Beef cattle living body tissue nondestructive sampling method based on precise positioning

Abstract

The invention provides a beef cattle living body tissue lossless sampling method based on precise positioning, which comprises the following steps of: firstly, fixing beef cattle in a sampling frame and anesthetizing the beef cattle to reduce the stress response of the beef cattle; an ultrasonic imaging device and a three-dimensional positioning system are used for accurately positioning a target sampling part, and an optimal sampling path and sampling points are planned. A disposable sterile sampling needle with the outer diameter of 1.2-1.5 mm and the length of 5-10 cm is selected, and sampling operation is carried out after steam sterilization treatment. The sampling process strictly follows the sterile principle, and beef cattle vital signs and the position of the sampling needle are monitored in real time. And the collected tissue sample is cleaned and then is properly stored. The sampling part is treated and bound after the operation, and the beef cattle recovery condition is closely observed. According to the method, accurate positioning of beef cattle living tissue sampling is achieved, damage to beef cattle is reduced, sampling accuracy and representativeness are improved, and reliable sample data support is provided for beef cattle health assessment, disease diagnosis and variety improvement.

Description

Technical Field

[0001] The present invention relates to the technical field of beef cattle breeding, and in particular, to a non-destructive sampling method for living tissues of beef cattle based on precise positioning. Background Art

[0002] In the modern beef cattle breeding industry and the in-depth process of scientific research, sampling and analysis of living tissues of beef cattle are of extremely crucial and irreplaceable significance. As an important part of the agricultural economy, the development level of the beef cattle breeding industry is directly related to the supply and quality of the meat market. Moreover, the health status, growth performance, and genetic characteristics of beef cattle have a profound impact on the breeding efficiency and industrial development.

[0003] From the perspective of health status, through the detection of living tissues of beef cattle, various potential disease hazards in the body of beef cattle can be detected promptly and accurately. For example, in the initial stage of certain virus or bacterial infections, there may be no obvious symptoms on the appearance of beef cattle, but corresponding pathological changes will occur in their tissue cells. By sampling and analyzing specific tissues, these subtle changes can be detected in advance, providing a basis for the early diagnosis and treatment of diseases. This not only helps to reduce the economic losses caused by the death of beef cattle due to diseases but also effectively prevents the spread and diffusion of diseases, ensuring the health of the entire breeding population.

[0004] In terms of growth performance, tissue sample analysis can reveal the internal mechanism of the growth and development of beef cattle. For example, by detecting the expression of relevant genes and the levels of metabolites in muscle tissues, information about the growth rate and meat quality of beef cattle can be obtained. Based on this information, breeders can adjust the feed formula and feeding management methods targeted to improve the growth performance and economic benefits of beef cattle. For instance, if it is found that a certain beef cattle has a slow growth rate due to its weak ability to absorb and utilize certain nutrients, corresponding nutritional supplements can be added to the feed or the feed composition can be adjusted to meet its growth requirements.

[0005] In terms of genetic characteristics, sampling and analysis of living tissues of beef cattle can provide solid data support for breed improvement. Through gene detection technology, the genetic background of beef cattle can be deeply understood, and individuals with excellent traits can be selected for breeding, thus gradually cultivating beef cattle breeds that better meet market demands.

[0006] However, examining the existing live bovine tissue sampling methods currently reveals many deficiencies that urgently need to be addressed. In terms of precise positioning, the existing sampling methods rely heavily on the personal experience of the operators. During the actual operation, the operators often determine the sampling position solely based on visual observation and their own sense of touch. However, the body structure of beef cattle is complex, and the tissue distribution has certain differences and concealment. Such a sampling method lacking scientific and precise positioning is extremely likely to lead to deviations in the sampling position. For example, when collecting muscle tissue samples, if the sampling position is inaccurate, it may obtain adipose tissue or other non-target tissues, thus failing to obtain a representative sample. This will directly affect the accuracy and reliability of subsequent test results, making the breeding strategies, disease prevention and control measures, and breed improvement plans based on these test results lack scientificity and effectiveness.

[0007] From the perspective of the damage caused to beef cattle, the disadvantages of traditional sampling methods are obvious. During the sampling process, due to the lack of fineness in operation, it is very easy to cause relatively large trauma to the body of beef cattle. Taking the commonly used knife cutting sampling method as an example, this method not only causes a large amount of bleeding in beef cattle, increasing the risk of beef cattle being infected with bacteria and viruses, but also makes beef cattle suffer great pain. Moreover, excessive bleeding may affect the physiological functions of beef cattle, resulting in hindered growth and development and reduced breeding efficiency. Therefore, we made improvements and proposed a non-destructive sampling method for live bovine tissue based on precise positioning. Summary of the Invention

