JPH06508522A - Manipulation of intestinal structure and enzymes in animals - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Manipulation of intestinal structure and enzymes in animals! Five Attempts to manipulate the gut microbiome of animals have continued to increase over the years. . Appropriate manipulation of the gut microbiome can have a positive effect on animal growth rates. It's clear. One means of influencing the microbiome is through the use of antimicrobial agents. The use of antimicrobials in animal feed is well documented in the literature. However, the effects of changes in the microbial populations of the small and large intestines are not fully understood. do not have.

Antimicrobial compounds used include forage steel, which is effective in improving growth rates. It has been reported that there is. Braude, R., “Copper is a Growth Stimulant in Pigs, -, Animal xm Fukiryo 3: 69-75 (1967). Antibiotics are also pathogenic in the gastrointestinal tract. It is used to eliminate microbial flora. However, the effects of antibiotics on the human body The agricultural industry is reluctant to use antibiotics because their definitive effects are not well established. This indicates a move to refrain from using the product. It is not recommended to add antibiotics to livestock. Growing public awareness has led to a move away from the use of antibiotics and a shift away from the use of antibiotics to favor the gut microbiome. Efforts continue to develop alternative measures that will have a positive effect.

Both consumers and manufacturers believe that the use of antibiotics as therapeutic agents is associated with side effects. In order to improve the There is a lot of interest. Symbiotics are generally supplements in feed by living microorganisms. Footwear (a　ive　m1crobial　feed　supplement) This improves the balance of intestinal microorganisms, and as a result, Mainly has a beneficial effect. Fuller, R, -Probiotics i n Man and Animals, -, AJIL Bacteriol, 66:365-378 (19119).

A major problem with symbiotic agents is how such substances act. There is a lack of information available. The beneficial effect on growth is due to the heavy use of interactions with other growth-promoting substances in the feed, and It is influenced by the perception of living organisms. Therefore, the growth of such formulations While there is data suggesting that it can affect animal growth, The extent to which this formulation has an effect and the mechanism of action of this formulation remain unclear. and remains unclear. Understanding this process will help you optimize your beneficial effects. A reasonable basis for the amount and frequency of dosing required is considered to be available. Ru. Having such knowledge will help you understand the need for treatment using symbiotic agents and its response. Physiological or environmental stresses that influence the balance of gut microorganisms by response may successfully address the growth stages and gastrointestinal tract development of susceptible animals.

Animals with large numbers of animals tend to lose weight after weaning and are particularly prone to infection.

Many studies have been conducted on pigs regarding this effect; This is because it is not uncommon for livestock to lose weight after weaning. This after weaning This delayed growth is recognized as a phenomenon that occurs in the "early weaning" production system. o Lecce, J.G., ArmStrong, W.D. ,Cr avford, P.C., and Ducharme, G.A., -Nutri tiOn　and　Manal<ea+ent　oτEarly　Weaned 　Piglets-Liquid　versus　Dry　Feeding,- J, Anim, Sci, 48:1007-1014 (1989).

It has been observed that the structure of the intestine changes after weaning.

When looking at cross-section specimens of the gastrointestinal tract, all regions within the tract are structurally similar. , Beneath the outer serous covering is a layer of muscle tissue. The innermost part of the gastrointestinal tract is , a mucosal barrier consisting of two layers of smooth muscle, a thick layer of connective tissue, and three layers of mucosa. It is composed of The mucosa, or the luminal surface of the gastrointestinal tract, is composed of columnar epithelial cells. This is the case in all areas except the esophagus.

The entire small intestine has a depression, and the Lieberchini protrudes into the connective tissue (lamina propria). They form areas called crypts.

The surface of the small intestine is further composed of the mucosa, submucosa folds, and the epithelial cells that line it. Continuing with their crypts. Weaning affects the structure and function of this epithelial cell. It has been noted that this can have a significant impact. Miller, B.G., J. ames, P.S., 5IIlith, N.W., and Bourne, F. , J, - Effective Weaning on the Capaci ty of Pig Industrial Virus to Diges t and Absorb Nutrients, -MUEMU LSet, Cam b., 107:579-589 (1986). After weaning, villus length decreases However, the complexity of the structure increases. After weaning, villus height decreases and crypt cell production increases. Due to the increased depth of the crypts, more Migration of young intestinal cells to the villus surface occurs. This allows the digestion and absorption of epithelial tissue. Yield capacity is limited. This results in reduced digestive and absorptive capacity in the intestinal tissue. may be the main cause of growth arrest in pigs seen after weaning.

In addition, deterioration of intestinal function due to changes in the composition of the gastrointestinal microbiota, i.e., E. coli In particular, acute intestinal infections occur after weaning due to an increase in enteropathogenic systems. can be obtained.

Therefore, in order to use and apply symbiotic substances more effectively, symbiosis in the animal intestine should be investigated. We need to better understand the mechanisms of action of substances. Furthermore, various animals, especially pigs, To overcome the adverse effects that occur in the intestinal tract after weaning in domestic animals and other domestic animals. There is also a need for

The present invention increases digestive enzymes in the intestinal tract, improves intestinal epithelial cell structure, and increases digestive power. in the intestinal tract, which improves absorption capacity and reduces the growth of the animal after weaning. Concerning the application of probiotics to reduce the adverse effects of.

Therefore, one of the objects of the present invention is to change the digestive enzymes of animals and thereby The goal is to provide a means for animals to benefit.

A further object of the present invention is to change the mucosal structure of the intestinal tract of an animal, and thereby to It's about providing ways to benefit animals.

