Method for assessing stress resistance in pigs

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science and agriculture. The method provides pig's blood analysis for the following values: malondialdehyde MDA, antioxidant activity AOA, total oxidative activity of blood plasma TOA, catalase activity CT, superoxide dismutase SOD. The two-month pigs having MDA 6.02±0.13 absorption unit/ml and less, AOA 60.86±0.31% and more, TOA 71.64±0.16% and less, CT 57.30±1.05 absorption unit/ml min and more, SOD 14.57±0.44 absorption unit/ml and more are considered to be stress-resistant PSS "-"; the values of MDA 7.63±0.07 absorption unit/ml and more, AOA 50.22±0.42% and less, TOA 79.21±0.83% and more, CT 47.16±1.08 absorption unit/ml min and less, SOD 10.49±0.49 absorption unit/ml and less enables stating the pigs as stress-sensitive PSS "+". The six-month pigs having MDA 7.37±0.17 absorption unit/ml and less, AOA 61.93±0.25% and more, TOA 41.86±0.19% and less, CT 76.68±0.74 absorption unit/ml min and more, SOD 22.80±0.68 absorption unit/ml and more are considered to be stress-resistant PSS "-"; the values of MDA 8.05±0.09 absorption unit/ml and more, AOA 59.72±0.17% and less, TOA 46.21±0.09% and more, CT 71.76±0.51 absorption unit/ml min and less, SOD 19.91±0.59 absorption unit/ml and less enables stating the pigs as stress-sensitive PSS "+".

EFFECT: method enables the high-accuracy stress resistance assessment of the two- and six-month pigs.

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The invention relates to the field of agriculture, the pig.

In terms of industrial cultivation of animals experiencing stress, causing significant economic damage. Pigs are particularly sensitive to stressors compared to other species of farm animals.

To successfully dealing with stress it is necessary to have reliable methods for determining stressfulness in pigs. Currently-proposed methods for diagnosing stress-state in pigs is not always available, unbiased and beneficial.

There is a method of determining stressoustojchivosti animals, in particular pigs, using turpentine test method Aiguaviva and Passagalia (1989). The essence of turpentine test consists in determining a response to the introduction into the body terpentine oil (purified turpentine) (Kuznetsov A.I., Sunagatullin FA Who gentler? J. Pig, 1991, No. 2. - P.28-29).

The disadvantage of this method is to a large error (inaccuracy) of the results.

Known for using the method to determine stressfulness pigs, in which piglets distribute stresstesting (positively reacting to halothane) and stressoustoicivij (not responding to halothane) (Wittmann M meat Quality in pigs sensitive to halothane gas. - International S.-H. journal, 195, No. 2. - S-86; Kovalenko V.A., Goldman ID, A.S. Orlov guidelines for the assessment of meat production, meat quality and fat pigs. 1987. - 64 S.).

The disadvantage is the case when testing stresstesting pigs up to 4% of their goals, the need for special equipment for inhalation narcosis, damage to the health of the operator carrying out animal testing.

There is a method of determining stressoustojchivosti pigs by setting polymerase chain reaction (PCR) according to method K. Mullis (1985), advanced R.Boometal. (1990) and modified Navlog (2002). It allows you to identify the genetic mutation in the gene for the receptor ryanodine (RYR-1) and install the genotypes NN, Nn - strasssteine, nn - stresstesting. For this purpose it is necessary to sample the ear (with an area of 1 cm²), which for the storage and transportation are placed in a Dewar vessel (with liquid nitrogen), or citrate blood (kowaluk NV Use in breeding pigs genetic markers stressoustojchivosti and multiple pregnancy. Abstract. dis. Kida. Biol. Sciences. - Borovsk, 2002. - 26 S.).

The disadvantage is that the laboratory setting polymerase chain reaction agriculture cannot have because of their high cost, they are in major cities (Krasnodar, Moscow and others), which requires high costs to tawki samples.

In terms of studying the biochemical mechanisms underlying the adaptive reactions, reduction resistance and pathology, currently, more and more attention of researchers is attracted processes peroxide (free radical) oxidation of lipids and the status of the antioxidant defense system of the organism.

The purpose of the invention is to provide low-cost methods available for assessing stressoustojchivosti pigs, using indicators of antioxidant defenses and lipid peroxidation.

To determine stressoustojchivosti (PSS) in pigs at the age of two or six months, depending on the requirements of the economy before feeding take blood from tail or ear artery and determine the parameters characterizing the status of the antioxidant defense system (AOD) and lipid peroxidation (LPO) (Methodical recommendations for the diagnosis, therapy and prevention of metabolic disorders in productive animals. Voronezh, 2005. - 94 C.):

the activity of superoxide dismutase (SOD) - method ..Misra, I.Fridovich (1972);

the activity of catalase (cat) Korolyuk, Lievanos, Igumenova, Westonaria (1988);

- the total oxidative activity of blood plasma (OOA) - Lperational, Averagenova (1998);

- total antioxidant activity (AOA) for the inhibition of oxidized Tween-80 to noted that a well known the CSOs dialdehyde according to Harutyunyan, Eby, ZnO (2000);

the content of malondialdehyde (MDA) by reaction with 2-thiobarbituric acid on Idestroy, Aggarwala (1977).

On the basis of the results of the studies make conclusions about PSS pigs on a scale of assessment of stressoustojchivosti and determine their future use.

Stressoustoicivij (PSS) when pigs used in further breeding to derive stressoustoicivij lines. Stresstesting (PSS "+") to increase stressoustojchivosti hybrid PSS "-" pigs, or sent to fattening.

Studies were conducted on farms in the Rostov region on the gilts two and six months. Based on these series of experiments were derived scale for measuring stressoustojchivosti pigs at the age of two and six months on indicators of antioxidant defense (AOD) and lipid peroxidation (LPO) (table 1, 2).

