A method and apparatus for collecting earthworm burrow wall soil

Abstract

This invention provides a method and apparatus for collecting earthworm burrow wall soil. The method includes collecting raw soil, adjusting the particle composition of the raw soil to obtain culture soil. A mesh screen is laid in a culture pot, and the culture soil is placed inside the mesh screen. Earthworms are placed in the culture soil, and the culture pot is placed in an incubator. The environmental parameters in the incubator are adjusted to allow the earthworms to construct burrows within the culture soil. A preset time is checked; if so, the culture soil is removed, and the burrow wall soil is peeled off. The culture soil balances plasticity and permeability to meet the needs of earthworm growth and burrow building during long-term cultivation. By adjusting the temperature, humidity, and light intensity in the incubator, the earthworms can fully construct burrows while maintaining stable burrow wall soil properties. A hard, pointed spoon is used to precisely peel off the burrow wall soil, ensuring that the burrow wall soil sample is free from cross-contamination and structurally intact.

Description

Technical Field

[0001] This invention relates to the fields of soil stripping and earthworm ecology, and particularly to a method and apparatus for collecting soil from earthworm burrow walls. Background Technology

[0002] In soil ecology and earthworm ecology research, earthworm burrow wall soil is an important research object characterizing the impact of earthworm surface contact on soil. When earthworms move in the soil and form burrows, their body surface undergoes physical, chemical, and biological interactions with the burrow wall soil. This results in significant differences between the burrow wall soil and the surrounding ordinary soil in terms of particle composition, structural characteristics, and nutrient content. These differences are important research evidence for revealing the mechanisms by which earthworms improve soil and their influence on soil nutrient cycling.

[0003] However, current methods for collecting earthworm burrow wall soil face several technical bottlenecks: First, earthworm species vary greatly, with different species exhibiting different body sizes and lifestyles, resulting in significant differences in the burrow morphology (such as burrow diameter, depth, and curvature). Existing collection methods are ill-suited to the burrow characteristics of different earthworm species. Second, the soil itself is fragile, and earthworm burrow structures are relatively weak. Even slight external intervention during collection can easily cause the burrow walls to collapse, resulting in the burrow wall soil mixing with the surrounding ordinary soil, making precise separation impossible. Third, existing collection methods lack systematic control over the earthworm's growth environment (such as temperature, humidity, soil conditions, and container compatibility) and cultivation cycle, leading to insufficient burrow construction and poor-quality burrow wall soil formation. Furthermore, improper soil extraction and impurity removal methods further increase the difficulty of collection. Fourth, the large number and scattered distribution of earthworm burrows in the soil can damage their structure during soil removal.

[0004] Currently, there is no standardized method for collecting earthworm burrow wall soil that can comprehensively solve the above problems. Existing methods mostly involve simple manual digging and stripping, resulting in low collection efficiency and poor purity. This seriously affects the accuracy and reliability of relevant research data and hinders research progress in earthworm ecology and soil ecology. Therefore, developing a method for accurately and efficiently collecting earthworm burrow wall soil has become an urgent technical problem to be solved in this field. Summary of the Invention

[0005] To overcome the problems existing in related technologies, the purpose of this invention is to provide a method and apparatus for collecting earthworm burrow wall soil. The method can accurately and efficiently collect earthworm burrow wall soil, and under the premise of systematically controlling the earthworm growth environment, completely peel off the earthworm burrow body in different areas to prevent damage to the structure of the earthworm burrow body.

[0006] A method for collecting soil from earthworm burrow walls includes:

[0007] Collect raw soil, adjust the particle composition of the raw soil, and obtain culture soil;

[0008] Lay the mesh in the culture pot and fill the mesh with the culture soil;

[0009] Earthworms are placed in the culture soil, the culture pot is placed in the incubator, and the environmental parameters in the incubator are adjusted so that the earthworms construct earthworm burrows in the culture soil.

[0010] If the preset time has been reached, remove the culture soil and peel off the soil from the earthworm burrow wall.

[0011] In a preferred embodiment of the present invention, the step of regulating the particle composition of the original soil to obtain cultured soil includes:

[0012] Impurities are removed from the original soil to obtain pretreated soil; the impurities include stones and tree roots.

[0013] Sand, soil, and clay particles were separated by sieving.

[0014] The culture soil was prepared by mixing clay, loam, and clay in a ratio of 30%, 60%, and 10% using a layered mixing method.

[0015] In a preferred embodiment of the present invention, the size of the sand particles is greater than 0.05 mm and less than 2 mm; the size of the soil particles is less than or equal to 0.05 mm and greater than or equal to 0.002 mm; and the size of the clay particles is less than 0.002 mm.

[0016] In a preferred embodiment of the present invention, the step of laying the mesh in the culture pot and filling the culture soil into the mesh includes:

[0017] Lay the mesh flat in the culture pot, so that the edge of the mesh extends upward along the pot wall and beyond the pot opening by 5-8cm;

[0018] The culture soil is filled into the mesh of the culture pot; the filling height of the culture soil is 4 / 5 of the height of the culture pot.

[0019] In a preferred embodiment of the present invention, adjusting the environmental parameters within the incubator includes:

[0020] The temperature range inside the incubator is set to 20-25℃, and the temperature range inside the incubator is kept constant during the cultivation of earthworms.

[0021] The soil moisture content in the incubator was measured during the early stage of cultivation, and water was added to the soil in the incubator. The early stage of cultivation lasted from the start of cultivation to 7 days before the end of cultivation.

