A method for measuring the quality of Dezhou donkey colostrum

Abstract

The present invention is applicable to the technical field of dairy product quality detection and provides a method for measuring the quality of colostrum from a Texas donkey, comprising the following steps: S1, collecting colostrum; S2, immediately measuring the Brix value of the collected colostrum using an optical Brix refractometer; measuring the collected colostrum using a 0%-32% optical Brix refractometer and recording the resulting value x; S3, calculating the concentration y of immunoglobulin G in the colostrum and determining the quality of the colostrum; y = 1.022x-10.36; S4, calibration. Thus, the present invention rapidly measures the colostrum of a Texas donkey using an optical Brix refractometer, and corrects the Brix value based on the measurement time and ambient temperature to determine the quality grade of the colostrum. This allows for targeted colostrum selection, ensuring that the donkey foal receives high-quality colostrum within two hours of birth, improving the foal's resistance, and ensuring the foal's survival rate.

Description

Technical Field

[0001] The invention relates to the technical field of milk product quality detection, in particular to a method for determining the quality of Dezhou donkey colostrum. Background Art

[0002] Donkeys and horses have similar placentas, with numerous microplacentas and a six-layer placental barrier. This prevents the mother donkey from transferring immunoglobulins to the foal, resulting in the foal being born with hypogammaglobulinemia. Therefore, donkey foals require prompt intake of high-quality colostrum after birth to develop immunity against disease during the first few months of life. In addition to the nutrients necessary for the foal's growth, such as protein, lipids, minerals, and vitamins, the mother donkey's colostrum also contains immunoglobulins, antimicrobial factors, and immunomodulatory factors that help the foal resist invasion by pathogens. Immunoglobulin (IgG), in particular, is crucial for the foal's resistance.

[0003] Currently, colostrum quality and composition vary significantly between donkey breeds, making it particularly important to establish specific parameters for different donkey breeds. The Dezhou donkey is one of China's five most elite donkey breeds and the primary breed currently raised on large-scale donkey farms. Years of research and clinical practice have revealed that the quality and timing of colostrum intake after birth are closely linked to morbidity and, indirectly, survival rates. If colostrum is consumed within two hours of birth, survival rates are very high; otherwise, they are lower. Therefore, rapid testing of colostrum quality is crucial.

[0004] Many methods can assess IgG content in colostrum, including radial immunodiffusion (the gold standard), enzyme-linked immunosorbent assay (ELISA), immunoturbidimetry (TIA), glutaraldehyde agglutination test, and refractometry. However, many of these tests require longer than the 24-hour optimal colostrum feeding time. Therefore, a rapid and accurate method is urgently needed to diagnose colostrum quality on-site.

[0005] In summary, the existing technology has obvious inconveniences and defects in actual use, so it is necessary to improve it. Summary of the Invention

[0006] In response to the above-mentioned defects, the object of the present invention is to provide a method for determining the quality of Texas donkey colostrum. The method determines the quality grade of the colostrum by rapidly determining the immunoglobulin content in the colostrum of the Texas donkey, so as to select the colostrum in a targeted manner, ensure that the donkey foal can smoothly consume high-quality colostrum within two hours after birth, improve the resistance of the donkey foal, and ensure the survival rate of the donkey foal.

[0007] In order to achieve the above object, the present invention provides a method for determining the quality of Texas donkey colostrum, comprising the following steps:

[0008] S1. Collection of colostrum;

[0009] Colostrum was collected from Dezhou donkeys within two hours after giving birth, and a portion of the colostrum was stored at -20 degrees Celsius.

[0010] S2, measuring the Brix value of the collected colostrum using an optical Brix refractometer;

[0011] The collected colostrum was measured using an optical Brix refractometer ranging from 0% to 32%, and the resulting value x was recorded;

[0012] S3. Calculate the concentration y of immunoglobulin G in colostrum and determine the quality of the colostrum;

[0013] y=1.022x-10.36;

[0014] When y≥8, the quality of colostrum is judged as: Grade 1;

[0015] When 5≤y<8, the quality of colostrum is judged as: Grade II;

[0016] When 2.8≤y<5, the quality of colostrum is judged as: Grade III;

[0017] When y<2.8, the quality of colostrum is judged as: Grade 4;

