A method for regulating the food-derived targeted proliferation of intestinal microorganisms in red-crowned cranes

Abstract

A method for regulating the targeted proliferation of food sources for the intestinal microorganisms of red-crowned cranes belongs to the field of wetland ecological engineering technology, and solves the restrictive problems of low efficiency of the coordinated cultivation of food source biological resources for red-crowned cranes and the lack of pathways for regulating the intestinal microorganisms of red-crowned cranes in a near-natural way. The method includes: 1. Improvement of the substrate environment and micro-geomorphological construction; 2. Zoning replenishment of water resources and addition of microbial agents; 3. Introduction and combination of food source organisms; 4. Sectional regulation of hydrological conditions; 5. Comparative evaluation of intestinal microorganisms of red-crowned cranes. The present invention is suitable for the targeted proliferation of food source organisms with the goal of regulating the intestinal microorganisms of red-crowned cranes.

Description

Technical Field

[0001] The invention belongs to the technical field of wetland ecological engineering, and specifically relates to a food source targeted proliferation technology. Background Art

[0002] The red-crowned crane (Grus japonensis) is a large wading bird of the genus Grus in the family Gruidae. It is widely distributed in Northeast China, eastern Mongolia, Russia, North Korea and South Korea. It is one of the most sensitive indicator organisms to changes in wetland environments.

[0003] The Red List of Vertebrates of China (2020 Edition) lists it as an endangered species, and the red-crowned crane is a first-class protected wild animal in my country. However, due to the combined influence of factors such as human interference, the problem of food source shortage for red-crowned cranes has become increasingly prominent, and the resulting change in diet has seriously interfered with the health and ecological security of the red-crowned crane population. Therefore, achieving targeted proliferation of food sources with the goal of maintaining the health of the red-crowned crane population has become a key scientific issue that needs to be urgently addressed in the fields of wetland ecology and rare bird protection.

[0004] Intestinal microorganisms are "acquired organs" that indicate the health quality of living organisms. Influenced by the life history activities of animals, they are widely involved in many physiological activities of the host, such as signal transduction, immune response, and nutrient metabolism. At present, studies on the threat of potential pathogenic bacteria in the intestines of red-crowned cranes to the life and health of red-crowned cranes have been reported many times. Regulating the intestinal microorganisms of red-crowned cranes has become a necessary measure to maintain the health of the red-crowned crane population.

[0005] At present, the methods for regulating the intestinal microbial flora of cranes are mainly taking probiotic preparations and feeding specific foods. The principle of taking probiotic preparations is clear, and the intestinal flora is improved by directly ingesting live Bacillus licheniformis and triple live bifidobacterium, but the application process is inconvenient and cannot be applied to large-scale populations; feeding specific foods can take into account both the food supply of red-crowned cranes and the optimization of intestinal microbial components, but such methods are costly and unsustainable. Therefore, how to achieve optimal regulation of intestinal microorganisms while taking into account the sustainable supply of food sources for red-crowned cranes has become a practical problem that needs to be urgently solved in the field of wetland biodiversity conservation. Summary of the invention

[0006] The present invention proposes a method for regulating the targeted proliferation of food-source microorganisms in the intestines of red-crowned cranes, which aims to solve the limiting problems of low efficiency of co-cultivation of food-source biological resources for red-crowned cranes and lack of pathways for regulating the intestinal microorganisms of red-crowned cranes in a near-natural way.

[0007] A method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes, comprising:

[0008] S1: Basement environment improvement and micro-geomorphic structure, including:

[0009] In late April to early May of the year of regulation, target plots are selected in the historical breeding habitat of red-crowned cranes to build food source targeted proliferation areas; the surface soil of the food source targeted proliferation areas is stripped, and earth ridges are piled around the food source targeted proliferation areas; microbial fertilizer is applied to the surface of the treated soil in the food source targeted proliferation areas and the soil is turned over, and circular islands are randomly constructed on the food source targeted proliferation areas, with a construction density of 2-3 places / 100m2, and discontinuous earth ridges are constructed in the unconstructed parts of the food source targeted proliferation areas, with a spacing of 30-45 cm between the earth ridges, an end distance of 30-40 cm, a height of 5-10 cm, and a length of 50-100 cm;

[0010] S2: Water resource zoning replenishment and microbial agent addition, including:

[0011] In early to late May of the year of regulation, water from nearby natural wetlands is used to replenish water in the targeted food source proliferation area; the substrate of the circular island is artificially sprinkled until the soil moisture content of the circular island is 80%-90%; the unconstructed part of the targeted food source proliferation area is replenished with water to a flooding depth of 8-10 cm, and microbial agents are applied;

[0012] S3: Foodborne organism introduction kit, including:

[0013] Selecting food-source plants and food-source organisms, the food-source plants include: Tripterygium wilfordii and Echinochloa chinensis; the food-source organisms include: food-source shrimps, food-source snails, and food-source fish; planting Echinochloa chinensis corms and rooted young plants of Tripterygium wilfordii in the non-slope area of ​​the circular island and the top of the soil ridge, wherein the planting density is 3-5 stems / square meter and the planting interval is 10-15 cm; after planting, mixing the food-source organisms in proportion and releasing them;