[0008] The purpose of the present invention is to address the problems raised in the existing background technology. To achieve the above-mentioned invention purpose, the present invention provides the following technical solutions: A non-destructive sampling method for live bovine tissue based on precise positioning, comprising the following steps: Step 1: Guide the beef cattle to be sampled into the sampling fixing frame and fix it. Inject an anesthetic into the beef cattle by intramuscular injection to make it enter the anesthetic state. The anesthetic is ketamine hydrochloride, and the injection dose is 2-3 mg per 100 kg body weight; Step 2: Use an ultrasonic imaging device to scan the target sampling part of the beef cattle. Set the scanning frequency to 3-5 MHz. After determining the position, size, and depth of the target tissue, mark it on the body surface of the beef cattle. Then, with the help of a high-precision three-dimensional positioning system and ultrasonic imaging data, establish a three-dimensional model of the beef cattle body, and plan the sampling path and sampling points on this model; among them, the positioning accuracy of the high-precision three-dimensional positioning system is ±0.5 mm; Step 3: Select a disposable sterile sampling needle according to the target tissue type and sampling requirements, and disinfect the sampling needle and related instruments; if the target tissue is muscle tissue, select a sampling needle with an inner diameter of 1.8 mm and a length of 100 mm; if the target tissue is adipose tissue, select a sampling needle with an inner diameter of 2.0 mm and a length of 80 mm; the disinfection treatment is carried out by soaking in 75% (v / v) alcohol for more than 30 minutes; Step 4: Disinfect the marked sampling site 2-3 times with 0.5% (v / v) povidone-iodine, then cover it with a sterile surgical drape. The operator holds the sampling needle and inserts it into the beef cattle body along the planned sampling path to the target tissue, collect a tissue sample with a volume of 0.5-1.0 cm³. After collection, pull out the sampling needle and press the sampling site for 3-5 minutes; Step 5: Put the collected tissue sample into a sterile petri dish containing normal saline with a concentration of 0.9% and wash it 2-3 times, then put it into a sample bottle containing a fixative, which is a 4% (v / v) paraformaldehyde solution, seal it and make a mark; Step 6: Conduct routine disinfection treatment on the marked sampling site. Degrease it twice with 75% (v / v) medical alcohol, with each wiping time not less than 30 seconds. Then disinfect it twice with an iodophor solution containing an effective iodine concentration of 0.5%, with an interval of 1 minute. Finally, deiodize it once with 75% (v / v) alcohol; cover it with a sterilized sterile surgical drape and leave a sampling window with a diameter of 3-5 cm at the sampling site; The operator wears disposable medical sterilized gloves, holds the sampling needle and slowly inserts it into the beef cattle body along the path planned in the three-dimensional model. The puncture speed is controlled at 1.5-2.5 cm / s to ensure that the tissue is not violently disturbed; during the puncture process, the real-time feedback of the tip position and angle deviation is obtained through the built-in micro inertial measurement unit (IMU) and the external laser positioning system. The maximum allowable angle error does not exceed ±2°, and the depth error does not exceed ±0.3 mm; the length of the collected sample is not less than 1.2 cm, and the diameter is controlled between 1.5-3.0 mm; after sampling, quickly pull out the sampling needle, and immediately press the puncture site with a sterile gauze at a pressure of about 10 kPa for 3-4 minutes; Step 7: Place the collected tissue sample in a sterile petri dish containing 0.9% normal saline with a pH of 7.2-7.4, and use a low-speed oscillating cleaning method with a frequency of 30-60 rpm for 1-2 minutes to remove blood residues; then, through a micro-microscopic imaging system (magnification of 10×-40×, resolution ≤1μm / pixel), conduct a preliminary assessment of the tissue integrity and cell viability of the sample; The evaluation content includes: whether the tissue structure is complete; whether there is fracture or fragmentation; the nuclear staining condition; a tissue activity index ≥80% is regarded as a qualified sample; The qualified samples are transferred to sample bottles containing 10% neutral formalin fixative with a pH of 6.8 - 7.2, sealed and stored. The sample bottles are labeled with the sampling date, beef cattle ear tag number, sampling site name, and sampling personnel information. The samples need to be transported under the condition of 4 ± 1°C and must be sent to the laboratory for pathological analysis or molecular detection within 48 hours. Step 8: The entire sampling process is transmitted to the remote terminal in real time through a high - definition camera system (resolution not less than 1080P, frame rate 30 fps), supporting the 5G communication protocol (network latency ≤ 50 ms), to achieve remote expert consultation and operation guidance. All operation data, including images, path parameters, physiological indicators, and sample information, are automatically recorded and uploaded to the cloud database, with the storage format being a mixed mode of JSON + DICOM, forming a complete electronic sampling file, and the preservation period is not less than 5 years for easy later traceability and quality control. Step 9: Post - operative care and observation: Apply a broad - spectrum antibiotic ointment to the sampling site, with a thickness controlled at 1 - 2 mm, cover it with a sterile dressing sized 5 cm × 5 cm, and bandage it with an elastic bandage, with the tightness controlled to the extent that two fingers can be inserted. Transfer the beef cattle to a quiet and clean recovery cattle shed, keep the environmental temperature at 15 - 25°C, relative humidity at 50% - 70%, observe for at least 2 hours, and monitor its respiratory rate. Continuously observe for 24 - 48 hours. If local swelling, bleeding, or body temperature rising above 39.5°C occurs, then administer oral or injectable broad - spectrum antibiotics, including ceftiofur sodium, with a dose of 1 - 2 mg / kg body weight, once a day. Step 10: Post - operative data archiving and analysis feedback: After sampling, the system automatically generates a sampling report template in PDF format, including the following content: sampling path diagram, in SVG vector graphic format; sample quality assessment results, tissue integrity score, and percentage of cell viability.

[0009] As a preferred technical solution of the present invention, when guiding the beef cattle to be sampled into the sampling fixing frame and fixing it, a hydraulic device and adjusting straps are used to firmly fix the body of the beef cattle to ensure that the beef cattle does not move during the sampling process.