[0022] In the later stage of cultivation, the water control program is initiated, active water replenishment is stopped, and the soil moisture content in the cultivation chamber is reduced to 30% using the ventilation system of the cultivation chamber; the later stage of cultivation is from 7 days before the end of cultivation to the end of cultivation.

[0023] The incubator is set to a periodic light cycle of 12 hours of light and 12 hours of darkness, with a light intensity of 500-800 lux.

[0024] In a preferred embodiment of the present invention, the step of detecting the soil moisture content in the incubator during the early stage of cultivation and replenishing the soil in the incubator with water includes:

[0025] The soil moisture content in the incubator was monitored every 3 days by gravimetric method.

[0026] If the moisture content is below 40%, water is sprayed onto the surface of the culture soil in the culture box to maintain the soil moisture content at 40-60% of field capacity.

[0027] In a preferred embodiment of the present invention, the step of peeling off the soil from the earthworm burrow wall includes:

[0028] The culture soil was subjected to CT scanning to obtain a diagram of the soil's internal structure.

[0029] Connectivity analysis was performed on the soil internal structure diagram to obtain the final connected regions; the final connected regions were used to characterize the earthworm burrow.

[0030] Select the starting and ending surfaces of the culture soil, wherein the culture soil is cylindrical in shape;

[0031] Along the direction from the starting surface to the ending surface, the final connected region is divided into N layers of pit wall soil; N≥2;

[0032] An earthworm hole wall soil collection device was used to sequentially peel off the first layer of hole wall soil to the Nth layer.

[0033] In a preferred embodiment of the present invention, the step of performing connected component analysis on the soil internal structure diagram to obtain the final connected region includes:

[0034] The soil internal structure map is binarized according to a preset pixel threshold to obtain a binarized image;

[0035] A point with a value of 1 in the region corresponding to the binarized image and the starting surface is randomly selected as the first seed point;

[0036] The first seed point is seed-filled to obtain the first connected region;

[0037] Randomly select a point with a value of 1 in the region corresponding to the binarized image and the termination surface as the second seed point;

[0038] Seed filling is performed on the second seed point to obtain the second connected region;

[0039] The final connected region is obtained by taking the union of the first connected region and the second connected region.

[0040] In a preferred embodiment of the present invention, the step of seed filling the first seed point to obtain a first connected region includes:

[0041] Take a point in the neighborhood of the first seed point as the current point. If the value of the current point is 1, replace the current point with other points in the neighborhood with a value of 1. If the value of the current point is 0, count the total number of points with a value of 1 in the neighborhood of the current point. If the total number is greater than the total number threshold, set the value of the current point to 1. The neighborhood is four adjacent pixels or eight adjacent pixels.

[0042] Update the position of the current point until the position of the current point remains unchanged for M consecutive times, and obtain the first connected region.

[0043] The present invention also provides an earthworm hole wall soil collection device for implementing an earthworm hole wall soil collection method. The device includes a spoon body and a spoon handle, which are detachably connected or integrally formed. The spoon body has a peeling opening, and a first flange and a second flange are respectively provided on both sides of the peeling opening. Insert strips are provided on the side walls of the spoon body near the first flange and the second flange, and the insert strips include multiple insertion holes. A baffle is connected to one of the insertion holes.

[0044] The beneficial effects of this invention are as follows:

[0045] This invention provides a method for collecting earthworm burrow wall soil, comprising: collecting raw soil; adjusting the particle composition of the raw soil to obtain culture soil; laying a mesh screen in a culture pot and filling the mesh screen with the culture soil; placing earthworms into the culture soil; placing the culture pot in a culture box; adjusting the environmental parameters in the culture box to allow the earthworms to construct burrows within the culture soil; detecting whether a preset time has been reached; if so, removing the culture soil and peeling off the burrow wall soil. The particle composition of the raw soil is precisely controlled to a ratio of 30% clay, 60% loam, and 10% sand to obtain the culture soil. The culture soil balances plasticity and permeability, meeting the needs of earthworm growth and burrow building during long-term cultivation. The size of the culture pot is designed according to the different body sizes of earthworms, and the mesh screen is designed to wrap the pot accordingly. This solves the problem of container compatibility for earthworms of different body sizes and allows for complete soil extraction, avoiding damage caused by soil adhering to the pot wall after long-term cultivation. After obtaining the culture soil and the mesh-wrapped culture pot, the culture soil was first placed into the mesh-wrapped culture pot, followed by the earthworms. Finally, the culture pot containing the earthworms was placed in an incubator. The temperature, humidity, and light intensity within the incubator were adjusted, and the optimal culture time was determined to be 30 days, ensuring that the earthworms could fully construct burrows and that the burrow wall soil properties remained stable. After 30 days of earthworm culture, the culture soil was completely removed by lifting the mesh. A stiff, pointed spoon was used to precisely peel away the burrow wall soil from the earthworm burrows, ensuring that the soil sample was free of cross-contamination and structurally intact. This method allows for precise and efficient collection of earthworm burrow wall soil, and under the premise of systematically controlling the earthworm growth environment, it completely separates the earthworm burrows from different areas, preventing damage to the burrow structure. Attached Figure Description

[0046] Figure 1 This is a flowchart of the earthworm hole wall soil collection method of the present invention;

[0047] Figure 2 This is a flowchart of the process for peeling the soil from the earthworm burrow wall in this invention;

[0048] Figure 3 This is a schematic diagram of the earthworm hole wall soil collection device of the present invention;

[0049] Figure 4 yes Figure 3 Enlarged view of point A in the middle;

[0050] Figure 5 This is a photograph of an earthworm burrow in the culture soil of this invention;

[0051] Figure 6 This is a diagram of the internal structure of soil obtained by CT scanning according to the present invention.

