PL239583B1 - The use of plasma water to improve the breeding performance of sows - Google Patents

Abstract

The subject of the application is the use of plasma water in a flow system with a capacity of 1000 l/min and parameters of 5x10-2 mbar, 800 - 1000 V to improve the reproductive performance of sows.

Description

Description of the invention

The subject of the invention is the use of plasma water to improve the breeding performance of sows.

Until now, plasma water has not been used to improve the breeding performance of sows.

The essence of the invention is the use of plasma water with the parameters of 5 x 10-2 mbar, 800-1000 V to improve the breeding performance of sows in a flow system with a capacity of 1000 l / min.

Research was carried out on the assessment of the effect of tap water treated with plasma generators in improving / improving the breeding performance of sows.

An apparatus was installed on the farm, in which the tap water was subjected to plasma generators and then it was fed to one row of pens, into which the sows after weaning are introduced on the farm.

Every week, "Nano" test sows (group 2) and control sows (group 1) were alternately included in the study group.

The sows from both groups, due to the inflow of water treated with plasma generators in the entire row of single pens, were located in two different divisions of the breeding sector on the farm. If the sows were not effectively covered, they were excluded from further study, even if they were returned to the observation animals.

On the day of insemination, the condition of the sows was assessed using the BCS method on a 5-point scale, then an image of the vagina of each sow was taken with a thermal imaging camera. The examination of the sexual organ was carried out with the Honda Electronics apparatus using a sectoral, abdominal convex probe with a frequency of 5 MHz. Ultrasound examinations were carried out in pens where the females were kept, with the preservation of veterinary medicine and animal welfare. A small amount of gel was applied to the ultrasound head, the probe was placed on the sow's body under the knee fold, and above the mammary gland, in the cephalic direction on the animal's spine. During the first examination, which was performed before the artificial insemination procedure, the condition of the ovaries was assessed. After locating the ovary, the number and diameter of ovarian follicles were recorded on it. Bubbles were classified into the group: small, medium and large. The test result of each sow was properly coded and recorded in the device's memory in the form of a separate graphic file (Fig. 3, 4). The duration of the ultrasound examination of one sow at this stage was approximately 7-8 minutes. After the ultrasound's own memory was full, all files were downloaded to a separate flash drive for the purpose of further evaluation and verification of the collected data.

Then, from 10 sows from one technological group (in 4 replications) blood was collected from the zygomatic vein for laboratory analyzes. Next, artificial insemination was performed by a farm worker.

The second ultrasound examination of each sow participating in the experiment was performed 28-30 days after insemination. This study assessed the effectiveness of insemination, namely the presence of developing embryonic vesicles with the embryos. The number of normal embryos in each uterine corner was assessed. Moreover, all pathologies such as dying off of embryos and abortions were noted. Each sow was examined for 5 to 10 minutes.

After the effectiveness of insemination was confirmed, the sows were taken to group pens where they only had access to regular tap water. The third examination of the sows' sexual organs was performed on day 60 after insemination. This study was intended to further confirm the presence and development of previously confirmed pregnancy (Fig. 5). The number of fetuses and their viability were determined. Each sow was examined for 5 to 10 minutes.

After the initial analysis of the first hormone determinations, it was decided to additionally collect blood for the determination of progesterone concentration on the day of pregnancy confirmation (from successfully covered sows), i.e. after the implantation process was completed. The first sow in the control group was born on October 27, and the last sow in the "Nano" group on November 26, 2018.

Laboratory analyzes

In the laboratory of the Department of Reproduction with the Farm Animal Clinic of the University of Life Sciences in WROCLAW, serum concentrations of progesterone and estradiol were determined using commercial Biomerieux tests: VIDAS PROGESTERONE and VIDAS ESTRADIOL. Other laboratory determinations were carried out in the Laboratory of the Department of Immunology and Veterinary Prevention. The concentration of fibrinogen in whole blood was determined by the capillary method. In the serum

PL 239 583 Β1 blood was determined using commercial kits: haptoglobin (ImmunoGen Polska HAPT-9); serum amyloid A (ImmunoGen Polska TP-802); leptin (Biokom ELISA Kit for Leptin), antioxidant status (Biokom, Antioxidant Assay), catalase (Biokom, Catalase Assay Kit).

