Method for predicting risk of bovine dystocia

Abstract

To provide a method for predicting the risk of dystocia of a cow by detecting the amount of inflammatory cytokine in cervical mucus as a biomarker indicating the progress of cervical ripening in a pregnant cow, a kit therefor, a composition containing an antibody against the biomarker, and the like.SOLUTION: It has been found that the difference in biological changes between a difficult-to-calve cow and an easy-to-calve cow at an earlier stage than the start of calving, that is, the fact that the amount of each of IL-8 and / or TNFα in the uterine cervix mucus becomes less than the reference value due to the absence of the onset of cervical ripening (physiological inflammatory response) in a difficult-to-calve cow, which has not been clarified so far, allows the risk of difficult-to-calve cow to be predicted based on the amounts of IL-8 and / or TNFα in the uterine cervix mucus.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for predicting the risk of dystocia in pregnant cows, a composition for predicting the risk of dystocia, and a kit for the same. [Background technology]

[0002] In recent years, demand for meat has been increasing both domestically and internationally, and there is a particular need to improve the productivity of Wagyu cattle, including Japanese Black, Japanese Brown, Japanese Shorthorn, and Japanese Polled. Of these, Japanese Black cattle are a superior meat breed unique to Japan and are attracting particular attention.

[0003] A cow normally gives birth about once a year, and during parturition, there is a high risk of accidents occurring, such as difficult labor due to birth canal abnormalities, weak labor pains, or a narrow birth canal, an oversized fetus due to a long gestation period, or stillbirth, and there is also a risk of losing the cow due to difficult labor. For this reason, in order to ensure a stable supply of beef cattle and other cattle, it is extremely important to accurately determine the risk of difficult labor in cows before parturition, take measures to reduce the risk of difficult labor, and provide appropriate assistance to prevent accidents from occurring.

[0004] One method for predicting difficulty in calving in cows is to estimate the difficulty of calving before calving by referring to photographs of the test cow (Publication No. 2021-157428). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Re-tabled publication No. 2021-157428 Summary of the Invention [Problem to be solved by the invention]

[0006] Approximately 10% of dystocia in cattle is caused by abnormalities in the birth canal, but until now, there has been no established method for predicting dystocia in cattle, and it has been particularly impossible to predict dystocia caused by abnormalities in the birth canal. Therefore, there has been a need to develop a method for predicting the risk of dystocia in cattle, and in particular a means for predicting dystocia caused by abnormalities in the birth canal.

[0007] Therefore, the present invention aims to provide a method and kit for confirming the progress of cervical ripening and predicting the risk of dystocia in cows by detecting the concentration of inflammatory cytokines in cervical mucus, which serves as a marker for the progress of cervical ripening in pregnant cows, as well as a composition containing an antibody specific to an inflammatory cytokine or a functional fragment thereof. [Means for solving the problem]

[0008] The inventors discovered that there is a difference in biological changes between cows with difficult labor and cows with easy labor at an earlier stage than the onset of labor (the start of the opening period, which is the first stage of labor), namely, that in cows with difficult labor, the onset of cervical ripening (physiological inflammatory response) does not occur, which results in the concentration of inflammatory cytokines in cervical mucus being below the standard value, and this led to the completion of the present invention.

[0009] The present invention provides a method and kit for predicting the risk of dystocia in pregnant cows by detecting the concentration of inflammatory cytokines in cervical mucus, which serves as a marker for the progression of cervical ripening. [1] 1. A method for predicting a risk of dystocia in a cow, comprising: (i) measuring the amount of IL-8 and / or TNFα in cervical mucus of pregnant cows; A method for predicting the risk of dystocia in cattle, including: [2] and (ii) determining that the pregnant cow is at risk of dystocia if the amount of IL-8 and / or TNFα measured in the step (i) is less than a cutoff value. The method according to [1], comprising: [3] the cutoff value is a value determined by the concentration of the IL-8 relative to the total protein concentration in the cervical mucus and / or the concentration of the TNFα relative to the total protein concentration in the cervical mucus, the cutoff value of IL-8 is 50 to 290 pg / mg, the cutoff value of TNFα is 4 to 11 pg / mg; [2] The method described in [2]. [4] The method according to [1] or [2], wherein the pregnant cow is a cow for which 270 days or more have passed since the date of artificial insemination. [5] The method according to [1] or [2], wherein the difficult labor is caused by an abnormality in the birth canal.

