Application of Bombella intestini in improving 10-HDA content of royal jelly

Abstract

The invention belongs to the technical field of bee breeding, and particularly relates to application of Bombella intestini in improving the 10-HDA content of royal jelly. According to the invention, it is found that the ability of worker bees to synthesize 10-HDA can be significantly improved by exogenously supplementing Bombella intestini with a specific concentration. Meanwhile, the method is safe, efficient and sustainable, does not influence bee colony health and bee product safety, can be used in combination with a traditional nutrition regulation and control method, meets the requirement of the market for high-quality royal jelly, and has broad market prospects. And a feasible technical support and a solution are provided for realizing healthy breeding of bees, standardized production of royal jelly and development of green organic bee products.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of bee breeding, and particularly relates to a honeybee intestinal bacterium Bombella intestini In the application of improving the 10-HDA content in royal jelly. BACKGROUND

[0002] Royal jelly is a kind of milk-like substance secreted by the pharyngeal gland and maxillary gland of worker bees, which is not only the exclusive food of queen bees but also the early nutritional source of bee larvae. It has important nutritional and health-care effects on human body, and has multiple biological activities such as antibacterial, antioxidant and immune regulation. The functions of royal jelly are closely related to the content of its main active ingredient, 10-hydroxy-2-decenoic acid (10-HDA). The content of 10-HDA is not only a key index for judging the quality and function of royal jelly, but also an important factor determining the commodity value of royal jelly. Therefore, improving the content of 10-HDA in royal jelly has become a core technical problem for improving the product quality, added value and market competitiveness of royal jelly.

[0003] The annual output of royal jelly in China is about 3000 to 4000 tons, accounting for more than 90% of the total global output, and the export volume is between 600 to 800 tons, which makes China the main supplier of royal jelly on the international market. The content of 10-HDA is the core standard for classifying the grade of royal jelly and conducting export inspection. According to the national standard, the 10-HDA content of qualified royal jelly products should not be less than 1.4%, and the superior product should reach more than 1.8%; the standard for exporting royal jelly is more stringent, requiring the 10-HDA content to be not less than 1.8%, especially for the European Union, the United States, most countries in Southeast Asia, and Japan, which have the highest requirements for the quality of exported royal jelly from China, requiring the 10-HDA content to be more than 2.0%. At present, among the royal jelly produced in China, 60% to 70% of the products have a 10-HDA content between 1.4% and 1.8%, only 30% to 40% of the products have a 10-HDA content exceeding 1.8%, and less than 5% of the products have a 10-HDA content exceeding 2.0%. Obviously, the content of 10-HDA has become a key factor restricting the quality and export volume of royal jelly.

[0004] Although the current methods for improving the 10-HDA content of royal jelly mainly focus on bee breeding, nutritional regulation and production period management, these methods generally have problems such as long cycle, complex operation, insignificant and unstable effect. In beekeeping production, there is still a lack of an efficient, safe, simple and widely applicable method to significantly improve the 10-HDA content in royal jelly. SUMMARY

[0005] In view of the above problems, the purpose of the present application is to provide a honeybee intestinal bacterium Bombella intestini The application of the honeybee intestinal bacterium in improving the 10-HDA content in royal jelly solves the problems of the prior art.

[0006] The object of the present application can be achieved by the following technical solutions: In a first aspect of the present application, a honeybee intestinal bacterium is provided. Bombella intestini In the application of improving the content of 10-HDA in royal jelly.

[0007] In a second aspect of the present application, a method for improving the content of 10-HDA in royal jelly is provided, characterized by comprising the following steps: S1, feeding the honeybee intestinal bacterium Bombella intestini , a saccharide compound and water to obtain a feeding liquid; S2, collecting royal jelly after feeding the feeding liquid obtained in step S1 to a bee colony; The concentration of the honeybee intestinal bacterium Bombella intestini in the feeding liquid is 1×10 5 ~1×10 7 cfu / mL; and the concentration of the saccharide compound is 35~45wt%.

[0008] The above technical solution has the following beneficial effects compared with the prior art: (1) The present application discovers that exogenous supplementation of a specific concentration of Bombella intestini The content of 10-HAD in royal jelly can reach an average of 2.427±0.0959%, which can significantly improve the ability of worker bees to synthesize 10-HDA.

