CN104215774B - A kind of method utilizing FGF21 factor diagnosis of milk cow sub-clinical ketosis - Google Patents

Abstract

A method for diagnosing recessive ketosis in dairy cows using FGF21 factor. FGF21 factor is positively correlated with recessive ketosis in dairy cows. ELISA kits and biochemical analyzers are used to measure the concentration of FGF21 in bovine serum and serum indicators of dairy cows during ketosis and recessive ketosis. Through statistical analysis, FGF21 is positively correlated with recessive ketosis. Significant correlation, no correlation with clinical ketosis; when 335ng/ml<FGF21<443ng/ml, it can be diagnosed as cow recessive ketosis. This method determines that FGF21 is positively correlated with recessive ketosis in dairy cows, and dairy cow serum FGF21 can replace BHBA in diagnosing recessive ketosis in dairy cows, so early detection of recessive ketosis will reduce the loss of dairy farming industry.

Description

A method for diagnosing recessive ketosis in dairy cows using FGF21 factor

technical field

The invention relates to a method for diagnosing recessive ketosis in dairy cows, in particular to a method for diagnosing recessive ketosis in dairy cows by using FGF21 factor.

Background technique

Dairy cattle breeding is one of the pillars of my country's animal husbandry industry, but dairy cattle ketosis has brought serious economic losses to the dairy industry. Especially the recessive ketosis of dairy cows, because the clinical symptoms of recessive ketosis are not obvious, which brings great difficulties to diagnosis and prevention, so early detection of recessive ketosis will reduce the loss of dairy farming industry .

FGF21 is a member of the fibroblast growth factor family. The latest research shows that FGF21 stimulates glucose uptake by fat cells, but not other types of cells, and that this effect is actually equivalent to enhancing insulin activity. Glucose transporter 1 was significantly increased in adipose tissue after FGF21 injection in obese mice. FGF21 is protective in diabetic mice under diet-induced obesity, hypoglycemia, and high triglyceride levels. Further research results showed that the injection of FGF21 in animals led to increased energy expenditure, accelerated fat utilization and lipid excretion. And there are reports that FGF21 can be used to diagnose fatty liver.

At present, the monitoring of recessive ketosis is mainly based on the change of BHBA concentration. When 1.2mmol/L<BHBA<1.6mmol/L, it can be diagnosed as recessive ketosis of cows. However, this method has low sensitivity and may cause missed diagnosis. Therefore, it is necessary to find a factor with higher sensitivity for the monitoring of recessive ketosis in dairy cows.

Contents of the invention

The invention provides a method for diagnosing recessive ketosis in dairy cows by using the FGF21 factor. The FGF21 is more sensitive to the change of ketone bodies during cow ketosis, especially in the stage of recessive ketosis in dairy cows. The relationship between FGF21 indicated that FGF21 was positively correlated with recessive ketosis in dairy cows.

The technical scheme adopted in the present invention is: a method for diagnosing recessive ketosis in dairy cows by using FGF21 factor, FGF21 factor is closely related to recessive ketosis in dairy cows, and the concentration of FGF21 factor in bovine serum is measured by using ELISA kit and biochemical analyzer. The serum index of dairy cows during ketosis and recessive ketosis, through statistical analysis, FGF21 is significantly correlated with recessive ketosis, and has no correlation with clinical ketosis; when 335ng/ml<FGF21<443ng/ml, it can be diagnosed as dairy cows. Ketosis.

The analysis of serum FGF21 metabolic indicators in the above scheme and the main detection results of the clinical ketosis group, recessive ketosis group, and healthy group showed statistical significance for serum FGF21 in the clinical ketosis group, recessive ketosis group, and healthy group, P<0.05 ; Serum BHBA in healthy group, recessive ketosis group and clinical ketosis group was statistically significant, P<0.05; serum FGF21 factor was positively correlated with BHBA in recessive ketosis, r=0.659, p=0.005; clinical ketosis There was no correlation between serum FGF21 factor and BHBA during illness r=0.005, p=0.979.

The beneficial effects of the present invention are as follows: based on the above background, the present invention can determine the level of dairy cows in recessive ketosis through the reaction of FGF21 in the blood of dairy cows to clinical ketosis, recessive ketosis, and the level of BHBA, an important intermediate metabolite in dairy cows in ketosis. Changes of FGF21 in the body, measuring metabolic indicators and FGF21 factors in cattle samples, and confirming that FGF21 is positively correlated with recessive ketosis in dairy cows, dairy cow serum FGF21 can replace BHBA to diagnose recessive ketosis in dairy cows, when 335ng/ml<FGF21<443ng/ It can be determined that the dairy cow is recessive ketosis at the time of ml, so early detection of recessive ketosis will reduce the loss of the dairy farming industry.

