to 500 ml of the same species of fecal microbiota can be applied to 100 liters of water.

The feed composition of the present invention may additionally contain other nutritional ingredients within a range that can be expected by those skilled in the art.

The homologous fecal microbiota of the present invention can be incorporated into commercially conventional animal feed compositions and fed to, for example, cattle, pigs, sheep, poultry, etc. For this purpose, the homologous fecal microbiota of the present invention can be mixed with conventional animal feed components and, if necessary, processed into a purified form. Conventional animal feed components are, for example, corn, barley, oats, soybeans, fish meal, bran, soybean oil, minerals, trace elements, amino acids, and vitamins.

According to another aspect of the present invention, the present invention provides a livestock mass increase method comprising the steps of: (a) obtaining feces from livestock; (b) mixing the feces of step (a) with an electrolyte and glycerol to prepare a fecal mixture containing fecal microbiota of the same species; and (c) administering the fecal mixture of step (b) to a livestock in need thereof.

In a specific example of the present invention, it is preferable that the livestock of step (a) is a livestock that satisfies at least one condition selected from the group consisting of the following conditions.

(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;

(ii) There shall be no record of administration of antibiotics, antifungal agents or antiviral agents.

(iii) When livestock feces are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, feces corresponding to stages 3 and 4 should be excreted.

(iv) The diagnostic multiplex PCR will not be positive for one or more of the following pathogens: group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus , bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli, shigatoxigenic Escherichia coli , enterohemorrhagic Escherichia coli , and Eimeria zuernii.

In a specific example of the present invention, the livestock in step (c) may be a livestock showing diarrhea symptoms, and in an example of the present invention, it was experimentally confirmed that the method of the present invention exhibits an excellent weight gain effect even in an individual showing diarrhea symptoms.

In a specific embodiment of the present invention, the administration in step (c) is preferably oral administration, but is not limited thereto.

According to another aspect of the present invention, the present invention provides a feed composition for improving meat quantity and meat quality comprising a homogeneous fecal microbiota.

In a specific example of the present invention, it is preferable that the meat quality improvement is an improvement in intramuscular fat and sirloin area.

In the present invention, the intramuscular fat grade is one of the factors that distinguishes beef, and is also called marbling. The intramuscular fat grade is comprehensively evaluated from 1 to 9 levels by graders belonging to the Livestock Product Quality Evaluation Institute by checking the degree of fat distribution, meat color, maturity, etc. within the muscle at each slaughterhouse, and the higher the number, the higher the grade.

In the present invention, the sirloin cross-sectional area refers to a numerical value measuring the size of the sirloin cross-section of a slaughtered cow. The higher the sirloin cross-sectional area of a slaughtered cow, the higher the price at which it is traded.

The feed composition for improving meat quantity and meat quality according to the present invention is excellent in improving meat quantity (i.e., carcass weight) as well as meat quality, and thus can be usefully utilized in the livestock industry.

Duplicate contents are omitted in consideration of the complexity of this specification, and terms not otherwise defined in this specification have meanings commonly used in the technical field to which the present invention belongs.

Hereinafter, the present invention will be described in more detail through examples. It will be apparent to those skilled in the art that these examples are intended only to illustrate the present invention, and the scope of the present invention is not to be construed as being limited by these examples.

Example 1. Selection of healthy fecal donor calves

Calves were selected for fecal donor transplantation. The specific selection steps were as follows.

(1) Among calves 21-50 days old, calves that were not suffering from Class 1 and Class 2 livestock infectious diseases (foot-and-mouth disease, contagious bovine pleuropneumonia, anthrax, or brucellosis) as prescribed by the Ministry of Agriculture, Food and Rural Affairs, had no discharge from the eyes, nose, ears, or rump, had good color, and were active were first selected.

(2) Among calves aged 2 weeks to 1 month, calves without a record of administration of drugs that could affect the microbial community, including antibiotics, antifungals, or antivirals, were selected for a second time.

(3) Based on the fecal consistency score, the calf feces were divided into stages 1 (hard) to 7 (diarrhea), and feces corresponding to stages 3 and 4 were selected in three stages.

(4) Using the diagnostic multiplex PCR technique, RNA viruses (group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus, bovine nebovirus, or bovine viral diarrhea virus), bacterial pathogens ( Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli , shigatoxigenic Escherichia coli , or enterohemorrhagic Escherichia coli ), and protozoa ( Eimeria zuernii ) reported to cause diarrhea in calves were tested, and if a positive reaction was found, all of the feces were discarded.

The experiments described below were performed according to the schedule in Fig. 1.