[0010] The present invention further relates to a composition comprising an antibody or a functional fragment thereof specific to an inflammatory cytokine, and a kit comprising the composition. [6] A composition for predicting the risk of dystocia in cattle, comprising an antibody specific to bovine IL-8 or a functional fragment thereof, or an antibody specific to TNFα or a functional fragment thereof. [7] The composition described in [6], wherein the difficult labor is caused by an abnormality in the birth canal. [8] A kit for predicting the risk of dystocia in cattle, comprising at least one antibody selected from the group consisting of an antibody specific to bovine IL-8 or a functional fragment thereof, and an antibody specific to TNFα or a functional fragment thereof. [9] A kit for predicting the risk of dystocia in cows, comprising a substance capable of specifically detecting a gene encoding IL-8 and / or a substance capable of specifically detecting a gene encoding TNFα in the cervical mucus of pregnant cows. [Effects of the Invention]

[0011] According to the present invention, it is possible to predict the risk of dystocia in cattle. Furthermore, according to the present invention, it is possible to provide a composition comprising an antibody specific to an inflammatory cytokine or a functional fragment thereof. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is a graph showing the concentrations of inflammatory cytokines in bovine cervical mucus in Test Example 1. [Figure 2] 1 shows a graph of the receiver operating characteristic (ROC) curve for IL-8 in Test Example 2. [Figure 3] 1 shows a graph of the ROC curve for TNFα in Test Example 2. [Figure 4] 1 shows a graph in which the concentrations of IL-8 and TNFα are plotted for each individual cow in each group in Test Example 2. DETAILED DESCRIPTION OF THE INVENTION

[0013] In the cervical mucus of pregnant cows, the levels (quantities) of interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-10, and TNFα were found to increase from 260±3 days of pregnancy onwards, with IL-8 and TNFα levels in particular being more significantly increased in cows with easy births than in cows with difficult births. Therefore, by monitoring the appearance and quantity of IL-8 and / or TNFα in the cervical mucus, it is possible to predict the risk of difficult births in cows and, ultimately, the timing for taking measures to prevent difficult births.

[0014] The present invention also relates to biomarkers that enable prediction or assessment of the risk of dystocia in cattle, kits for predicting dystocia in cattle, and compositions containing these biomarkers.

[0015] As used herein, cattle includes, but is not limited to, beef cattle including Wagyu cattle such as Japanese Black Cattle, Japanese Brown Cattle, Japanese Shorthorn Cattle, and Japanese Polled Cattle.

[0016] As used herein, pregnant cattle include, but are not limited to, for example, calves and / or heifers.

[0017] In this specification, a pregnant cow includes, for example, a pregnant cow for which the number of days since the date of artificial insemination has been 260 days or more, 265 days or more, 270 days or more, 275 days or more, 280 days or more, 285 days or more, 290 days or more, 295 days or more, 300 days or more, 305 days or more, or 310 days or more.

[0018] Pregnant cows include, for example, pregnant cows that are one day or more, two days or more, three days or more, four days or more, five days or more, six days or more, seven days or more, etc. from the expected date of delivery.

[0019] In this specification, the expected delivery date is, for example, 285 days for Wagyu cattle and 280 days for dairy cattle calculated from the date of artificial insemination, but is not limited to these.

[0020] [Method for predicting the risk of dystocia in cattle] One aspect of the present invention relates to a method for predicting the risk of dystocia in a cow, comprising the step of (i) measuring the amount of IL-8 and / or TNFα in the cervical mucus of a pregnant cow.

[0021] As used herein, bovine dystocia includes, but is not limited to, bovine birth accidents caused by birth canal abnormalities, fetal enlargement, fetal dislocation, abnormal labor pains, multiple births, etc. Dystocia refers to, for example, a case in which delivery does not occur within 9 hours from the onset of labor, regardless of the gestational age of the fetuses, and / or a condition in which human assistance or midwifery is required.

[0022] The risk of dystocia in cattle includes, but is not limited to, the possibility that cattle will experience dystocia due to abnormalities in the birth canal, large fetuses, fetal dislocation, abnormal labor, and / or multiple births.

[0023] Cervical mucus can be collected at any time using methods known to those skilled in the art and then analyzed, or collected and stored in a refrigerator or freezer before being analyzed. The amount of cervical mucus collected is preferably 0.1 to 6 g, more preferably 0.2 to 5.5 g, even more preferably 0.5 to 5 g, even more preferably 1 to 4.5 g, and particularly preferably 2 to 4 g.