Yet another object of the present invention is to reduce the adverse effects on the intestinal tract of animals. It is the production of changes in the intestinal tract and its chemical structure.

A further object of the invention is to provide a method for improving the health status of post-weaning animals. That's true.

Further detailed objects of the present invention will become apparent from the following disclosure.

Beans and rice cakes The present invention relates to Lactobacillus and Streptococcus. Digestive enzymes for animals, including administering probiotic strains of P. ptococcus Concerning how animals can be benefited from changes in animals. The present invention It is also important to note that changes caused by administering such probiotic strains may also benefit the animal. The present invention relates to a method for changing the intestinal mucosal structure of an animal, such as causing a change in the intestinal mucosal structure of an animal. The present invention Improve the health status of animals after weaning by administering strains such as provide a method.

! 1 year old and 1 year old Figure 1 shows the control group, tyrosine Showing alkaline phosphatase activity in treatment group and Probios treatment group. This is a diagram.

Figure 2 shows the control group, tyrosine It is a figure showing sucrase activity in a treatment group and a Probios treatment group. .

Figure 3 shows the control group, the terrothorn FIG. 2 is a diagram showing lactase activity in treatment groups and Probios treatment groups. .

Figure 4 shows control, tyrosine-treated pigs at 7, 17, 42 and 80 days of age. This is a diagram showing tripeptidase activity in the treated group and the Probios treated group. be.

Figure 5 shows the control group, the Diagram showing dipeptidase activity in Probios-treated group and Probios-treated group It is.

Figure 6 shows the control group, Prob FIG. 2 is a diagram showing alkaline phosphatase activity in the ios treatment group.

Figure 7 shows the control group, Prob It is a figure showing the lactase activity in the ios treatment group.

Figure 8 shows the control group, Prob FIG. 3 is a diagram showing tripeptidase activity in the ios treatment group.

Figure 9 shows the control group, Prob It is a figure showing the dipeptidase activity in the ios treatment group.

The amount of days! 8 The mode of action of probiotics in animals, especially how they act on the animal's intestinal tract. How it is used is not well understood. As mentioned above, this effect Increasing our understanding of when, how, and which probiotics to administer to animals This will greatly contribute to understanding.

The surprising discovery of this invention is that the symbiotic substance alters the digestive enzymes and is associated with improved ability to digest nutrients. The inventors also The mucosal structure of the intestinal epithelial surface inside the intestinal tract has a higher ability to digest nutrients than younger cells. They also found that changes occur as more mature cells are exposed. That's it When given to animals before weaning, the symbiont improves their ability to digest nutrients. post-weaning by increasing such enzymes and changing mucosal structure to improve It has further been found that adverse effects can be reduced.

The digestive enzymes studied in this application include sucrase (sucrose α-D-glucohydrate). dolorase, lactase (α-D- is lactocytogalactohydrolase), zype Peptidase (base JtL-leucylglycine) and tripeptidase (substrate L-leucylglycylglycine). These enzymes transport nutrients into the body. Active during the final stages of digestion before absorption. Among these, carbohydrates are It is located in the brush border membrane of the inner cell, and dipeptidase is located in the cytosol. tripe Putidase activity occurs primarily, but not exclusively, in the brush border region of the cell. are doing. Among these studied enzymes, when treated with probiotics, It was found that all enzymes other than peptidase had significant effects. The experiment described below As shown, in piglets that started receiving the symbiotic on day 17 after birth, the control group compared to the symbiotic-added group or even in the antibiotic-added control group. It was found that less enzyme was present. In addition, enzymes from before weaning to after weaning The typical decrease in activity was found to be significantly reduced by administration of the probiotic.

Additionally, the administration of the probiotics caused changes in the morphology of the intestines. Throw symbiont When given, the villus height and the ratio of villus height to crypt depth are There was no significant decrease between before and after weaning. When pigs are given a symbiotic, especially Prevents reduction in villus height and crypt elongation typically between 7 and 17 days after birth This influenced changes in mucosal structure before weaning.

While not wishing to be bound by any theory, this increase in enzyme activity The stronger effect is due to changes in epithelial cell turnover, with more numerous cells on the villi. and/or are believed to produce functionally mature intestinal cells. pig Postnatal changes in villous structure that occur in humans can be influenced by the composition of the endogenous microbial population. Ru. Administration of probiotics can improve the microbial balance and metabolic patterns of microorganisms in the gastrointestinal tract. changes in intestinal tissue and enzyme activity observed in Experiment 1. It is theorized that this occurs.

Administering the symbiotic before weaning has a positive effect on the growth rate of the animal. experiment As shown in Figure 1, the growth rate increases from day 26 when compared to the control group. . In comparison with control animals, an increased growth rate was observed compared to the control group; This is from the 36th day.

The probiotics of the present invention can be used in various forms as long as they improve enzyme activity and intestinal structure. Possible. The first formulation in this application is Lactobacillus 11 (Lactobacillus Illus acidophilus (3 strains), Lactobacillus casei (L, Ca5e) i) (1 strain), L, PlantariuI (4 strains) and fecal chain method m ( Streptococcus faecium) (2 strains). Ru. The 5 strain types include Lactobacillus acidophilus, Lactobacillus casei, L, and Planta. two strains of S. rium and fecal streptococcus. In Germany, these are Deut sche Sammlong von Mikroorganismen Un d Zell has been deposited with ulturen Ga+bh, and any and all requests have been America 0 symbiotic substances deposited in nType Culture Co11ection The mode of administration has not yet been clearly established. For livestock, in feed It is typically administered by mixing in the form of granules. These granules are usually stressed Administered at the time of outbreak, upon arrival at the facility, and when changing feed rations. administering the symbiotic Gel-like compositions may be used to supplement the diet, and granules may be added to the feed. gel at birth, between 7 and 10 days after birth, at weaning, or during other stress events. It is typical to administer The administration of symbiotic substances can reduce the enzyme reduction that occurs after weaning and the intestinal Administration before weaning has a significant effect on the unfavorable effects of structural changes in It is very effective to do so.