Example 2. Transplantation of fecal masses

Feces were collected from the calves that donated feces selected in the above Example 1. The obtained 5 g of feces were mixed with 50 ml of a solution containing 4% by volume of electrolyte and 10% by volume of glycerol to prepare a feces mixture. The electrolyte is a powder for oral administration that prevents dehydration in livestock, and its main components are sodium, potassium, and glucose. Before use, it is mixed in warm water and fed to calves with symptoms of dehydration and metabolic acidosis. The composition of the electrolyte used in this experiment is as follows.

[4.68% sodium citrate (w/v), 2.29% sodium propionate (w/v), 3.92% sodium acetate (w/v), 5.59% sodium chloride (w/v), 3.56% potassium chloride (w/v), 75.2% glucose (w/v), 0.1% sunset yellow (w/v), 3.0% silicon dioxide (w/v), 1.62% potassium phosphate (w/v)].

The prepared fecal mixture was rapidly frozen. The 10% glycerol prevents the destruction of microbial cells in a low-temperature environment.

For fecal mass transplantation, frozen fecal mixture was sufficiently thawed by boiling in hot water (1 hour) and oxygen contact was minimized. After that, it was administered orally three times at 12-hour intervals to calves showing diarrhea symptoms. If necessary, additional administration was given on the 8th and 48th days of diarrhea.

Calves with diarrhea were randomly selected and administered fecal mixture, electrolyte mixture, and antibiotics three times orally at 12-hour intervals to 20 calves receiving fecal mass transplantation, 14 calves in the electrolyte administration control group, and 23 calves in the antibiotic administration control group, respectively. The electrolyte administration control group (CON) was orally administered the same volume of electrolyte/glycerol mixture to confirm the effect of the 50 ml solution containing 4% by volume electrolyte and 10% by volume glycerol used for fecal suspension in the process of manufacturing the FMT mixture. The antibiotic administration control group (ABX) was orally administered 1 g of neomycin in powder form mixed with the same volume of electrolyte/glycerol solution to reflect the administration of antibiotics to diarrheal calves on farms.

Example 3. Measurement of calf weight

The body weights of 20 calves that received fecal mass transplantation, 12 calves in the electrolyte-administered control group (2 calves excluded due to death), and 19 calves in the antibiotic-administered control group (4 calves excluded due to death) were measured at 6, 12, and 24 months of age, respectively. The body weight measurement results are shown in Fig. 2.

As shown in Figure 2, there was little difference in the 6-month body weight in all groups. The 12-month and 24-month fecal transplant calf groups (FMT) showed higher body weights compared to the electrolyte administration group (CON) and the antibiotic administration group (ABX) in both males and females.

Based on the above weight measurement results, the weight gain at 6-12 months and the weight gain at 12-24 months were calculated. The results of calculating the weight gain are shown in Fig. 3.

As shown in Figure 3, both male and female fecal transplant groups (FMT) showed superior weight gain compared to the electrolyte administration group (CON) and antibiotic administration group (ABX).

Example 4. Analysis of slaughter performance

The animals used in the experiment were slaughtered at the Farm Story Korean Refrigerated LPC slaughterhouse located in Cheongju, Chungcheongbuk-do, and the dressed weight, intramuscular fat grade, and sirloin area were calculated. The dressed weight refers to the weight excluding by-products such as the head, legs, skin, fat, and internal organs of the slaughtered cattle. In addition, the intramuscular fat grade is a term commonly referred to as marbling. It is a comprehensive evaluation from 1 to 9 levels by graders belonging to the Livestock Product Quality Evaluation Institute by checking the degree of fat distribution, meat color, and maturity within the muscles at each slaughterhouse, with a higher number indicating a higher grade. The sirloin area is a number measuring the size of the sirloin cross-section, and a wider cross-sectional area is traded at a higher price. The results of analyzing the slaughter performance of cattle are shown in Fig. 4.

As shown in Fig. 4, the fecal mass transplant (FMT) group was confirmed to be superior to the electrolyte administration group (CON) and the antibiotic administration group (ABX) in terms of carcass weight, loin cross-sectional area, and intramuscular fat grade, regardless of gender. In particular, in terms of carcass weight, the male individuals in the fecal mass transplant (FMT) group had an average of 473 kg, which was significantly superior to the average shipping carcass weight of 442 kg of the top 10% farms nationwide based on slaughter performance. In terms of loin cross-sectional area, the male individuals in the fecal mass transplant (FMT) group had an average of 113 cm ² , which was significantly superior to the average shipping loin cross-sectional area of 98 cm ² of the top 10% farms nationwide. In terms of intramuscular fat grade, the male individuals in the fecal mass transplant (FMT) group had an average of 7.4, which was significantly superior to the average intramuscular fat grade of 6.5 of the top 10% farms nationwide.

