CN107260754B - Method for resisting eimeria coccidium by using agaricus blazei murill polysaccharide in livestock and poultry breeding - Google Patents
Method for resisting eimeria coccidium by using agaricus blazei murill polysaccharide in livestock and poultry breeding Download PDFInfo
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Abstract
The invention relates to a method for resisting eimeria coccidium by using agaricus blazei polysaccharide in livestock and poultry breeding, in the method, the agaricus blazei polysaccharide is taken into bodies of livestock and poultry at a dose of 100mg/kg body weight every day in the breeding process of the livestock and poultry so as to resist the infection of the eimeria coccidium, and the specific method for taking the agaricus blazei polysaccharide into the bodies of the livestock and poultry comprises the following steps: the agaricus blazei polysaccharide is stirred in the feed to be fed, and is dissolved in water to be drunk or is manually administrated more accurately. The agaricus blazei murill polysaccharide is taken into bodies of livestock and poultry at a dose of 100mg/kg of body weight every day, so that the livestock and poultry can effectively resist infection of eimeria coccidium, and meanwhile, the agaricus blazei murill polysaccharide plays a good role in protecting growth and development of immune organs of livestock and poultry.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to an application of agaricus blazei murill polysaccharide in resisting parasites in livestock and poultry breeding.
Background
Agaricus Blazei Murrill (ABM) is produced in Brazil, has good taste, can enhance the immunoregulatory function of organism, has antiviral, antibacterial, antiparasitic infection, and antitumor effects, and is edible and medicinal fungi with health promotion function.
The agaricus blazei polysaccharide can effectively stimulate T cells and macrophages to release interleukin-1 β (IL-1 β) and interleukin-6 (IL-6), and further improve specific antibody production of organisms aiming at xenoantigens.
In vitro experiments show that the agaricus blazei murill polysaccharide can induce macrophage of mouse bone marrow to secrete tumor necrosis factor- α (TNF- α), interleukin-8 (IL-8) and Nitric Oxide (NO), and inhibit Vero cytopathy caused by WEEV, so as to show an anti-virus infection effect.
The Agaricus blazei Murill extract also has antibacterial effect on bacteria and fungi. The agaricus blazei murill polysaccharide can stimulate human mononuclear cells to capture candida albicans and produce proinflammatory factors.
The researches show that the hot water extract of the agaricus blazei murill can reduce the mycoderm forming capability of the pseudomonas aeruginosa, reduce the generation of pyocin, reduce the movement capability of thalli and obviously reduce the virulence factor of the pseudomonas aeruginosa.
The agaricus blazei polysaccharide can resist the infection of plasmodium, enhance the oxidation resistance of a host, reduce the occurrence of hematuria, increase the survival rate of the host, reduce the loss of weight, generate protective immune response to cerebral malaria, reduce the secretion of tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β) and interleukin-6 (IL-6) and reduce the damage to the brain, and the extract of the agaricus blazei polysaccharide is used as a Th1 type vaccine adjuvant and can enhance the immune response to visceral Leishmania littoralis DNA vaccine.
Eimeria falciparum (Eimeria lobata) and Eimeria tenella (Eimeria tenella) are both members of the phylum Apicomplexa (Apicompplexia), the class Sporozoasida (Sporozoasida), the subclass Coccidia (Coccidia), the order Eucoccoidioida (Eimeria), the family Eimeriaceae (Eimeria), the genus Eimeria (Eimeria), which infect the caecal mucosal tissues of mice and poultry, respectively. Wherein the cryptoperiod of the Eimeria falcate is 7 days, and the period with obvious symptoms is 10-16 days; the cryptoperiod of Eimeria tenella is 5.5 days, and the period with obvious symptoms is 8-14 days. Eimeria causes great loss to agricultural production, and the prevention of drug-preventive conjugate vaccine is mainly used as a means for preventing and treating coccidia. At present, because the use of anticoccidial chemical drugs easily causes the generation of drug-resistant type insect strains and causes the harm of drug residues after long-term use, the agaricus blazei murrill polysaccharide has good use safety and has important research significance and application value as a candidate anticoccidial drug.
In recent years, more and more studies have proved that the polysaccharide of Agaricus blazei has the effect of resisting endoparasite infection, but the immunoregulatory effect of the polysaccharide of Agaricus blazei on other endoparasites has not been studied in detail, such as the influence of the polysaccharide of Agaricus blazei on the body weight and immune organs of mice infected with Eimeria sickle, the influence of the polysaccharide of Agaricus blazei on the oocyst elimination of chicken infected with Eimeria tenella, and the like are continuously sought by those skilled in the art.