If animals values of the studied parameters are related boundaries PSS+and PSS "-", then they are referred to "conditionally sensitive to stress.

Example 1. One series of experiments to derive the scale of stressoustojchivosti pigs at the age of two months. We selected a group of animals-analogues of the origin, growth and development of the 20 goals by the age of two months. The selected pigs before feeding took the blood from the tail artery. Pigs according to the method of polymerase chain reaction were divided into two groups: I - strasssteine, II - stresstesting. For PCR samples citrate blood was sent to SKNIIG Krasnodar. In parallel, in selected blood was determined by the activity of superoxide dismutase (SOD), catalase (cat), antioxidant AK is Yunosti (AAO) and total oxidative activity of blood plasma (AOA) and the content of malondialdehyde (MDA) (table 3).

On the basis of the received data series of such experiments, we suggest a rating scale of stressoustojchivosti pigs at two months of age on indicators of antioxidant defense and lipid peroxidation (table 1).

Example 2. Determined stressoustojchivosti pigs (age two months) on indicators of antioxidant defense and lipid peroxidation. For this selected 10 pigs at the age of two months. In piglets before feeding took the blood from the tail artery in an amount of 5 ml of blood was determined by the activity of superoxide dismutase (SOD), catalase (cat), antioxidant activity (AOA), total oxidative activity of blood plasma (AOA), the content of malondialdehyde (MDA). The research results are reflected in table 4.

To determine stressoustojchivosti pigs at two months of age used the Lee scale (table 1). Analysis of the obtained data showed that gilts under the numbers 1, 2, 3, 4, 8, 10 have indices in the range typical for stressoustoicivij animals, and on the basis that they were related to PSS "-" animals. Number 5, 6 and 7 AOA, AAO, CT, MDA, SOD, respectively, within 79,21±0,83% and above, 50,22±0,42% and below, 47,16±1,EU/ml and below, 7,63±0,07 EA/ml and above, 10,49±0,49 EA/ml and below is stresstesting animals. At gilt at number 9 values of the studied parameters are related boundaries PSS+and PSS "-" animals, therefore, considered to be "conditionally" sensitive to stress.

Example 3. One series of experiments to derive the scale of stressoustojchivosti pigs at the age of six months. We selected a group of animals-analogues of the origin, growth and development of the 22 goals by the age of six months. The selected pigs before feeding took the blood from the tail artery. Pigs according to the method of polymerase chain reaction were divided into two groups: I - strasssteine, II - stresstesting. For setting the NDP citrate sample of blood was sent to SKNIIG Krasnodar. In parallel, in selected blood was determined by the activity of superoxide dismutase (SOD), catalase (cat), antioxidant activity (AOA) and total oxidative activity of blood plasma (AOA) and the content of malondialdehyde (MDA) (table 5).

On the basis of the received data series of such experiments, we suggest a rating scale of stressoustojchivosti swine six months of age on indicators of antioxidant defense and lipid peroxidation (table 2).

Example 4. Determined stressoustojchivosti pigs (age six months) on indicators of antioxidant defense and lipid peroxidation. For this selected 10 pigs at the age of six months. In piglets before feeding took the blood from the tail artery in an amount of 5 ml of blood was determined by the activity of superoxide dismutase (SOD), catalase (cat), antioxidant activity (AOA), total oxidative activity of blood plasma (AOA), the content of malondialdehyde (MDA). The research results are reflected in table 6.

To determine stressoustojchivosti swine six months of age used the scale (table 2). The analysis of the obtained data we can see that the pigs under the numbers 1, 2, 3, 4, 6, 8 and 9 are indicators of AOA, AAO, CT, MDA and SOD within 41,86±0,19% and below, 61,93±0.25% and above, 76,68±0,74 EA/ml min and above, 7,37±0,17 EA/ml and below, 22,80±0,68 EA/ml and above and on the basis of the years of this refer them to stressoustojchivye animals and number 5 and 7 within typical PSS"+", respectively is stresstesting animals. At gilt at number 10 values of the studied parameters are related boundaries PSS+and PSS "-" animals, therefore, considered to be "conditionally" sensitive to stress.

A method of evaluating stressoustojchivosti pigs, including the selection of blood from tail or ear artery, conducting her research, characterized in that in the blood of pigs two months and/or six months of age to determine the parameters characterizing the status of the antioxidant defense system and lipid peroxidation: the content of malondialdehyde MDA, antioxidant activity, antioxidant activity, total oxidative activity of blood plasma, OOA, the activity of catalase CT, superoxide dismutase SOD; pigs at two months of age when the indicators MDA 6,02±0,EU/ml and below, the AOA 60,86±0.31% and above, OOA 71,64±0,16% or lower, CT 57,30±1,05 EA/ml min and higher SOD of 14.57±0,44 EA/ml and above are referred to stressoustojchivye PSS "-"when the indicators MDA 7,63±0,07 EA/ml and above, the AOA 50,22±0,42% and below, OOA 79,21±0,83% and higher, CT 47,16±1,08 EA/ml min and below, SOD 10,49±0,49 EA/ml and below refer to stressfully PSS "+"; swine six months of age when the indicators MDA 7,37±0,17 EA/ml and below, the AOA 61,93±0.25% and above, OOA 41,86±0,19% or lower, CT 76,68±0,74 EA/ml min and above, SOD 22,80±0,68 EA/ml and above are referred to stressoustojchivye PSS is-", and when indicators MDA 8,05±0,09 EA/ml and above, the AOA 59,72±0,17% and below, OOA 46,21±0,09% and higher, CT 71,76±0,51 EA/ml min and below, SOD of 19.91±0,59 EA/ml and below refer to stressfully PSS "+".