[0052] Reference numerals: 1. Spoon body; 2. Spoon handle; 3. Peeling opening; 4. First flange; 5. Second flange; 6. Insert bar; 7. Insertion hole; 8. Baffle. Detailed Implementation

[0053] Preferred embodiments of the invention will now be described in more detail with reference to the accompanying drawings. While preferred embodiments of the invention are shown in the drawings, it should be understood that the invention can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to make the invention more thorough and complete, and to fully convey the scope of the invention to those skilled in the art.

[0054] Example 1

[0055] like Figure 1 As shown, this embodiment provides a method for collecting soil from the walls of earthworm burrows, including:

[0056] S1: Collect raw soil, adjust the particle composition of the raw soil to obtain culture soil;

[0057] S2: Lay the mesh in the culture pot and fill the mesh with the culture soil;

[0058] S3: Place earthworms into the culture soil, place the culture pot into the incubator, and adjust the environmental parameters in the incubator so that the earthworms can construct earthworm burrows in the culture soil;

[0059] S4: Check if the preset time has been reached. If so, remove the culture soil and peel off the soil from the earthworm hole wall.

[0060] Raw soil is collected from the field. The raw soil contains clay, loam, sand and various impurities. According to the living habits of earthworm species, the particle composition of the raw soil needs to be precisely adjusted to a ratio of 30% clay, 60% loam and 10% sand to obtain culture soil that is suitable for the living habits of earthworm species.

[0061] The process of regulating the particle composition of the original soil to obtain cultured soil includes:

[0062] S11: Remove impurities from the original soil to obtain pretreated soil; the impurities include stones and tree roots;

[0063] S12: Sand, soil and clay particles are separated by sieving.

[0064] S13: Using a layered mixing method, sand, loam, and clay particles are mixed in a ratio of 30% clay particles, 60% soil particles, and 10% sand particles to obtain the culture soil.

[0065] To regulate the particle composition of the original soil, it is first pretreated to remove impurities such as stones and plant debris, resulting in pretreated soil. Since sand, loam, and clay particles have different diameters (sand particles 2mm-0.05mm, loam particles 0.05mm-0.002mm, and clay particles less than 0.002mm), the pretreated soil is then separated into sand, loam, and clay particles using a sieving method. This method utilizes a set of sieves with decreasing aperture sizes from top to bottom. The pretreated soil is placed on the top sieve, and shaking or vibration causes particles smaller than the sieve apertures to fall, while larger particles are retained. This separates the pretreated soil according to particle diameter, resulting in sand, loam, and clay particles. Then, using a layered mixing method, the three types of particles are thoroughly mixed in a ratio of 30% clay particles, 60% loam particles, and 10% sand particles. First, the sand and loam particles are mixed, then the clay particles are added and stirred to ensure uniform particle distribution, thus obtaining the potting soil. This potting soil not only has good plasticity, allowing earthworms to form stable burrows through the synergistic effect of the clay and loam particles, but also possesses suitable permeability. The sand particles provide pores to meet the long-term respiratory needs of earthworms, while preventing the burrow walls from sticking together due to excessive stickiness or the burrow from collapsing due to excessive looseness.

[0066] The step of laying the mesh in the culture pot and filling the mesh with the culture soil includes:

[0067] S21: Lay the mesh flat in the culture pot, so that the edge of the mesh extends upward along the pot wall and beyond the pot opening by 5-8cm;

[0068] S22: The culture soil is filled into the mesh of the culture pot; the filling height of the culture soil is 4 / 5 of the height of the culture pot.

[0069] When designing the size of the cultivation pot, the diameter and height of the pot should be determined based on the body length of the target earthworm species. The diameter of the pot should be no less than 1.5 times the earthworm's body length, and the height no less than 2 times the earthworm's body length. For example, for earthworms with a body length of 8cm-10cm, a circular cultivation pot with a diameter of 15cm-18cm and a height of 20cm-25cm should be selected. When designing the drainage holes, 5-7 drainage holes should be evenly drilled in the bottom of the pot, with a hole diameter of 3mm-5mm. A layer of 200-mesh nylon mesh should be laid inside the drainage holes to prevent soil loss while ensuring excess water drainage, avoiding waterlogging and oxygen deficiency in the earthworms during long-term cultivation. When designing the square mesh covering, the side length of the mesh should be determined based on the height of the cultivation pot, and the side length should be 5 times the height of the pot. For example, if the cultivation pot is 20cm high, the side length of the mesh should be 100cm. The mesh is made of 80-100 mesh nylon, which is both breathable and supportive.

[0070] Lay the designed mesh flat inside the culture pot, extending the edge of the mesh upwards along the pot wall and beyond the rim by 5-8 cm. Then, fill the mesh-wrapped culture pot with culture soil, filling it to four-fifths of its height, ensuring the entire soil mass is completely covered by the mesh. After cultivation, the soil mass can be completely removed by lifting the edge of the mesh, avoiding direct contact with the pot wall and preventing adhesion and damage, thus providing a complete soil sample for subsequent pit wall soil collection. This design, employing both shape adaptation and mesh wrapping, ensures both container compatibility and ease of soil extraction.