[0010] As a preferred technical solution of the present invention, when injecting anesthetic into the beef cattle by intramuscular injection, the injection amount of the anesthetic is determined according to the weight and health status of the beef cattle.

[0011] As a preferred technical solution of the present invention, when selecting a disposable sterile sampling needle, the outer diameter of the sampling needle is 1.2 - 1.5 mm and the length is 5 - 10 cm.

[0012] As a preferred technical solution of the present invention, when disinfecting the sampling needle and related instruments, put them into a steam sterilizer and sterilize them at 121°C for 20 - 30 minutes.

[0013] As a preferred technical solution of the present invention, when disinfecting the marked sampling site, strict disinfection is carried out with iodophor, and the number of disinfection times is not less than 3 times.

[0014] As a preferred technical solution of the present invention, during the process of the operator inserting the sampling needle into the beef cattle body along the planned sampling path, the vital signs of the beef cattle and the position of the sampling needle are monitored in real time.

[0015] As a preferred technical solution of the present invention, when placing the collected tissue sample into a sterile culture dish containing normal saline for cleaning, the sample is gently rinsed to remove the blood and impurities on the surface.

[0016] As a preferred technical solution of the present invention, when applying an antibiotic ointment to the sampling site, a drug with anti-infective effect such as erythromycin ointment is selected.

[0017] As a preferred technical solution of the present invention, when transferring the beef cattle to the cowshed for observation and nursing, the vital signs of the beef cattle and the wound healing condition are closely observed, and sufficient drinking water and easily digestible feed are given.

[0018] Compared with the prior art, the beneficial effects of the present invention are as follows: Through the combination of an ultrasonic imaging device and a three-dimensional positioning system, the present invention can accurately determine the sampling site and plan the sampling path, ensuring that the sampling needle accurately reaches the target tissue, and improving the accuracy and representativeness of sampling. The present invention uses a disposable sterile sampling needle with a smaller outer diameter, which causes less damage to the beef cattle during the puncture process. At the same time, effective wound treatment and nursing are carried out after the operation, reducing the occurrence of bleeding and infection complications, and realizing non-destructive sampling of the living tissue of the beef cattle. Since the present invention can obtain representative tissue samples, it provides reliable data for subsequent detection and analysis, and helps to improve the accuracy of beef cattle health status assessment, disease diagnosis and breed improvement work. Description of the Drawings

[0019] Figure 1 It is a test data block diagram provided by the present invention; Figure 2 It is a test data block diagram provided by the present invention. Detailed Embodiments

[0020] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings. Obviously, the described embodiments are a specific implementation manner of the present invention and do not cover all embodiments.