[0014] S4: Hydrological conditions are divided into different areas for segmented regulation, including:

[0015] From mid-July to mid-October of the regulation year, the moisture content of the non-flooded soil of the circular island is maintained at 80%-90%; from mid-July to mid-August of the regulation year, the surface flooding depth of the soil ridge area is maintained at 8-10 cm; from mid-August to mid-October of the regulation year, the surface flooding depth of the soil ridge area is maintained at 15-20 cm; from mid-October to mid-November of the regulation year, the surface flooding depth of the soil ridge area is maintained at 25-30 cm; in winter, after the red-crowned crane population migrates and there are no red-crowned cranes feeding, water is replenished to the soil ridge area to make its surface flooding depth reach 40-45 cm.

[0016] Furthermore, a preferred solution is provided: the requirements for the historical breeding habitat of the red-crowned crane are: the rest period of the wild red-crowned crane population is greater than 5 years, and the single population habitat size is greater than 500.

[0017] Furthermore, a preferred solution is provided: the area of ​​the target plot is 1000-2500 square meters.

[0018] Furthermore, a preferred solution is provided: in S1, the microbial fertilizer is: a microbial fertilizer obtained by tissue culture of Gram-positive bacteria, Bacillus coagulans, and Bacillus megaterium; and the application amount of the microbial fertilizer is 0.06-0.09 kg / m2.

[0019] Furthermore, a preferred solution is provided: the height of the circular island is 30-40 cm, the width of the slope is 40-50 cm, the angle of the slope is 8°-15°, and the base area is 20-25 square meters.

[0020] Furthermore, a preferred solution is provided: in S2, the ammonia nitrogen content of the water body adjacent to the natural wetland is lower than 1.25 mg / L, the total nitrogen content is lower than 10 mg / L, and the total phosphorus content is lower than 0.25 mg / L.

[0021] Furthermore, a preferred solution is provided: in S2, the components of the microbial agent include: Rhodopseudomonas palustris, Bacillus clausii, and Bacillus licheniformis; and the application concentration of the microbial agent is 0.10-0.12 g / L.

[0022] Furthermore, a preferred solution is provided: the food-source shrimp organisms include: Macrobrachium nipponense and Macrobrachium rosenbergii; the food-source snail organisms include: Hemispheric multi-veined flat snail, flat disc snail, and bright septate flat snail; the food-source fish organisms include: grass carp, silver carp, silver carp, and Lingyuan minnow.

[0023] Furthermore, a preferred scheme is provided: in said S3, the food-source organisms are mixed and released in proportion, specifically: food-source shrimp organisms are mixed and released at a density of 500-1000 / m2 and a gender ratio of 30-35% male shrimps and 65-70% female shrimps; food-source snail organisms are mixed and released at a density of 200-450 / m2 and a gender ratio of 30-35% male snails and 65-70% female snails; food-source fish organisms are mixed and released at a density of 50-100 / m2 and a gender ratio of 40-50% male fish and 50-60% female fish.

[0024] Furthermore, a preferred scheme is provided: the method also includes: S5, comparative evaluation of the intestinal microorganisms of red-crowned cranes: after the red-crowned crane population has continuously fed in the targeted proliferation area of ​​the food source for 7-10 days, the excretion behavior of the red-crowned cranes is observed at a distance, and fresh fecal samples are collected immediately after the red-crowned cranes finish feeding and 16SrDNA high-throughput sequencing is performed, with Firmicutes as the dominant phylum and Catelliococcus as the dominant genus as the criteria for regulating the intestinal microbial components and determining the health of the red-crowned cranes.

[0025] Compared with the prior art, the advantages of the present invention are:

[0026] 1. The method of the present invention has clear principles, is easy to implement and has low cost.

[0027] The method described in the present invention involves multiple processes and factors such as micro-topography construction, plant zoning configuration, biological quantitative placement, hydrological zoning boundary management, and microbial component regulation, and can follow the principle of niche differentiation to achieve the benign and rapid targeted proliferation of red-crowned crane food source biological resources. In addition, the method described in the present invention can provide food sources for red-crowned cranes and attract red-crowned crane populations to forage in clusters in the same year of implementation, and can effectively improve the intestinal microbial component structure of red-crowned cranes.

[0028] The method described in the present invention is suitable for promotion and application in the development of the food source replenishment project for red-crowned cranes and the biological population health maintenance industry.

[0029] 2. The method described in the present invention can effectively regulate the intestinal microbial flora structure of the red-crowned crane population.