[0018] Grade 1 is the best and grade 4 is the worst, and the quality grades are marked on the outside of the frozen samples;

[0019] S4, verification;

[0020] After the colostrum stored in S1 was thawed, the concentration of immunoglobulin G in the colostrum of the donkey was determined using a radial immunodiffusion test kit for horse IgG detection. ， and compared with y to verify the quality level of colostrum;

[0021] When y ， When it is at the same level as y, the colostrum quality grade is correct;

[0022] When y ， is at the adjacent level to y, and |y ， -y|≤0.1, the colostrum quality is treated according to the higher quality of the two;

[0023] When y ， is at the adjacent level to y, and |y ， If -y|>0.1, the colostrum quality will be treated as the lower of the two.

[0024] The method for determining the quality of colostrum from a Texas donkey according to the present invention further comprises step S2.1, correcting the Brix value x based on the time t from colostrum collection to measurement:

[0025] When 0 < t ≤ 30 min, the corrected Brix value x ， = x;

[0026] When 30 < t ≤ 50 min, the corrected Brix value x ， = 1.01x;

[0027] When 50 < t ≤ 80 min, the corrected Brix value x ， = 1.03x; L>

[0028] When 80 < t ≤ 120 min, the corrected Brix value x ， = 1.04x.

[0029] According to the method for measuring the quality of Dezhou donkey colostrum of the present invention, it further includes step S2.2 of measuring the room temperature C during sampling, calculating the correction coefficient F of the Brix value x according to the room temperature C, F = 1 + (C - 15.6) / 800; the corrected Brix value x ″ = F * x ， .

[0030] According to the method for measuring the quality of Dezhou donkey colostrum of the present invention, in step S2, 3 drops of colostrum are dropped on the prism, the cover plate is covered to fill the entire mirror surface, and the automatically generated value is recorded. Before measuring each new sample, the refractometer is calibrated with ultrapure water.

[0031] According to the method for measuring the quality of Dezhou donkey colostrum of the present invention, in step S4, a preliminary estimate is made based on the concentration y of immunoglobulin G in the colostrum. For colostrum samples with a possible immunoglobulin G concentration greater than the detection range specified by the manufacturer, they are diluted 1:10 with physiological saline; 5 μl of the sample is added to the well of the plate and incubated at room temperature of 22°C - 24°C for more than 48 h, and the diameter of the precipitation ring is measured using a vernier caliper.

[0032] According to the method for measuring the quality of Dezhou donkey colostrum of the present invention, in step S4, 3 standard products are added to each plate, and the average value of the 3 repeated measurements of the standard products is used to establish a calibration curve to determine the concentration y of immunoglobulin G in the colostrum sample ， .

[0033] The beneficial effects of the present invention are as follows:

[0034] The present invention rapidly measures the colostrum of Dezhou donkeys through an optical Brix refractometer, corrects the Brix value according to the measurement time and environmental temperature, determines the quality grade of the colostrum, selects the colostrum targeted, ensures that the foal can successfully eat high-quality colostrum within two hours after birth, improves the resistance of the foal, and ensures the survival rate of the foal. Specific embodiments

[0035] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0036] The present invention provides a method for determining the quality of Texas donkey colostrum. The method comprises:

[0037] S1. Collection of colostrum;

[0038] Colostrum is collected immediately after giving birth, within two hours at the latest, and a portion of the collected colostrum is stored at -20 degrees Celsius for later use.

[0039] S2. Immediately measure the Brix value of the collected colostrum sample using an optical Brix refractometer;

[0040] The collected colostrum samples were measured using an optical Brix refractometer ranging from 0% to 32%, and the resulting value x was recorded;

[0041] S3. Calculate the concentration y of immunoglobulin G in the colostrum based on the Brix value x, and determine the quality of the colostrum;

[0042] y=1.022x-10.36;

[0043] When y≥8, the quality of colostrum is judged as: Grade 1;

[0044] When 5≤y<8, the quality of colostrum is judged as: Grade II;

[0045] When 2.8≤y<5, the quality of colostrum is judged as: Grade III;

[0046] When y<2.8, the quality of colostrum is judged as: Grade 4;

[0047] Grade 1 is the best, and grade 4 is the worst. The quality grade is marked on the outside of the frozen sample. Colostrum of grade 1 or 2 can be fed directly to the foal. Grade 3 colostrum is determined based on the actual colostrum reserves. If sufficient colostrum is available, use the colostrum from grade 1 or 2; otherwise, use grade 3 colostrum. Grade 4 colostrum should be used for feeding.