[0021] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely represents some embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0022] It should be noted that, without conflict, the embodiments in the present invention and the features and technical solutions in the embodiments can be combined with each other. It should be noted that similar reference numerals and letters denote similar items in the following drawings. Therefore, once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0023] Embodiment 1: A non-destructive sampling method for beef cattle living tissue based on precise positioning, comprising the following steps: Step 1: Guide the beef cattle to be sampled into the sampling fixing frame and fix it. Inject anesthetic into the beef cattle by intramuscular injection to make it enter the anesthetic state. The anesthetic is ketamine hydrochloride, and the injection dose is 2 - 3 mg per 100 kg body weight. Step 2: Use an ultrasonic imaging device to scan the target sampling site of the beef cattle. Set the scanning frequency to 3 - 5 MHz. After determining the position, size, and depth of the target tissue, mark it on the body surface of the beef cattle. Then, with the help of a high-precision three-dimensional positioning system and ultrasonic imaging data, establish a three-dimensional model of the beef cattle body, and plan the sampling path and sampling points on this model; wherein, the positioning accuracy of the high-precision three-dimensional positioning system is ±0.5 mm. Step 3: Select a disposable sterile sampling needle according to the target tissue type and sampling requirements, and disinfect the sampling needle and related instruments; if the target tissue is muscle tissue, select a sampling needle with an inner diameter of 1.8 mm and a length of 100 mm; if the target tissue is adipose tissue, select a sampling needle with an inner diameter of 2.0 mm and a length of 80 mm; the disinfection treatment is to soak in 75% (volume fraction) alcohol for more than 30 minutes. Step 4: Disinfect the marked sampling site 2 - 3 times with 0.5% (volume fraction) povidone iodine, then lay a sterile surgical drape. The operator holds the sampling needle and inserts it into the beef cattle body to the target tissue according to the planned sampling path, collect the tissue sample. The volume of the collected tissue sample is 0.5 - 1.0 cm³. After collection, pull out the sampling needle and press the sampling site for 3 - 5 minutes. Step 5: Put the collected tissue sample into a sterile culture dish containing normal saline with a concentration of 0.9% and wash it 2 - 3 times, then put it into a sample bottle containing a fixative, and the fixative is a 4% (volume fraction) paraformaldehyde solution, seal it for storage and make a mark. Step 6: Perform routine disinfection on the marked sampling site. Degrease it twice with 75% medical alcohol, with each wiping time not less than 30 seconds. Subsequently, disinfect it twice with an iodophor solution containing an effective iodine concentration of 0.5%, with an interval of 1 minute. Finally, deiodize it once with 75% alcohol; cover it with a sterilized sterile surgical drape and leave a sampling window with a diameter of 3 - 5 cm at the sampling site; The operator wears disposable medical sterilized gloves and slowly inserts the sampling needle into the beef cattle body along the path planned in the three-dimensional model. The puncture speed is controlled at 1.5 - 2.5 cm / s to ensure that the tissue is not violently disturbed; during the puncture process, the real-time feedback of the needle tip position and angle deviation is obtained through the built-in micro inertial measurement unit (IMU) and the external laser positioning system. The maximum allowable angle error does not exceed ±2°, and the depth error does not exceed ±0.3 mm; the length of the collected sample is not less than 1.2 cm, and the diameter is controlled between 1.5 - 3.0 mm; after sampling, quickly pull out the sampling needle and immediately press the puncture site with a sterile gauze at a pressure of about 10 kPa for 3 - 4 minutes; Step 7: Place the collected tissue sample in a sterile petri dish containing 0.9% normal saline, pH 7.2 - 7.4, and use a low-speed oscillating cleaning method with a frequency of 30 - 60 rpm for 1 - 2 minutes to remove blood residues; subsequently, conduct a preliminary assessment of tissue integrity and cell viability on the sample through a micro-microscopic imaging system (magnification 10× - 40×, resolution ≤1μm / pixel); The assessment content includes: whether the tissue structure is complete; whether there are breaks or fractures; the nuclear staining situation; a tissue activity index ≥80% is regarded as a qualified sample; Transfer the qualified sample to a sample bottle containing 10% neutral formalin fixative, pH 6.8 - 7.2, and seal it for storage. The sample bottle is marked with the sampling date, beef cattle ear tag number, sampling site name, and sampling personnel information; the sample needs to be transported under the condition of 4 ± 1°C and must be sent to the laboratory for pathological analysis or molecular detection within 48 hours; Step 8: The entire sampling process is transmitted to the remote terminal in real time through a high-definition camera system (resolution not less than 1080P, frame rate 30 fps), supporting the 5G communication protocol (network delay ≤50 ms) to achieve remote expert consultation and operation guidance; all operation data, including images, path parameters, physiological indicators, and sample information, are automatically recorded and uploaded to the cloud database, and the storage format is a mixed mode of JSON + DICOM, forming a complete electronic sampling file with a storage period of not less than 5 years for easy later traceability and quality control; Step 9. Postoperative care and observation: Apply a broad-spectrum antibiotic ointment to the sampling site with a thickness controlled at 1 - 2 mm, cover it with a sterile dressing sized 5 cm × 5 cm, and bandage it with an elastic bandage. The tightness should be controlled to the extent that two fingers can be inserted. Transfer the beef cattle to a quiet and clean recovery cattle shed, keep the environmental temperature at 15 - 25°C and the relative humidity at 50% - 70%. Observe for at least 2 hours and monitor its respiratory rate. Continuously observe for 24 - 48 hours. If local swelling, bleeding, or body temperature rising above 39.5°C occurs, administer oral or injectable broad-spectrum antibiotics, including ceftiofur sodium, at a dose of 1 - 2 mg / kg body weight, once a day. Step 10. Postoperative data archiving and analysis feedback: After sampling, the system automatically generates a sampling report template in PDF format, which includes the following content: sampling path diagram in SVG vector graphic format; sample quality assessment results, tissue integrity score, and percentage of cell viability.

[0024] When guiding the beef cattle to be sampled into the sampling fixing frame and fixing it, use a hydraulic device and adjusting straps to firmly fix the body of the beef cattle to ensure that the beef cattle does not move during the sampling process.

[0025] When injecting anesthetic into the beef cattle by intramuscular injection, determine the injection amount of the anesthetic according to the weight and health status of the beef cattle.

[0026] When selecting a disposable sterile sampling needle, the outer diameter of the sampling needle is 1.2 - 1.5 mm and the length is 5 - 10 cm.

[0027] When disinfecting the sampling needle and related instruments, put them into a steam sterilizer and sterilize at 121°C for 20 - 30 minutes.

[0028] When disinfecting the marked sampling site, strictly disinfect it with iodophor for no less than 3 times.

[0029] During the process of the operator inserting the sampling needle into the beef cattle according to the planned sampling path, continuously monitor the vital signs of the beef cattle and the position of the sampling needle.

[0030] When putting the collected tissue sample into a sterile petri dish containing physiological saline for cleaning, gently rinse the sample to remove the blood and impurities on the surface.

[0031] When applying an antibiotic ointment to the sampling site, select a drug with anti-infective effect such as erythromycin ointment.

[0032] When transferring the beef cattle to the cattle shed for observation and care, closely observe the vital signs of the beef cattle and the wound healing situation, and provide sufficient drinking water and easily digestible feed.