Disclosure of Invention
The invention aims to provide a method for resisting eimeria coccidium by using agaricus blazei polysaccharide in livestock and poultry breeding aiming at the defects of the application technology of the agaricus blazei polysaccharide.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
a method for resisting Eimeria coccidium by using Agaricus blazei Murill polysaccharide in livestock and poultry breeding comprises taking Agaricus blazei Murill polysaccharide into livestock and poultry body at a dosage of 100mg/kg body weight daily to resist infection of Eimeria coccidium.
Further, the specific method for taking the agaricus blazei murrill polysaccharide into the bodies of livestock and poultry comprises the following steps: the agaricus blazei polysaccharide is stirred in the feed to be fed, and is dissolved in water to be drunk or is manually administrated more accurately.
Advantages and effects of the invention
1. The invention determines the optimal infection dose of the Eimeria sickle and the Eimeria tenella to be 500 per unit and 5 multiplied by 103One/only.
2. The invention finds out the good effect of the agaricus blazei murrill polysaccharide on resisting the eimeria infection of domestic animals represented by mice by drenching the agaricus blazei murrill polysaccharide at the dose of 100mg/kg every day.
3. The invention determines the good effect of the agaricus blazei polysaccharide on resisting the eimeria infection on poultry represented by chicks when the agaricus blazei polysaccharide is mixed with the feed and fed at the dose of 100 mg/kg.
4. The invention confirms that the agaricus blazei murill polysaccharide feed with the dosage of 100mg/kg has good effect on the growth and development of animal immune organs in livestock and poultry breeding.
Drawings
FIG. 1 is a graph showing the daily survival rates of mice infected with different doses of Eimeria species: 4 groups of test animals, 10 per group, 1-10 days, were compared for each group at the same time point.
FIG. 2 is a graph showing the results of measurement of body weight of mice in each group: 4 groups of test animals, 0-17 days, 10 animals per group, were compared for each group at the same time point.
FIG. 3 is a graph showing the results of measurement of OGP in feces of mice of each group: 4 groups of 10 animals were tested and compared to each group at the same time point.
FIG. 4 is a graph showing the measurement results of organ indexes of mice in each group: 4 groups of 10 animals were tested, and the groups were compared for differences at the same time point, where P < 0.050.
FIG. 5 is a graph showing the daily survival rates of mice infected with different doses of Eimeria tenella: 4 groups of test animals, 10 per group, 1-10 days, were compared for each group at the same time point.
FIG. 6 is a graph showing the results of the OGP measurement in the feces of chickens of each group: 4 groups of 10 animals were tested and compared to each group at the same time point.
FIG. 7 is a graph showing the results of measurement of immune organ indexes of chickens in each group: 4 groups of 10 animals per group, with differences in each group at the same time point, wherein, P < 0.050; p < 0.010.
Detailed Description
The present invention will be further described with reference to the following drawings, which are illustrative, not restrictive, and the scope of the invention is not limited thereto.
In the first step, in order to find out an application method of agaricus blazei polysaccharide, the invention firstly determines the optimal infection dose of eimeria falcate, and the specific method comprises the following steps:
(1)40 female Kunming mice of 30 days old were adaptively fed for 3 days, randomly divided into 4 groups of 10 mice each, and the mice of group 1, group 2 and group 3 were orally infected with 500/one, 5000/one and 50000/one Eimeria sickle at the dose, and group 4 was drenched with the same amount of physiological saline.
(2) The mice were observed 10 days after infection and the daily clinical signs of mental state, activity, fur luster, appetite excretion, death, etc. were recorded.
The mice of the 1 st group and the 4 th group have no death condition in the test, and the clinical symptoms of the two groups of mice, such as spirit, activity, fur, appetite, excretion and the like, are not abnormal; as shown in fig. 1, mice in group 2 began to die 7 days post infection, for a total of 4 mice by day 10 of the experiment; mice in group 3 began to die at day 5 post-infection and all mice died by day 10 of the experiment, indicating that mice infected with 500/E.falciparum did not result in death, whereas E.falciparum lethal dose was 50000/E.falciparum. Thus, the test group of mice was infected with Eimeria sickle at a dose of 500/mouse.
Secondly, determining the effect of the agaricus blazei murill polysaccharide on the growth and development of the mice and the resistance to the infection of the Eimeria sickle, wherein the specific determination process is as follows:
(1) mice were acclimatized for 2 days and randomized into 4 groups of 10 mice each. The group 1 was drenched with 100mg/kg of agaricus blazei murill polysaccharide solution, the group 2 was drenched with 40mg/kg of agaricus blazei murill polysaccharide solution for 7 days continuously, and the groups 3 and 4 were drenched with the same amount of physiological saline, and the weight change of each group of mice was recorded. As shown in FIG. 2, the body weight of each group of mice showed a gradual increase from the 1 st day of the test, and the average body weight of the group 1 mice was higher than that of the groups 2, 3 and 4, indicating that 100mg/kg of the polysaccharide was superior to the effect of increasing the weight of the mice of 40mg/kg of the polysaccharide and the mice that did not drink the polysaccharide.