The collected numerical material was statistically analyzed using the analysis of variance using the STATISTICA 10.0 statistical package, the significance of the differences was estimated with the NIR test, verifying the results with the Tukey test.

Production results

The sows entering the experiment were on average 2.74 absentees. There were 13 elements in the control group, and 12 in the experimental group. Artificial insemination was usually performed on the 5-6 day after weaning. On the day of insemination, all sows were first photographed with a thermal imaging camera, and then their condition was assessed on a 5-point scale. Measurements at the hottest point of the vulva were taken using the Flir Tools computer program. Two examples of thermograms from a sow that became pregnant (Fig. 1) and a sow where the insemination procedure was unsuccessful (Fig. 2) are presented below. The results of the thermal imaging studies confirm the occurrence of estrus in the sows. All sows with a temperature reading less than 37.50 did not become pregnant. The thermal imaging camera examination can be used immediately before the artificial insemination procedure to confirm the validity of its conduct.

All sows were in very good breeding condition - on average 2.84 points. The difference between the control group (1) and the experimental group (2) was only 0.14 points. (Ex. 1).

Sp1______ measurements

Parameters

Emissivity 0.9

Temp Reflected ^ _ _22

Geo-location

Compass

location

0 ° N

25.2?

Comment

Sow in heat

Photo 1. Thermogram of a sow that has been successfully inseminated.

PL 239 583 Β1

Measurements _________X sleeps [37.4 ................

Parameters

Emissivity _! 0.9

Teny. reflected ~ i 22 ^S C '

........___________________

Compass j o Pn t ^ c ^ za ^ .....

Comment________________

No heat

Photo 2. Thermogram of a sow that was not successfully inseminated on the day of the study.

Finished Average condition of the sows on the day of insemination (points).

3.8

3.6

3.4

3.2 3.0 m c CD

2.6

2.4 2.2

2.0

1.8

Group □ Mean □ Mean ± Constant error

Z Average 8 ± 0dch.std

PL 239 583 Β1

The tolerance reflex during searching for sows in oestrus (on the farm, the search and the insemination procedure are performed in the presence of a boar standing in front of the sows) was shown by 100% of the sows from the "Nano" group, and 86.36% of the sows in the control group. The difference of 13.63% between the groups turned out to be statistically significant. The chart shows the percentage share of inseminated sows from both groups in the entire population covered by the observation (Graph 2). 3 and 5 mm, and small ones less than 3 mm, hemorrhagic corpuscles are also included. This assessment should be treated only as illustrative as it is not possible to physically assess all bubbles in typical field conditions. Their visibility / exposure depends, among others from the posture of the sow, the filling of the gastrointestinal tract and the bladder. Nevertheless, based on the evaluation performed, preliminary conclusions can be drawn regarding the upcoming or already ongoing / completed ovulation and the sow's readiness for insemination (Figures 3-8).

In the post-weaning period, heterogeneity of the follicles is observed in sows, expressed by the variability of their size (diameter) and their number on the ovaries. This heterogeneity of ovarian follicles is mainly influenced by the ambient temperature, the energy balance during lactation, the number of births and the general condition of the organism. The correlation of the abovementioned zoohygienic and production aspects with the number and size of ovarian follicles in sows has been proven. The negative energy balance during lactation, stress, first childbirth, abnormal temperature in the environment and poor body condition are responsible for the smaller diameter and smaller number of large ovarian bladders in this species. As a consequence, the size of the ovarian follicle significantly affects the pigs' reproductive rates: length from weaning to heat, fertilization rate, duration of estrus and the time of ovulation. Heterogeneity of ovarian follicles, manifested by a low number of follicles with a diameter of more than 5 mm and a high number of up to 2 mm in diameter, may manifest itself in the diversified development and maturation of oocytes, a variable response to the secretion of gonadotropic hormones (FSH and LH), poor development of the embryo, its differentiation and survival, decreased fertilization rate, development of cysts on the ovaries, and production of lowered yellow bodies. Proper hormonal regulation on the hypothalamic - pituitary - ovaries axis is manifested by the presence of medium and large follicles on the gonads. The presence of these follicles is a guarantee of estrus, ovulation, fertilization and nation (implantation) of the embryos. In the sows in the control group, both ovaries showed a total of 298 "large" follicles (diameter over 5 mm), which is 11.5 per sow, and 86 medium "medium" follicles (diameter from 3 to 4 mm), which is makes up 3.3 bubbles per sow. Hemorrhagic cells were found in this group of 93 females, which is 3.5 per animal.