[0024] When comparing the amounts of IL-8 and / or TNFα in cervical mucus, it is preferable that the conditions for collecting and measuring the cervical mucus from each pregnant bovine are the same or similar. It is preferable that the same amount of cervical mucus is collected from each pregnant bovine at as similar a time as possible. It is also preferable that the amount of IL-8 and / or TNFα is measured as soon as possible after collection.

[0025] The pregnant cow from which cervical mucus is collected is not particularly limited as long as the effects of the present invention are achieved. For example, the number of days since artificial insemination is 260 days or more, 265 days or more, 270 days or more, 275 days or more, 280 days or more, 285 days or more, 290 days or more, 295 days or more, 300 days or more, 305 days or more, or 310 days or more, preferably 265 days or more, 270 days or more, 275 days or more, 280 days or more, 285 days or more, 290 days or more, 295 days or more, or 300 days or more, and more preferably 270 days or more, 275 days or more, 280 days or more, 285 days or more, 290 days or more, or 295 days or more. Furthermore, the pregnant cow from which cervical mucus is collected is not particularly limited as long as the effects of the present invention are achieved, but for example, it may be one day or more, two days or more, three days or more, five days or more, or seven days or more after the expected date of delivery.

[0026] Before measuring the amount of IL-8 and / or TNFα in cervical mucus, a procedure including concentration or dilution can be carried out as a pretreatment.

[0027] One aspect of the method for predicting the parturition period of the present invention relates to a method for predicting the risk of dystocia in cows, comprising: (i) measuring the amount of IL-8 and / or TNFα in the cervical mucus of a pregnant cow; and (ii) determining that the pregnant cow is at risk of dystocia if the amount of IL-8 and / or TNFα measured in step (i) is below a cutoff value.

[0028] The cutoff value in the prediction method of the present invention refers to a value that distinguishes between the presence or absence of a risk of dystocia in pregnant cows.

[0029] The cutoff value for the amount of IL-8 in the prediction method of the present invention can be a value determined by the concentration of IL-8 relative to the total protein concentration in cervical mucus.

[0030] The cutoff value for the amount of TNFα in the prediction method of the present invention can be a value determined by the concentration of TNFα relative to the total protein concentration in cervical mucus.

[0031] In the prediction method of the present invention, when the value of the amount of IL-8 or TNFα in the cervical mucus of a cow is less than the cutoff value for IL-8 or TNFα, the cow is determined to have a risk of dystocia.On the other hand, when the value of the amount of IL-8 or TNFα in the cervical mucus of a cow is equal to or greater than the cutoff value for IL-8 or TNFα, the cow is determined to have no risk of dystocia.

[0032] In the prediction method of the present invention, the total protein concentration in cervical mucus is measured by a protein quantification method using a spectrophotometer. The spectrophotometer is not particularly limited, but for example, a NanoDrop microspectrophotometer (NanoDrop Eight, manufactured by Thermo Scientific) can be used.

[0033] In the prediction method of the present invention, the concentration of IL-8 or TNFα in cervical mucus is measured using a measurement method that utilizes a multi-parameter simultaneous measurement device that employs magnetic fluorescent microbeads. The magnetic fluorescent microbeads are not particularly limited, but examples that can be used include MILLIPLEX® Bovine Cytokine / Chemokine Magnetic Bead Panel 1 (manufactured by Millipore). The multi-parameter simultaneous measurement device is also not particularly limited, but examples that can be used include Bio-Plex MAGPIX, a multi-parameter simultaneous measurement device manufactured by BIO-RAD.

[0034] The cutoff values ​​for the amount of IL-8 are, for example, 50 to 500 pg / mg, 50 to 480 pg / mg, 50 to 285 pg / mg, 50 to 220 pg / mg, 50 to 215 pg / mg, 50 to 180 pg / mg, 50 to 155 pg / mg, 55 to 500 pg / mg, 55 to 480 pg / mg, 55 to 285 pg / mg, 55 to 220pg / mg, 55~215pg / mg, 55~180pg / mg, 55~155pg / mg, 70~500pg / mg, 70~480pg / mg, 70~2 85pg / mg, 70~220pg / mg, 70~215pg / mg, 70~180pg / mg, 70~155pg / mg, 80~500pg / mg, 80~48 0pg / mg, 80~285pg / mg, 80~220pg / mg, 80~215pg / mg, 80~180pg / mg, 80~155pg / mg, 155~5 00pg / mg, 155~480pg / mg, 150~290pg / mg, 155~285pg / mg, 155~220pg / mg, 155~215pg / mg, The IL-8 level can be set within ranges such as 155 to 180 pg / mg, 180 to 500 pg / mg, 180 to 480 pg / mg, 180 to 285 pg / mg, 180 to 220 pg / mg, 180 to 215 pg / mg, 215 to 500 pg / mg, 215 to 480 pg / mg, 215 to 285 pg / mg, and 215 to 220 pg / mg. That is, if the amount of IL-8 in the cervical mucus of a bovine subject to screening is less than a numerical value within these ranges, the bovine subject to screening can be considered to be at risk of dystocia.

