TWI494432B - Poultry water soluble additives and its manufacturing method - Google Patents

Description

Poultry drinking water additive and manufacturing method thereof

The present invention relates to a poultry drinking water additive; in particular, a fermented product of Cordyceps militaris obtained by solid state fermentation of cereals, and a poultry drinking water additive for regulating immunity.

In recent years, the ban or restriction of the use of antibiotics for animal feed has become a global trend, and the addition of antibiotics to animal feed may lead to public health problems such as drug-resistant strains and drug residues. The production of green and safe animal feed has become a trend in today's livestock and poultry industry. A number of studies have suggested that the addition of Chinese herbal medicines to livestock and poultry diets has the effect of replacing antibiotics. However, due to the unstable sources of Chinese herbal medicines, high prices, heavy metals and pesticides Remaining doubts and other issues.

" Cordyceps" is a general term for Cordyceps fungi, classified as Ascomycotina, Pyrenomycetes, Lavicipitales, Clavicipitaceae, Cordyceps (Cordyceps). There are more than 350 species of Cordyceps found in the world, the most famous being Cordycepssinensis and Cordycepsmilitaris.

The intracellular and extracellular polysaccharides of Cordyceps militaris can significantly increase the phagocytic ability of mouse peritoneal macrophages (Jiang et al., 2002); it can also activate macrophages, stimulate antibody production, enhance cellular immunity and humoral immunity, thereby enhancing the human body. Immunity (Fang et al., 2000; Yu et al., 2006). Cordyceps polysaccharide can promote lymphocyte transformation, increase the antibody content of serum IgG and the immune function of antibodies, and enhance the body's ability to fight cancer and inhibit cancer.

Cordyceps militaris, a secondary metabolite of Cordyceps militaris, can increase the expression of IL-10 mRNA and has a significant positive regulatory ability. Stimulation of macrophages via lipopolysaccharide and simultaneous administration of cordycepin can inhibit the gene expression of cytokines and chemokines. The amount of IL-10, IL-1ra, and TGF-β was increased (Shin et al., 2009). Cordycepin can be expressed by negative regulation of cyclooxygenase-2 (COX-2) and gene expression of inducible nitric oxide synthase (iNOS), as well as NF-κB activity, AKT and p38 phosphorylation. Inhibition to antagonize the production of NO products (Kim et al., 2006b).

The research and development of green feed additives such as Chinese herbal medicines allows livestock and poultry farmers to have more alternatives to non-drugs and reduce the shadow of drug residues in livestock products. The present invention uses poultry-chicken as a test animal and uses scorpion as a solid-state fermentation. The substrate is adjusted and the nitrogen source is adjusted, and the obtained fermentation product of Cordyceps militaris is dried and pulverized, and then added to drinking water to achieve the application of regulating the immune response of the poultry.

Figure 1 shows the effect of different doses of Cordyceps militaris fermentation on the weight of chickens.

Figure 2 shows the expression of iNOS mRNA in the spleen lymphoid tissues of chickens.

Figure 3 shows the amount of COX-2 mRNA in the spleen lymphoid tissues of chickens.

Figure 4 shows the expression of INF-γ mRNA in the lymphoid tissue of chicken spleen.

Figure 5 shows the expression of IL-4 mRNA in the spleen lymphoid tissues of chickens.

Figure 6 shows the amount of iNOS mRNA expression in the chicken sac.

Figure 7 shows the amount of COX-2 mRNA expression in the chicken sac.

Figure 8 shows the expression of INF-γ mRNA in chicken Fahrenheit.

Figure 9 shows the expression of IL-4 mRNA in the chicken sac.

The present invention uses a Cordycepsmilitaris strain to carry out solid state fermentation culture of cereals. The Cordyceps sinensis strain was subcultured with PDA (potato dextrose agar) plate and cultured at 22 ° C for 7 days, then at a constant temperature of 22 ° C, the rotation speed was controlled at 150 rpm, cultured for 5 days to activate the bacteria, and brown rice was used as a solid fermentation substrate, which contained 5% fish meal was used as a nitrogen source, and 10 mL of pre-activated bacterial solution was inoculated into brown rice solid medium, cultured at 22 ° C, protected from light for 3 days, lighted for 16 hours per day for 16 hours, dark for 8 hours, and dried for 42 days after fermentation. Milling.

