KR20190014255A - Synbiotic composition comprising lactulose and probiotic - Google Patents

Abstract

The present invention relates to a synbiotic composition. A probiotic and prebiotic according to the present invention surprisingly exhibit a synergistic effect, have an excellent antibacterial effect against strains causing diarrhea of piglets, and can improve the productivity of piglets without side effects.

Description

≪ Desc / Clms Page number 1 > SYNBIOTIC COMPOSITION COMPRISING LACTULOSE AND PROBIOTIC COMPOSITION CONTAINING LACTOTROUS AND PROBIOTIC LACTIC ACID BACTERIA [

The present invention relates to the field of feed additives. More specifically, the present invention relates to the development of a mass production process for a lactic acid bacteria feed additive and a feed additive thereof, which are effective for improving intestinal immune activity and total intestinal bacteria in a piglet disease model, To a feed additive. More particularly, the present invention relates to the use of the probiotic Pediococcus acidilactici GB-U15 strain (Accession No .: KCCM11856P) and lactulose, and a mass production process thereof.

Synbiotics is a mixture of lactic acid bacteria and lactic acid bacteria. That is, a mixture of probiotic and prebiotic is a synbiotic. Examples of the prebiotics include oligosaccharides such as raffinose, soy oligosaccharide, fructooligosaccharide, and galactooligosaccharide, and other lactulose, lactitol, xylitol, and the like. However, it is not known which prebiotic is most suitable for the purpose of synbiotics and the relationship with the combination lactic acid bacteria.

For example, Korean Patent Laid-Open Publication No. 10-2013-0089560 discloses that the conventional prebiotics used in conventional synbiotics do not efficiently function as a nutritional source of probiotics due to the progress of digestion in the host's digestive organs And lack of species specificity for probiotics, which is not effective when it is desired to increase the activity of a specific lactic acid bacterium. However, it is a prebiotic suitable for Bifidobacterium, Lactobacillus, and Pediococcus, Suggest fermented product of oats. Korean Patent Publication No. 10-2013-0089560 discloses that the synbiotics of the combination are effective in enhancing the immune system of the human body.

Korean Patent Publication No. 10-2017-0004897

The present inventors contemplated the development of a synbiotics suitable for the feed of piglets. In particular, the present inventors have paid attention to the topicality of synbiotics useful as a feed additive capable of suppressing E. coli causing diarrhea of piglets and improving the productivity of piglets. To this end, attention has focused on new probiotics and their probiotics, Therefore, it is an object of the present invention to provide a combination of suitable prebiotics and a process design capable of mass production thereof.

The present inventors have been able to solve the above-described problems by the following means.

1. A novel Pediococcus acidilactici GB-U15 strain (Pediococcus acidilactici GB-U15, accession number: KCCM11856P) having antimicrobial activity against E. coli and Salmonella,

2. A synbiotic composition comprising a strain and lactulose according to 1 above.

3. A feed additive for pigs comprising the synbiotic composition according to 2 above.

4. A method for producing a synbiotic composition according to the above 2, wherein the strain according to the above 1 and the lactulose are mixed together in a soybean meal / wheat flour / molasses mixed medium to perform solid fermentation.

5. The production method according to the above 4, wherein the initial moisture content of the fermentation is 50% or more.

The feed additive comprising the synbiotic composition according to the present invention can improve the productivity of the weanling pig, improve the survival rate, show no side effects, and help recover the E. coli infection. Also, according to the process according to the present invention, it is possible to mass-produce the gastric synbiotic composition.

Fig. 1 shows the results of evaluation of antibacterial activities of strains GB-U15, GB-U17 and GB 1-3.
FIG. 2 shows the fermentation characteristics of the strains according to the liquid fermentation composition.
FIG. 3 shows the fermentation characteristics of each strain according to the solid fermentation medium composition.

Lactic acid bacteria strains with antimicrobial activity that can effectively inhibit enteropathogenic bacteria, especially E. coli causing diarrhea of E. coli were selected and strains usable as feed additives were selected by identification.

The bacterial isolates were isolated from BCP medium (5 g / L Casein Enzymic Hydrolysate, 2.5 g / L Yeast Extract, 1 g / L Dextrose, L, Agar 15 g / L) were used to isolate highly probable strains based on organic acid production ability and morphological characteristics. Each sampled sample was diluted with sterile physiological saline (0.85% NaCl), plated on BCP medium, and cultured at 33 ° C for more than 24 hours. The color change in the medium due to the production of organic acid After the biochemical and morphological characteristics of colonies were evaluated, strains determined to be lactic acid bacteria were selected.