[0071] Place healthy, undamaged, and viable earthworms of the target species into soil wrapped in mesh. The number of earthworms in each pot depends on the size of the pot and the size of the earthworms. For example, place 3-4 earthworms with a body length of 8cm in a 15cm diameter pot.

[0072] The cultivation time was adjusted because the formation of earthworm burrows and the stability of the burrow wall soil require long-term adaptation and action. A 30-day cultivation cycle can ensure that earthworms can fully create burrows, with each pot containing 12-15 burrows. The differences between the burrow wall soil and ordinary soil (such as organic matter enrichment and structural compaction) will reach a stable state. This avoids the problems of insufficient number of burrows and thin, easily damaged burrow walls caused by too short a cultivation time. Therefore, the optimal cultivation time was determined to be 30 days.

[0073] Adjusting the environmental parameters within the incubator includes:

[0074] S31: Set the temperature range inside the incubator to 20-25℃, and keep the temperature range inside the incubator constant during the cultivation of earthworms;

[0075] S32: Detect the soil moisture content in the incubator during the early stage of cultivation and replenish the soil in the incubator with water; the early stage of cultivation is from the start of cultivation to 7 days before the end of cultivation;

[0076] S33: In the later stage of cultivation, start the water control program, stop active watering, and use the ventilation system of the incubator to reduce the soil moisture content in the incubator to 30%; the later stage of cultivation is from 7 days before the end of cultivation to the end of cultivation.

[0077] S34: Set the incubator to a periodic light cycle of 12h light / 12h dark, with a light intensity of 500-800 lux.

[0078] The step of detecting the soil moisture content in the incubator during the early stage of cultivation and replenishing the soil in the incubator with water includes:

[0079] S321: Monitor the soil moisture content in the incubator every 3 days by gravimetric method;

[0080] S322: If the moisture content is below 40%, water is sprayed onto the surface of the culture soil in the culture box to maintain the soil moisture content at 40-60% of field capacity.

[0081] The environmental parameters within the incubator were adjusted, including temperature, humidity, and lighting settings. First, the temperature was controlled. During earthworm cultivation, the temperature was maintained at a constant 20-25℃ using the incubator's thermostat system to prevent sudden temperature changes and reduce burrowing activity due to temperature stress. Next, humidity was controlled. In the early stages of cultivation (from the start to 7 days before the end), soil moisture content was monitored every 3 days using a gravimetric method. When the soil moisture content fell below 40%, active watering was performed by spraying water onto the surface of the soil in the incubator, avoiding water flow that could damage the soil, thus maintaining a soil moisture content of 40%-60% of field capacity. During the later stages of cultivation, specifically 7 days before the end of cultivation, a water control program was initiated, and active watering was stopped. The ventilation system of the cultivation chamber was used to slowly reduce the soil moisture content, ensuring that the field water holding capacity of the soil dropped to 30% by the end of the 30-day cultivation period. At this point, the soil moisture was sufficient to ensure the stability of the burrow structure after long-term cultivation, while also reducing the adhesion between the burrow wall soil and ordinary soil, making it easier to separate. During the 30-day cultivation period, the cultivation chamber was set to a 12-hour light / 12-hour dark cycle with a light intensity of 500-800 lux, simulating the natural diurnal rhythm. This avoided direct sunlight stimulating the earthworms, as earthworms are negatively phototactic and will actively move away from the light source, tending to stay in the dark environment. This ensured the normal burrowing and growth activities of the earthworms during the 30-day cultivation period.

[0082] This embodiment describes a method for collecting earthworm burrow wall soil, including collecting raw soil, adjusting the particle composition of the raw soil to obtain culture soil; laying a mesh screen in a culture pot, filling the mesh screen with the culture soil; placing earthworms into the culture soil, placing the culture pot in a culture box, and adjusting the environmental parameters in the culture box to allow the earthworms to construct burrows within the culture soil; detecting whether a preset time has been reached, and if so, removing the culture soil and peeling off the burrow wall soil. The particle composition of the raw soil is precisely adjusted to a ratio of 30% clay, 60% loam, and 10% sand to obtain the culture soil. The culture soil balances plasticity and permeability, meeting the needs of earthworm growth and burrow construction during a 30-day long-term culture. The size of the culture pot is designed according to the different body sizes of earthworms, and the mesh screen is designed to wrap the pot according to the size of the culture pot. This solves the problem of container adaptation for earthworms of different body sizes and allows for complete soil extraction, avoiding damage caused by soil sticking to the pot wall after long-term culture. After obtaining the culture soil and the mesh-wrapped culture pot, the culture soil was first placed into the mesh-wrapped culture pot, followed by the earthworms. Finally, the culture pot containing the earthworms was placed in an incubator. The temperature, humidity, and light intensity within the incubator were adjusted, and the optimal culture time was determined to be 30 days, ensuring that the earthworms could fully construct burrows and that the burrow wall soil properties remained stable. After 30 days of earthworm culture, the culture soil was completely removed by lifting the mesh. A stiff, pointed spoon was used to precisely peel away the burrow wall soil from the earthworm burrows, ensuring that the soil sample was free of cross-contamination and structurally intact. This method allows for precise and efficient collection of earthworm burrow wall soil, and under the premise of systematically controlling the earthworm growth environment, it completely separates the earthworm burrows from different areas, preventing damage to the burrow structure.