It is known to include other ingredients in such gel compositions, and additives include enzymes. This poses no problem as long as it does not affect increased activity or structural changes in the intestines. supply Additives for improvement include yeast, vegetable oil, silicon dioxide, titanium dioxide, and vitamins. This includes ingredients such as pigments, colorants, and preservatives.

The lactic acid bacteria content is sufficient to produce the desired enzymatic and structural changes in the intestine. It must be in sufficient quantity. An effective amount used in the present experiments is 1 gram of the composition. Contains approximately 107 viable bacteria per ram.

Examples are shown below, but the scope of the present invention is not limited thereby.

(Margin below) Fruit 11-1 The results of measuring the growth rate that occur from birth to 80 days of age are measured using a test that does not contain growth promoters. growth rate of animals fed feed (negative control group), antibiotics used in the pig industry Growth rate of pigs treated with growth-promoting substances (positive control group) and the aforementioned probiotics. Comparisons were made between treated pigs. 7.17. Microscopicity at 42 and 80EI ages Biological aspects and development of the gastrointestinal tract were studied as shown below.

On the ninth desk 11 The probiotic preparation administered during this study was named Probios■ (Pion eer Hi-Bred International, Inc., John ston,, Iova, U, S, A,). This includes dry and ac tobacillus pJ3 landlord rum, fecal streptococcus, casei and reed It contained 12 bacterial strains, including P. dophylles as well as their fermentation products.

The formulations used in this experiment were of two types: '2LIPrObiO5' Rogel Composition: dried fermentation products and lactic acid bacteria (7 lactic acid bacteria subspecies and 5 chain species) (22 yeast cultures, vegetable oil, sucrose, diacid Silicon oxide, titanium dioxide, bicumino A, D, E and BSF, D and C Yellow #6 rake, Palysorbate 80, TBHQ and Etkinki Preservatives. Total lactic acid bacteria = 107 Number of countries of birth 72g0 Recommended dosing! = 1 Igo at the birth of a new pig or the first administration 1 from 7 days 1 g was added after 0 days.

Weaning/starting pigs - Administer 2 g at weaning, during periods of ration changes and other environmental changes. Ta.

Pig ProbiO5I Composition: dried fermentation products and lactic acid bacteria (7 lactic acid bacteria subspecies and 5 chain species) Yeast cultures, calcium carbonate, corn Chickpea cob, vegetable oil, TB) IQ and Etkinquin preservative. Total lactic acid bacteria = 1 07 Viable bacteria count/g.

Recommended dosage: Weaning and starting pigs (up to 11 kg) - environmental change, on arrival , or during the period of ration change, the feeder should administer granules at a rate of 1 kg/ton of feed. did. Adult pigs (up to 18-45 kg) - environmental changes, arrival or ration changes The granules were administered at a rate of 500g/) of total feed during the period. Completed pig (45 kg to market weight) - during periods of environmental change, arrival, or ration change. Granules were administered at a rate of 250g/)ton of feed.

During the 5EI period until sow parturition, granules were added to the feed at a rate of 10g for 7 pigs and 7 days. . During the lactation period, granules were administered at a rate of 500 g/ton of total feed.

Adding tyrosine to the teruguchi of Tyrosine is 5tretomces fradiae (Brander, G, L, -Chemicals for Animal Health Co ntrol-Taylor and Francis Ltd, London (1986)) is a gram-positive macrolide antibiotic. Active against living organisms. subtherapeutic level Tyrosine is widely used as a growth promoter in pigs and poultry.

In this study, tyrosine was added to the [positive control] diet in Tylamix Prem. ix IQOg/kg (product approval number 0006/4055, Elanco Pr products Ltd, Basingstcke, England). Included. Each kilogram of this preparation contains phosphoric acid equivalent to loog of tyrosine bases. Contains tyrosine.

廼1 The test consisted of three treatments = (1) no growth promoters and nutrient levels; (2) basic feed and the antibiotic growth promoter tyrosine (E lanco Products Ltd, Basingstoke, Hant s), isolated from 5tre tow CeS fradiae, a Gram-positive organism Macrolide antibiotics with activity against (Brander, supra, 40a +g/kg): (3) Basal feed and probiotic preparation Pr.

bios (Pioneer H4-Bred International L td, Johnston, lowa, LISA) 24 hours after birth ( 1 g), again administered as an oral gel at 7 days of age (1 g) and at weaning (2 g) did. This symbiotic substance can be fed as granules in feed and follo-o-o. n) contained in both feeds at a rate of 1 kg/l. 107 colony forming mono Both forms of the symbiont, including Lactobacillus sp. Diversity of Lactobacillus acidophilus, Lactobacillus casei, and Liaohongdan faecium. contained a mixture of strains.