The antimicrobial activity of selected strains was evaluated by agar well diffusion method. Diarrhea causing E. coli was used under the pre-sale outdoor strains that may cause the actual disease on a farm in the pathology laboratory of Seoul National University veterinary Salmonella strain was used as an indicator strain S. entritidis strain. The indicator strains of the two species (diarrhea-producing E. coli, Salmonella strains) were inoculated into the LB liquid medium, cultured at 37 ° C in a shaking incubator at 200 rpm for 24 hours, and then added 1% to the LB agar medium, Agar medium was prepared. The prepared culture medium was punctured with a perforator using a perforator, and the antimicrobial activity against the indicator strains of each strain was evaluated by adding the culture medium of each selected strain. The culture of the selected strain was cultured for 24 hours at a temperature of 37 ° C using MRS broth for each of the selected strains, and the culture broth was prepared. To confirm the antimicrobial activity of the culture broth as well as the antimicrobial activity The culture solution prepared by adjusting the pH of the culture solution to 7.0 was also prepared, and 200 μl of the sterilization solution which had been sterilized with a 0.45 μm filter was added to the perforated medium containing the harmful microorganisms, followed by refrigerated storage at 4 ° C. for 12 hours to sufficiently spread the culture solution on the agar After that, the cells were cultured in a 37 ° C incubator for 12 hours to confirm the production of transparent rings.

Based on the results of the antimicrobial activity evaluation, strains showing antimicrobial activity against the indicator strain, yeast pneumoniae-causing E. coli or Salmonella were selected as the first candidate strains. The results of antimicrobial activity evaluation of representative strains GB-U15, GB-U17 and GB-1-3 are shown in FIG.

Biochemical characterization of strains with antimicrobial activity was carried out using API 50CHL kit. The results were as follows.

Sample Strain GB-U 15 Pediococcus pentosaceus  One GB-U 17 Lactobacillus plantarum  One GB 1-3 Lactobacillus plantarum  One

After confirming the biochemical characteristics of the strain using the PI 50CHL kit, 16S rRNA sequencing was performed on the selected strain for the identification of the strain. The results of strain identification are as follows.

Sample Strain GB-U 15 Pediococcus acidilactici GB-U 17 Lactobacillus plantarum GB 1-3 Lactobacillus plantarum

The pH stability was investigated in order to investigate the possibility of using three selected strains as feed additive.

To confirm the pH stability, each strain was inoculated into MRS liquid medium and cultured at 37 ° C for 24 hours at 200 rpm for 24 hours. The culture was adjusted to pH 3, 4, 6, 8 using 1N HCl and 1N NaOH MRS broth was inoculated with 1% of each strain and cultured at 37 ° C and 200 rpm for 24 hours. The stability of each strain was confirmed by analysis of viable cell counts.

As a result of evaluation of pH stability, three strains were found to be stable to acid resistance and bile resistance.

Strain pH 3 pH 4 pH 6 pH 8 0 hour 24 hours 0 hour 24 hours 0 hour 24 hours 0 hour 24 hours GB-U 15 8.90E + 07 1.10E + 08 6.20E + 07 9.00E + 08 1.67E + 08 4.30E + 09 1.34E + 08 7.10E + 09 GB-U 17 1.02E + 07 8.34E + 06 4.76E + 07 8.31E + 08 5.36E + 07 2.42E + 09 4.12E + 07 5.12E + 08 GB 1-3 3.88E + 07 9.83E + 07 6.24E + 07 7.91E + 08 8.00E + 07 6.17E + 09 9.00E + 07 4.80E + 09

The units in Table 3 are CFU / mL. In the following table, the units for the number of bacteria are the same.

The thermal stability of the strain was also evaluated. For the evaluation of the thermal stability, each strain was inoculated into the MRS liquid medium and cultured at 37 ° C. for 24 hours with shaking at 200 rpm. The culture broth was inoculated into the MRS liquid medium at a rate of 1% at 37 ° C. and 200 rpm for 24 hours After culturing, the degree of decrease in the number of viable cells was confirmed by heat treatment at 60 ° C, 70 ° C and 80 ° C for 5 minutes and 10 minutes.

As a result of evaluation of heat stability, all of the strains showed a tendency to be stable to heat resistance generally at 70 ° C for up to 5 minutes.

