CN112772558A - Method for improving stress and meat quality of live pigs by using protein acetylation inhibitor - Google Patents
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Abstract
The invention discloses a method for improving stress and meat quality of live pigs by using a protein acetylation inhibitor, which comprises the following steps: injecting curcumin into abdominal cavity half an hour to 1 hour before slaughtering, with the dosage of 15-20mg/kg, can significantly reduce stress level of live pig, and is manifested by reduced concentration of creatine kinase, lactate dehydrogenase, cortisol and heat shock protein in blood; the meat quality of main meat cuts such as shoulder meat, longissimus dorsi, psoas major, semimembranosus and semitendinosus is obviously improved, and the meat cuts show that the activity of creatine kinase and hexokinase is reduced, the activity of pyruvate kinase is increased, the content of glycogen and ATP is high, the content of lactic acid is low, the pH reduction of muscle is slowed down, and the occurrence rate of PSE meat is reduced. The method can not only reduce animal stress, but also improve meat quality.
Description
Technical Field
The invention relates to a method for improving stress of live pigs and meat quality by applying a protein acetylation inhibitor, belonging to the technical field of food processing.
Background
Pork is one of indispensable animal source foods for most people in China and is an important source for people to obtain high-quality protein, heme iron, zinc and B vitamins. In recent years, as people have more and more high requirements for good life, high-quality pork products are not lacked.
In the industrialized slaughtering and processing process of live pigs, due to the influence of factors such as long-distance transportation, high-temperature and high-humidity climate, low-temperature and cold air temperature, artificial improper drive and the like, pre-slaughtered animals are seriously stressed, pork with poor appearance quality and eating quality such as PSE (Meat by soy), and the like, has high incidence, influences the purchase desire of consumers, often causes serious consequences such as return of goods, lost sales, complaints and the like, and brings huge economic loss and social influence to the Meat industry. Although a great deal of research shows that the occurrence rate of PSE meat can be reduced to a certain extent by appropriate resting and resting before slaughtering, in the actual operation process, the effect of resting and resting measures before slaughtering is greatly different due to the fact that the management specifications are not uniform, the quality of operators is inconsistent and the like. Therefore, the method for relieving animal stress and improving meat quality, which is simpler, more effective and convenient to operate, becomes a key technology which is urgently needed by pig slaughtering enterprises.
Existing studies show that PSE pork is mainly related to energy metabolism, particularly anaerobic glycolysis, in early muscle before and after slaughter of animals, and pH is reduced too fast, and is related to phosphorylation or dephosphorylation, acetylation or deacetylation of energy metabolism enzymes (Zou et al. jafc,2020), so that from the biochemical point of view, acetylation of energy metabolism enzymes in muscle can be properly controlled, and the anaerobic glycolysis process and lactic acid generation in pork can be theoretically controlled, and the pH reduction rate is further reduced.
Curcumin is considered to be a high-efficiency protein acetylation inhibitor, and has been researched and applied to treatment and prevention of human diseases to a certain extent. In the production of pigs, research reports on short-term treatment of various diseases and long-term stress prevention by using curcumin are also reported. From the standpoint of animal physiology, many oceans (2010, master paper of Nanjing university of agriculture) have found that oral administration of 8mg/kg body weight of curcumin per day 3 weeks prior to transport can alleviate plasma cortisol elevation caused by transport stress, and can reverse or alleviate pig hippocampal NO/NOS system activation caused by transport stress, but the effect of curcumin treatment on meat quality has not been studied. Wang bin et al (2010, livestock and veterinary, volume 45, period 7: 47-49) and Zhu Guo Qiang (2013, feed industry, volume 34, period 16: page 9-12) report that feeding curcumin at 400mg/kg per day five weeks before slaughter can significantly increase slaughter live weight, lean meat percentage and eye muscle area, reduce backfat thickness, and show good production performance, but inhibit the post-slaughter pH drop (the pH 24h after slaughter is still 6.34, the meat color score is significantly lower, and is the character of DFD meat), the meat quality is reduced, and the purpose of improving meat quality cannot be achieved. Zhou Ming et al (2014, reported in the food and oil sciences of China, volume 29, No. 3: 67-73) report that curcumin treatment at the same dosage has no influence on pork pH, but causes obvious change of meat color (brightness value and yellowness value are increased, meat color is deteriorated), contradict with the research result, and indicate that the production performance is improved at the cost of sacrificing the quality of meat by high-dosage curcumin feeding before slaughter. Today, where meat quality is an increasing concern, this technique should be used with caution. Patent applications (201310607656.5) and (201911007241.8) mention that two curcumin-containing mixed feed additives have certain improvement effect on pork quality, but both need to be mixed with other components for use, and the effect of curcumin is not clear. Meanwhile, the additives are used in the production link, so that the use time is long, the comprehensive cost is high, the effect is unstable, and the technical popularization has certain limitations.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a method for improving stress and meat quality of live pigs, which has a clear action mechanism, is simple and easy to operate, and has low cost, so as to meet the important technical requirements of live pig slaughtering and processing enterprises on reducing the stress of animals before slaughtering and improving the meat quality after slaughtering, particularly reducing the incidence rate of white muscle (PSE meat) and producing high-quality pork.