detailed description

Below in conjunction with experiment the present invention is further described:

Animal experiments: The cows were all from a cattle farm in Mishan, Heilongjiang Province. There was no statistically significant difference in age and parity among the cows. All cows were fed with TMR during the peripartum period. BHBA>1.2mmol/L is used as the standard for diagnosing dairy cow ketosis, if BHBA>1.6mmol/L in serum of dairy cows, and show obvious clinical symptoms, it is diagnosed as clinical ketosis group; if 1.2mmol/L<BHBA<1.6mmol/L , and no obvious clinical symptoms, were diagnosed as the recessive ketosis group; BHBA<1.2mmol/L and no clinical symptoms, as the healthy group. Among them, 30 were healthy, 30 were recessive, and 30 were clinical. In addition, cows in which diseases such as mastitis, metritis, miscarriage, liver disease due to drugs, fever, etc. were excluded from this experiment.

Sample Collection and Processing

All cows were fasted in the morning to draw tail vein blood, without adding anticoagulant, the sampling tube was placed in the ice box, brought back to the laboratory within 1 hour, centrifuged at 3000g for 15 minutes, and the separated serum was stored at -20°C for later use.

Index measurement method: FGF21 was measured using RD bovine fibroblast factor ELISA kit, and other metabolic indexes were measured on an automatic biochemical analyzer.

Data analysis: All data were analyzed using IBM SPSS 19.0 software, and the measurement data was measured with the mean Indicates that the significance of the mean difference between groups was analyzed by one-way ANOVA, and the correlation analysis was conducted by Pearson linear correlation analysis. p<0.05 was considered significant correlation.

Experimental results:

Table 1 shows the results of FGF21, blood glucose, BHBA, and NEFA of 90 dairy cows by one-way ANOVA.

Table 1 Main detection indicators of dairy cow serum

Person correlation analysis The correlation analysis between FGF21 metabolic index and ketosis is shown in Table 2. Serum FGF21 was positively correlated with NEFA and positively correlated with BHBA.

Table 2 Correlation analysis between dairy cow FGF21 and main serum indexes

The relationship between different types of ketosis FGF21 and ketosis metabolic indicators was analyzed by one-way ANOVA. Among the 90 cows, there were 30 cases of clinical ketosis, 30 cases of recessive ketosis, and 30 cases of healthy cows. The parity and age of the three groups of cattle are similar, and the main detection results of serum FGF21 metabolic index and dairy cow ketosis are shown in Table 3. Serum FGF21 in clinical ketosis group, recessive ketosis group and healthy group (P<0.05), with statistical significance; comparison of serum NEFA and BHBA in healthy group, recessive ketosis group, clinical group (P<0.05), with statistical significance Statistical significance.

Table 33 Comparison of FGF21 and serum main detection indicators in dairy cows

Person linear correlation analysis The correlation analysis between the serum FGF21 metabolic index and ketosis index in the three groups of cattle is shown in Table 4. Serum FGF21 in the recessive ketosis group was positively correlated with BHBA; there was no correlation between serum FGF21 and BHBA in the clinical ketosis group.

Table 4 Correlation analysis between FGF21 metabolic indicators and ketosis indicators in different groups

FGF21 is positively correlated with BHBA, an important detection indicator of recessive dairy cow ketosis. FGF21 can be used to replace BHBA as a new factor for diagnosing dairy cow recessive ketosis. When 335ng/ml<FGF21<443ng/ml, it can be diagnosed as dairy cow recessive ketosis.

Claims (2)

1. the purposes in milk cow recessive ketoacidosis diagnostic reagent prepared by the reagent measuring FGF21, it is characterized in that: the FGF21 factor and milk cow recessive ketoacidosis closely related, all the other metabolic index of cow serum when application ELISA kit and Biochemical Analyzer measure FGF21 factor concentration and ketoacidosis, sub-clinical ketosis in cow serum, by statistical analysis FGF21 and sub-clinical ketosis in obviously relevant, with clinical ketoacidosis non-correlation; Milk cow recessive ketoacidosis can be diagnosed as 335ng/ml<FGF21<443ng/ml.

2. purposes according to claim 1, it is characterized in that: serum analysis FGF21 metabolic index and clinical ketoacidosis group, sub-clinical ketosis group, health organize main testing result, clinical ketoacidosis group, sub-clinical ketosis group and healthy group serum FGF21 have statistical significance, P<0.05; Healthy group, sub-clinical ketosis group, clinical ketoacidosis group compare, and in serum, BHBA has statistical significance, P<0.05; During sub-clinical ketosis, the serum FGF21 factor and BHBA are proportionate, r=0.659, p=0.005; During clinical ketoacidosis, the serum FGF21 factor and BHBA are without relevant, r=0.005, p=0.979.