Strain 60 ° C 70 ℃ 80 ℃ 5 min 10 min 5 min 10 min 5 min 10 min GB-U 15 2.50E + 09 1.40E + 08 3.50E + 08 2.17E + 08 5.00E + 07 1.67E + 07 GB-U 17 1.85E + 09 8.78E + 08 1.33E + 09 6.74E + 08 5.32E + 08 7.45E + 07 GB 1-3 2.71E + 09 3.10E + 08 5.30E + 08 1.50E + 07 5.76E + 08 1.00E + 07

To investigate the effect of the combination of pre - and pre - biotics, we investigated the bacterial numbers of the selected strains and E. coli.

For the evaluation of the selection and prebiotic combination, each strain was inoculated into the MRS liquid medium and cultured at 37 ° C. for 24 hours with shaking at 200 rpm. After culturing, 1% of the candidate culture medium and LB liquid Escherichia coli cultured in the medium for 24 hours was added thereto, and 0.5% prebiotic was mixedly inoculated. After incubation at 37 ° C and 200 rpm for 24 hours, the changes of lactic acid bacteria and E. coli bacteria were investigated.

The combination of GB-U 15 and lactulose showed the highest growth rate of lactic acid bacteria in the growth evaluation, and the inhibitory effect of E. coli was also excellent.

Synbiotics Lactobacillus Escherichia coli Probiotic Prebiotic 0 hour 24 hours 0 hour 24 hours GB-U 15 - 8.10E + 07 8.96E + 09 1.10E + 07 ND Lactulose 1.20E + 07 7.56E + 09 8.00E + 06 ND Fructo-oligosaccharide (FOS) 1.50E + 07 6.84E + 09 1.20E + 07 ND Monosaccharide 1.60E + 08 9.60E + 09 1.50E + 07 ND

ND in Table 5 indicates that no ND was detected.

We evaluated the efficacy of lactulose for other strains. However, the combination of GB-U 15 and lactulose showed the best effect on the rate of lactic acid production increase.

Synbiotics Lactobacillus Escherichia coli Probiotic Prebiotic 0 hour 24 hours 0 hour 24 hours GB-U 17 Lactulose 1.40E + 07 4.00E + 09 1.90E + 07 ND GB 1-3 Lactulose 5.40E + 07 3.53E + 09 6.00E + 06 ND

In the case that the tendency of increase of lactic acid bacteria growth is not high even if the lactic acid bacteria growth and the inhibition effect of Escherichia coli are shown, the growth rate of the bacteria is too slow in the initial culture, and it takes a long time to cultivate, have. In other words, although the exact cause is not known, only the combination of GB-U 15 and lactulose exhibits mutual synergistic effect, excellent growth rate of lactic acid bacteria, and excellent effect of inhibiting E. coli. Thus, It was most suitable for actual production such as production possible.

In vivo. Efficacy was also assessed. The effects of synbiotics - induced diarrhea recovery after coliform challenge were investigated in rats without prior feeding of synbiotics before challenge with E. coli.

In this experiment, each strain (GB-U 15, GB-U 17, GB 1-3) was inoculated into the MRS liquid medium and cultured at 37 ° C. for 24 hours with shaking for 24 hours. / L, KCl 0.2 g / L, Na ₂ HPO ₄ 1.42 g / L, KH ₂ PO ₄ 0.24 g / L, DW 1 L)) with 0.5% lactulose and 5.0 × 10 ⁷ CFU / mL or more And diluted as much as possible to prepare a synbiotics. Finally, the number of probiotic bacteria when orally administered to rats was 5.0 × 10 ⁶ CFU / mL or more.

3 weeks old BALB / cAnNTac was infected with Lactobacillus cultures having antimicrobial activity in a control group (healthy mouse group) and an experimental group (a group of mice that caused diarrhea after challenge with E. coli) after about one week of adaptation to the surrounding environment The in vivo recovery efficacy of neurobiotics between groups fed lactulose was evaluated. On day 1, E. coli was inoculated inoculated and the body weight change was observed while feeding synbiotics for a total of 12 days.