[0030] The method described in the present invention involves the linkage structure replenishment of the soil, hydrology, food plants, and food animals in the food source area of ​​the red-crowned crane, and integrates the method of adding microbial fertilizers and microbial agents to the basic environment for the growth and development of food source organisms. It can achieve the near-natural existence and efficient transmission of targeted microbial flora in the hydrology-soil-food plants-food animals ecological cascade process, thereby regulating the intestinal microbial flora structure of the red-crowned crane in the form of food source provision and optimizing its individual health status.

[0031] In addition, the bacterial fertilizer mainly composed of Metamycetes used in the present invention can promote the decomposition of soil organic matter and nutrient circulation through heterotrophic metabolism and autotrophic metabolism pathways, thereby realizing the carrying of targeted microbial flora of food-source organisms and the coordinated transfer and enrichment of nutrients, thereby effectively regulating the intestinal microbial flora structure of red-crowned cranes.

[0032] 3. The method described in the present invention can effectively enhance the linkage characteristics of targeted proliferation of food sources and improvement of intestinal microorganisms in red-crowned cranes.

[0033] The method described in the present invention combines the construction of a suitable habitat for red-crowned crane food source organisms with a method for efficient population expansion and propagation, and the targeted application of microbial agents can effectively improve the soil microbial component structure and promote the enrichment of beneficial microbial flora in food source organisms.

[0034] In addition, the content of the present invention covers food source habitat improvement measures such as microhabitat creation, hydrological zoning and hierarchical regulation, which can effectively enhance the level of self-fertilization and mutualism of food source organisms, the degree of nutrient absorption and improve the probiotic characteristics of food source organisms, thereby directly promoting the proliferation of beneficial flora in the intestines of red-crowned cranes and the formation of a benign flora structure, thereby achieving the linkage improvement of targeted proliferation of food sources for red-crowned cranes and improvement of intestinal microorganisms.

[0035] 4. The method described in the present invention can significantly improve the coordinated targeted proliferation of food-borne organisms.

[0036] The method described in the present invention is based on ecological, botanical and biological principles. On the premise of clarifying the suitable hydrological habitat requirements of different food plants and the spatial gaps of the optimal populations of different food animals, the self-reproduction niche space of food organisms is constructed in multiple ways from time and space dimensions. In this way, the rapid increase in production and sustainable breeding of food materials for red-crowned cranes can be promoted through near-natural regulation of plant material production regulation strategies and animal population polymorphism amplification methods, thereby effectively achieving the targeted proliferation of food organisms.

[0037] 5. The method described in the present invention follows the concept of near-natural restoration and does not involve any potential safety hazards or ecological safety issues.

[0038] The food-borne organisms selected in the present invention are all the main food sources of red-crowned cranes and are widely distributed in natural wetland environments. The microbial agent components used are all safe strains, and there are no risks of ecological safety hazards such as invasion of alien species and invasion of pathogenic microorganisms.

[0039] In summary, the method of the present invention has a clear principle and strong operability. It can create a suitable feeding habitat for red-crowned cranes in multiple dimensions and improve the targeted proliferation efficiency of target food source organisms, thereby achieving effective regulation of the structure of the intestinal microbial flora of red-crowned cranes in a near-natural way, and there is no risk of any biosafety hazards and interference with the stability of the natural wetland ecosystem structure. The method described in the present invention is used to carry out targeted proliferation of food sources for red-crowned cranes, and the probability of attracting and feeding the red-crowned crane population reaches 100%, and the success rate of regulating the intestinal microorganisms of red-crowned cranes reaches 90%. The present invention is used for the targeted proliferation of food source organisms with the goal of regulating the intestinal microorganisms of red-crowned cranes. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0041] Figure 1 This is a flow chart of a method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to a specific embodiment of the present invention;

[0042] Figure 2 This is a schematic diagram of the germination result of the water chestnut according to the third specific embodiment of the present invention;

[0043] Figure 3 This is a schematic diagram of the germination results of the flat-stalked thorn thorn described in the third specific embodiment of the present invention;

[0044] Figure 4 This is a schematic diagram of the survival rate results of food-borne shrimp organisms according to the third specific embodiment of the present invention;

[0045] Figure 5 This is a schematic diagram of the survival rate results of food-borne snail organisms according to the third specific embodiment of the present invention;

[0046] Figure 6 This is a schematic diagram of the survival rate results of food-borne fish organisms described in the third specific implementation mode of the present invention. DETAILED DESCRIPTION

[0047] In the following description, specific details such as specific system structures and technologies are provided for the purpose of illustration rather than limitation, so as to provide a thorough understanding of the embodiments of the present application. However, it should be clear to those skilled in the art that the present application may also be implemented in other embodiments without these specific details. In other cases, detailed descriptions of well-known systems, devices, circuits, and methods are omitted to prevent unnecessary details from obstructing the description of the present application.

[0048] It should be understood that when used in this specification and the appended claims, the term "comprising" indicates the presence of described features, integers, steps, operations, elements and / or components, but does not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components and / or groups thereof.

[0049] It should also be understood that the terms used in this application specification are only for the purpose of describing specific embodiments and are not intended to limit the present application. As used in this application specification and the appended claims, the singular forms "a", "an" and "the" are intended to include plural forms unless the context clearly indicates otherwise.