Eating belly lt This study is based on Large White/Landrace/Duroc formation. ) was performed on a litter of piglets. And because there was a limit to the size of the farrowing shed, there were three It was planned in phases, and two treatments were carried out in each phase. At each stage All four sows were farrowed within 48 hours of each other, with no time between stages. It was two weeks. Pair the sows according to their farrowing date and The piglets from the same litter were divided into two groups, and the sex ratio was divided into two groups, with half male and half male.

Therefore, each sow has half her offspring and half her foster piglets. had a cohort of Replacement parenting process (CR).

ss-fostering process) includes growth stunted pigs. , X.

Selection and allocation occurred in three epochs (i.e., this experiment was conducted in three stages). Ta). Treatments represented by specific stages were randomly assigned to four cohorts and Within each half of the pair, one fohoto was assigned to each treatment. Inside the farrowing shed In (X), the fuchots are penned separately, while the animals after weaning (in Two cohorts of treatment groups were defined from each treatment in the study. at all points in time Therefore, care had to be taken not to mix up the treatment groups.

Kei Dan Kei Prisoner 1 The feed and follow-on samples contained 253 g/kg of crude tannin and 200g/kg (nitrogen x 6.2s), lysine content 0.133g/kg and 0 ．． It was blended so that it would be 11g/kg. The feed formulation and appropriate chemical analysis are shown in Table 1. Shown below. Both diets contained nutrient levels of copper (20-70 mg/kg).

Feed the littermates from the age of 7 days, and remove the cubs from the farrowing shed at the age of 21 days. I moved it to a tJ\ya with a flat floor. - Weigh all pigs weekly Ta.

Sample sample Group 1u Aya Tori At 7.17.42 and 80 days after birth, each One pig was randomly selected from the cohort.

At the time of birth (J), care was taken to maintain a balance between gender and biological mother/adoptive mother. to this method Thus, for each treatment, four pigs had tissue harvested at each of four ages. The result was Restrain the animal to obtain tissue that has not undergone mucosal slough; Halothane (4%)-oxygen m-nitrogen oxide (nttrous oxide) with an open mask. de) was administered for anesthesia. Next, the abdomen was opened and valpitone sodium was injected through the portal vein. They went in and killed the animals. The gastrointestinal tract was immediately removed and the small intestine was separated from the mesentery. small intestine The length was taken at 0.05.0.1.0.3.0°6 and 0.8 along Samples of mucosal tissue were isolated from 11 50 ml intestines. Open the intestine sections lengthwise and cool them in water. Wash with saline solution (9 g/l sodium chloride), blot dry and cool. It was placed on top of a porcelain tile. Scrape the mucous membrane away from the stone layer using a microscope slide. then quickly frozen in liquid nitrogen before wrapping in tin foil and storing at -70 °C. I let it happen.

LL 25 Emperors For all assays, approximately 1 g of tissue was thawed and placed in a 12-stroke Teflon glass holder. 50IIIM nomannitol and 2mM Tris-salt using a modinizer. The tissue was homogenized in 15 mL of a solution containing an acid salt (pH 7.1). It was performed using a homogenate of mucosal tissue. Add the same buffer to the homogenate and 5ml and stored at -70'C until assayed.

For sucrase and lactase, see Dahlqvist, Arne, -M method for assay of 1ntestinal disacc haridases-■■■-Biochem, 7:18-25 (1964 ), dipeptidase (substrate Hiromi/luglycine) and and tripeptidase (substrate is L-loinulgurin), N 1cholson, J.T., and Kim, Y.S., -A one-ste p L-aminoacidoxidase assay for 1ntes tinal peptide hydrolase activetty-AnL LLL Biochem, 63:110-117 (1975) I went there. They are incorporated herein by reference. In these assays t′! , Hydrolysis of di- or tripeptides to release amino acids can be associated with the oxidation of loin by amino acid oxidase, resulting in peracid Oo-dianisidine, which generates hydrogen oxide, is oxidized to hydrogen peroxide, and the level level continues to increase at L%. Due to the action of oxidase, the specific end of the reaction with absorption at 530 nm, reach Using loinulglycine and loinulguri/luglycine as substrates and the activities of these two hydrolases can be distinguished from each other. Tripep The final product of the tidase assay, i.e., grinlglycine, is a dipeptidase If it is hydrolyzed by activity, it will not react in this assay system. Everything All assays are performed by comparing the volume of homogenate added and the response of substrate concentration versus time. and adjusted to maintain a linear ratio.

Punishment 'S, E1M Changes in overall intestinal morphology due to age are observed in sections S, E, and M. It was evaluated by From tissue fixed in 4% neutral buffered formalde Collect small sections (3+am x 3mm) and add graded concentrations of ethanol solution. (25, 50, 70, 9G, 95 and 100% V/V) for 30 seconds each. I was dehydrated. Next 1. % of the tissue section, the Department of Plant Biology, University of Nevcast It was treated with leupon Tyne. Each section was washed with 50% amyl acetate:50% ethyl acetate. in ethanol for 15 minutes, followed by 100% amyl acetate (compounded). , dryly distort the tissue section to a critical point (as in A), S, E, M, (Jeo l St S, E, M, Jeol (UK) Ltd, Lond.

n) “splat coated” with carbon and gold prior to inspection under ter-coated).

Egasahada feces irises View tissue sections under a light microscope to quantitatively study changes in intestinal morphology. I guessed it. First, 3% glutaraldehyde RS was collected under an M69 dissecting microscope. and stored overnight in phosphate buffer (pH 7, 4). The tissue section was then exposed to t he Department of Anatomy. Medical Sch ool. Treated with Nevcastle upon Tyne.