Experimental results showed that body weight loss due to inoculation of Escherichia coli on day 1 was up to day 3. From the third day to the sixth day, the recovery effect according to the synbiotics appears. From the 6th day onwards, it is hard to judge the increase in recovery due to synbiotics benefits due to the natural ability of the mice to recover. Therefore, we observed intensively the change of body weight of mice from 3 days to 6 days after challenge with E. coli. In the case of GB 1-3, the antimicrobial activity was observed, but in vivo test results showed that the GB 1-3 and lactulose-fed rats showed a slight decrease in body weight compared with the rats not fed. However, the weight gain of rats fed with GB-U 15 and lactulose showed a weight difference of 15.7 g (based on the 6th day of feeding) compared to the mice infected with E. coli without the addition of synbiotics, and the recovery effect on clear coliform diarrhea . Thus, even a probiotic having an antibacterial activity may not have an effect on the actual E. coli diarrhea, but it has been confirmed that the combination of GB-U 15 and lactulose shows a large recovery effect on E. coli.

Treatment E. coli
Whether 1st day tea 2nd day car 3 day car 4th day car 5th day car 6 day car Negative PBS 100.00 101.70 107.70 113.77 117.10 122.83 E. coli 100.00 88.68 89.53 92.90 99.65 108.18 GB-U 15 + lactulose PBS 100.00 101.80 105.20 111.10 118.17 108.18 E. coli 100.00 89.18 89.70 97.70 103.33 123.87 GB-U 17 + lactulose PBS 100.00 101.77 102.00 106.37 112.60 141.37 E. coli 100.00 89.48 89.30 96.28 102.58 110.53 GB 1-3 + lactulose PBS 100.00 98.60 101.07 107.50 111.20 115.57 E. coli 100.00 88.70 86.20 90.35 99.90 106.35

The units in Table 7 are g.

In addition, each strain (GB-U15, GB-U17, GB1-3) was inoculated into the MRS liquid medium and cultured at 37 ° C with shaking at 200 rpm for 24 hours. L, KCl 0.2 g / L, Na ₂ HPO ₄ 1.42 g / L, KH ₂ PO ₄ 0.24 g / L, DW 1 L) was diluted to 0.5% lactulose and 5.0 × 10 ⁷ CFU / mL or higher To prepare a synbiotics. Finally, when the mice were orally administered, the number of probiotic bacteria was 5.0 × 10 ⁶ CFU / mL or more.

After 3 weeks of age, BALB / cAnNTac was applied to the control group (healthy mice group) and experimental group (lactic acid bacteria cultured with antimicrobial activity and lactulose Were evaluated for prevention and recovery efficacy by synbiotics.

The experimental results are as follows.

The results of this experiment were statistically analyzed using the statistical program SAS. From the third day (22 days) after the inoculation of Escherichia coli, most of the mice showed a tendency to decrease in body weight. On the 5th day (24th day) after the Escherichia coli inoculation, the body weight started to increase again. The subjects with the lowest relative weight and the highest individual were selected and euthanized after euthanasia. After the inoculation of Escherichia coli at the 5th day (24th day), the mouse body weight was almost constantly increased until the 12th day (31 days) after inoculation of Escherichia coli. in During the vivo experiment, mouse death was not induced.

Experimental results showed that the body weight gain of GB-U 15 and lactulose-fed rats was significantly improved compared to rats infected with E. coli without synbiotic feeding.

Primary Positive Control GB-U15
+ Lactulose GB-U17
+ Lactulose GB 1-3
+ Lactulose P value Average Standard error Average Standard error Average Standard error Average Standard error 0 days 100.00 ^A 0.00 100.00 ^A 0.00 100.00 ^A 0.00 100.00 ^A 0.00 3.0000 1 day 100.75 ^B 0.42 102.63 ^BA 0.69 103.46 ^A 0.51 101.49 ^B 0.18 0.0040 2 days 101.55 ^B 0.42 105.08 ^A 1.14 106.85 ^A 1.01 105.23 ^A 0.45 0.0015 3 days 91.31 ^B 0.42 94.03 ^BA 0.72 94.42 ^A 1.29 92.72 ^BA 0.14 0.0400 4 days 90.52 ^A 0.71 91.94 ^A 0.84 91.85 ^A 1.52 90.15 ^BA 0.79 0.5055 5 days 96.91 ^A 0.99 98.53 ^A 0.91 98.27 ^A 1.52 94.87 ^A 0.58 0.0878 6 days 100.15 ^B 0.17 103.03 ^A 0.78 102.82 ^A 0.73 99.77 ^B 0.28 0.0016 7 days 102.97 ^B 0.25 105.70 ^A 0.72 105.63 ^A 0.84 102.26 ^B 0.39 0.0022 8 days 103.17 ^B 0.63 107.96 ^A 1.13 107.51 ^A 1.42 102.71 ^B 0.50 0.0037 9th 105.41 ^A 0.67 109.82 ^A 1.75 109.79 ^A 1.55 106.51 ^A 1.73 0.1296 10 days 108.17 ^BA 0.63 112.12 ^A 1.62 111.10 ^BA 1.45 106.67 ^B 0.49 0.0205 11th 107.25 ^BA 0.65 112.54 ^A 1.50 111.01 ^BA 1.89 106.43 ^B 0.57 0.0153 12th 109.19 ^B 0.56 114.84 ^A 1.81 114.60 ^BA 1.62 109.47 ^BA 0.76 0.0115