[0035] The cutoff values ​​for the amount of TNFα are, for example, 3 to 11 pg / mg, 3 to 10.5 pg / mg, 3 to 10 pg / mg, 3 to 9.5 pg / mg, 3 to 9 pg / mg, 3 to 8.5 pg / mg, 3 to 8 pg / mg, 3 to 7.5 pg / mg, 3 to 7 pg / mg, 3.5 to 11 pg / mg, 3.5 to 10.5 pg / mg, 3.5 to 10 pg / mg, and 3.5 to 9.5 pg / mg. / mg, 3.5~9pg / mg, 3.5~8.5pg / mg, 3.5~8pg / mg, 3.5~7.5pg / mg, 3.5~7pg / mg, 4~11pg / mg, 4~10.5pg / mg, 4~10pg / mg, 4~9.5pg / mg, 4~9pg / mg, 4~8.5pg / mg, 4~8pg / mg, 4~7.5pg / mg, 4~7pg / mg, 4.5~11 pg / mg, 4.5~10.5pg / mg, 4.5~10pg / mg, 4.5~9.5pg / mg, 4.5~9pg / mg, 4.5~8.5pg / mg, 4.5~8pg / mg, 4.5~7.5pg / mg, 4.5~7pg / mg, 5~11pg / mg, 5~10.5pg / mg, 5~10pg / mg, 5~9.5pg / mg, 5~9pg / mg, 5~8.5 The TNFα level can be set within ranges such as 5 to 8 pg / mg, 5 to 7.5 pg / mg, 5 to 7 pg / mg, 5.5 to 11 pg / mg, 5.5 to 10.5 pg / mg, 5.5 to 10 pg / mg, 5.5 to 9.5 pg / mg, 5.5 to 9 pg / mg, 5.5 to 8.5 pg / mg, 5.5 to 8 pg / mg, 5.5 to 7.5 pg / mg, 5.5 to 7 pg / mg, etc. It can also be set within ranges such as 9 to 11 pg / mg, 9.5 to 11 pg / mg, 10 to 11 pg / mg, etc. In other words, if the amount of TNFα in the cervical mucus of a bovine subject to screening is less than a numerical value within these ranges, it can be determined that there is a risk of dystocia.

[0036] The prediction method of the present invention can also use cutoff values ​​for both the amount of IL-8 and TNFα, or a cutoff value for either the amount of IL-8 or TNFα.

[0037] The prediction method of the present invention can be used in combination with one or more of these, or can be used in appropriate combination with conventional prediction methods using blood or body temperature.

[0038] The cause of bovine dystocia determined by the prediction method of the present invention is not limited, but from the viewpoint of significantly achieving the effects of the present invention, it is preferable that the dystocia be caused by one or more causes selected from the group consisting of birth canal abnormalities, fetal enlargement, fetal dislocation, abnormal labor, and multiple births, and it is more preferable that the dystocia be caused by birth canal abnormalities.

[0039] [Composition for predicting the risk of bovine dystocia] One aspect of the present invention relates to a composition used for predicting the risk of dystocia in cattle, comprising one or more antibodies or functional fragments thereof selected from the group consisting of antibodies or functional fragments thereof that specifically bind to IL-8 in cervical mucus, or antibodies or functional fragments thereof that specifically bind to TNFα.

[0040] The conditions and methods for the antibody of the present invention, its functional fragment, or a composition containing them are as described in [Method for predicting the risk of dystocia in cattle].

[0041] [Cattle dystocia risk prediction kit] One aspect of the kit of the present invention relates to a kit for predicting the risk of dystocia in cattle, comprising one or more antibodies or functional fragments thereof selected from the group consisting of antibodies or functional fragments thereof that specifically bind to IL-8, and antibodies or functional fragments thereof that specifically bind to TNFα.