(2) On the 8 th day of the experiment, the 1 st, 2 nd and 3 th groups were each administered with 500/one, 0.2 mL/one, and the 4 th group was administered with the same amount of physiological saline, and the mice in each group were observed daily for clinical symptoms after infection with Eimeria falciparum, and the change in body weight of the mice in each group was recorded. As shown in FIG. 2, the weight average of the body weight of each group of mice increased slowly after the infection with Eimeria falciparum, i.e., on days 8-14 of the test, and the weight average of the body weight of each group of mice decreased significantly on day 15 of the test, wherein the weight decrease of group 1 was lower than that of group 2 and group 3, indicating that 100mg/kg of the polysaccharide group of Agaricus blazei Murill was superior to 40mg/kg of the polysaccharide group of Agaricus blazei Murill and the polysaccharide group of Agaricus blazei Murill.
(3) On the 14 th day of the experiment, padding of each group of mice is removed, feces of 15 th to 17 th days are collected, each group of feces is uniformly stirred, 2g of feces is weighed into a plastic small beaker, 60mL of saturated saline is added, the mixture is uniformly stirred, the mixture is filled into a counting chamber of a modified Macama plate, the counting chamber is arranged under a microscope after the mixture is kept still for 3min, and the number of oocysts in each gram of feces is calculated (reading obtained by counting is multiplied by 200). As shown in FIG. 3, the OPG groups are 300 for group 1 and 6X 10 for group 24 Group 3 is 6X 105And group 4 is 0, indicating that 100mg/kg agaricus blazei polysaccharide group has better effect on mice against eimeria sickle infection than 40mg/kg agaricus blazei polysaccharide group and agaricus blazei polysaccharide group without drinking.
(4) At day 17 after the test, the organ lesions of the mice in each group were observed, and organ indexes (weight of each organ to body weight) of the liver, kidney and spleen were calculated. After the 17 th day of the test, the shape, size and color of the parenchymal organ of each group of mice are normal, and no pathological changes are found. As shown in FIG. 4, the difference between the liver index and the kidney index of each group of mice is not significant, and the difference between the spleen index group 1 and the spleen index group 3 is significant, which indicates that 100mg/kg of agaricus blazei murill polysaccharide can promote the growth and development of the spleen of the mice and enhance the resistance of the mice to the infection of the Eimeria sickle.
Thirdly, in order to find out an application method of the agaricus blazei polysaccharide, the optimal infection dose of the eimeria tenella needs to be determined, and the specific method comprises the following steps:
(1) feeding 40 AA broilers of 4 days age for 3 days, randomly dividing into 4 groups of 10 broilers, and respectively feeding chicks of 1 st group, 2 nd group and 3 rd group at a ratio of 5 × 103One/one, 5 x 104One/one, 5 x 105The dose of Eimeria tenella/E.tenella was infected orally and group 4 was drenched with an equal amount of physiological saline.
(2) The chicks were observed 10 days after infection and recorded daily clinical signs of mental status, activity, gloss of coat, appetite excretion, death, etc. for each group of chicks.
The chicks in the groups 1 and 4 have no death condition in the test, and the clinical symptoms of the chicks in the two groups, such as spirit, activity, feather, appetite, excretion and the like, are not abnormal; as shown in fig. 5, group 2 chicks began to die on day 9 post infection, for a total of 2 chicks died by day 10 of the experiment; the chicks in group 3 began to die at day 6 after infection, and all chicks died by day 10 of the experiment, indicating that the dose of infection was 5X 103Eimeria tenella/Eimeria tenella does not cause death of chicks, and the lethal dose of Eimeria tenella is 5X 105One/only. Thus, the test group chicks were tested at 5X 103The dose/body was infected with Eimeria tenella.