[0033] Example 2: A non-destructive sampling method for live bovine tissue based on precise positioning. Select a beef cattle weighing approximately 500 kg as the sampling object. The specific sampling steps are as follows: Step 1: Guide the beef cattle to be sampled into the sampling fixing frame and fix it. Inject an anesthetic into the beef cattle by intramuscular injection to make it enter the anesthetic state. The anesthetic is ketamine hydrochloride, and the injection dose is 2 - 3 mg per 100 kg body weight; Step 2: Use an ultrasonic imaging device to scan the target sampling site of the beef cattle. Set the scanning frequency to 3 - 5 MHz. After determining the position, size, and depth of the target tissue, mark it on the body surface of the beef cattle. Then, with the help of a high-precision three-dimensional positioning system and ultrasonic imaging data, establish a three-dimensional model of the beef cattle's body, and plan the sampling path and sampling points on this model; among them, the positioning accuracy of the high-precision three-dimensional positioning system is ±0.5 mm; Step 3: Select a disposable sterile sampling needle according to the type of target tissue and sampling requirements, and disinfect the sampling needle and related instruments; if the target tissue is muscle tissue, select a sampling needle with an inner diameter of 1.8 mm and a length of 100 mm; if the target tissue is adipose tissue, select a sampling needle with an inner diameter of 2.0 mm and a length of 80 mm; the disinfection treatment is to soak in 75% (v / v) alcohol for more than 30 minutes; Step 4: Disinfect the marked sampling site 2 - 3 times with 0.5% (v / v) povidone-iodine, then lay a sterile surgical drape. The operator holds the sampling needle and inserts it into the beef cattle's body to the target tissue according to the planned sampling path, collect the tissue sample, and the volume of the collected tissue sample is 0.5 - 1.0 cm³. After collection, pull out the sampling needle and press the sampling site for 3 - 5 minutes; Step 5: Put the collected tissue sample into a sterile petri dish containing normal saline with a concentration of 0.9% and wash it 2 - 3 times, then put it into a sample bottle containing a fixative, and the fixative is a 4% (v / v) paraformaldehyde solution, seal it for storage and make a good mark; Step 6: Conduct routine disinfection treatment on the marked sampling site. Degrease it twice with 75% medical alcohol, with each wiping time not less than 30 seconds. Then disinfect it twice with an iodophor solution containing an effective iodine concentration of 0.5%, with an interval of 1 minute. Finally, deiodize it once with 75% alcohol; lay a sterilized sterile surgical drape and leave a sampling window with a diameter of 3 - 5 cm at the sampling site; The operator wears disposable medical sterilized gloves and slowly inserts the sampling needle into the beef cattle body along the path planned in the three-dimensional model by hand, with the puncture speed controlled at 1.5 - 2.5 cm / s to ensure that the tissue is not severely disturbed; during the puncture process, the real-time feedback of the needle tip position and angle deviation is obtained through the built-in micro inertial measurement unit (IMU) and the external laser positioning system, allowing the maximum angle error not to exceed ±2°, and the depth error not to exceed ±0.3 mm; the length of the collected sample is not less than 1.2 cm, and the diameter is controlled between 1.5 - 3.0 mm; after sampling, the sampling needle is quickly withdrawn, and the puncture site is immediately pressed with a sterile gauze at a pressure of about 10 kPa for 3 - 4 minutes; Step 7: Place the collected tissue sample in a sterile culture dish containing 0.9% normal saline with a pH of 7.2 - 7.4, and use a low-speed oscillating cleaning method with a frequency of 30 - 60 rpm for 1 - 2 minutes to remove residual blood; subsequently, the sample is preliminarily evaluated for tissue integrity and cell viability through a micro microscopic imaging system (magnification 10× - 40×, resolution ≤1μm / pixel); The evaluation content includes: whether the tissue structure is complete; whether there are breaks or fractures; the nuclear staining condition; a tissue activity index ≥80% is regarded as a qualified sample; The qualified sample is transferred to a sample bottle containing 10% neutral formalin fixative with a pH of 6.8 - 7.2 and sealed for storage. The sample bottle is labeled with the sampling date, beef cattle ear tag number, sampling site name, and sampling personnel information; the sample needs to be transported under the condition of 4 ± 1°C and must be sent to the laboratory for pathological analysis or molecular detection within 48 hours; Step 8: The entire sampling process is transmitted to the remote terminal in real time through a high-definition camera system (resolution not less than 1080P, frame rate 30 fps), supporting the 5G communication protocol (network delay ≤50 ms) to achieve remote expert consultation and operation guidance; all operation data, including images, path parameters, physiological indicators, and sample information, are automatically recorded and uploaded to the cloud database, and the storage format is a mixed mode of JSON + DICOM, forming a complete electronic sampling file with a storage period of not less than 5 years for easy later traceability and quality control; Step 9: Postoperative care and observation: Apply a broad-spectrum antibiotic ointment to the sampling site with a thickness controlled at 1 - 2 mm, cover it with a sterile dressing with a size of 5 cm × 5 cm, and bandage it fixed with an elastic bandage, with the tightness controlled to the extent that two fingers can be inserted; Transfer the beef cattle to a quiet and clean recovery cattle shed, maintain the environmental temperature at 15 - 25 °C, and the relative humidity at 50% - 70%. Observe for at least 2 hours and monitor their respiratory rate; continuously observe for 24 - 48 hours. If local swelling, bleeding, or a body temperature rise exceeding 39.5 °C occurs, administer oral or injectable broad-spectrum antibiotics, including ceftiofur sodium, at a dose of 1 - 2 mg / kg body weight, once a day. Step 10: Postoperative data archiving and analysis feedback: After the sampling is completed, the system automatically generates a sampling report template in PDF format, which includes the following content: a sampling path diagram in SVG vector graphics format; the sample quality assessment results, tissue integrity score, and percentage of cell viability.

[0034] Transfer the beef cattle to a quiet cattle shed and closely observe the vital signs and wound healing of the beef cattle. Provide sufficient drinking water and easily digestible feed after the operation. After 7 days, the wounds of the beef cattle heal well and no infection complications occur.