[0042] In another embodiment, the kit of the present invention may comprise a composition used for predicting the risk of dystocia in cattle, comprising one or more antibodies or functional fragments thereof selected from the group consisting of antibodies or functional fragments thereof that specifically bind to IL-8, or antibodies or functional fragments thereof that specifically bind to TNFα. Another embodiment of the kit of the present invention may be a PCR kit comprising one or more primers selected from the group consisting of primers capable of detecting IL-8 and primers capable of detecting TNFα.

[0043] Another embodiment of the kit of the present invention is a kit for predicting the risk of dystocia in cattle, comprising a substance capable of specifically detecting genes encoding IL-8 and / or TNFα in cervical mucus from pregnant cattle. The kit of the present invention preferably comprises, but is not limited to, a collection container, a dissolving solution for preparation immediately after use, a needle, and / or an instruction manual, in addition to the substance capable of specifically detecting genes encoding IL-8 and / or TNFα. The conditions and details of the kit of the present invention are as described in the "Method for predicting the risk of dystocia in cattle." [Example]

[0044] Next, the present invention will be specifically explained by way of examples, but the present invention is not limited to the following examples.

[0045] The statistical analysis of the test results below was carried out under the following statistical analysis conditions. <Statistical analysis conditions> · Kruskal-Wallis test was performed and Holm method was used as a post-hoc test - Draw ROC curves and calculate optimal cutoff values Results are expressed as mean ± standard error All significance levels <0.05 EZR is used as the statistical analysis software (EZR is a statistical software that extends the functionality of R and R Commander).

[0046] [Collection of bovine cervical mucus and sample preparation] The study subjects were 145 Japanese Black cows (multiparous cows, average age: 6.72 years) that had been requested for house calls by the Miyazaki Prefectural Agricultural Mutual Aid Association due to long-term gestation, among those raised in Miyazaki Prefecture. Here, long-term gestation is defined as cows that have not given birth even after 295 days of gestation, according to the Ministry of Agriculture, Forestry and Fisheries' Livestock Mutual Aid Association's criteria for illness and injury accident benefits.

[0047] Of the 145 cows that requested the above-mentioned house calls, they were divided into three groups based on the status of veterinarian assistance in childbirth: easy birth, difficult birth, and Caesarean section. The easy birth group consisted of cows that did not require veterinarian assistance, accounting for 121 of the total. The difficult birth group consisted of cows that received veterinarian assistance, accounting for 22 of the total. The Caesarean section group consisted of cows that received veterinarian assistance and then underwent a Caesarean section, accounting for 2 of the total.

[0048] The vulva of each of the 145 cows, which were at least 295 days (mean ± SD: 296.72 ± 2.08) after artificial insemination, was disinfected with 0.02% mono, bis (trimethylammonium methylene chloride)-alkyltoluene. Afterwards, a hand wearing a glove (Fujidaira Kogyo Co., Ltd.) disinfected with 70% ethanol was inserted into the vagina, and approximately 3 g of cervical mucus was collected using a palm-sized finger. The glove containing the collected mucus was turned inside out, and the glove was transported to the clinic at 4°C within a maximum of 5 hours of collection and stored at -20°C or below. The mucus was then transported to the University of Miyazaki at -15°C or below and stored at -30°C or below until cytokine concentration measurements were made.

[0049] [Test Example 1: Confirmation test of inflammatory cytokine concentrations in bovine cervical mucus] The collected mucus was thawed on ice, and 800 μL of phosphate-buffered saline (PBS) was added to 0.2 g of the thawed mucus to prepare a mixed solution. The mixed solution was then subjected to an automatic tissue dispersion and homogenization apparatus (gentleMACS TM The cells were disrupted using a Dissociator (Miltenyi Biotec) and centrifuged at 4°C, 4700 rpm, and 20 minutes to obtain the supernatant.

[0050] The total protein concentration in the bovine cervical mucus was measured using a NanoDrop microspectrophotometer (NanoDrop Eight, Thermo Scientific). Magnetic fluorescent microbeads (MILLIPLEX® Bovine Cytokine / Chemokine Magnetic Bead Panel 1, Millipore) were added to the supernatant, and the concentrations of IL-1α, IL-1β, IL-6, IL-8, IL-10, and TNFα were quantified using a multiparameter analyzer (Bio-Plex MAGPIX, BIO-RAD).

[0051] The concentrations of each cytokine in the cervical mucus of each group of cows measured above were divided by the total protein concentration, and the results are shown in Figure 1. Here, the units on the vertical axis are all pg / mg.