The first purpose of the invention is to provide a method for improving stress and meat quality of live pigs by using a protein acetylation inhibitor, wherein the protein acetylation inhibitor is curcumin; the method comprises injecting curcumin into pig body by one-time intraperitoneal injection before pig slaughtering.
Further, the curcumin is injected for 0.5-1.0 h before slaughtering the live pigs.
Further, the injection dosage of the curcumin for one-time intraperitoneal injection is 15-20mg/kg of body weight. In a specific embodiment, the curcumin mother liquor is injected in an amount of 1800-2400 mg based on 120kg of commercial pigs.
Furthermore, the one-time intraperitoneal injection is to dissolve curcumin in dimethyl sulfoxide to prepare curcumin mother liquor for intraperitoneal injection, and the concentration of the curcumin mother liquor is 80-100 mg/mL.
Further, the dosage of the curcumin mother liquor for intraperitoneal injection is 18-30 mL.
A second object of the invention is to provide the use of the aforementioned method for reducing stress levels in live pigs and/or for improving the quality of meat.
Further, the meat quality is improved by reducing the rapid reduction of pH, the risk of PSE meat is avoided, and the white muscle occurrence rate is reduced.
Further, the application is to reduce the stress level of the live pigs by reducing the concentration of creatine kinase, lactate dehydrogenase, cortisol and heat shock protein in blood,
further, the application is to improve the meat quality by reducing the activity of creatine kinase and hexokinase, reducing the content of lactic acid, slowing down the pH drop of muscle and increasing the activity of pyruvate kinase, the content of glycogen and ATP.
The third object of the present invention is to provide the use of the aforementioned method for reducing creatine kinase and lactate dehydrogenase activity in live pig blood and/or for reducing cortisol and heat shock protein concentrations in live pig blood.
The fourth object of the present invention is to provide the use of the aforementioned method for reducing creatine kinase and/or hexokinase activity in live pig muscle, and/or reducing lactic acid content in live pig muscle, and/or slowing down the rate of pH drop in live pig muscle, and/or increasing pyruvate kinase activity in live pig muscle, and/or increasing glycogen and ATP content in live pig muscle, reducing the incidence of white muscle PSE meat.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
(1) the curcumin is used as an acetylation inhibitor for the first time and is applied to the production of animal husbandry, and the curcumin is injected through the abdominal cavity before slaughter, so that the curcumin injection has the advantages of definite injection dosage, short action time, simple operation steps and the like. And the experimental result proves that the energy metabolism related index which plays a decisive role in the formation of the meat quality after slaughtering has a significant improvement effect compared with a control group after curcumin treatment, so that the curcumin-modified meat quality control agent has a positive effect on reducing the incidence rate of heterogeneous meat and improving the meat quality.
(2) By adopting the method determined by the invention, the stress level of the live pig can be obviously reduced by using curcumin treatment, which is shown by the reduction of creatine kinase activity, lactate dehydrogenase activity, cortisol and heat shock protein concentration in blood;
(3) by adopting the method determined by the invention, the meat quality of the main divided meat of shoulder meat, longissimus dorsi, psoas major, semimembranosus, semitendinosus muscle and the like of the live pig is obviously improved, and the meat is expressed by the reduction of the activity of creatine kinase and hexokinase, the increase of the activity of pyruvate kinase, the high content of glycogen and ATP, the low content of lactic acid and the slowing of the pH reduction of the muscle.
(4) The method is implemented within half an hour to 1 hour before slaughtering, is simple to operate, low in cost, obvious in meat quality improvement and wide in application prospect.