[0083] Example 2

[0084] This embodiment provides a method for collecting soil from the walls of earthworm burrows. This embodiment describes the differences from Embodiment 1, and the method includes:

[0085] S1: Collect raw soil, adjust the particle composition of the raw soil to obtain culture soil;

[0086] S2: Lay the mesh in the culture pot and fill the mesh with the culture soil;

[0087] S3: Place earthworms into the culture soil, place the culture pot into the incubator, and adjust the environmental parameters in the incubator so that the earthworms can construct earthworm burrows in the culture soil;

[0088] S4: Check if the preset time has been reached. If so, remove the culture soil and peel off the soil from the earthworm hole wall.

[0089] This embodiment takes the soil from the burrow walls of *Euthymus spp.* as an example. First, the particle composition of the original soil is adjusted. The pretreated soil is separated into sand, loam, and clay particles by sieving. The sand, loam, and clay particles are mixed in a ratio of 10%, 60%, and 30%, respectively. During the mixing process, a layered stirring method is used. The sand and loam particles are mixed for 20 minutes first, and then the clay particles are added and stirred for another 15 minutes to ensure that the particle uniformity is ≥90%, thereby obtaining the culture soil.

[0090] Next, the culture container was designed, employing a two-pronged approach of "body shape adaptation + mesh wrapping." Since the body length of *Pleurotus ostreatus* is approximately 16cm, a cylindrical culture pot with a diameter of 50cm and a height of 60cm was chosen to fit the body shape of the worm. Seven drainage holes, each 1cm in diameter, were drilled at the bottom of the pot. The square mesh was made of 100-mesh nylon, with a side length five times the height of the pot. Since the pot's height is 60cm, the mesh side length was designed to be 300cm. The mesh was laid inside the pot, and culture soil was added, with the mesh edge extending upwards along the pot wall and beyond the rim by 5-8cm. The soil filling height was four-fifths of the pot's height; that is, for a pot height of 60cm, the soil filling height was 48cm.

[0091] After designing the culture containers, three healthy *Pleurotus ostreatus* were placed in each mesh-wrapped culture pot. The *Pleurotus ostreatus* weighed 10.0g-12.0g and had a body diameter of approximately 10mm. The mesh-wrapped culture pots were then placed in the incubator. Based on the time required for *Pleurotus ostreatus* to form burrows and for the soil properties of the burrow walls to stabilize, the optimal culture time was determined to be 30 days.

[0092] After placing the mesh-wrapped culture pots into the incubator, the temperature, humidity, and light were adjusted. The incubator temperature was controlled at 24℃ (fluctuation ±1℃), and the humidity was controlled at 100%. During the first 23 days of cultivation, water was added every 3 days using a weight method to maintain the soil moisture content at 55% of field capacity. During the later stage of cultivation, from the end of day 23 to the end of day 30, a water control program was initiated, and watering was stopped from day 24 onwards. By the end of day 30, the soil field capacity had decreased to 30%. The light was set to a periodic light cycle of 12 hours of light / 12 hours of darkness, with a light intensity of 700 lux. After 30 days of cultivation, the number of burrows of *Pteris vittata* was observed to be stable, and the burrow walls were intact and of uniform thickness (0.6mm-0.8mm). The soil was then completely removed by lifting the mesh.

[0093] Finally, the soil from the pit walls was collected. The first step involved selecting the appropriate collection tool: a custom-made stainless steel peeling spoon with an oval head, 6.4mm wide, and a hard tip (30° angle, HRC42 hardness). The second step involved sifting out impurities from the pit walls. The mesh screen was cut open to expose the soil, and a soft-bristled brush (5mm bristles) was used to sweep away surface soil in one direction. The collection area was marked, and the surrounding area was then thoroughly cleaned with the soft brush. The third step involved collecting the soil from the pit walls. Holding the hard-bristled peeling spoon, the tip was inserted into the gap in the pit wall, peeling along the inner wall from top to bottom, controlling the thickness to 0.8mm. The peeled soil was placed into numbered sterile centrifuge tubes. The tools were disinfected with 75% alcohol after each pit collection. Ultimately, the success rate of collecting the pit wall soil was 100%.

[0094] Example 3

[0095] This embodiment provides a method for collecting soil from the walls of earthworm burrows. This embodiment describes the differences between it and Embodiment 2, and the method includes:

[0096] S1: Collect raw soil, adjust the particle composition of the raw soil to obtain culture soil;

[0097] S2: Lay the mesh in the culture pot and fill the mesh with the culture soil;

[0098] S3: Place earthworms into the culture soil, place the culture pot into the incubator, and adjust the environmental parameters in the incubator so that the earthworms can construct earthworm burrows in the culture soil;

[0099] S4: Check if the preset time has been reached. If so, remove the culture soil and peel off the soil from the earthworm hole wall.

[0100] This embodiment takes the burrow wall soil of *Pheretima filamentosa* as an example. First, the particle composition of the original soil is adjusted. After separating the pretreated soil into sand, loam, and clay particles by sieving, the soil is mixed in a ratio of 10% sand, 60% loam, and 30% clay. The mixture is then stirred using the layered stirring method described in Example 2 to obtain the culture soil.

[0101] Next, the culture container was designed. The body length of *Eupolyphaga sinensis* is approximately 8cm. A circular culture pot with a diameter of 50cm and a height of 60cm was selected. Five drainage holes with a diameter of 1cm were drilled in the bottom of the pot. A square mesh made of 100-mesh nylon was used, with a side length five times the height of the pot, i.e., 300cm. Following the method in Example 2, the mesh was laid in the pot and filled with culture soil. The soil filling height was four-fifths of the pot height, i.e., 48cm.