[0009] (2) The Bombella intestini strain in the present application is a natural symbiotic bacterium in the intestinal flora of honeybees, and its abundance in the queen bee intestine is higher than that in the worker bee. It has good colonization ability and stable metabolic characteristics in the body of honeybees. The strain is non-toxic and harmless to individual honeybees and bee colonies, and has no pollution and no residue to bee products and the environment in the application process, and has good biological safety. It has good effect and stability in improving the quality of royal jelly, especially in improving the content of 10-HDA. The strain is simple to operate and has strong adaptability, without the need for special equipment and operation technology, and is convenient for popularization and application in bee farms. At the same time, the strain is easy to culture and has stable metabolism under laboratory conditions, and has good basis for industrial production.

[0010] (3) The method of the present application is simple to operate, has strong feasibility, is safe and non-toxic, and has significant effect, without affecting the health of the bee colony and the safety of the bee product. It can be used in combination with traditional nutritional regulation methods to improve the content of 10-HDA in royal jelly and significantly improve the quality and commodity value of royal jelly, and provides practical technical support and solutions for realizing healthy breeding of honeybees, standardized production of royal jelly and development of green and organic bee products. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1A diagram showing the statistical results of 10-HAD content in royal jelly of the feeding probiotic group and the control group in the embodiments of the present application. DETAILED DESCRIPTION

[0012] The present application will be further described in conjunction with the specific embodiments. The embodiments given herein are only to illustrate the present application, and are not intended to limit the scope of the present application. The embodiments provided below can serve as a guide for further improvement by those of ordinary skill in the art, and do not in any way constitute a limitation on the present application.

[0013] In the following examples, the experimental methods are conventional methods, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained from commercial channels, unless otherwise specified.

[0014] As described above, the methods for increasing the content of 10-HDA in royal jelly in the prior art are mostly focused on bee breeding, nutrition regulation, and management during the royal jelly production period, etc. However, these methods generally have problems such as long cycle, complex operation, insignificant effect, and instability. In view of this, the present application provides a method for increasing the content of 10-HDA in royal jelly by feeding bees with a sugar solution containing a natural symbiotic bacteria in the intestines of honeybees. Bombella intestini application in increasing the content of 10-HDA in royal jelly.

[0015] In one specific embodiment of the present application, a method for increasing the content of 10-HDA in royal jelly by feeding bees with a sugar solution containing a natural symbiotic bacteria in the intestines of honeybees is provided. Bombella intestini application in increasing the content of 10-HDA in royal jelly.

[0016] a natural symbiotic bacteria in the intestines of honeybees Bombella intestini is a natural symbiotic bacteria in the intestines of honeybees, belonging to Acetobacteraceae, and has a significantly higher relative abundance in the intestines of queen bees than in the intestines of worker bees. The symbiotic bacteria has a significant effect on regulating intestinal health, and can improve the balance of intestinal flora and optimize the intestinal mucosa structure, thereby improving the digestion and absorption and metabolic efficiency of nutritional substances.

[0017] In some embodiments of the present application, the application is feeding a bee colony with a sugar solution containing a natural symbiotic bacteria in the intestines of honeybees. Bombella intestini In some embodiments of the present application, the natural symbiotic bacteria in the intestines of honeybees

[0018] In some embodiments of the present application, the natural symbiotic bacteria in the intestines of honeybees Bombella intestini is a live bacteria, and its activity must be ensured during feeding, and it is prepared on demand.

[0019] In some embodiments of the present application, the feeding time is 10-20 days before the royal jelly production, and the feeding is continued until the end of the royal jelly production, so as to ensure that the bee colony has the most basic nutrition for the royal jelly production during the royal jelly production period.

[0020] In some embodiments of the present application, the concentration of the natural symbiotic bacteria in the intestines of honeybees in the sugar solution is 1x10 5~1×10 7 cfu / mL.

[0021] In some embodiments of the present invention, the sugar includes sucrose, white sugar, fructose or beet sugar, preferably sucrose.