In summary, the inventors of the present invention confirmed that transplanting fecal microbiota from healthy calves to calves with diarrhea through fecal transplantation had a weight gain effect during the growing period (12 months) and fattening period (24 months). In addition, it was confirmed that the carcass weight, loin area, and intramuscular fat grade were excellent after slaughter, which can be utilized in various ways in the livestock industry.

Above, specific parts of the present invention have been described in detail. It will be obvious to those skilled in the art that such specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. Accordingly, it will be said that the actual scope of the present invention is defined by the appended claims and their equivalents.

Claims (13)

In a feed composition for livestock weight gain containing fecal microbiota of homogeneous origin,
The above livestock are cattle in the growing or fattening stage,
The composition increases body weight, improves intramuscular fat grade, and increases sirloin area.
The composition comprises a mixture of electrolyte and glycerol containing the homogeneous fecal microbiota,
The above fecal microbiota is fecal microbiota derived from a healthy cow, and the above healthy cow is a cow satisfying the following conditions, a livestock feed composition for weight gain:
(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;
(ii) No record of administration of antibiotics, antifungal agents or antiviral agents;
(iii) When the feces of livestock are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, the feces corresponding to stage 3-4 shall be excreted; and
(iv) group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus, bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli , shigatoxigenic Escherichia coli , enterohemorrhagic Must not be positive on diagnostic multiplex PCR for Escherichia coli and Eimeria zuernii . delete delete delete In the first paragraph,
A feed composition for livestock weight gain, wherein the livestock is a livestock showing diarrhea symptoms. delete A composition of feed additive for livestock weight gain comprising fecal microbiota of the same species,
The above livestock are cattle in the growing or fattening stage,
The composition increases body weight, improves intramuscular fat grade, and increases sirloin area.
The composition comprises a mixture of electrolyte and glycerol containing the homogeneous fecal microbiota,
The above fecal microbiota is fecal microbiota derived from a healthy cow, and the above healthy cow is a cow satisfying the following conditions, a livestock feed additive composition for weight gain:
(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;
(ii) No record of administration of antibiotics, antifungal agents or antiviral agents;
(iii) When the feces of livestock are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, the feces corresponding to stage 3-4 shall be excreted; and
(iv) group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus, bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli , shigatoxigenic Escherichia coli , enterohemorrhagic Must not be positive on diagnostic multiplex PCR for Escherichia coli and Eimeria zuernii . (a) a step of obtaining feces from livestock;
(b) a step of mixing the feces of step (a) with electrolyte and glycerol to prepare a fecal mixture containing homogeneous fecal microbiota; and
(c) a step of administering the fecal mixture of step (b) to livestock in need thereof;
The livestock of the above step (a) is a livestock increasing method that satisfies the following conditions:
The above livestock weight gain method is for increasing the carcass weight of cattle in the growing or fattening period, improving the intramuscular fat grade, and increasing the sirloin area:
(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;
(ii) No record of administration of antibiotics, antifungal agents or antiviral agents;
(iii) When the feces of livestock are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, the feces corresponding to stage 3-4 shall be excreted; and
(iv) group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus, bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli , shigatoxigenic Escherichia coli , enterohemorrhagic Must not show positive diagnostic multiplex PCR for Escherichia coli and Eimeria zuernii . delete In Article 8,
A livestock increasing method, wherein the livestock of the above step (c) shows symptoms of diarrhea. In Article 8,
A livestock weight gain method in which the administration of the above step (c) is oral administration. In a feed composition for improving meat quantity and meat quality containing fecal microbiota of the same species,
The composition comprises a mixture of electrolytes and glycerol containing the homologous fecal microbiota, which increases the body weight of cattle in the growing or fattening period,
The above fecal microbiota is fecal microbiota derived from a healthy cow, and the above healthy cow is a cow that satisfies the following conditions, a feed composition for improving meat quantity and meat quality:
(i) No history of infection with foot-and-mouth disease, tuberculosis, contagious bovine pleuropneumonia, anthrax or brucellosis;
(ii) No record of administration of antibiotics, antifungal agents or antiviral agents;
(iii) When the feces of livestock are divided into stages 1 (hard) to 7 (diarrhea) based on the fecal consistency score, the feces corresponding to stage 3-4 shall be excreted; and
(iv) group A rotavirus, group B rotavirus, group C rotavirus, bovine coronavirus, bovine torovirus, bovine norovirus, bovine enteric Nebraska-like calicivirus, bovine nebovirus, bovine viral diarrhea virus, Clostridium perfringens , Salmonella enterica , Salmonella enterica typhimurium , enterotoxigenic Escherichia coli , shigatoxigenic Escherichia coli , enterohemorrhagic Must not be positive on diagnostic multiplex PCR for Escherichia coli and Eimeria zuernii . In Article 12,
The above meat quality improvement is a feed composition for improving meat quantity and meat quality by improving intramuscular fat grade and sirloin area.