Photo 3. Ovary with large follicles

Photo 4. Ovary with medium follicles

Among the sows of the experimental group, 421 large ovarian follicles, 292 medium ones and 28 hemorrhagic bodies were found. This gives, respectively: 11.01; 7.68 and 0.77 of the mentioned physiological formations for the sow.

A smaller number or even the absence (right ovary) of small follicles in sows fed with water treated with plasma generators may be a good predictor of the proper development of the aforementioned physiological processes. The greater number of haemorrhagic cells on the sow's ovaries in the control group indicates premature or accelerated ovulation, which may lead to a decreased

PL 239 583 Β1 of the number of fertilized eggs and, consequently, a reduction in reproductive efficiency. In the sows from the experimental group, the ovulation process was more homogeneous, as evidenced by the lower number of haemorrhagic cells on the ovaries on the day of artificial insemination.

Fig. 3-8 Number of large (d), medium (d) and small (m) follicles on the left (JL) and right (JP) ovaries in groups 1 (control) and 2 (experimental - "Nano").

11.5

11.0

105

10.0 in 9.0

6.58.0

Λ

Chart 3

Group

Chart 4

group

group

Chart 5

Chart 6

Graph 7

Geqja

Chart 8

PL 239 583 Β1

The assessment of its effectiveness carried out between 28 and 30 days after insemination showed that 86.36% and 98% of the sows from the control and "nano" groups were pregnant (Graph 9). On the occasion of this assessment, using the ultrasound machine, attempts were made to count normal embryonic vesicles in the right and left corner of the uterus (Graph 10-11). In the control group, on average, 17.18 embryos were found in one sow in the uterus, and 16.08 embryos in the experimental group. The difference between the groups was only 1.1. This result should be treated as indicative, with such a difference, it is more important to verify on the basis of the number of piglets born.

The effectiveness of petrification (Graph 9), assessed after the completion of petitions, was 73% and 84%, respectively, in the control and "Nano" groups. In the control group, from the time the group was formed after weaning the piglets to the day of farrowing, 32% of the sows left the group due to the lack of oestrus or miscarriage (both at the early and late stages of pregnancy), and in the "Nano" group it was almost twice as small (17%) and all "losses" in this group occurred after the pregnancy was confirmed and the sows were transferred to group pens where all animals were treated the same, ie no longer had access to nanowater. Around the 75th day of pregnancy, the sows' fertility was again verified using an ultrasound machine (Fig. 5).

Photo 5. Visible fetus on day 75 of pregnancy

The average fertility of the sows covered by the observations was 14.11 animals (Tab. 1). In the litters of the sows from the control group, there were on average less than half of the head more and the same number of dead piglets were born in this group. The results obtained in the experimental group seem to be more desirable. Smaller differentiation of this parameter (the number of live born piglets) expressed as a standard deviation within the group, as well as a smaller spread between the minimum and the maximum are associated with a smaller differentiation of piglet body weight within the litter, which translates into a greater probability of their survival during further rearing. with mothers. When no more than the sow has mammary glands, providing them with good conditions during rearing with their mothers usually does not pose any major problems, even if the litter standardization procedure has to be performed. Nowadays, with more and more fertile sows, the main goal is not just to increase the number of piglets born in the litter, but to obtain vital piglets weighing more than 1 kg at birth.

Table 1 Mean number of live and stillborn piglets in the observation population.

Number of piglets control group Nano group Overall - live X 14.36 13.92 14.11 born + 3.5 2.24 2.82 min-max 8-22 9-19 8-22 - dead X 1.94 1.44 1.69 born + 1.1 0.51 0.88 min-max 1-4 1-2 1-4