Observe the intestinal sections using a Vickers V80 stereomicroscope (x50). Accurately using the box part (camera lucida attachment) I drew it. Draw a scale (0-50um) from the micrometer on the stage ( x50), to overlay a clear drawing of the camera by copying it onto an acetate sheet. It was used to enable accurate measurement of tissue sections. The measurements taken included villus height; (distance from tip of villus to villus/crypt junction) and depth of crypt (distance from villus tip to villus/crypt junction) The distance from the crypt junction to the anterior mucosal plate) was included. Each parameter for the intercept The villus height:crypt depth ratio was calculated by taking the average of six measurements.

Naokogosho This data is stored in the GLM statistical package (statistical packag). e) (Baker. R.J. and Nelder. J.A., -GLM S. ysteIIIRelease- 3Numerical Algorithm s Group, Oxford (1978)). this method can sequentially match all factors that can influence the variables being analyzed. the first The analysis included studies of all ages using data summed for all parts of the intestine. age and treatment, or stage, gender, which is a “blocking” factor. and whether mothering has a significant effect. We planned to consider the following. The latter factor may be due to the fact that some pigs are more likely to be cared for by their adoptive mothers than by their biological mothers. It was established to confirm the effects caused by the fact that they were raised. difference Additionally, all factor interactions are matched and the residual sum of squares is adjusted across the null hypotheses considered. was used to calculate the error term for

As a result of this initial analysis, gender and mothering factors were It was found that there was no significant effect on either the enzyme activity, the stage, or the treatment. We combined the analyzes to determine the interaction effects of Ta.

The phase is due to the unbalanced nature of the experimental design (that Although not all procedures are included in each stage, The stage was left in the analysis because it was found to be significant in many cases.

This analysis also separated the age effect into pre-weaning and post-weaning (7+i7 days vs. 42+80 days). , and treatment effects were determined for control vs. tyrosine + Probios and tyrosine vs. Pr The study was conducted separately for obios. A summary of the variance analysis table is shown in Table 2.

The data analyzed for each of the four pig ages studied included data from five intestinal regions. Expanded to include values for each. factor, stage, treatment, intestinal site and and treatment Considering analysis of vs. Tyrosine-I-ProbiOS and vs. Tyrosine-I-Probios divided to do. For intestinal sites, also proximal (Q.05, 0, 10 and 0 ．． Considering the effect of 30) site versus distal (0.60 and 0.80) site Analyzed again. A summary of the variance analysis table is shown in Table 3.

Mt Sha (Margin below) U Gakuang Wλ hill sex 107 g mucosal wet weight in 5 intestinal regions of pigs alkaline phosphatase (Figure 1), as indicated by the measured value of (Figure 2), lactase (Figure 3), tripeptidase (Figure 4) and dipeptidase The mean values (± SEM) of the activity of 7.17. The treatment for this is shown below.

Table 4 shows all intestines subjected to each treatment at 7.17.42 and 80 days of age. Effects of treatment and age on each mucosal parameter studied for site and pigs. Summarize the impact.

MaP(0,05H”P(0,OIH”su〈O,OOl.

The activities of alkaline phosphatase, lactase and dipeptidase after weaning Compared to animals, it was significantly higher in unweaned pigs. On the contrary, Scrap -se activity increased between the pre-weaning period and the post-weaning period. Tripeptidase activity Regarding sex, there was no significant change between before and after weaning.

Dipeptida at certain ages of pigs treated with Probios and antibiotics. There was a significant effect for all enzymes except for enzymes.

ヱ” in the mouth At this age, alkaline phosphatase activity shows no significant interaction between treatment and site. showed increased activity from the proximal to the distal small intestine compared to treated animals. It was significantly lower in pigs (IU/g!ffWte is p<o, os, specific activity p<0.01) At the age of C, other enzymes showed no treatment effect. proximal Alkaline phosphatase, tripeptidase and dipeptidase enzymes in tissues activity was significantly lower compared to the activity at the distal region (specificity of tripeptidase All other values were p<o, 001), except for activity, which was p<0.05. this On the contrary, sucrase activity was significantly higher in the proximal tissue <p<o, 001). There was no significant effect of site on lactase activity.

17th day 1± With the exception of dipeptidase, the activities of all enzymes studied compared to treated animals. In the target pigs, it was found to be significantly lower across all regions (scraper For enzyme IU/gMWW and lactase activity, p<o, 001: alkaline phosphatide p<0.01 for sphatase IU/gMWW oyohiscrase specific activity; p<o, os for calyphosphatase-specific activity and tripeptidase activity) . Except for tripeptidases and dipeptidases expressed as III/gMWW , the activity of all enzymes studied was higher in animals treated with Probios. was also significantly higher in pigs treated with Tyrone (lactase activity o, 001; pro for alkaline phosphatase activity, 01; scler p<0.0 for enzyme activity and tripeptidase and dipeptidase specific activity. 5).

Apart from the effects of treatment, sucrase and lactase activities are more likely to occur proximally than distally. (p<o for IU/g!ffW of both enzymes, 0 01, and p<0.01 for the specific activities of both enzymes. On the contrary, the ponds considered The activities of all enzymes were significantly lower in the proximal region compared to the distal region (in All activities were positive, except for lipeptidase IU/gMWW, where p<O, O5. p<o, ool).