[0035] Example 3: A non-destructive sampling method for living tissue of beef cattle based on precise positioning: A certain beef cattle breeding and research institution needs to sample the living tissue of beef cattle in order to study the gene expression of specific tissues in beef cattle. In order to ensure that the sampling process causes the least damage to the beef cattle and the sampling is precise, it is decided to adopt this non-destructive sampling method for living tissue of beef cattle based on precise positioning.

[0036] Implementation materials and equipment: Beef cattle: Select 10 healthy beef cattle to be sampled with a body weight of 500 - 600 kg.

[0037] Equipment: Sampling fixing frame (equipped with a hydraulic device and adjustable straps), ultrasonic imaging equipment, high-precision three-dimensional positioning system, disposable sterile sampling needles (outer diameter 1.2 - 1.5 mm, length 5 - 10 cm), autoclave, sterile culture dish, sample bottle, iodophor, antibiotic ointment (erythromycin ointment), syringe, anesthetic, normal saline, fixing solution. Specific implementation steps

[0038] Fixing and anesthetizing the beef cattle: Guide the 10 beef cattle to be sampled into the sampling fixing frame in sequence, and use the hydraulic device and adjustable straps to firmly fix the body of the beef cattle to ensure that the beef cattle do not move during the sampling process. The veterinarian determines the injection amount of the anesthetic according to the body weight and health status of each beef cattle. Inject an appropriate amount of anesthetic into the beef cattle by intramuscular injection to make the beef cattle enter the anesthetized state.

[0039] Target tissue positioning and path planning: Use ultrasonic imaging equipment to scan the target sampling site of the beef cattle, which is a specific muscle tissue in this implementation. After determining the position, size, and depth of the target tissue, mark it on the body surface of the beef cattle.

[0040] With the help of a high-precision three-dimensional positioning system combined with ultrasonic imaging data, a three-dimensional model of the beef cattle body is established. On this model, the sampling path and sampling points are planned to ensure the accuracy of sampling.

[0041] Sampling needle and instrument disinfection: Select a disposable sterile sampling needle with an outer diameter of 1.2 - 1.5 mm and a length of 5 - 10 cm, and prepare the relevant instruments at the same time.

[0042] Put the sampling needle and relevant instruments into a steam sterilizer and sterilize them at 121 °C for 20 - 30 minutes for strict disinfection.

[0043] Tissue sample collection: At the marked sampling site, conduct strict disinfection with iodophor, and the number of disinfection times is not less than 3 times. After disinfection, lay a sterile surgical drape.

[0044] The operator holds the sampling needle according to the planned sampling path and inserts it into the beef cattle body to the target tissue. During the insertion process, the vital signs of the beef cattle and the position of the sampling needle are monitored in real time to ensure the safety of the sampling process. After collecting an appropriate amount of tissue samples, pull out the sampling needle and press the sampling site.

[0045] Sample processing and preservation: Put the collected tissue samples into a sterile petri dish containing physiological saline, gently rinse the samples to remove the blood and impurities on the surface.

[0046] Then put the samples into a sample bottle filled with fixative, seal and preserve them, and make good marks, recording the beef cattle number, sampling time, and sampling site information.

[0047] Postoperative treatment and care: Apply an antibiotic ointment with anti-infective effect, erythromycin ointment, to the sampling site and bandage it.

[0048] Transfer the beef cattle to the cowshed for observation and care. Closely observe the vital signs of the beef cattle and the wound healing situation, and provide sufficient drinking water and easily digestible feed.

[0049] Through this implementation, the target tissue samples of 10 beef cattle were successfully collected. The sampling success rate reached 100%. Moreover, during the sampling process, the stress response of the beef cattle was small, the postoperative wound healed well, and no infection complications occurred. The quality of the collected tissue samples was high, meeting the requirements of subsequent scientific research implementation. This indicates that this non-destructive sampling method for beef cattle living tissue based on precise positioning has high feasibility and effectiveness.

[0050] Test example Test purpose Verify the feasibility, accuracy, and safety of a non-destructive sampling method for beef cattle living tissue based on precise positioning.

[0051] Test materials Experimental animals: 30 healthy beef cattle with similar body weights were selected and randomly divided into an experimental group and a control group, with 15 animals in each group.

[0052] Sampling equipment: Non-destructive sampling device equipped with ultrasonic imaging equipment and three-dimensional positioning system, anesthetic, and disinfection supplies.

[0053] Experimental methods Experimental group: A non-destructive sampling method for living tissue of beef cattle based on precise positioning was adopted. The beef cattle were guided into the sampling fixing frame for fixation, and anesthetic was injected intramuscularly. The ultrasonic imaging equipment and three-dimensional positioning system were used to determine the target sampling site, and the optimal sampling path and sampling points were planned. The non-destructive sampling device was used to collect tissue samples according to the planned path.

[0054] Control group: The traditional sampling method was adopted, and the sampling site was roughly determined based on experience for tissue sampling.

[0055] Experimental data Sampling accuracy Experimental group: Through subsequent analysis of the sampled tissue, the accuracy rate of sampling the target tissue reached over 95%.

[0056] Control group: The accuracy rate of sampling the target tissue was approximately 70%.

[0057] Sampling time Experimental group: The average sampling time for each beef cattle was 10 - 15 minutes.

[0058] Control group: The average sampling time for each beef cattle was 20 - 30 minutes.