[0050] The following is a clear and complete description of the technical solutions in the embodiments of the present application in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0051] In the following description, many specific details are set forth to facilitate a full understanding of the present application, but the present application may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present application. Therefore, the present application is not limited to the specific implementation methods disclosed below.

[0052] Implementation method 1:

[0053] Reference Figure 1 This embodiment will be described.

[0054] A method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes, comprising:

[0055] S1: Basement environment improvement and micro-geomorphic structure, including:

[0056] From late April to early May, a 1,000-2,500 square meter plot was selected in the historical breeding habitat where the wild red-crowned crane population rested for more than 5 years and the single population size was more than 500 to build a food source targeted proliferation area. In the early stage and process of regional construction, the surface soil with a depth of 0-10 cm was stripped and a 50 cm high closed soil ridge was piled around the plot without the premise of the wild red-crowned crane population resting. Gram-positive bacteria, coagulant Bacillus, and nitrogen-fixing Bacillus megaterium were applied to the surface of the stripped soil with a specification of 0.06-0.09 kg / square meter. The soil was turned over 2-3 times at a depth of 0-50 cm. Circular islands with a height of 30-40 cm, a slope width of 40-50 cm, a slope angle of 8-15°, and a base area of ​​20-25 square meters were randomly constructed with a specification of 2-3 places / 100 square meters. In non-construction areas, discontinuous soil ridges with a height of 5-10 cm and a length of 50-100 cm are constructed with a spacing of 30-45 cm and an end distance of 30-40 cm.

[0057] S2: Water resource zoning replenishment and microbial agent addition, including:

[0058] From early to late May, water from nearby natural wetlands with ammonia nitrogen content below 1.25 mg / L, total nitrogen content below 10 mg / L, and total phosphorus content below 0.25 mg / L were used to replenish the targeted food source proliferation area. The circular island matrix was artificially sprinkled until the soil moisture content was 80-90%, and the non-construction area was replenished with water to a flooding depth of 8-10 cm, and then a microbial agent composed of Rhodopseudomonas palustris, Bacillus Clausii, and Bacillus licheniformis was applied at a concentration of 0.10-0.12 g / L.

[0059] S3: Foodborne organism introduction kit, including:

[0060] Selected flat-stalked thorny water chestnut and big-based water chestnut as food plants, selected Japanese swamp shrimp and Macrobrachium rosenbergii as food shrimp organisms for red-crowned cranes, selected hemispherical multi-veined flat snails, flat disc snails, and bright septate flat snails as food snail organisms for red-crowned cranes, and selected grass carp, silver carp, silver carp, and Lingyuan goby as food fish organisms. After planting big-based water chestnut corms and flat-stalked thorny water chestnut rooted young plants at 3-5 stems / square meter and 10-15 cm intervals in the non-slope area of ​​the circular island and the top of the soil ridge, the food shrimp organisms were mixed and released at a density of 500-1000 / square meter and a sex ratio of 30-35% male shrimp and 65-70% female shrimp. The food snail organisms were mixed and released at a density of 200-450 / square meter and a sex ratio of 30-35% male snails and 65-70% female snails. The food source fish are mixed and released at a density of 50-100 per square meter and a gender ratio of 40-50% males and 50-60% females.

[0061] S4: Hydrological conditions are divided into different areas for segmented regulation, including:

[0062] From mid-July to mid-October, the soil moisture content in the non-flooded area of ​​the circular island was kept constant at 80-90%. From mid-July to mid-August, mid-August to mid-October, and mid-October to mid-November, the surface flooding depth in the ridge construction area was maintained at 8-10 cm, 15-20 cm, and 25-30 cm, respectively. In winter, when the red-crowned crane population migrated and there were no red-crowned cranes feeding, artificial water was added until the surface flooding depth in the ridge construction area reached 40-45 cm.

[0063] S5, Comparative evaluation of the gut microbiota of red-crowned cranes, including:

[0064] After the red-crowned crane population had been feeding continuously in the targeted food proliferation area for 7-10 days, the excretion behavior of the cranes was observed at a distance of 500-700 meters away. Fresh fecal samples were collected immediately after the cranes finished feeding and 16S rDNA high-throughput sequencing was performed. Firmicutes was used as the dominant phylum and Catelliococcus was used as the dominant genus as the criteria for regulating the intestinal microbial composition and determining health of the red-crowned cranes.

[0065] Implementation method 2:

[0066] This embodiment is a further example of the method for regulating the food-based targeted proliferation of intestinal microorganisms of red-crowned cranes described in Embodiment 1.