The magnitude of the change in activity between the proximal and distal regions is significantly lower in control pigs compared to animals treated with enzyme and dipeptidase. (p<0.001 for tripeptidase specific activity; sucrase 10/g p for MWW, tripeptidase IU/gM*W and dipeptidase specific activity. <0.01; p for sucrase-specific activity and dipeptidase IU/gMWW ro, 05). Sucrase activity was observed in the proximal and distal small intestines in control animals. activity increased in the group treated with tyrosine and Probios, whereas decreased. In contrast, tripe between these two sites in control animals Treatment with tyrosine and Prob ios decreased putidase activity. Activity increased in treated animals. Compared to Probios treatment, Tyrosine treatment The increase was greater (p<0.05) in pigs exposed to Control and treated pigs dipeptidase activity increases between the proximal and distal small intestine, and dipeptidase activity in the An increase in the specific activity of this enzyme for animals treated with Probios (p<0.01). alkaline phosphatase or for lactase activity, there is no significant interaction between site and treatment at this age. activity in animals treated with tyrosine, all of which were treated with Probios. (sucrase IU/gMWW pr o, 001; p<o, 01 for sucrase-specific activity; alkaline phosph pro, 05) for atase IU/gMlfW.

At this age, alkaline phosphatase-specific activity, There was no significant treatment effect on peptidase or dipeptidase activity . Alkaline phosphatase IU/g! ffW and sucrase activities were determined in control pigs. There were no significant differences between the treated pigs and the treated pigs. Lactase activity in control pigs sex was significantly higher than in treated animals (pro, 05).

at the level of alkaline phosphatase activity or tripeptidase IU/gMWW. The site was found to have no effect. Sucrase activity in proximal tissues and Tripeptidase-specific activity was significantly lower than in the distal region (pro, 001). On the contrary, proximal activity is more important than distal activity for lactase (p<0. 001) and dipeptidase IU/gMWW (p<0.05> significant) It was expensive. There was no significant difference in dipeptidase-specific activity between the proximal and distal regions. It was. The magnitude of the increase in tripeptidase-specific activity between the proximal and distal regions is There were no significant differences in membrane pigs when compared to treated animals. However, Ciro This increase in Syn-treated animals was greater than in the Probios-treated group. was also significantly lower (p<0.05>).

dan0dan10- Sucrase and dipeptidase activities were significantly more significant in the proximal region examined than in the distal region. was low (sucrase activity was pro.

001; p<o for dipeptidase IU/gMWW, 01 and dipeptidase p<0.05 for enzyme-specific activity). Conversely, lactase activity occurs distally The activity was significantly higher in the proximal region when compared to the activity of (pro, 001). alkaliho The level of sphatase or tripeptidase activity differs between the proximal and distal small intestines. There was no significant difference between the two.

Control pigs at 17 days of age for all enzymes studied except dipeptidase. The activity in the treatment group was statistically greater than the activity in the other treatment groups across all sites. showed lower values and was treated with tyrosine than animals treated with Probios. The activity in animals showed statistically high values (but expressed on the basis of wet mucosal weight). and this was not the case for tripeptidase).

The specific activity of dipeptidase in 17-day-old pigs treated with Probios was higher in the tyrosine-treated animals than in the control group and in the control group. The Probios and Probios groups showed similar activity. Dipeptidase activity ([U/g There was no statistical effect of treatment on Probios (MWW); was higher in the tyrosine-treated pigs than in the control group. It was the lowest. Adding either antibiotics or symbiotics to the feed It affected the change in enzyme activity from pre-weaning to post-weaning.

(Margin below) 6 days and days For pigs at 7 days, 17 days, 42 days, and 80 days after birth, each treatment was performed. Mean value (±SE: standard deviation), crypt depth at 0.10 and 0.60 intestinal sites. The results for villus height:crypt depth ratio are presented in Table 5 below.

060 Utsutachibana O7- Place 1 °c - h9. Q'τ-1-shi'7+? -P:QbLO5- of villi Height: Initial analysis of crypt depth revealed significant differences during treatment and mothering. interaction, and the latter is therefore included as a “pro, king” factor in the following analysis. was included. At a given age of the animal, treatment increases villus height or crypt size. There was a significant effect on depth.

Tyrosine or Probios administered to pigs during this experiment reduced villus height. was associated with the prevention of larvae, and was also observed between 7 and 17 days of age in control animals. There was also an association with increased crypt depth at intestinal sites 0 and 10. At 0.60 part , only pigs treated with antibiotics showed significant changes in villus height between 7 and 17 days of age. showed no decrease. At the 0.10 site, all animals were between 17 and 42 days old. There was no decrease in villus height. For 0.60 sites, between 17 and 42 days of age In control pigs, no changes occurred in villus height or crypt depth. There wasn't. However, tyrosine-treated and Probios-treated A decrease and an increase in these rosettes were evident in both animals, respectively. It was.

Probiotics or antibiotics should not be given to pigs before weaning, especially in the proximal intestine. , the pattern of changes in epithelial structure changed. These changes are related to these treated movements. Epithelial cell turnover is slower in the This reflects the theory that there is an expansion of the population of functionally mature intestinal cells. intestine Among the potential benefits conferred by such changes in function, this treatment The results are reflected in improved digestion and increased growth rate after weaning. Both Biological treatments also address consumer concerns associated with antibiotic treatments. There are advantages to avoiding

Moxibustion belly λ The experiment also involved feeding pre-weaned and post-weaned animals with symbiotic substances. clearly demonstrate the effects of

The method used is that this probiotic preparation is effective against four Lactobacillus subspecies and one streptococcus. Essentially the same as experiment 1, with the main difference that it only contained bacterial subspecies. It was like that.