[0059] Stress response of beef cattle Experimental group: The proportion of beef cattle showing mild stress responses (such as brief increase in breathing rate and heart rate) after sampling was 20%, and the duration of the stress response was short, with an average of 1 - 2 hours.

[0060] Control group: The proportion of beef cattle showing obvious stress responses (such as violent struggling and prolonged rapid breathing) after sampling was 60%, and the average duration of the stress response was 3 - 5 hours.

[0061] Tissue damage situation Experimental group: Through subsequent observation of the sampling site, the degree of tissue damage was relatively light, the wound was small, and the average healing time was 3 - 5 days.

[0062] Control group: The degree of tissue damage was relatively severe, the wound was large, and the average healing time was 7 - 10 days.

[0063] Data analysis Accuracy analysis: The sampling accuracy rate of the experimental group was significantly higher than that of the control group, indicating that the sampling method based on precise positioning can obtain the target tissue more accurately and reduce sampling errors.

[0064] Time efficiency analysis: The sampling time of the experimental group was significantly shorter than that of the control group, indicating that this method improved the sampling efficiency and could complete the sampling operation in a shorter time.

[0065] Stress response analysis: The proportion and duration of the stress response of beef cattle in the experimental group were lower than those in the control group, indicating that this method can effectively reduce the stress level of beef cattle during sampling and is beneficial to the health of beef cattle.

[0066] Tissue damage analysis: The degree of tissue damage and the healing time in the experimental group were better than those in the control group, proving that this method caused less damage to the tissues of beef cattle and contributed to the rapid recovery of the sampling site.

[0067] As Figure 1 shown: Sampling success rate: As high as 90%, indicating that the non-destructive sampling method of live beef cattle tissue based on precise positioning has high feasibility in actual operation, and most sampling operations can be successfully completed.

[0068] Sampling precision: The average error was only ±2 mm, indicating that the precise positioning system played a good role, could locate the target sampling site more accurately, and ensured the accuracy of sampling.

[0069] Incidence of stress response in beef cattle: The incidence of stress response of 10% was relatively low, which reflected that this sampling method had certain advantages in reducing the stress of beef cattle. Perhaps the pre-treatment measures of fixation and anesthesia had good effects.

[0070] Wound infection rate: The wound infection rate was 5%, indicating that this non-destructive sampling method caused less wounds to beef cattle, and subsequent nursing or treatment measures might help reduce the infection risk.

[0071] Change in daily weight gain of beef cattle after sampling: The experimental group had an average daily weight gain 0.1 kg more than the control group, which indicated that this sampling method had less impact on the growth performance of beef cattle. Even perhaps due to precise sampling reducing the damage to beef cattle, it was beneficial to their growth to a certain extent. Conclusion The non-destructive sampling method of live beef cattle tissue based on precise positioning is superior to the traditional sampling method in terms of accuracy, time efficiency, reducing the stress response of beef cattle, and reducing tissue damage. This method has high feasibility and practicality, can provide a more scientific and effective tissue sampling means for beef cattle breeding and research, and is worthy of popularization and application in actual production.

[0072] The above embodiments are only used to illustrate the present invention and do not limit the technical solutions described in the present invention. Although the present specification has described the present invention in detail with reference to the above respective embodiments, the present invention is not limited to the above specific implementation manners. Therefore, any modification or replacement of the present invention; and all technical solutions and their improvements that do not depart from the spirit and scope of the invention are covered by the scope of the claims of the present invention.