The fermentation product of Cordyceps militaris was analyzed by HPLC for HPLC. The HPLC conditions were as follows: mobile phase was 0.02 M KH2PO4 and 15% methanol, sample injection HPLC volume was 20 μL, flow rate was controlled at 1 mL/min, UV detection wavelength was 254 nm. The analytical column was a high-efficiency reverse phase column RP-18 endcapped (5 μm, Merck). The analysis showed that the content of cordycepin in the fermentation product of Cordyceps militaris was 30 ppm.

The dose test of the fermentation of the Cordyceps militaris was divided into four groups: the control group (the water was not added to the fermentation of the Cordyceps militaris), and the concentration of the cordycepin after the conversion of the drinking water of the Cordyceps militaris was 30 ppm, 60 ppm, and 90 ppm. For each three replicates, 4 chickens were repeated, and the test period was 28 days, and the weight of the chickens was weighed. The results shown in Figure 1 show that the final body weight of the fermented Cordyceps militaris was 96.5%, 100.6%, and 97.8% of the weight of the control group. Therefore, the concentration of Cordyceps sinensis fermented in the drinking water of the chicken was adjusted to 60 ppm, which could promote the chicken. Weight gain.

Regulatory immunization test, the test was divided into three groups: control group, lipopolysaccharide group and lipopolysaccharide plus Cordyceps militaris fermented product (the concentration of cordycepin was 60 ppm). Each group of triple-over each 6-day-old white broiler was tested. After 12 days, after 7 days of acclimatization, the drinking water containing 60ppm of cordycepin was given for three consecutive days (added to the chicken drinking water with the powder of the Cordyceps militaris fermentation product, and the water was added to the water containing 60 ppm of cordycein). Day, lipopolysaccharide group and lipopolysaccharide plus Cordyceps sinensis fermentation The group was intraperitoneally injected with 1 mg of lipopolysaccharide per kilogram of body weight, sacrificed 3 hours after the injection, and the spleen lymphoid tissue and the Fahrenheit sac were taken to detect the inflammatory cytokines (iNOS, COX-2, INF) by real-time quantitative polymerase chain reaction. - γ, IL-4) mRNA expression amount.

From the results of Figures 2-9, the relative performance of inflammatory cytokines iNOS, COX-2, INF-γ, and IL-4 in the spleen increased when the expression level of inflammatory cytokines was 100 in the lipopolysaccharide group. 4.25 times, 4.5 times, 3.66 times, 4.28 times, the relative inflammatory cytokines iNOS, COX-2, INF-γ, IL-4 in the lymphoid tissue of Fahrenheit increased by 6.5 times, 3.6 times, 10 times, 4.7 times. Therefore, the solid fermentation product of Cordyceps militaris can be added to poultry drinking water, and its cordycepin content can be used as an additive index to adjust the immune response of poultry, and the use of antibiotics can become a green feed additive for Chinese herbal medicine.

Claims (3)

A poultry drinking water additive for regulating immunity, characterized in that the cordycepin content is 60 ppm. According to the poultry drinking water additive of claim 1, wherein the regulating immune system refers to the relative expression amount of inflammatory cytokines when the expression amount of the lipopolysaccharide group is 100, and the inflammatory cytokines iNOS, COX-2, INF in the lymphoid tissue of the spleen. The relative performance of -γ and IL-4 increased by 4.25 times, 4.5 times, 3.66 times, and 4.28 times, respectively. According to the poultry drinking water additive of the scope of claim 1, wherein the regulating immunity refers to the relative expression of inflammatory cytokines when the amount of the lipopolysaccharide group is 100, and the inflammatory cytokines iNOS, COX-2, INF- in the Fahrenheit capsule. The relative performance of γ and IL-4 increased by 6.5 times, 3.6 times, 10 times, and 4.7 times, respectively.