[0102] After designing the culture containers, three healthy *Pheretima spp.* were placed in each mesh-wrapped culture pot. The *Pheretima spp.* weighed 1.5g-2.0g and had a body diameter of approximately 5mm. The mesh-wrapped culture pots were then placed in the incubator. Based on the time required for *Pheretima spp.* burrow formation and the stabilization of the burrow wall soil, the optimal culture time was determined to be 30 days.

[0103] Temperature, humidity, and light were controlled in the incubator. After placing the mesh-wrapped culture pots in the incubator, the temperature was maintained at 22℃ (fluctuation ±1℃), and humidity at 100%. For the first 23 days of cultivation, water was added every 3 days using a weight method to maintain soil moisture content at 48% of field capacity. From day 24 onwards, a water control program was initiated, and watering was stopped. By the end of cultivation on day 30, the soil field capacity had decreased to 30%. Light was set to a periodic 12h light / 12h dark cycle, with a light intensity of 600 lux. After 30 days of cultivation, the number of burrows of *Pheretima villonifera* was observed to be stable, with intact and uniform burrow walls. The intact soil was then removed by lifting the mesh.

[0104] Finally, the soil from the pit walls was collected. The first step involved selecting the appropriate collection tool: a custom-made, 4mm wide, hard-tipped stainless steel peeling spoon with a 30° tip angle and a hardness of HRC40. The second step involved sifting out impurities from the pit walls. First, the mesh screen was cut to expose the soil. A soft brush was used to sweep away surface soil in one direction. After marking the collection area, the area around the marked area was thoroughly cleaned again with a soft brush. The third step involved collecting the soil from the pit walls. Holding the hard-tipped peeling spoon, the tip was inserted into the gap in the pit wall, peeling along the inner wall from top to bottom, controlling the thickness to 0.6mm. The peeled soil was placed into numbered sterile centrifuge tubes. The tool was disinfected with 75% alcohol after each pit collection. Ultimately, the success rate of collecting the pit wall soil was 100%.

[0105] Example 4

[0106] This embodiment provides a method for collecting soil from the walls of earthworm burrows. This embodiment describes the differences from Embodiment 1, and the method includes:

[0107] S1: Collect raw soil, adjust the particle composition of the raw soil to obtain culture soil;

[0108] S2: Lay the mesh in the culture pot and fill the mesh with the culture soil;

[0109] S3: Place earthworms into the culture soil, place the culture pot into the incubator, and adjust the environmental parameters in the incubator so that the earthworms can construct earthworm burrows in the culture soil;

[0110] S4: Check if the preset time has been reached. If so, remove the culture soil and peel off the soil from the earthworm hole wall.

[0111] like Figure 2 As shown, the process of peeling the soil from the earthworm burrow wall includes:

[0112] S42: Perform a CT scan on the culture soil to obtain a diagram of the soil's internal structure;

[0113] S43: Perform connected component analysis on the soil internal structure diagram to obtain the final connected region; the final connected region is used to characterize the earthworm burrow.

[0114] S44: Select the starting and ending surfaces of the culture soil, wherein the culture soil is cylindrical in shape;

[0115] S45: Divide the final connected region into N layers of cavity wall soil along the direction from the starting surface to the ending surface; N≥2;

[0116] S46: Use an earthworm hole wall soil collection device to sequentially peel off the first layer of hole wall soil to the Nth layer of hole wall soil.

[0117] Before step S42, there is also step S41: if the earthworm culture has reached the preset time. Soil CT scanning is a non-destructive testing method for soil using modern imaging technology. It can observe the internal conditions of the soil and is widely used in agriculture, environmental monitoring, and geological exploration. X-ray CT or synchrotron radiation CT scans soil samples to generate high-resolution tomographic images, and connected component analysis is used to identify the connectivity, size, and spatial distribution of pores in the soil.

[0118] When earthworms are placed in potting soil, their burrowing activity creates numerous burrows (up to 800 burrows / m²), significantly improving soil aeration and aggregate structure. Earthworm excrement (vermicompost) further optimizes water and fertilizer management, and different ecological groups of earthworms, such as surface-dwelling and deep-dwelling types, develop differentiated porosity systems. 2

[0119] The potting soil is cylindrical, with its two bottom surfaces serving as the starting and ending surfaces. In this embodiment, the side of the potting soil furthest from the ground is used as the starting surface, and the side closest to the ground is used as the ending surface; that is, the starting surface is located above the ending surface. The final connected region within the potting soil may extend in different directions; therefore, the final connected region is divided into N layers of pit wall soil from top to bottom. The potting soil is as follows... Figure 5 As shown, the soil internal structure obtained from CT scans is as follows: Figure 6 As shown, the burrow formed by earthworms is Figure 6 The white area in the middle, the other soil areas are Figure 6 The black area in the text.

[0120] The connection domain analysis of the soil internal structure map yields the final connected regions, including:

[0121] S431: The soil internal structure map is binarized according to a preset pixel threshold to obtain a binarized image;

[0122] S432: Randomly select a point with a value of 1 in the region corresponding to the binarized image and the starting surface as the first seed point;

[0123] S433: Fill the first seed point with a seed to obtain the first connected region;

[0124] S434: Randomly select a point with a value of 1 in the region corresponding to the binarized image and the termination surface as the second seed point;

[0125] S435: Fill the second seed point with a seed to obtain the second connected region;

[0126] S436: Take the union of the first connected region and the second connected region to obtain the final connected region.