PL 239 583 Β1

Laboratory results

Hormones: estradiol and progesterone

In the blood serum collected on the day of insemination, the concentration of estradiol (Graph 12) and progesterone (Graph 13) was determined to estimate the “readiness” of the sows for fertilization. After ovulation, a yellow body is formed from the follicle, which is responsible for the synthesis and secretion of progesterone, which in turn determines the implantation of the embryo in the uterine mucosa. In the first stage of pregnancy diagnosis (the floating embryo phase - from fertilization to implantation), pig embryos secrete large amounts of estrogens whose task is to protect the corpus luteum against the luteolytic effect of prostaglandin F2 alpha. By marking these hormones on the day of insemination, higher estradiol and lower progesterone concentrations were expected. The concentration of estradiol was similar in both groups, within ranges similar to those recommended in the literature on the subject. The concentration of progesterone was lower in the "Nano" group as compared to the control group, which may be related to the fact that in these sows ovulation was just about to begin or was just beginning. This is confirmed by the higher effectiveness of insemination noted in the Nano group.

To confirm these observations, additionally, the determination of progesterone concentration after the end of the implantation period was introduced (usually it takes place between the 16th and 25th day after fertilization), on the day when the effectiveness of insemination was confirmed (Graph 14). The determined mean concentrations of this hormone were similar in both groups, a slightly higher value was recorded in the "Nano" group, which may be a proof of the higher functionality of the yellow bodies on the ovaries in this group of sows.

Graph 12. Average concentration of estradiol in the observation population on the day of insemination ί ----------- 40 33 30 -

Control group Nano group

Graph 13 Average concentration of progesterone in the population under observations on the day of insemination

control group

Nano Group "

PL 239 583 Β1

Exhibit 14 Average progesterone concentration in the observation population on day 30 after insemination.

Acute phase proteins

To assess the health status and immune stimulation in whole blood (fibrinogen - Fb) and serum (haptoglobin - Hp, serum amyloid A - SAA), the concentrations of selected acute phase proteins were determined (Figures 15-16). The average concentrations of all proteins (slow and fast reacting) were within the reference range, which confirms the good health of the sows. A very large individual variation was noted for the concentration of SAA (the value of the standard deviation significantly exceeds the average concentration of this protein) in both experimental groups. The fact that in individual animals the concentration of this protein was above the reference range, and in many it was not possible to determine it, may confirm the greater immunological stimulation of these sows due to ongoing or just completed ovulation. SAA is a fast-reacting protein, its concentration increases as early as 3 hours after the stimulus that induces its synthesis, which may also be the beginning of ovulation. The ascertained situation is a physiological phenomenon, and especially with regard to the determined concentrations of slow-reacting proteins, it confirms the presented explanations.

Ex. 15. The mean concentration of Fb and Hp in the observation population on the day of insemination

S Citupa Łoutidua □ Nauo Group

PL 239 583 Β1

Exercise 16. The mean concentration of AAS in the observation population on the day of insemination

Oxidative status

Maintaining the reduction-oxidative balance is a very important aspect when assessing the homeostasis of the organism, which is expressed, among others, by the proper course of life processes (such as the biosynthesis of certain hormones, regulation and differentiation of many cells) and the ability to defend against various pathogens. This balance may be disturbed when too many reactive oxygen species (ROS), including oxygen free radicals, are formed as a result of oxygen transformations. The body protects itself against this excess by creating various barriers (enzymatic or non-enzymatic, using substances of both endogenous and exogenous origin), preventing the reaction of ROS with biologically active compounds, interrupting these reactions or repairing their effects. It is impossible to fully assess the antioxidant status in field conditions, hence attempts to use various tests measuring the oxidative status based on the assessment of the degree of lipid peroxidation or directly the concentration of malonyldialdehyde. The enzymes involved in enzymatic defense can also be determined: superoxide dismutase, catalase or glutathione peroxidase.

The own research used the Cayman Chemical antioxidant test, which determines the equivalent of the antioxidant potential of trolox in a reaction with the free radical ABTS. This method consists in extinguishing the characteristic, long-term absorption band of the radical ABTS (diammonium salt) by antioxidants. Unfortunately, it was not possible to obtain reliable results (absorbance readings), despite the fact that all determinations were carried out in accordance with the methodology recommended by the manufacturer of the test. The determinations were repeated three times.