[0067] The method comprises:

[0068] 1. Basement environment improvement and micro-geomorphic structure:

[0069] From late April to early May, a 1,000-2,500 square meter plot of land was selected within the historical breeding habitat of wild red-crowned cranes to construct a targeted food source proliferation area. The wild red-crowned crane population in the selected area must have a historical rest period of more than 5 years and a single resting population size of more than 500 cranes. No wild red-crowned crane populations were required to rest in the early and during construction of the area to avoid interfering with the rest and foraging activities of the cranes. The 0-10 cm surface soil in the designated area was stripped and a 50 cm high closed earth ridge was built around the plot to separate the root tissue of native plants and construct local negative terrain for later habitat construction and biological cultivation management. Microbial fertilizer was applied to the surface of the stripped soil in the area at a specification of 0.06-0.09 kg / m2 to regulate the soil microbial component structure in the area. The applied microbial fertilizer contained Gram-positive bacteria ( Bacillus subtilis )、Bacillus coagulans( Bacillus coagulans )、Bacillus megaterium( Bacillus megaterium ), in order to improve the soil microbial structure in a targeted manner to optimize the microbial components carried by food organisms; it is required to turn the surface soil at a depth of 0-50 cm 2-3 times after the application of microbial fertilizer to fully mix the microbial fertilizer to promote the absorption of the soil and the root tissue of later food plants; it is required to randomly pile up 2-3 circular islands with a height of 30-40 cm, a slope width of 40-50 cm, a slope angle of 8-15°, and a base area of ​​20-25 square meters in units of 100 square meters, and to construct discontinuous soil ridges with a height of 5-10 cm and a length of 50-100 cm with a spacing of 30-45 cm and an end distance of 30-40 cm in non-construction areas to meet the growth and expansion of food plants and the benthic activities of food animals.

[0070] 2. Water resource zoning replenishment and microbial agent addition:

[0071] From early to late May, targeted zoning water replenishment is carried out in the targeted proliferation area of ​​food sources, and probiotics are added to the water after replenishment. The replenishment water is required to come from the adjacent natural wetland water body, and the ammonia nitrogen content of the water body is less than 1.25 mg / L, the total nitrogen content is less than 10 mg / L, and the total phosphorus content is less than 0.25 mg / L, so as to avoid eutrophication of the water body and contamination of food source organisms; it is required to replenish the circular island with water by artificial sprinkling until the soil moisture content is 80-90%, so as to avoid the loss of matrix resources of the circular island caused by runoff replenishment and provide a suitable environment for the growth of food source plant bulbs; it is required to replenish water in non-construction areas to a depth of 8-10 cm to the top of the soil ridge, so as to form a hydrological habitat that promotes the development of food source plant stems and the survival of food source fish and food source snails. It is required to apply 0.10-0.12 g / L of Rhodopseudomonas palustris ( Rhodop seudanonas palustris ), Bacillus clausii ( Bacillus clausii ), Bacillus licheniformis ( Bacillus licheniformis ) to improve water quality and promote photosynthetic activity of food-borne organisms; it is required to carry out the introduction of food-borne organisms 5-7 days after the addition of the microbial agent to ensure that the soil and water absorb the microbial agent.

[0072] 3. Introduction of food source organisms:

[0073] From early June to mid-July, select red-crowned crane food plants, fish, shrimps and snails to introduce them into the food source targeted proliferation area in different time periods. Bolboschoenus planiculmis )、Daji water chestnut( Eleocharis kamtschatica ) as a food source for red-crowned cranes; it is required to plant water chestnut bulbs at a density of 3-5 stems / square meter in the non-slope area of ​​the circular island in early June, and plant young plants of flat-stalked thorny thorns with roots at intervals of 10-15 cm at the top of the soil ridge to meet the suitable hydrological habitat requirements of various food source plants and reserve space for the later expansion and development of root systems and bulb tissues; it is required to select Japanese swamp shrimp ( Macrobrachium nipponense )、Macrobrachium rosenbergii( Macrobrachium rosenbergii ) are food-source shrimp organisms; after the food-source plants are planted, the food-source shrimp organisms are mixed and released at a density of 500-1000 / m2 and a gender ratio of 30-35% male shrimps and 65-70% female shrimps to ensure the growth of individual food-source shrimps and achieve intergenerational reproduction of the population; select the hemispheric multi-veined flat snail ( Polypylis hemisphaerula )、Flat disc snail( Valuata cristata )、Bright septum flat snail( Segmentina nitida ) as food source snails; after the food source plants are planted, the food source snails are mixed and released at a density of 200-400 per square meter and a gender ratio of 30-35% male snails and 65-70% female snails to reserve the ecological niche required for the subsequent expansion of the snail population; grass carp ( Ctenopharyngodon idella )、Silver Carp(Silver prussian )、Silver Carp( Hypophthalmichthys molitrix )、Lingyuan goby( Gobio lingyuanensis ) as food source fish organisms; it is required that in mid-July, the fry of food source fish organisms be mixed and released at a density of 50-100 fish / square meter and a gender ratio of 40-50% male fish and 50-60% female fish, so as to reserve time for the growth and development of food source plant seedlings, food source shrimp and food source snail larvae, and avoid problems such as gnawing and capturing food source plants and organisms caused by the expansion of food source fish populations.