[0022] In some embodiments of the present invention, the concentration of the sugar solution is 35-45 wt%, more preferably 40 wt%.

[0023] In some embodiments of the present invention, when the bee colony is a single hive, the feeding amount is 250-350 mL / time, and the feeding frequency is once every 2 days.

[0024] In some embodiments of the present invention, when the bee colony is a single hive, the feeding amount is 300 mL / time, the feeding frequency is once every 2 days, and the feeding location is to add 300 mL in full to the feed box of the hive box at one time.

[0025] In some embodiments of the present invention, when the bee colony is a double-hive, the feeding amount is 450-550 mL / time, and the feeding frequency is once every 2 days.

[0026] In some embodiments of the present invention, when the bee colony is a double-box structure, the feeding amount is 500 mL / time, the feeding frequency is once every 2 days, and the feeding location is 300 mL in the feeding box of the brood box and 200 mL in the feeding box of the super box.

[0027] In some embodiments of the present invention, the bee colony species may be Italian honeybees or Italian-Caniolar hybrid honeybees.

[0028] In some embodiments of the present invention, the bee colony is an Italian honeybee.

[0029] In another specific embodiment of the present invention, a method for increasing the 10-HDA content in royal jelly is provided, comprising the following steps: S1, Including bee gut bacteria Bombella intestini Sugar compounds and water are mixed to obtain a feeding solution; S2. After feeding the bee colony with the feeding solution obtained in step S1, collect the royal jelly. The feeding solution contains bee gut bacteria Bombella intestini The concentration is 1×10 5 ~1×10 7 cfu / mL; the concentration of the carbohydrate compound is 35~45wt%.

[0030] In some embodiments of the present invention, the feeding solution contains bee gut bacteria. Bombella intestini The concentration is 1×10 6cfu / mL; the concentration of the carbohydrate compound is 40 wt%.

[0031] In some embodiments of the present invention, the feeding time is from 10 to 20 days before the production of plasma to the end of the production period, ensuring that the bee colony has the most basic nutritional supply for plasma production.

[0032] In some embodiments of the present invention, the feeding cycle is 2 months.

[0033] In this invention, the feeding time can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 days before plasma production, and feeding can continue until plasma production results are achieved. During the plasma production period, the bee colony is guaranteed to have the most basic nutritional supply for plasma production.

[0034] In some embodiments of the present invention, the method is applicable to multiple royal jelly producing areas with different primary nectar sources, and its effectiveness is not limited by a single nectar source or geographical location.

[0035] In some embodiments of the present invention, the implementation areas include spring nectar source areas, such as the rapeseed and citrus flowering season in the Yangtze River Basin and areas to the south, and the locust flowering season in North China.

[0036] In some embodiments of the present invention, the implementation area includes summer flowering areas, such as the flowering period of Vitex negundo and Jujube in North China and Northeast China, and the flowering period of Linden in Northeast China.

[0037] In some embodiments of the present invention, the implementation area includes areas with summer and autumn flowering seasons, such as the rapeseed and sunflower flowering seasons in the Northwest Plateau, and the buckwheat and perilla flowering seasons in the Northeast region.

[0038] It should be further noted that the method described in this invention can be applied as long as there is sufficient external nectar and pollen to meet the needs of bee colony reproduction and production. During periods of poor nectar supply, the bee colony's ability to nurse bees can be maintained by supplementing their diet with high-quality pollen; however, complete reliance on white sugar for royal jelly production should be avoided.

[0039] As an example, in one embodiment of the present invention, the bee colony is raised in Tai'an, Shandong Province, and the feeding period is from May to June of the same year. The main nectar sources for the bee colony are locust, vitex, and jujube flowers.

[0040] In this invention, the bee gut bacteria Bombella intestiniIt is a gut symbiotic bacterium native to the honeybee's intestines, strain number LMG28161. Purchased from the Belgian National Culture Collection (BCCM / LMG Bacteria Collection, Faculty of Sciences, Ghent University, KL Ledeganckstraat 35, B-9000Ghent, Belgium). Growth conditions: 28℃, microaerophilic.