The oxidative status was also assessed by determining the activity of catalase (Graph 17). Catalase is mainly involved in the inactivation of hydrogen peroxide and transforms it into molecular oxygen and water, and plays a special role in the metabolism of erythrocytes, which are exposed to high concentrations of oxygen. Many studies have shown that the excessive production of ROS has a negative impact on the course of pregnancy and the health of the mother and the fetus. It can cause abortions, death of embryos and their resorption, and consequently the birth of weak newborns, often with defects. The average activity of catalase expressed in nmol / min / ml in the control group was 30.7, and in the experimental group it was 63.6. The difference of 32.65 between the groups turned out to be highly statistically significant (P <0.01). The obtained result (serum from the day of insemination) clearly correlates with the effectiveness of insemination found in the presented studies (also statistically significant differences) and the maintenance of low pregnancy (until the day of sows regrouping and transfer to group pens). The higher activity of catalase can be directly explained by the presence of more free radicals, which could be a disadvantageous phenomenon. The cumulative interpretation of the results of our own research prompts me to explain this higher activity in experimental group 2 ("Nano") somewhat differently. The presence of antioxidant substances inhibits the progress of oxidative processes, allowing for further proper development of oocytes, and later embryos. Such an interpretation is based on the higher effectiveness of insemination in the experimental group ("Nano"), as found in the authors' own research.

Leptin is a multifaceted hormone, primarily it affects the centers that control the animals' appetite and reproductive processes. Its serum concentration is normal

The amount of adipose tissue deposited is directly proportional to the amount of adipose tissue deposited, but its concentration is also influenced by even a short-term change in diet / feed intake, which usually occurs during the stimulation of flushing of the sows after weaning. Along with the decrease in leptin concentration, the secretion of LH and GH hormones also decreases, although these changes occur after a few days. The effect of leptin on the resumption of ovarian cyclicity after weaning was confirmed.

In our own research, the mean serum leptin concentration on the day of insemination in control sows was 4.51 ng / ml (Graph 18). In the experimental group it was 2.31 ng / ml more. This difference was not confirmed statistically, probably due to the greater individual diversity of the tested females from the experimental group. However, the higher concentration of this hormone may reflect better absorption / absorption of nutrients by the sows in the "Nano" group. This, in turn, may be due to the properties of the water treated with plasma generators. Illustratively, it can be said that this water causes greater / stronger adherence of the contents of the gastrointestinal tract to the intestine, which may facilitate / accelerate / increase the absorption of nutrients. This fact is illustrated by an experiment that was carried out to confirm the difference between ordinary tap water and such water treated with plasma generators. An equal volume of water ("Nano" or ordinary tap water) was poured into two beakers and a hematocrit capillary was inserted into each. After 7-10 min, the difference in the volume of water sucked in by the capillary was visible, its bar was always greater with water treated with plasma generators. On this basis, it can be concluded / expected that in animals drinking such water, it will support better adhesion of the contents of the digestive tract to its walls.

The conducted research seems to confirm the beneficial effect of water splashing with water treated with plasma generators on the results of the breeding performance of sows. Undoubtedly, this brings measurable economic benefits resulting from a better balance with feed for the sows on the farm and from the reduction of unproductive days, i.e. days when the sow is not pregnant or lactating. A significant increase in the effectiveness of insemination and reduction of early miscarriages contributes to better management of technological groups on the farm (shorter sterilization period, better use of insemination doses, reduction of workload for additional regrouping of sows and possible re-insemination). The conducted research does not allow to explain the mechanism of action of water treated with plasma generators on the animal body, although the effects seem to be visible. The attempt to explain its effects by improving / supporting the absorption processes seems worth considering and perhaps pharmacokinetic studies in laboratory conditions should be carried out. It remains a question of how this water supports / influences better use of nutrients, or, as we have tried to describe it, using the observed properties of this water in the capillary, or maybe it is related to better / faster dissolution / decomposition of nutrients from the feed.

The obtained results allow us to state that the assessed experimental factor influenced, first of all, the effectiveness of insemination and the maintenance of pregnancy in the first period, i.e. until embryo implantation. It does not seem that the flushing of water treated with plasma generators in the period after weaning could significantly affect the number of born piglets, so that the expectation of such an effect, sows should have access to it already during the lactation period. If access to such water significantly improves the availability of nutrients obtained from feed, lactation time is the most desirable period to recommend access to such water.

Claims (2)

1. The use of plasma water with the parameters of 5 x 10-2 mbar, 800-1000 V to improve the breeding performance of sows. 2. Use of the water according to claim 1 in a flow system with a capacity of 1000 l / min.