[0127] Points with values ​​greater than or equal to a preset pixel threshold in the soil internal structure image are set to 1, and points with values ​​less than the preset pixel threshold are set to 0, resulting in a binarized image. Points with values ​​of 1 in the starting surface (the surface of the cultivation soil furthest from the ground) are selected to form the first point set. A point is randomly selected from the first point set as the first seed point, and this seed point is added to the second point set. Starting from the first seed point, all points with values ​​of 1 near the first seed point are added to the second point set. Then, points with values ​​of 1 are detected centered on the current point with values ​​of 1 near the first seed point. This process is repeated, traversing in the direction from the starting surface to the ending surface, until no other points with values ​​of 1 are detected. At this point, the points in the second point set are considered as the first connected region.

[0128] The process begins by selecting points with a value of 1 on the termination surface (the surface of the soil closest to the ground), resulting in a third set of points. A point is randomly selected from this third set as a second seed point, which is then added to the fourth set. Starting from the second seed point, all nearby points with a value of 1 are added to the fourth set. Each nearby point with a value of 1 is then used as the current point, and points with a value of 1 are detected around this current point. This process is repeated, traversing the direction from the termination surface to the starting surface, until no other points with a value of 1 are detected. At this point, the points in the fourth set form the second connected region.

[0129] Traverse the earthworm burrow in two opposite directions, and take the union of the first and second connected regions. This ensures that the first and second connected regions complement each other, preventing any omissions of earthworm burrow areas due to breakpoints. This comprehensively reflects the earthworm's burrowing situation and burrow structure in the cultivation soil.

[0130] The step of seed filling the first seed point to obtain the first connected region includes:

[0131] S4331: Take a point in the neighborhood of the first seed point as the current point. If the value of the current point is 1, replace the current point with other points in the neighborhood with a value of 1. If the value of the current point is 0, count the total number of points with a value of 1 in the neighborhood of the current point. If the total number is greater than the total number threshold, set the value of the current point to 1. The neighborhood is four adjacent pixels or eight adjacent pixels.

[0132] S4332: Update the position of the current point until the position of the current point remains unchanged for M consecutive times, thus obtaining the first connected region.

[0133] During the top-down traversal, the first seed point is selected first, and then the values ​​of the pixels in its four or eight neighboring regions are checked. For example, if the coordinates of the first seed point are (x, y, z), and the current point (x, y-1, z) has a value of 1, then no processing is done on the current point (x, y-1, z), and it is replaced with (x, y+1, z), which is the point to the right of the first seed point. If the value of the current point (x, y+1, z) is 0, the total number of points with a value of 1 in the eight neighboring regions of the current point (x, y+1, z) is counted. If the total number is 5, and the threshold is 4, the total number is greater than the threshold, so the value of the current point (x, y+1, z) is set to 1. The coordinates of the current point are continuously updated, with the z-coordinate value decreasing. If the coordinates of the current point remain unchanged after M checks, the traversal stops, and the first connected region is obtained. This embodiment uses M=4 as an example. The process of traversing from bottom to top to obtain the second connected region is the same and will not be repeated here.

[0134] Example 5

[0135] like Figures 3-4 As shown, this embodiment provides an earthworm hole wall soil collection device for implementing an earthworm hole wall soil collection method. The device includes a spoon body 1 and a spoon handle 2, which are detachably connected or integrally formed. The spoon body 1 has a peeling opening 3, and a first flange 4 and a second flange 5 are respectively provided on both sides of the peeling opening 3. Insert strips 6 are provided on the side walls of the spoon body 1 near the first flange 4 and the second flange 5, and the insert strips 6 include multiple insertion holes 7. A baffle 8 is connected to one of the insertion holes 7.

[0136] The sidewalls of the spoon body 1, near the first flange 4 and the second flange 5, are provided with insert strips 6. The extension direction of the peeling opening 3 is a first direction, and the insert strips 6 extend along a second direction perpendicular to the first direction. The insert strips 6 include multiple insertion holes 7, which are adapted to the baffle 8. The first end of the baffle 8 is inserted into the first insertion hole 7 of the first insert strip 6, and the second end is inserted into the second insertion hole 7 of the second insert strip 6. The first insertion hole 7 and the second insertion hole 7 are arranged opposite to each other.

[0137] The baffle 8 can move up and down, inserting into different holes 7 to adjust the depth of the peeling opening 3, thereby adjusting the sampling thickness of the culture soil and achieving precise peeling of the earthworm burrow soil. For example, if the soil internal structure map obtained from the CT scan shows frequent earthworm activity and numerous earthworm burrows scattered throughout the culture soil, the baffle 8 is moved closer to the bottom of the peeling opening 3, resulting in a smaller thickness of culture soil peeled each time. If the soil internal structure map obtained from the CT scan shows less earthworm activity and fewer earthworm burrows concentrated in a few locations within the culture soil, the baffle 8 is moved further away from the bottom of the peeling opening 3, resulting in a larger thickness of culture soil peeled each time.

[0138] The earthworm burrow wall soil collection device of this embodiment operates on the culture soil in step S4 of Examples 1-4, peeling off the burrow wall soil from the culture soil. Since both the culture pot and the culture soil are cylindrical, based on the soil internal structure diagram obtained from CT scanning and the N layers of burrow wall soil, the burrow wall soil is peeled off layer by layer from the starting surface of the culture soil towards the ending surface, i.e., peeling from top to bottom. Alternatively, it can start from the side of the cylindrical culture soil, peeling off layer by layer from the outer layer towards the center of the culture soil.