Fecal index was also evaluated. There was no significant difference in fecal indices between treated group fed with synbiotics after challenge with E. coli and control group fed only with E. coli without attack.

An autopsy was performed to visually observe the changes in digestive organs (stomach or intestine) due to synbiotic treatment. Euthanasia proceeded with dislocation of cervical vertebrae. At autopsy, whole organs were extracted from airway (esophagus) to anus, and rectum and ileum were cut into approximately 0.5 ㎝ each and fixed to 10% formalin. All but the rectum and the ileum were fixed at 10% formalin. Autopsy revealed no abnormal signs associated with the synbiotic treatment of GB-U 15 and lactulose combinations.

After the autopsy, the excised ileum and rectum were fixed in 10% formalin, and two were randomly selected for each group to perform a biopsy on AbionCRO. As a result of the biopsy, inflammation and mucosal proliferation of the treated group treated with the synbiotics and the control group treated with only the Escherichia coli without the synbiotics were weak, and the significant difference in the inflammation and mucosal proliferation between the treated group and the control group Were not observed. This confirms that there is no side effect due to synbiotics of GB-U 15 and lactulose combinations.

In order to investigate the optimum culture conditions of GB-U 15, the changes of bacterial counts were investigated in liquid and solid phase fermentation conditions.

Liquid medium composition was investigated in three commonly used media. Fermentation experiments were conducted on raw materials that can be easily obtained for solid fermentation medium composition, and optimum fermentation medium composition was selected.

Medium composition  ( % ) L-1 L-2 L-3 Glucose 2.0 4 15 Yeast extract 1.5 One 5 peptone 2 NaCl 0.5 0.5 K ₂ HPO ₄ 0.2 5 MgSO ₄ 0.01 0.75 molasses 5

Medium composition  ( % ) S-1 S-2 S-3 S-4 S-5 S-6 Soybeans / wheat flour 95 50 50 bran 95 50 20 Main berth 95 45 45 25 molasses 5 5 5 5 5 5

As a result, as shown in FIG. 2 and FIG. 3, the composition of the liquid fermentation broth for seed culture was the best in the L-2 composition for 24 hours, and the solid fermentation broth for mass production was the soybean / The molasses mixed culture medium was selected as the optimum medium for the growth.

Solid fermentation conditions at lab scale were evaluated by evaluating the fermentation characteristics of GB-U 15 and lactulose by single and multiple fermentation at laboratory level in feed grains.

Under lab scale conditions, solid fermentation seedlings were inoculated with MRS broth in GB-U 15 and incubated at 37 ° C in a shaking incubator at 200 rpm for 24 hours.

In order to establish the lab-scale solid fermentation condition, the water content of the initial raw material was adjusted to 40%, 50% and 60% by S-1 medium composition, sterilized at 121 ° C for 15 minutes, sufficiently cooled at room temperature, 1% (initial number of bacteria: 1.0 x 10 ⁴ CFU / g). After inoculation, the fermentation temperature was changed to 30 ℃, 35 ℃ and 40 ℃, respectively. In addition, in order to check the changes such as the degree of growth of bacteria through the mixed fermentation of prebiotics in the solid fermentation stage, 1% of prebiotic was added and the fermentation was proceeded under the same conditions to investigate the number of bacteria and pH.

The fermentation temperature was 35 ℃ and the initial moisture content was 60% for the optimum solid fermentation. At initial water content of more than 50% The addition of lactulose to GB-U 15 also confirmed the growth of bacteria by solid fermentation. Based on the above results, the optimum conditions of solid fermentation were 50 ~ 60% initial moisture content and 35 ~ 36 ℃ fermentation temperature.