[0052] Figure 1 confirms that the concentrations of IL-1α, IL-1β, IL-6, IL-8, IL-10, and TNFα were lower in the dystocia and cesarean section groups compared to the easy birth group. Furthermore, the differences in the concentrations of IL-8 and TNFα between the easy birth and difficult birth groups were significantly large and significant (P<0.01).

[0053] [Test Example 2: Examination of specificity and sensitivity in IL-8 and TNFα] Graphs of the ROC curves for IL-8 and TNFα, for which significant differences in concentration were observed between the easy delivery group and the difficult delivery group in Test Example 1, are shown in FIGS.

[0054] Furthermore, the areas under the ROC curves for IL-8 and TNFα were calculated and the results are shown in Tables 1 and 2.

[0055] [Table 1]

[0056] [Table 2]

[0057] As shown in Figure 2 and Table 1, the area under the ROC curve for IL-8 was 0.712 (95% confidence interval 0.611-0.814), and the optimal cutoff value for predicting the risk of dystocia in cows was 54.44 pg / mg (specificity 0.711, sensitivity 0.667) as the IL-8 concentration / total protein concentration ratio. When sensitivity was emphasized, the optimal cutoff value was 220.68 pg / mg (specificity 0.41, sensitivity 0.92).

[0058] As shown in Figure 3 and Table 2, the area under the receiver operating characteristic curve for TNFα was 0.709 (95% confidence interval 0.613-0.805), and the optimal cutoff value for predicting the risk of dystocia in cows was 4.4 pg / mg (specificity 0.65, sensitivity 0.67) as the TNFα concentration / total protein concentration ratio. When sensitivity was emphasized, the optimal cutoff value was 10.26 pg / mg (specificity 0.40, sensitivity 1.00).

[0059] Figure 4 shows the results of a graph plotting the concentrations of IL-8 and TNFα for each individual cow in each group. The units of the vertical and horizontal axes in Figure 4 are pg / mg. By setting the cutoff values ​​for IL-8 at 150 pg / mg and TNFα at 10 pg / mg, it is believed that it is possible to predict the risk of dystocia with a probability of 80% for the cows screened. Furthermore, by setting the cutoff values ​​for IL-8 at 290 pg / mg and TNFα at 11 pg / mg, it is believed that it is possible to predict the incidence of dystocia due to incomplete cervical ripening in cows not screened as zero.

[0060] The present invention is not limited to the above-described embodiments and examples, and various modifications are possible within the scope of the claims. The technical scope of the present invention also includes embodiments obtained by appropriately combining the technical means disclosed in different embodiments. Furthermore, all academic literature and patent documents described in this specification are incorporated herein by reference.

Claims

1. 1. A method for predicting a risk of dystocia in a cow, comprising: (i) measuring the amount of IL-8 and / or TNFα in cervical mucus of pregnant cows; A method for predicting the risk of dystocia in cattle, including:

2. Furthermore, (ii) determining that the pregnant cow is at risk of dystocia when the amount of IL-8 and / or TNFα measured in the step (i) is less than a cutoff value; The method of claim 1 , comprising:

3. the cutoff value is a value determined by the concentration of the IL-8 relative to the total protein concentration in the cervical mucus and / or the concentration of the TNFα relative to the total protein concentration in the cervical mucus, the cutoff value of the IL-8 is 50 to 290 pg / mg; the cutoff value of TNFα is 4 to 11 pg / mg; The method of claim 2.

4. The method according to claim 1 or 2, wherein the pregnant cow is a cow for which 270 days or more have passed since the date of artificial insemination.

5. The method according to claim 1 or 2, wherein the difficult labor is caused by an abnormality in the birth canal.

6. A composition for predicting the risk of dystocia in cattle, comprising an antibody specific to bovine IL-8 or a functional fragment thereof, or an antibody specific to TNFα or a functional fragment thereof.

7. The composition according to claim 6, wherein the difficult labor is caused by an abnormality in the birth canal.

8. A kit for predicting the risk of dystocia in cattle, comprising at least one antibody selected from the group consisting of an antibody specific to bovine IL-8 or a functional fragment thereof, and an antibody specific to TNFα or a functional fragment thereof.

9. A kit for predicting the risk of dystocia in cows, comprising one or more substances selected from the group consisting of substances capable of specifically detecting a gene encoding IL-8 and substances capable of specifically detecting a gene encoding TNFα in the cervical mucus of pregnant cows.

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