[0139] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, apparatus, article, or method that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, apparatus, article, or method. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, apparatus, article, or method that includes that element. The above descriptions are merely preferred embodiments of the present invention and do not limit the scope of the patent. Any equivalent structural or procedural modifications made based on the description and drawings of this invention, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of this invention.

[0140] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention's specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A method for collecting soil from the walls of earthworm burrows, characterized in that, include: Collect raw soil, adjust the particle composition of the raw soil, and obtain culture soil; Lay the mesh in the culture pot and fill the mesh with the culture soil; Earthworms are placed in the culture soil, the culture pot is placed in the incubator, and the environmental parameters in the incubator are adjusted so that the earthworms construct earthworm burrows in the culture soil. If the preset time has been reached, remove the culture soil and peel off the soil from the earthworm burrow wall. The process of regulating the particle composition of the original soil to obtain cultured soil includes: Impurities are removed from the original soil to obtain pretreated soil; the impurities include stones and tree roots. Sand, soil, and clay particles were separated by sieving. The culture soil was prepared by mixing clay, loam and clay in a ratio of 30% clay, 60% loam and 10% sand using a layered mixing method. Adjusting the environmental parameters within the incubator includes: In the early stage of cultivation, the soil moisture content in the cultivation box is measured, and water is added to the soil in the cultivation box to maintain the soil moisture content at 40-60% of the field capacity. The early stage of cultivation is from the start of cultivation to 7 days before the end of cultivation. In the later stage of cultivation, the water control program is initiated, active water replenishment is stopped, and the soil moisture content in the cultivation chamber is reduced to 30% using the ventilation system of the cultivation chamber; the later stage of cultivation is from 7 days before the end of cultivation to the end of cultivation. The process of stripping the soil from the earthworm burrow wall includes: The culture soil was subjected to CT scanning to obtain a diagram of the soil's internal structure. Connectivity analysis was performed on the soil internal structure diagram to obtain the final connected regions; the final connected regions were used to characterize the earthworm burrow. Select the starting and ending surfaces of the culture soil, wherein the culture soil is cylindrical in shape; Along the direction from the starting surface to the ending surface, the final connected region is divided into N layers of pit wall soil; N≥2; An earthworm hole wall soil collection device was used to sequentially peel off the first layer of hole wall soil to the Nth layer of hole wall soil. The connection domain analysis of the soil internal structure map yields the final connected regions, including: The soil internal structure map is binarized according to a preset pixel threshold to obtain a binarized image; A point with a value of 1 in the region corresponding to the binarized image and the starting surface is randomly selected as the first seed point; The first seed point is seed-filled to obtain the first connected region; Randomly select a point with a value of 1 in the region corresponding to the binarized image and the termination surface as the second seed point; Seed filling is performed on the second seed point to obtain the second connected region; The final connected region is obtained by taking the union of the first connected region and the second connected region. The earthworm hole wall soil collection device includes a spoon body and a spoon handle, which are detachably connected or integrally formed; the spoon body has a peeling opening, and a first flange and a second flange are respectively provided on both sides of the peeling opening; the side walls of the spoon body near the first flange and the second flange are provided with inserts, and the inserts include multiple insertion holes; the two ends of the baffle are respectively inserted into two oppositely arranged insertion holes, and the baffle can move up and down.

2. The method for collecting soil from earthworm burrow walls according to claim 1, characterized in that, The size of the sand particles is greater than 0.05 mm and less than 2 mm; the size of the soil particles is less than or equal to 0.05 mm and greater than or equal to 0.002 mm; the size of the clay particles is less than 0.002 mm.

3. The method for collecting soil from earthworm burrow walls according to claim 1, characterized in that, The step of laying the mesh in the culture pot and filling the mesh with the culture soil includes: Lay the mesh flat in the culture pot, so that the edge of the mesh extends upward along the pot wall and beyond the pot opening by 5-8cm; The culture soil is filled into the mesh of the culture pot; the filling height of the culture soil is equal to the height of the culture pot. .

4. The method for collecting soil from earthworm burrow walls according to claim 1, characterized in that, Adjusting the environmental parameters within the incubator includes: The temperature range inside the incubator is set to 20-25℃, and the temperature range inside the incubator is kept constant during the cultivation of earthworms. The incubator is set to a periodic light cycle of 12 hours of light and 12 hours of darkness, with a light intensity of 500-800 lux.

5. The method for collecting soil from earthworm burrow walls according to claim 1, characterized in that, The step of detecting the soil moisture content in the incubator during the early stage of cultivation and replenishing the soil in the incubator with water includes: The soil moisture content in the incubator was monitored every 3 days by gravimetric method. If the moisture content is less than 40%, water is sprayed onto the surface of the culture soil in the incubator using a spraying method.

6. The method for collecting soil from earthworm burrow walls according to claim 1, characterized in that, The step of seed filling the first seed point to obtain the first connected region includes: Take a point in the neighborhood of the first seed point as the current point. If the value of the current point is 1, replace the current point with other points in the neighborhood with a value of 1. If the value of the current point is 0, count the total number of points with a value of 1 in the neighborhood of the current point. If the total number is greater than the total number threshold, set the value of the current point to 1. The neighborhood is four adjacent pixels or eight adjacent pixels. Update the position of the current point until the position of the current point remains unchanged for M consecutive times, and obtain the first connected region.

Images