The feeding and rearing diets were changed somewhat from the original experiment. antibiotics The quality treatment is not included, but instead the control treatment and the symbiotic treatment are included in one added. They were various as follows.

(Margin below) (,,,PlcLvX　2s,o　-)9し(Muhege　48..

) 4ya” 24.0 Wheat ° 2S, Q Ka, Sa... 24.o Die waste ls, Q4% S'-+7. 15.0%, S Yuo,.

Lend, 10.5 - Mi) / Mi λ Ul Flash Obstacle 2.0 Bit / S8 U l Shizushi Squid 1.5d1 morning fee 3jojo (lucid 餠 i hair book): One / Saku'l (tl 23.40 20.40D, E, (MJ/kgl 15.20 13 ．． 10 ax strength (%1 1.29' 1.07 Cu (mg/kgl) 30.40 36.50 Tsukuda value (tl 7.8 engineering 1.ri 1 fl&t%+1-13 3-84 Go! SL Ka +'41 0.57 0.45 Slepniso (t) 0.72 0. 54Ca ('kl 1-19 1.13 p + zl O, 770, 71 (F-F-Kin υn There were four stages. All pigs were brought to 21 days of age according to standard 22-5 control methods. Weaned but not at 28 days of age for animals born in stages 1 and 2. Weaned. Samples were taken at 8 days of age: at this time the pigs were acclimated to sows and no food was provided. No feed had been taken yet; 20 days of age: At this time, the pigs were fed, but Not yet weaned; 28 days of age: At this time the pigs have been weaned for a week and have not been weaned yet. 35 days of age: At this point the pigs are recovering from the stress of weaning and Adapted to growing feed.

■ Five symbiotic strains showed significant recovery for both peptidase enzymes studied. produced an effect. Activities of all enzymes were determined in animals weaned by days 21 and 35. decreased between days 20 and 28, with both dipeptidase levels decreasing in treated animals. There was a clear improvement in the activity of the enzyme. Post-weaning fermentation of animals weaned at 28 days of age No decrease in protein was observed in animals weaned at 21 days of age. Seen in the first experiment The significant effect of treatment, as previously reported, was not repeated here. However However, there was a marked recovery of efficacy with treatment, and a significant increase in dipeptidase This was seen on the 8th day of the experiment. Animals in the control group had weight loss after weaning, indicating symbiosis. This was not evident in substance-treated animals.

The table shows that the villus height of Proooios treated animals compared to control pigs. between 8 and 20 days of age. 2 Between 0 and 28 days of age, there is a decrease in cilia height, i.e. villus height: crypt depth. The change in the proportion of was greater in symbiotic-treated pigs than in the control group.

At the 0.60 site, the effect of treatment was evident at the difference between 28 and 35 days of age. . This observation was accomplished by performing crypt depth measurements on one pig per treatment. It was done. Symbiotic treatment parameters for the structure studied in 5 strain species There was no significant effect on the magnitude of change before and after weaning.

However, the decrease in villus height observed between 8 and 20 days of age in the control group The decrease in the villus height:crypt depth ratio is twofold for the elongation of the crypt depth. was not clearly affected by Probios administration during the day of age. However, it was shown that this treatment increased the size. Conversely, in control pigs , between 20 and 28 days of age, the observed decrease in villus height was due to Probios It was unclear which animals were treated.

(Margin below) to 7 L 徨 22 dumb J [branch Rigumi - Tsuchitaki 1 and 5 C) Su Senpoku Quri) The decrease in villus height between 20 and 28 days of age at the 0.10 site was 28 It was not evident in animals weaned on days. 0.60 parts for pigs weaned at 21 days of age At 28 days of age, the large decrease in villus height seen between 20 and 28 days of age This did not occur in weaned pigs.

In the first experiment, improvements in weight gain were observed in both the Probios and thyronone-treated groups. It was not evident until day 26 and day 36, respectively. Also, the latter experiment The animals found that if measurements were continued, there would be a greater correlation across treatment boundaries. It would have been shown. In this experiment, the control group experienced weaning-related weight loss. It is interesting to note that this was not evident in the Probios treatment group, and Biologically treated animals showed higher weight gain at 34 days of age. seen at weaning The effect of this treatment on recovery from the decrease in growth rate and enzyme activity caused by was evident in both animals tested in this study.

Therefore, these experiments demonstrate that Probios does not affect intestinal structure and enzyme reduction after weaning. It has been shown that beneficial changes are correlated with mitigating effects.

Administration of Probios is associated with an increase in the activity of digestive enzymes; are alkaline phosphatase, sucrase, lactase, and tripeptidase may be included. Its effects vary throughout the intestinal region.

A decrease in the villus height:crypt depth ratio also demonstrated the effect of Probios administration. do. the reorientation of enzyme activity and villus height that typically occurs after weaning; The associated increase in crypt depth is reduced by the use of symbiotic agents.

Therefore, it is believed that the present invention has achieved at least all of its objectives.

To small 4+=c~″r=　<’t　L to Even the ears are 1% dirty〒2/monme 1 Overheard L; U1 Huge 11 °−1 11・Iq%I- Scary I'a-L Garden 4 I''\12 months each 'togo H'+taL +1 (1%&+zs notice-, v-<-rbL'8@-Ml?"Iat\Lu=*- 91ffl -31 umbrella 2 meter mouth bite (1 weight and L1 footer (n -311i) ? ]M4+zu9,1呵CL-L 11 28 wa 滲子) Straw now Shijine 1r evening (Y3) 岨 5 Small portion-1: ShiW, Zui It no Oki 2 '　2 hard shiq, l guo shear ugly shi-t, 7-ta　(h 1 button 9) Continuation of front page (51) Int, C1,5 identification symbol Internal office reference number C12R1:245) (CI 2 N 1/20 C12R1:225) (C12N 1/20 C12R1:01) (81) Designated countries EP (AT, BE, CH, DE.