[0074] 4. Hydrological conditions zoning and segmentation control:

[0075] From mid-July to mid-November, hydrological regulation was implemented in the targeted proliferation area of ​​food sources. It was required that the soil moisture content in the non-flooded area of ​​the circular island be kept constant at 80-90% from mid-July to mid-October to provide a suitable hydrological environment for the development of water chestnut corms; the surface flooding depth in the ridge construction area was required to be 8-10 cm from mid-July to mid-August, and the surface flooding depth was required to be 15-20 cm from mid-August to mid-October, so as to regulate the biomass of food source plants by changing the flooding environment. The distribution strategy promotes the development of underground bulbs; it requires water to be replenished to a surface flooding depth of 25-30 cm in the ridge construction area from mid-October to mid-November to meet the multiple supply needs of the red-crowned cranes such as foraging and water diversion; it requires artificial water replenishment to a surface flooding depth of 40-45 cm in the ridge construction area after the red-crowned crane population migrates in winter and there are no red-crowned cranes feeding, so as to reserve sufficient water during the ice period, thereby providing a suitable hydrological environment for the red-crowned crane population to forage in the spring of the following year and preventing the death of food source organisms due to lack of water and freezing in winter.

[0076] 5. Comparative evaluation of intestinal microorganisms of red-crowned cranes:

[0077] Fecal samples were collected and tested during the feeding period of the red-crowned crane population, and the composition of the intestinal microorganisms of the red-crowned cranes was compared and analyzed. After the red-crowned cranes continuously fed in the targeted proliferation area of ​​the food source for 7-10 days, the excretion behavior of the red-crowned cranes was observed from a distance of 500-700 meters using a monocular telescope to avoid human activities interfering with the feeding activities of the red-crowned cranes; fresh fecal samples were collected immediately after the red-crowned cranes finished feeding and 16SrDNA high-throughput sequencing was performed, with Firmicutes as the dominant bacterial phylum and Catarrhalis as the dominant bacterial phylum. Catellicoccus ) is the dominant bacterial genus as a criterion for regulating the intestinal microbial composition and determining health of red-crowned cranes.

[0078] Implementation method three:

[0079] Reference Figure 2 , Figure 3 , Figure 4 ,Figure 5 , Figure 6 This embodiment will be described.

[0080] This embodiment is a further example of the method for regulating the food-based targeted proliferation of intestinal microorganisms of red-crowned cranes described in Embodiments 1 and 2.

[0081] This implementation method carries out targeted proliferation of food sources in typical habitats of red-crowned cranes with the goal of regulating the intestinal microorganisms of red-crowned cranes. In the process of targeted proliferation of food sources for red-crowned cranes, the following methods are implemented:

[0082] 1. Basement environment improvement and micro-geomorphic structure:

[0083] On April 20, 2022, three plots of 2,500 square meters were designated as targeted food source proliferation areas in areas where the wild red-crowned crane population has a historical rest period of more than 5 years and a single population rest scale of more than 500. A small agricultural bulldozer was used to strip the topsoil of the plot to a depth of 10 cm and then piled up to form a 50-cm high closed earth ridge. After the topsoil was stripped, microbial fertilizers cultured with Gram-positive bacteria, Bacillus coagulans and Bacillus megaterium were artificially spread on the exposed soil surface in the plot at a specification of 0.08 kg / square meter, and then a rotary tiller was used to turn the soil 3 times at a depth of 0-50 cm to allow the microbial fertilizer to be fully mixed with the soil.

[0084] On April 22, 2022, a small agricultural bulldozer was used to randomly pile up circular islands with a height of 38 cm, a slope width of 45 cm, a slope angle of 12°, and a base area of ​​24 square meters at a standard unit of 3 pieces / 100 square meters. In the non-construction area, intermittent soil ridges with a height of 8 cm and a length of 86 cm were constructed with a horizontal spacing of 40 cm and a ridge end distance of 38 cm.

[0085] 2. Water resource zoning replenishment and microbial agent addition:

[0086] On May 24, 2022, the water bodies of the adjacent natural wetlands were used to replenish water in the targeted proliferation area of ​​the food source. The ammonia nitrogen content in the adjacent water bodies was 1.01 mg / L, the total nitrogen content was 8 mg / L, and the total phosphorus content was 0.21 mg / L. The tubular water spraying equipment was used to spray the circular island with water until the soil moisture content was 85-87%. The pumping machinery was used to replenish water to a depth of 10 cm in the non-construction area and submerge the top of the soil ridge. After the soil spraying of the circular island and the replenishment of water in the non-construction area were completed, a composite microbial agent of Rhodopseudomonas palustris, Bacillus Clausii, and Bacillus licheniformis was spread into the water body at a concentration of 0.11 g / L.