Fourthly, determining the effect of the agaricus blazei polysaccharide on the growth and development of the chicks and the resistance to the infection of the Eimeria tenella, wherein the specific determination process is as follows:
(1)40 AA broilers aged 4 days were adaptively fed for 3 days, and were randomly divided into 4 groups of 10 broilers. Group 1 was prepared by blending Agaricus blazei Murill polysaccharide at a dose of 100mg/kg body weight, group 2 was prepared by blending Agaricus blazei Murill polysaccharide at a dose of 40mg/kg body weight, and the feed was continuously fed for 7 days, while the groups 3 and 4 were fed normally. On test day 8, Eimeria tenella sporulated oocysts 5X 10 were administered to groups 1, 2, and 3, respectively3 Group 4 was administered with an equal amount of physiological saline, and the clinical symptoms of each group of chicks after infection with Eimeria tenella were observed daily. Removing the feces of each group of chicks on the 13 th day of the experiment, collecting the feces of the 14 th to 17 th days, respectively and uniformly stirring the feces of each group, then respectively weighing 2g of the feces into a small plastic beaker, adding 60mL of saturated saline, uniformly stirring, filling into a counting chamber of a modified Macmajen plate, standing for 3min, counting under a microscope, and calculating the number of oocysts in each gram of feces (reading obtained by counting is multiplied by 200). As shown in FIG. 6, among the OPGs, group 1 is 800, and group 2 is 4.8X 104 Group 3 is 2.56X 105 Group 4 is 0, indicating that a 100mg/kg blend of agaricus blazei polysaccharide provides better resistance to eimeria tenella infection in chicks than 40mg/kg blend of agaricus blazei polysaccharide and no agaricus blazei polysaccharide.
(4) After the test day 17, the chickens in each group were examined for organ lesions, and organ indices (weight of each organ to body weight) of the thymus, spleen and bursa of Fabricius were calculated. After the 17 th day of the test, the chicks in each group had normal shapes, sizes and colors of their parenchymal organs and no pathological changes were observed. As shown in fig. 7, in the thymus index, the difference between 3 groups and 4 groups is very significant (P <0.01), indicating that eimeria tenella infection affects the development of thymus, thereby reducing the immunity of the organism; the difference of the group 1 is significant compared with the group 2 (P <0.05), the difference of the group 1 is significant compared with the group 3 (P <0.05), the difference of the group 2 is significant compared with the group 4 (P <0.01), the difference of the group 2 is insignificant compared with the group 3 (P >0.05), and the difference of the group 1 is insignificant compared with the group 4 (P >0.05), which indicates that the growth and development of thymus can be promoted by feeding the agaricus blazei murill polysaccharide 100mg/kg to the agaricus blazei murill polysaccharide 40mg/kg, and the infection of the eimeria tenella can be effectively resisted.
As shown in fig. 7, in the spleen index, 3 groups were significantly different from 4 groups (P <0.05), indicating that eimeria tenella infection affects the development of spleen, thereby reducing the immunity of the organism; the difference of the group 1 and the group 3 is significant (P <0.05), the difference of the group 2 and the group 3 is very significant (P <0.01), the difference of the group 2 and the group 4 is not significant (P >0.05), the difference of the group 1 and the group 4 is not significant (P >0.05), and the difference of the group 1 and the group 2 is not significant (P >0.05), which indicates that the agaricus blazei murill polysaccharide can promote the development of the spleen, and further effectively resist the infection of the eimeria tenella, and the difference of the development of the spleen is not significant when the agaricus blazei murill polysaccharide dressing of 100mg/kg and 40mg/kg is fed.
As shown in fig. 7, in the bursal index, the difference between 3 groups and 4 groups is very significant (P <0.01), indicating that the infection of eimeria tenella affects the development of bursal disease, thereby reducing the immunity of the organism; the difference of the group 1 is obvious compared with the group 3 (P <0.05), the difference of the group 2 is obvious compared with the group 4 (P <0.05), and the difference of the group 1 is not obvious compared with the group 4 (P >0.05), which shows that 100mg/kg of agaricus blazei murrill polysaccharide can more obviously promote the development of bursal disease than 40mg/kg of agaricus blazei murrill polysaccharide mixed material to feed, thereby effectively resisting the infection of Eimeria tenella.
Claims (2)
1. The application of agaricus blazei polysaccharide as a sole active ingredient in preparing a preparation for preventing eimeria coccidium in chicken breeding is characterized in that: the preparation is stirred in feed for administration, and Agaricus blazei Murill polysaccharide is dissolved in water for oral administration or administered by artificial irrigation.
2. The use of agaricus blazei murill polysaccharide as the sole active ingredient in the preparation of an anti-eimeria agent for chicken breeding according to claim 1, wherein: the preparation is administered by mixing with feed at a dose of 100mg/kg body weight every day, dissolving Agaricus blazei Murill polysaccharide in water, and administering by artificial irrigation.
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Application publication date: 20171020 Assignee: Tianjin kaisibo Biotechnology Co.,Ltd. Assignor: TIANJIN AGRICULTURAL University Contract record no.: X2023120000029 Denomination of invention: A Method for Using Agaricus blazei Polysaccharides to Resist Eimeria in Livestock and Poultry Breeding Granted publication date: 20200214 License type: Common License Record date: 20230713 |
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