Drawings
FIG. 1 is a graph showing the change of stress-related indices in sera of different treatment groups of example 1 (in the graph: C, control group; Ati, acetyltransferase inhibitor injection group of the present invention; DATi, deacetylase inhibitor injection group. a, b: different letters indicate that the difference is significant P < 0.05; n ═ 10); wherein,
FIG. 1A: creatine kinase and lactate dehydrogenase activity in the sera of the different treatment groups;
FIG. 1B: cortisol and Heat shock protein 70 levels in sera of different treatment groups.
FIG. 2 is a graph showing the effect of different treatments on glycolytic enzyme activity in example 1 (in the figure C, control; Ati: acetyltransferase inhibitor-injected group of the present invention; DATi: deacetylase inhibitor-injected group. A, B: different letters indicate that the difference between the three treatment groups is significant P < 0.05; a, B, C, d: different letters indicate that the difference between five sites in the same group is significant P < 0.05; and n ═ 10). Wherein,
FIG. 2A. Effect of different treatments on glycolytic enzyme hexokinase activity;
FIG. 2B is a graph of the effect of different treatments on glycolytic enzyme pyruvate kinase activity;
FIG. 2C is a graph of the effect of different treatments on the phosphofructokinase activity of the glycolytic enzyme;
FIG. 2D Effect of different treatments on the glycolytic enzyme creatine kinase activity.
FIG. 3 is a graph showing the effect of different treatments on postmortem energy metabolism in example 1 (in the graph, C: control group; Ati: acetyltransferase inhibitor-injected group of the present invention; DATi: deacetylaltransferase inhibitor-injected group. A, B: different letters indicate that the difference between the three treatment groups is significant P < 0.05; a, B, C: different letters indicate that the difference between five sites in the same group is significant P < 0.05; and n ═ 10). Wherein,
FIG. 3A Effect of different treatments on ATP metabolism;
FIG. 3B effects of different treatments on glycogen metabolism;
FIG. 3C Effect of different treatments on lactate metabolism.
Detailed Description
The following examples are intended to further illustrate the present invention, but are not intended to limit the scope of the invention.
Example 1
The processing of this embodiment is divided into three processing groups, specifically as follows:
(1) 30 pigs were divided into three groups, i.e., a control group, an acetyltransferase inhibitor-injected group, and a deacetylase inhibitor-removed group 30 minutes before slaughtering, in the following manner:
control group: preparing a solvent by the proportion of 20 percent of dimethyl sulfoxide and 80 percent of normal saline in volume ratio, wherein the intraperitoneal injection dose is 0.97ml/kg of body weight (a control group C, n is 10);
acetyltransferase inhibitor injection group: dissolving curcumin in dimethyl sulfoxide to obtain curcumin mother liquor with concentration of 80mg/mL, and performing intraperitoneal injection according to the dose of 20mg/kg body weight to obtain curcumin mother liquor with total amount of 30mL (Ati, n is 10);
③ Deacetyltransferase inhibitor injection group: dissolving trichostatin A injection in dimethyl sulfoxide to prepare trichostatin A injection with the concentration of 7.5 mg/mL; dissolving nicotinamide in normal saline to prepare 50mg/mL nicotinamide injection, wherein the injection dosage of trichostatin A is 1.25mg/kg, the injection dosage of nicotinamide is 38.9mg/kg, and the two injections are injected simultaneously (DATi, n is 10).
(2) 30min after injection, the live pigs are killed according to the national standard GB/T17236 live pig slaughtering operation rules.
The method comprises the following steps of centrifuging blood at 2000g and 4 ℃ for 10min (AvaniJ-E; Beckman, Brea, CA), taking supernatant (serum), determining creatine kinase activity by adopting a creatine kinase test box (product number A032) produced by Nanjing institute of bioengineering, determining lactate dehydrogenase activity by using a lactate dehydrogenase test box (product number A020-2) produced by the company, determining Cortisol content by using a pig skin alcohol (Cortisol) enzyme-linked immunoassay kit produced by the company, and determining heat shock protein 70 content by using a HSP70 enzyme-linked immunoassay kit produced by the company.