[0048] S4, verification;

[0049] After the colostrum stored in S1 was thawed, the concentration of immunoglobulin G in the colostrum of the donkey was determined using a radial immunodiffusion test kit for horse IgG detection. ， and compared with y to verify the quality level of colostrum;

[0050] When y ， is at the same level as y, the colostrum quality grade is judged correctly;

[0051] When y ， is at an adjacent level to y, and |y ， - y| ≤ 0.1, the colostrum quality is processed according to the quality mark of the higher grade of the two;

[0052] When y ， is at an adjacent level to y, and |y ， - y| > 0.1, the colostrum quality is processed according to the quality mark of the lower grade of the two.

[0053] The stored colostrum is used for backup. When it is subsequently found that the colostrum quality grade of the female donkey is low, the backup higher-grade colostrum is selected for substitute feeding.

[0054] Furthermore, it also includes step S2.1: According to the time t from colostrum collection to measurement, the Brix value x is corrected:

[0055] [[ID=?]]When 0 < t ≤ 30 min, the corrected Brix value x ， = x; When the colostrum is measured within half an hour after collection, the components of the colostrum basically do not change, and the Brix value x measured during this period is the same as the Brix value of the colostrum when it was just collected;

[0056] When 30 < t ≤ 50 min, the corrected Brix value x ， = 1.01x; When measured within 30 - 50 minutes after colostrum collection, the measured Brix value changes, and the measured Brix value during this period is lower than the actual Brix value of the colostrum when it was just collected. If judged and marked according to the measured Brix value, there will be a large error, affecting the accurate judgment of the colostrum quality grade;

[0057] When 50 < t ≤ 80 min, the corrected Brix value x ， = 1.03x; When measured within 50 - 80 minutes after colostrum collection, the measured Brix value changes significantly, and the measured Brix value during this period is lower than the Brix value of the colostrum when it was just collected; If judged and marked according to the measured Brix value, there will be a large error, affecting the accurate judgment of the colostrum quality grade;

[0058] When 80 < t ≤ 120 min, the corrected Brix value x ， It seems there is a missing tag in the original text for the "0 < t ≤ 30 min" part. I've translated it as best as possible with the given information.=1.04x; The Brix value measured within 60-120 minutes of colostrum collection changes significantly, resulting in a lower Brix value than the colostrum immediately after collection. Marking colostrum based on the measured Brix value can result in significant errors, affecting the accurate determination of colostrum quality. If the colostrum is measured more than two hours after collection, the data will be significantly inaccurate due to the effects of temperature and the surrounding environment, making the results meaningless. It is recommended to collect fresh colostrum for measurement.

[0059] Furthermore, the method further includes step S2.2, measuring the room temperature C during sampling, and calculating the quadratic correction coefficient F of the Brix value x according to the room temperature C, where F=1+(C-15.6) / 800; the corrected Brix value x ″ =F*x ， The estrus season for female donkeys is mostly from February to August, with May and June being the most common. The donkey's reproductive cycle is one year, and its gestation period is generally around 348 to 377 days, with an average of 360 days, and can take up to a year and a half. This is because multiparous donkeys generally have shorter gestation periods, first-time donkeys have longer gestation periods, and smaller donkeys have shorter gestation periods, while larger donkeys have longer gestation periods. This means that various factors can lead to significant differences in the seasons in which female donkeys give birth, and the natural external temperature can vary significantly. When measuring the Brix value using a Brix refractometer, the influence of the external temperature, in addition to the colostrum's own temperature, can also be significant, leading to errors in the colostrum's grade. Correcting the Brix value x using the room temperature C allows for more accurate judgment of the colostrum's quality grade.

[0060] Furthermore, in step S2, three drops of colostrum are dripped onto the prism, a cover is placed to fill the entire mirror surface, and the automatically generated value is recorded. Before measuring each new sample, the refractometer needs to be calibrated with ultrapure water to ensure the accuracy of the data.