[0041] In this invention, the experimental bee colony is the Italian honeybee. Apis mellifera The samples originated from the bee farm of the Animal Husbandry Science and Technology Station of Shandong Agricultural University, located in Tai'an City, Shandong Province (longitude: 117.08°E; latitude: 36.20°N).

[0042] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0043] Example Taking a bee farm in Tai'an, Shandong Province as an example, this technology was used in May and June. The main external nectar and pollen sources were locust, vitex, and jujube blossoms, ensuring abundant nectar. Ten healthy Italian honeybee colonies of similar size (double-hive, 14 frames) and queen bee age were selected and randomly divided into two groups of five colonies each. The control group was fed a 40% sucrose solution, starting 15 days before royal jelly production, every two days, 500 mL each time. The feeding locations were: 300 mL in the feeding box of the brood box and 200 mL in the feed box of the super. The experimental group was fed a solution prepared using 40 wt% sucrose as a solvent. 6 cfu / mL Bombella intestini Feed the bees every two days, with 500 mL of bacterial solution prepared fresh before each feeding to ensure bacterial activity and stability. Feeding is done manually by injecting the solution into a feeder located above the hive. The entire experiment lasted two months, until plasma production ended. Other interfering factors were strictly controlled during the experiment, and consistent feeding and management conditions were maintained before and after the experiment.

[0044] The 10-HDA content in royal jelly was determined using the method specified in the national standard GB9697-2008 "Royal Jelly". The results were analyzed using one-way ANOVA with SPSS software, and the significance level was set at p<0.05. Statistical results are as follows: Figure 1 As shown, the average 10-HDA content in royal jelly of the sugar water control group was 1.986±0.0651%, while the average 10-HDA content in royal jelly of the probiotic group was 2.427±0.0959%. Compared with the control group, the 10-HDA content in the royal jelly of the bee colony fed with sugar water was significantly higher. Bombella intestiniIt can increase the 10-HDA content in royal jelly by 0.441%.

[0045] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of them. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Enterococcus sp. of Apis mellifera Bombella intestini In the use of increasing the content of 10-HDA in royal jelly.

2. Use according to claim 1, characterized in that, The application is feeding a bee colony with a sugar solution containing intestinal bacteria of honey bees Bombella intestini of honey bees Preferably, the bee gut bacteria Bombella intestini are live bacteria.

3. Use according to claim 2, characterized in that, The concentration of the honeybee gut bacteria in the sugar solution is 1 x 10 5 ~1 x 10 7 cfu / mL.

4. The use according to claim 2, wherein the compound is ###0002### The sugar includes sucrose, white sugar, fructose or beet sugar, preferably sucrose; Preferably, the concentration of the sugar solution is 35-45 wt%, further preferably 40 wt%.

5. Use according to claim 2, characterized in that, The feeding is specifically: when the bee colony is single box, the feeding amount is 250-350 mL / time, preferably 300 mL / time, and the feeding frequency is once every 2 days; when the bee colony is double box, the feeding amount is 450-550 mL / time, preferably 500 mL / time, and the feeding frequency is once every 2 days.

6. Use according to claim 5, characterized in that, When the bee colony is single box, the feeding amount is 300 mL / time, and the feeding position is to add 300 mL in the nest box feeding box once; When the bee colony is double box, the feeding amount is 500 mL / time, and the feeding position is to add 300 mL in the nest box feeding box and 200 mL in the nest box feeding box.

7. Use according to any one of claims 1 to 6, wherein The bee colony is an Apis mellifera bee colony.

8. A method of increasing the content of 10-HDA in royal jelly, characterized by, The method comprises the following steps: S1, mixing the honeybee intestinal bacteria Bombella intestini , the saccharide compound and water to obtain a feeding liquid; S2, after feeding the bee colony with the feeding solution obtained in step S1, collecting royal jelly; The feeding solution contains bee gut bacteria Bombella intestini The concentration is 1×10 5 ~1×10 7 cfu / mL; the concentration of the carbohydrate compound is 35~45wt%.

9. The method of claim 8, wherein, The feeding time is to start feeding 10-20 days before jelly production and end when jelly production is completed.

10. The method of claim 8, wherein, The feeding cycle is 2 months.

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