[0087] 3. Introduction of food source organisms:

[0088] On June 1, 2022, large-based water chestnut corms were planted in the non-slope area of ​​the circular island in each plot at a density of 3-5 stems / square meter, 7-10 stems / square meter, and 12-15 stems / square meter, respectively, and young plants of flat-stalked thorny tree with roots were planted on the top of the earth ridge at intervals of 5-10 cm, 10-15 cm, and 15-20 cm, respectively. Among them, the germination rate was the highest when large-based water chestnut corms were planted at a density of 3-5 stems / square meter and young plants of flat-stalked thorny tree with roots were planted at intervals of 10-15 cm. On June 15, 2022, Macrobrachium nipponense and Macrobrachium rosenbergii were mixed and released at densities of 500, 1,000, and 1,500 per square meter, respectively, and at a sex ratio of 32% male shrimp and 68% female shrimp. The survival rate of food source shrimp at the release density of 500 and 1,000 per square meter was significantly higher than that at the density of 1,500 per square meter. On June 21, 2022, Hemispherical multi-veined flat snails, flat disc snails, and bright septate flat snails were mixed and released at densities of 200-400, 400-600, and 600-800 per square meter, respectively, and at a sex ratio of 30% male snails and 70% female snails. The survival rate of food source snails was the highest at a release density of 200-400 per square meter. On July 16, 2022, grass carp, silver carp, bighead carp and Lingyuan minnow seedlings were released at densities of 10-30, 50-100 and 100-200 per square meter, respectively, with a gender ratio of 50% males and 50% females. The survival rate of food fish was the highest at a density of 50-100 per square meter.

[0089] 4. Hydrological conditions zoning and segmentation control:

[0090] From July 15 to October 16, 2022, the soil moisture content in the non-flooded area of ​​the circular island will be maintained at 80-90% to create a suitable soil hydrological environment for the rapid development of water chestnut corms; from July 15 to August 15, from August 15 to October 16, and from October 16 to November 14, the surface flooding depth in the ridge construction area will be maintained at 8-10 cm, 15-20 cm and 25-30 cm, respectively, so as to promote the development of key feeding organs of food plants through flooding depth regulation and achieve coordinated optimization of hydrological recharge and foraging habitats for red-crowned cranes; after the red-crowned crane population stops feeding and migrates, the surface flooding depth in the ridge construction area will be artificially replenished to 40-45 cm to pre-replenish the infiltration volume of water in winter, thereby pre-setting a suitable foraging hydrological environment for red-crowned cranes in the spring of the following year and preventing frostbite and death of food organisms.

[0091] 5. Comparative evaluation of intestinal microorganisms of red-crowned cranes:

[0092] From October 16 to 29, 2022, a monocular telescope was used to continuously observe the feeding behavior of the red-crowned crane population 600 meters outside the targeted food source proliferation area. After the wild red-crowned cranes fed continuously for 8 days, fresh feces samples were collected immediately after the red-crowned cranes finished feeding, and feces samples from artificially raised red-crowned cranes and red-crowned cranes that did not feed in the targeted food source proliferation area were also collected. After artificial extraction of the DNA substrate in the laboratory, 16SrDNA high-throughput sequencing was carried out and the intestinal microbial composition structure was compared. The intestinal microbial health index was used as the reference, with Firmicutes, Proteobacteria and Bacteroidetes as the dominant phyla, and Caterpillarcocephala as the dominant genus as the standard for regulating the intestinal microbial components and judging the health of the red-crowned cranes. When the dominant phylum of the intestinal microorganisms of the red-crowned cranes is Firmicutes, and the dominant genus is Caterpillarcocephala, the food source targeted proliferation method is successfully achieved.

[0093] The test results showed that the dominant phylum of intestinal microorganisms of red-crowned cranes feeding in the targeted food proliferation area was Firmicutes, and the dominant genus was Caterpillarococcus; the dominant phylum of intestinal microorganisms of captive red-crowned cranes was Proteobacteria, and the dominant genus was Sphingolipidomonas; the dominant phylum of intestinal microorganisms of red-crowned cranes not feeding in the targeted food proliferation area was Proteobacteria, and the dominant genus was Sphingolipidomonas.

[0094] Those skilled in the art will appreciate that the above are only preferred embodiments of the present invention, and the various embodiments of the present disclosure and / or the features described in the claims may be combined or combined in various ways, even if such combinations or combinations are not explicitly described in the present disclosure. It is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art may still modify the technical solutions described in the aforementioned embodiments, or perform equivalent substitutions on some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

[0095] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0096] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

[0097] Finally, it should be noted that the above implementation modes are only used to illustrate the technical solutions of the present disclosure rather than to limit its protection scope. Although the present disclosure is described in detail with reference to the above implementation modes, ordinary technicians in the relevant field should understand that after reading the present disclosure, those skilled in the art can still make various changes, modifications or equivalent substitutions to the specific implementation modes of the invention, but these changes, modifications or equivalent substitutions are all within the protection scope of the disclosed claims to be approved.