[0061] Furthermore, in step S4, a preliminary estimate of the immunoglobulin G concentration y in the colostrum is made. For colostrum samples whose immunoglobulin G concentration may be greater than the manufacturer's specified detection range, the colostrum samples are diluted 1:10 with normal saline; 5 μl of the sample is added to the plate wells and incubated at room temperature of 22°C-24°C for more than 48 hours. The diameter of the precipitation ring is measured using a vernier caliper. The concentration of immunoglobulin G y in donkey colostrum is determined using a horse IgG detection radial immunodiffusion test kit. ， , directly examine the concentration of immunoglobulin G in colostrum ， More accurately, the calculated immunoglobulin G concentration y is verified to make correction marks, thereby improving the accuracy of the colostrum quality grade and ensuring that the donkey foal is fed with qualified colostrum in a short period of time. At the same time, the subsequent spare colostrum grade can be accurately corrected to improve the resistance and survival rate of the donkey foal.

[0062] Furthermore, in step S4, three standards are added to each plate, and the average value of the three standard replicates is determined to establish a calibration curve to determine the immunoglobulin G concentration y in the colostrum sample. ， , multiple measurements are taken to get the average value, and the data accuracy is higher.

[0063] In addition to containing proteins, lipids, minerals, and vitamins—nutrients essential for foal growth—a mother donkey's colostrum also contains immunoglobulins, antimicrobial factors, and immunomodulatory factors that help foals resist pathogenic microorganisms. Immunoglobulins, in particular, are crucial for a newborn foal's immunity, leading some to call colostrum a "talisman" bestowed upon the foal by the mother donkey. Therefore, colostrum quality is crucial for the foal's health. Furthermore, the timing of colostrum intake is crucial. At birth, the foal's small intestine has a specialized structure that allows it to directly absorb the immunoglobulins in colostrum. However, this specialized structure gradually disappears over time. Three hours after birth, the small intestine's ability to absorb immunoglobulins drops to 25% of its initial capacity. Therefore, it's important to ensure that foals receive high-quality colostrum within two hours of birth to boost their immunity.

[0064] To ensure colostrum quality and timely ingestion, colostrum should be graded based on its immunoglobulin G content. Levels above 8g / dL, 5-8g / dL, 2.8-5g / dL, and below 2.8g / dL are designated "Grade 1," "Grade 2," "Grade 3," and "Grade 4," respectively. For Grade 3 and 4 colostrum, higher-grade colostrum should be selected based on the amount and grade of spare colostrum available. When colostrum quality is good and lactation is high, any remaining colostrum should be frozen for future use. Furthermore, proper farrowing house management is crucial. Since most donkeys give birth at night, caregivers must be assigned to ensure that the foal receives colostrum as soon as possible. For the first six hours after birth, a colostrum intake of 200-250mL per hour should be ensured to ensure adequate immunoglobulin intake and meet the foal's nutritional needs.

[0065] In summary, the present invention includes the following steps: S1, collecting colostrum; collecting colostrum from a Texas donkey within one hour after giving birth, and storing part of the colostrum at -20 degrees Celsius; S2, measuring the Brix value of the collected colostrum immediately using an optical Brix refractometer; the collected colostrum is measured using a 0%-32% optical Brix refractometer, and the resulting value x is recorded; S3, calculating the concentration y of immunoglobulin G in the colostrum, and judging the quality of the colostrum; y = 1.022x - 10.36; when y ≥ 8, the quality of the colostrum is determined to be: Grade 1; when 5 ≤ y < 8, the quality of the colostrum is determined to be: Grade 2; when 2.8 ≤ y < 5, the quality of the colostrum is determined to be: Grade 3; when y < 2.8, the quality of the colostrum is determined to be: Grade 4, where Grade 1 is the best and Grade 4 is the worst; where Grade 1 is the best and Grade 4 is the worst, and the quality grades are marked on the outside of the frozen sample; S4 , verification; after thawing the colostrum stored in S1, the concentration y of immunoglobulin G in the colostrum of the female donkey is determined by using a horse IgG detection radial immune diffusion detection kit, and compared with y to verify the quality grade of the colostrum; when y, and y are at the same grade, the quality grade of the colostrum is judged correctly; when y, and y are at adjacent grades, and |y, -y|≤0.1, the quality of the colostrum is processed according to the higher grade of the two; when y, and y are at adjacent grades, and |y, -y|>0.1, the quality of the colostrum is processed according to the lower grade of the two; the present invention uses an optical Brix refractometer to quickly measure the colostrum of the Texas donkey, and corrects the Brix value according to the measurement time and ambient temperature to determine the quality grade of the colostrum, so as to select colostrum in a targeted manner, ensure that the donkey foal can smoothly eat high-quality colostrum within two hours after birth, improve the resistance of the donkey foal, and ensure the survival rate of the donkey foal.