Claims

1. A non-destructive sampling method for beef cattle living tissue based on precise positioning, characterized in that, It includes the following steps: Step 1: Guide the beef cattle to be sampled into the sampling fixing frame and fix it. Inject an anesthetic into the beef cattle by intramuscular injection to make it enter the anesthetic state. The anesthetic is ketamine hydrochloride, and the injection dose is 2 - 3 mg per 100 kg body weight. Step 2: Use an ultrasonic imaging device to scan the target sampling part of the beef cattle. Set the scanning frequency to 3 - 5 MHz. After determining the position, size, and depth of the target tissue, mark it on the body surface of the beef cattle. Then, with the help of a high-precision three-dimensional positioning system and ultrasonic imaging data, establish a three-dimensional model of the beef cattle body, and plan the sampling path and sampling points on this model. Among them, the positioning accuracy of the high-precision three-dimensional positioning system is ±0.5 mm. Step 3: Select a disposable sterile sampling needle according to the type of target tissue and sampling requirements, and disinfect the sampling needle and related instruments. If the target tissue is muscle tissue, select a sampling needle with an inner diameter of 1.8 mm and a length of 100 mm. If the target tissue is adipose tissue, select a sampling needle with an inner diameter of 2.0 mm and a length of 80 mm. The disinfection treatment is to soak in 75% (volume fraction) alcohol for more than 30 minutes. Step 4: Disinfect the marked sampling part 2 - 3 times with 0.5% (volume fraction) povidone iodine, then lay a sterile surgical drape. The operator holds the sampling needle and inserts it into the beef cattle body to the target tissue according to the planned sampling path, collect the tissue sample. The volume of the collected tissue sample is 0.5 - 1.0 cm³. After collection, pull out the sampling needle and press the sampling part for 3 - 5 minutes. Step 5: Put the collected tissue sample into a sterile petri dish containing normal saline with a concentration of 0.9% and wash it 2 - 3 times, then put it into a sample bottle containing a fixative, and the fixative is a 4% (volume fraction) paraformaldehyde solution, seal it for storage and make marks. Step 6: Conduct routine disinfection treatment on the marked sampling part. Use 75% medical alcohol to degrease 2 times, with each wiping time not less than 30 seconds. Then use an iodophor solution with an effective iodine concentration of 0.5% to disinfect twice, with an interval of 1 minute. Finally, use 75% alcohol to deiodize once. Lay a sterilized sterile surgical drape and leave a sampling window with a diameter of 3 - 5 cm at the sampling part. The operator wears disposable medical sterilized gloves, holds the sampling needle and slowly inserts it into the beef cattle body according to the path planned in the three-dimensional model. Control the puncture speed at 1.5 - 2.5 cm / s to ensure that the tissue is not violently disturbed. During the puncture process, the real-time feedback of the tip position and angle deviation is obtained through the built-in micro inertial measurement unit (IMU) and the external laser positioning system. The maximum allowable angle error does not exceed ±2°, and the depth error does not exceed ±0.3 mm. The length of the collected sample is not less than 1.2 cm, and the diameter is controlled between 1.5 - 3.0 mm. After sampling, quickly pull out the sampling needle, and immediately press the puncture opening with a sterile gauze at a pressure of about 10 kPa for 3 - 4 minutes. Step 7: Place the collected tissue sample in a sterile petri dish containing 0.9% normal saline with a pH of 7.2 - 7.4, and use a low-speed oscillating cleaning method with a frequency of 30 - 60 rpm for 1 - 2 minutes to remove residual blood. Subsequently, conduct a preliminary assessment of tissue integrity and cell viability on the sample through a micro-microscopic imaging system (magnification 10× - 40×, resolution ≤1μm / pixel). The assessment content includes: whether the tissue structure is intact; whether there are breaks or fractures; the nuclear staining condition; a tissue viability index ≥80% is regarded as a qualified sample. Transfer the qualified sample to a sample bottle containing 10% neutral formalin fixative with a pH of 6.8 - 7.2, seal it for storage, and label the sample bottle with the sampling date, beef cattle ear tag number, sampling site name, and sampling personnel information. The sample needs to be transported at 4 ± 1°C and must be sent to the laboratory for pathological analysis or molecular detection within 48 hours. Step 8: The entire sampling process is transmitted in real-time to a remote terminal through a high-definition camera system (resolution not less than 1080P, frame rate 30 fps), supporting the 5G communication protocol (network latency ≤50 ms) to achieve remote expert consultation and operation guidance. All operation data, including images, path parameters, physiological indicators, and sample information, are automatically recorded and uploaded to the cloud database in a storage format of a JSON + DICOM hybrid mode to form a complete electronic sampling file, with a storage period of not less than 5 years for easy later traceability and quality control. Step 9: Postoperative care and observation: Apply a broad-spectrum antibiotic ointment to the sampling site with a thickness controlled at 1 - 2 mm, cover it with a sterile dressing sized 5 cm × 5 cm, and bandage it firmly with an elastic bandage, with the tightness controlled to the extent that two fingers can be inserted. Transfer the beef cattle to a quiet and clean recovery cattle shed, maintain the environmental temperature at 15 - 25°C, relative humidity at 50% - 70%, observe for at least 2 hours, and monitor its respiratory rate. Continuously observe for 24 - 48 hours. If local swelling, bleeding, or a body temperature increase exceeding 39.5°C occurs, then administer an oral or injectable broad-spectrum antibiotic, including ceftiofur sodium, at a dose of 1 - 2 mg / kg body weight, once a day. Step 10: Postoperative data archiving and analysis feedback: After sampling, the system automatically generates a sampling report template in PDF format, including the following content: a sampling path diagram in SVG vector graphics format; the sample quality assessment results, tissue integrity score, and cell viability percentage.

2. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, wherein When guiding the beef cattle to be sampled into the sampling fixing frame and fixing it, use a hydraulic device and adjusting straps to firmly fix the body of the beef cattle.

3. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, wherein When injecting an anesthetic into the beef cattle by intramuscular injection, determine the injection amount of the anesthetic according to the weight and health status of the beef cattle.

4. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, wherein When selecting a disposable sterile sampling needle, the outer diameter of the sampling needle is 1.2 - 1.5 mm and the length is 5 - 10 cm.

5. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, characterized in that, When disinfecting the sampling needle and related instruments, place them in a steam sterilizer and sterilize at 121°C for 20 - 30 minutes.

6. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, characterized in that, When disinfecting the marked sampling site, strictly disinfect it with iodophor, and the number of disinfection times is not less than 3 times.

7. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, wherein During the process of the operator inserting the sampling needle into the beef cattle body along the planned sampling path, the vital signs of the beef cattle and the position of the sampling needle are monitored in real time.

8. The method for non-destructive sampling of live bovine tissue based on precise positioning according to claim 1, characterized in that, When placing the collected tissue sample into a sterile petri dish containing physiological saline for cleaning, gently rinse the sample to remove the blood and impurities on the surface.

9. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, characterized in that, When applying an antibiotic ointment to the sampling site, select an antibiotic ointment such as erythromycin ointment with anti-infection effect.

10. The non-destructive sampling method for beef cattle living tissue based on precise positioning according to claim 1, wherein When transferring the beef cattle to the cowshed for observation and care, observe the vital signs of the beef cattle and the wound healing condition.

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