Measuring creatine kinase activity by taking shoulder, longissimus dorsi, psoas major, semimembranosus and semitendinosus, measuring the creatine kinase activity by adopting a creatine kinase test box (product number A032) produced by Nanjing institute of bioengineering, measuring hexokinase activity by using a hexokinase test box (product number A077-1-1) produced by the Nanjing institute of bioengineering, measuring pyruvate kinase activity by using a pyruvate kinase test box (product number A076-1-1) produced by the Nanjing institute of bioengineering, measuring glycogen content by using a glycogen test box (product number A043) produced by the Nanjing institute of bioengineering, measuring the lactic acid content by using a lactic acid test box (product number A019-2) produced by the Nanjing institute of bioengineering, and measuring the ATP content by using an ATP test box (product number S0026) produced by the Nanjing institute of bioengineering.
Taking shoulder meat, longissimus dorsi, psoas major, semimembranosus and semitendinosus, and measuring the pH (mainly reflecting the early pH reduction rate after slaughter, closely related to the quality of the meat, wherein the lower the pH, the faster the reduction rate is, the worse the quality of the meat is, and the pH is less than or equal to 6.20, and is judged as PSE meat) and the meat color L a b after slaughter in accordance with the agricultural industry standard' meat food quality objective evaluation method NY/T2793-.
(3) The results show that: the creatine kinase, lactate dehydrogenase activity, cortisol and heat shock protein concentration in the blood of the curcumin treated group were reduced to some extent compared with the control group (fig. 1), indicating that the method of the invention can reduce the stress level of live pigs;
compared with a control group, the curcumin-treated group has the advantages that the activity of creatine kinase and hexokinase in shoulder meat, longissimus dorsi, psoas major, semimembranosus, semitendinosus and other main divided meat is reduced, the activity of pyruvate kinase is increased (figure 2), the content of glycogen and ATP is high, and the content of lactic acid is reduced (figure 3), which shows that the method can improve the meat quality while reducing the stress level of the live pigs.
The curcumin treated group developed a lower incidence of PSE meat in the five selected meat cuts than the control group, with a generally similar meat color index (table 1).
TABLE 1 Effect of different treatments on meat quality 45min post slaughter
Table 1 shows the effect of the different treatments on the meat quality 45min after slaughter in example 1 (C: control; Ati: acetyltransferase inhibitor-injected group of the present invention; DATi: deacetylase inhibitor-injected group. x, y, z: different letters in each column indicate significant differences P <0.05 between the different treatment groups; n: 10. L: brightness values, a: redness values, b: yellowness values).
Compared with the existing research reports, the obtained result of the invention has obvious benefits, obvious and definite effects on reducing the pre-slaughter stress and PSE meat incidence, low dosage and simple use.
The above description is only for the purpose of understanding 1 specific embodiment of the present invention by those skilled in the art, and is not intended to limit the scope of the present invention as claimed. All equivalent changes in the features, structures and principles of the invention as described in the claims should be embraced therein.
Claims (9)
1. A method for simultaneously improving stress and meat quality of live pigs by using a protein acetylation inhibitor is characterized in that the protein acetylation inhibitor is curcumin; the method comprises injecting curcumin into pig body by one-time intraperitoneal injection before pig slaughtering.
2. The method according to claim 1, wherein curcumin is injected for 0.5-1.0 h before slaughtering of live pigs.
3. The method as claimed in claim 1, wherein the curcumin is injected in a single intraperitoneal injection at a dose of 15-20mg/kg body weight.
4. The method as claimed in claim 3, wherein the one-time intraperitoneal injection is performed by dissolving curcumin in dimethyl sulfoxide to prepare curcumin mother liquor, and the concentration of the curcumin mother liquor is 80-100 mg/mL.
5. The method as claimed in claim 4, wherein the dose of the curcumin mother solution for intraperitoneal injection is 18-30 mL.
6. Use of the method of any one of claims 1 to 5 for reducing stress levels in live pigs and/or for improving meat quality.
7. Use according to claim 6, wherein the improvement in meat quality is a reduction in white muscle incidence.
8. Use of the method according to any one of claims 1 to 5 for reducing creatine kinase and lactate dehydrogenase activity in live pig blood and/or for reducing cortisol and heat shock protein concentrations in live pig blood.
9. Use of the method of any one of claims 1 to 5 for reducing creatine kinase and hexokinase activity in live pig muscle, and/or reducing lactic acid content in live pig muscle, and/or slowing the rate of pH drop in live pig muscle, and/or increasing pyruvate kinase activity in live pig muscle, and/or increasing glycogen and ATP content in live pig muscle, reducing the incidence of white muscle PSE meat.
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