[0066] Of course, the present invention may have many other embodiments. Without departing from the spirit and essence of the present invention, those skilled in the art may make various corresponding changes and modifications based on the present invention, but these corresponding changes and modifications should all fall within the scope of protection of the claims attached to the present invention.

Claims

1. A method for determining the quality of Dezhou donkey colostrum, characterized in that: It includes the following steps: S1. Collection of colostrum; Collect colostrum within two hours after the delivery of Dezhou donkeys, and留样保存部分初乳 at -20 °C; S2. Measure the Brix value of the collected colostrum with an optical Brix refractometer; Measure the collected colostrum with an optical Brix refractometer of 0%-32%, and record the generated value x; Drop 3 drops of colostrum on the prism, cover the cover plate to fill the entire mirror surface, and record the automatically generated value. Before measuring each new sample, calibrate the refractometer with ultrapure water; S2.

1. Correct the Brix value x according to the time t from the collection of colostrum to the measurement: When 0 < t ≤ 30 min, no correction is required; 30 < t ≤ 50 min, the corrected Brix value x ， = 1.01x; When 50 < t ≤ 80 min, the corrected Brix value x ， = 1.03x; 80 < t ≤ 120 min, the corrected Brix value x ， = 1.04x; When the measurement time exceeds two hours from the collection time of colostrum, at this time, the colostrum is affected by temperature and the surrounding environment, and the data is significantly inaccurate, and the measurement result is meaningless. Collect new colostrum for measurement; S2.

2. Measure the room temperature C during sampling and calculate the correction factor F for the Brix value x based on the room temperature C: F = 1 + (C - 15.6) / 800; the corrected Brix value x ″ =F*x ， ; S3. Calculate the concentration y of immunoglobulin G in the colostrum and judge the quality of the colostrum; y=1.022x ″ -10.36; When y ≥ 8 g / dL, judge the quality of the colostrum as: first grade; When 5 g / dL ≤ y < 8 g / dL, judge the quality of the colostrum as: second grade; When 2.8 g / dL ≤ y < 5 g / dL, judge the quality of the colostrum as: third grade; When y < 2.8 g / dL, judge the quality of the colostrum as: fourth grade; Among them, the first grade is the best and the fourth grade is the worst, and mark the quality grade on the outside of the frozen sample; S4. Verification; After the colostrum stored in S1 was thawed, the concentration of immunoglobulin G in the colostrum was determined using a radial immunodiffusion test kit for horse IgG detection. ， and compared with y to verify the quality level of colostrum; When y ， When it is at the same level as y, the colostrum quality grade is correct; When y ， is at the adjacent level to y, and |y ， -y|≤0.1, the colostrum quality is treated according to the higher quality of the two; When y ， is at the adjacent level to y, and |y ， If -y|>0.1, the colostrum quality will be treated as the lower of the two.

2. The method for determining the quality of the colostrum of a Texas donkey according to claim 1, wherein In step S4, make a preliminary estimate according to the concentration y of immunoglobulin G in the colostrum. Dilute the colostrum sample with a possible immunoglobulin G concentration greater than the manufacturer's specified detection range with physiological saline at a ratio of 1:10; Add 5 μl of the sample to the well of the plate, incubate at room temperature of 22 °C - 24 °C for more than 四十八 hours, and measure the diameter of the precipitation ring with a vernier caliper. It should be noted that there is an unclear expression "留样保存部分初乳" in the original text. I have translated it as "留样保存部分初乳" as it is, which may need to be further clarified in the actual context. Also, "四十八 hours" should be "48 hours" for correct English expression.

3. The method for determining the quality of the colostrum of Texas donkey according to claim 2, wherein Three standards were added to each plate, and the average value of the three standard replicates was used to establish a calibration curve to determine the concentration of immunoglobulin G in colostrum samples. ， .