DK, ES, FR, GB, GR, IT, LU, MC, NL, SE), 0A (BF , BJ, CF, CG, CI, CM, GA, GN, ML, MR, SN, TD, TG. ), AT, AU, BB, BG, BR, CA, CH, C3, DE.

DK, ES, FI, GB, HU, JP, KP, KR, LK, LU, MG, MN, MW, NL, No, PL, RO, RU, SD, SE, US I (72) Inventor: Collington, Josina Kay.

United Kingdom FE34 Newscastle upon Tyne, PE Nweal, Hodgkin Park Road

Claims (1)

[Claims] 1. digestive enzyme activity in an animal comprising administering to the animal a composition of a probiotic; A method of stimulating and benefiting an animal through changes in the same. 2. The composition of the probiotic used contains Lactobacillus and Streptococcus. The method described in claim 1. 3. The composition used may contain Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus llus plantorium and fecal streptococcus. How to put it on. 4. The composition of the probiotic used is ATCC number _, _, _, _, _, _, Lactobacillus strains identified by ＿ and ATCC numbers ＿, ＿, ＿, ＿, ＿, Identified 4. The method of claim 3, comprising a streptococcal strain. 5. a Lactobacillus strain in which the composition of the probiotic is identified by ATCC numbers _, _; and a streptococcal strain identified by ATCC number _. How to put it on. 6. The digestive enzymes include sucrase, lactase, alkaline phosphatase, and dipep. selected from the group consisting of tidases and tripeptidases and combinations thereof 2. The method according to claim 1, wherein: 7. the intestinal mucosal structure of the intestine of an animal, comprising administering to the animal a composition of probiotics; A method of changing the animal's structure and benefiting the animal by the change. 8. The composition of the probiotic used contains Lactobacillus and Streptococcus. The method described in claim 7. 9. The composition may contain Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus 9. The method according to claim 8, comprising C. plantorium and fecal streptococci. 10. The composition of the probiotic used is ATCC number _, _, _, _, _, _ , Lactobacillus strains identified by ___ and ATCC numbers ___, ___, ___, ___, ___ Risame 10. The method of claim 9, comprising a streptococcal strain determined. 11. milk in which the composition of said probiotic is identified by ATCC number ___, ___, ___; Claims containing Acidobacteria strains as well as Streptococcus strains identified by ATCC_number The method according to item 9. 12. 8. The method according to claim 7, wherein an increase in the villus height to crypt depth ratio is obtained. . 13. A reduction in the ratio of villus height to crypt depth that occurs after weaning was obtained. 8. The method according to claim 7, wherein: 14. 14. The method of claim 13, wherein the probiotic is administered to the animal prior to weaning. 15. the intestine of the animal after weaning, comprising administering to the animal a composition of probiotics; The structure of the animal and its enzymatic processing of ingested nutrients can be altered to benefit the animal. how to. 16. the probiotic composition used contains Lactobacillus and Streptococcus; 16. The method according to claim 15. 17. The composition may contain Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus The method according to claim 16, which contains plantorium and fecal streptococcus. Law. 18. The composition of the probiotic used is ATCC number _, _, _, _, _, _ , Lactobacillus strains identified by ___ and ATCC numbers ___, ___, ___, ___, ___ identification 18. The method of claim 17, comprising a streptococcal strain. 19. Milk in which the composition of said probiotic is identified by ATCC number ___, ___, ___, ___ Claims containing Acidobacteria strains as well as Streptococcus strains identified by ATCC number _ The method according to item 17. 20. 16. The method of claim 15, wherein the probiotic is administered prior to weaning. 21. 17. The method of claim 16, wherein the probiotic is administered to a pig. 22. said probiotic contains a composition of lactic acid bacteria, and per gram of composition. 22. The method of claim 21, wherein the method is administered at a dose of about 107 viable lactic acid bacteria. 23. 23. The method of claim 22, wherein the probiotic composition is administered in the form of a gel. Law. 24. The compound may be used during the neonatal period of the animal, from about 7 days to 10 days after birth, and until weaning. 24. The method of claim 23, wherein the method is administered at the time of stress or stress. 25. 23. The probiotic composition according to claim 22, wherein the probiotic composition is administered in granular form in the feed. How to put it on. 26. The composition of the symbiotic substance is present when the animal is stressed, arrives at the facility, or 26. The method of claim 25, wherein the method is administered at the time of a change in food ration. 27. Administering a composition of probiotics containing Lactobacillus and Streptococcus before weaning Changes in the structure and chemical composition of the animal's intestines after weaning, including A method that benefits the animal more. 28 The composition is identified by ATCC number ___, ___, ___, ___, ___, ___, ___. Lactobacillus strains and linkages identified by ATCC numbers ___, ___, ___, ___, ___ 28. The method of claim 27, comprising a coccus strain. 29. Milk in which the composition of said probiotic is identified by ATCC number ___, ___, ___, ___ Claims containing Acidobacteria strains as well as Streptococcus strains identified by ATCC number _ The method according to item 27. 30. Stimulates the animal's digestive enzymes and increases the depth of the crypts at the height of the villi after weaning. 28. The method of claim 27, comprising reducing a decrease in the ratio of