Claims

1. A method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes, characterized in that: The method comprises: S1: Basement environment improvement and micro-geomorphic structure, including: From late April to early May of the regulation year, target plots were selected in the historical breeding habitat of red-crowned cranes to build targeted food source proliferation areas; the surface soil of the targeted food source proliferation areas was stripped, and earth ridges were piled around the targeted food source proliferation areas; microbial fertilizer was applied to the surface of the soil in the treated targeted food source proliferation areas and turned over, and circular islands were randomly built on the targeted food source proliferation areas, with a construction density of 2-3 places / 100m 2 , constructing discontinuous soil ridges in the unconstructed part of the targeted food source proliferation area, wherein the spacing between the soil ridges is 30-45 cm, the end distance is 30-40 cm, the height is 5-10 cm, and the length is 50-100 cm; S2: Water resource zoning replenishment and microbial agent addition, including: In early to late May of the year of regulation, water from nearby natural wetlands is used to replenish water in the targeted food source proliferation area; the substrate of the circular island is artificially sprinkled until the soil moisture content of the circular island is 80%-90%; the unconstructed part of the targeted food source proliferation area is replenished with water to a flooding depth of 8-10 cm, and microbial agents are applied; S3: Foodborne organism introduction kit, including: Selecting food-source plants and food-source organisms, the food-source plants include: Tripterygium wilfordii and Echinochloa chinensis; the food-source organisms include: food-source shrimps, food-source snails, and food-source fish; planting Echinochloa chinensis corms and rooted young plants of Tripterygium wilfordii in the non-slope area of ​​the circular island and the top of the soil ridge, wherein the planting density is 3-5 stems / square meter and the planting interval is 10-15 centimeters; after planting, mixing the food-source organisms in proportion and releasing them; S4: Hydrological conditions are divided into different areas for segmented regulation, including: From mid-July to mid-October of the regulation year, the moisture content of the non-flooded soil of the circular island is maintained at 80%-90%; from mid-July to mid-August of the regulation year, the surface flooding depth of the soil ridge area is maintained at 8-10 cm; from mid-August to mid-October of the regulation year, the surface flooding depth of the soil ridge area is maintained at 15-20 cm; from mid-October to mid-November of the regulation year, the surface flooding depth of the soil ridge area is maintained at 25-30 cm; in winter, after the red-crowned crane population migrates and there are no red-crowned cranes feeding, water is replenished to the soil ridge area to make its surface flooding depth reach 40-45 cm.

2. A method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: The requirements for the historical breeding habitat of the red-crowned crane are: the rest period of the wild red-crowned crane population is greater than 5 years, and the single population size is greater than 500.

3. The method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: The area of ​​the target plot is 1,000-2,500 square meters.

4. The method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: In S1, the microbial fertilizer is: a microbial fertilizer obtained by tissue culture of Gram-positive bacteria, Bacillus coagulans, and Bacillus megaterium for nitrogen fixation; and the application amount of the microbial fertilizer is 0.06-0.09 kg / m2.

5. The method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: The height of the circular island is 30-40 centimeters, the width of the slope is 40-50 centimeters, the angle of the slope is 8°-15°, and the base area is 20-25 square meters.

6. The method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: In the S2, the ammonia nitrogen content of the water body adjacent to the natural wetland is lower than 1.25 mg / L, the total nitrogen content is lower than 10 mg / L, and the total phosphorus content is lower than 0.25 mg / L.

7. The method for regulating the food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: In the S2, the components of the microbial agent include: Rhodopseudomonas palustris, Bacillus clausii, and Bacillus licheniformis; the application concentration of the microbial agent is 0.10-0.12 g / L.

8. The method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: The food-source shrimp organisms include: Japanese swamp shrimp and Macrobrachium rosenbergii; the food-source snail organisms include: Hemispheric multi-veined flat snail, flat disc snail, and bright septate flat snail; the food-source fish organisms include: grass carp, silver carp, silver carp, and Lingyuan minnow.

9. A method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 8, characterized in that: In S3, the food-source organisms are mixed and released in proportion, specifically: food-source shrimps are mixed and released at a density of 500-1000 per square meter and a gender ratio of 30-35% male shrimps and 65-70% female shrimps; food-source snails are mixed and released at a density of 200-450 per square meter and a gender ratio of 30-35% male snails and 65-70% female snails; food-source fish are mixed and released at a density of 50-100 per square meter and a gender ratio of 40-50% male fish and 50-60% female fish.

10. The method for regulating food-source targeted proliferation of intestinal microorganisms of red-crowned cranes according to claim 1, characterized in that: The method also includes: S5, comparative evaluation of the intestinal microorganisms of red-crowned cranes: after the red-crowned crane population has continuously fed in the targeted proliferation area of ​​the food source for 7-10 days, the excretion behavior of the red-crowned cranes is observed at a distance, and fresh fecal samples are collected immediately after the red-crowned cranes finish feeding and 16SrDNA high-throughput sequencing is performed, with Firmicutes as the dominant phylum and Catelliococcus as the dominant genus as the criteria for regulating the intestinal microbial components and determining the health of the red-crowned cranes.

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