Fermentation temperature 30 ℃ 35 ℃ 40 ℃ Singer 40% 50% 60% 40% 50% 60% 40% 50% 60% GB-U 15 1.54E + 09 2.31E + 09 3.48E + 09 1.81E + 09 3.11E + 09 5.20E + 09 9.20E + 08 2.35E + 09 3.43E + 09 GB-U 15
+ lactulose 1.61E + 09 2.14E + 09 3.51E + 09 2.10E + 09 5.20E + 09 7.10E + 09 1.2E + 09 3.14E + 09 5.12E + 09

Establishment of the pilot scale fermentation process for establishing the final mass production process was carried out at the level of 3 tons / batch and the variation of the fermentation quality due to the characteristics of solid fermentation was large,

The pilot scale fermentation process was established based on the results of establishing lab scale fermentation conditions for the combination of GB-U 15 and lactulose. The total fermentation temperature was 35 ℃ for 36 ~ 40 hours and the fermentation temperature was 50% and the fermentation temperature was 35 ℃. As a result, the growth of 10 ⁹ CFU / g or more was confirmed at 1.02 × 10 ⁹ CFU / g at 18 hours of fermentation, and the growth of the bacteria was confirmed up to 36 hours after fermentation. As a result of fermentation experiments for 6 batches, it was confirmed that the samples were stable at a concentration of 10 ⁹ CFU / g or more from 24 hours to 36 hours after fermentation. The final fermentation process was established by obtaining stable results with an average of 1.78 × 10 ⁹ CFU / g.

Processing date Raw material amount  (kg) Number of bacteria  ( CFU / g) Yield ( % ) April 11 3,010 3.20E + 08 88.9 May 12 3,011 5.90E + 08 90.2 May 18 3,015 1.90E + 09 90.1 May 26 3,100 9.80E + 08 90.2 June 7 3,017 2.50E + 09 89.8 June 20 3,100 3.10E + 09 90.5 Average 3,042 1.78E + 09 89.95

GB-U15 and yiyujadon in vivo of new probiotic formulations that combines lactulose Efficacy was also assessed. Productivity, delay in standing, feeding delay, fecal index and survival rate.

(Landrace × Yorkshire) × Duroc] Weighed 50 dogs were weighed and weighed 5.33 ± 0.60 kg at the start of the test. Specimens were tested for 4 weeks. The test design consists of 1) CON (basic feed); 2) CON + 0.05% synbiotics; 3) CON + 0.1% synbiotics; 4) CON + 0.1% probiotic; 5) CON + 0.05% prebiotic; 5 treatments, 2 replicates per treatment, and 5 replicates per replicate. In addition, PBS (phosphate buffered saline) was used for one repetition and E. coli (2 x 10 < ⁹ > CFU / ml).

The test was conducted at Dankook University test farm. The test diets were fed with corn-soybean meal based on NRC (2012) requirements, and water was freely fed using an automatic water dispenser.

Mix 50% of lactulose in soybean meal and wheat mixed cereal medium, add water to 50% of initial water content and mix well. After heat treatment at 121 ° C for 30 minutes, the solid medium is sufficiently cooled for inoculation and then 1% of GB-U15 strain is inoculated. After inoculation, the mixture was fermented at 35 ° C for 35 hours, dried with hot air at 40 ° C and finally pulverized to prepare a prototype for the specification test. Among the above test samples, the probiotic is GB-U15 and the prebiotic is lactulose. 0.0 > 0.05% < / RTI > in COM + 0.05% Shinbarotics represents the treatment in which 0.05% of the synbiotic sample (GB-U15 + lactulose) according to the present invention is added to the control feed CON. The percentages in the remaining samples are equivalent.

Body weight was measured at initiation, at 2 weeks and at the end (4 weeks) by treatment. Feed intake was calculated by subtracting the remaining amount from the feed amount during body weight measurement. Feed efficiency was calculated by dividing the feed intake by the body weight gain.

E. coli was cultured in LB (Luria-Bertani) broth, and PBS was added to each treatment for each treatment at the 2nd week, and E. coli (2 x 10 ⁹ CFU / ml) Respectively.

All data were analyzed using Duncan's multiple range test (Duncan, 1955) to determine the efficacy of the additive using the General Linear Model procedure of SAS (2013).

The effect of the addition of the synbiotics according to the present invention on the productivity of weaned piglets is shown in the table below. On the 2nd and 4th weeks of each treatment, TRT1 and TRT2 treatments were higher in TRT1 and TRT2 treatments than in control and probiotics treatments. The TRT1 and TRT2 treatments were significantly higher (p <0.05) than the control treatments (p <0.05). Also, the TRT1 and TRT2 treatments were significantly higher than the control and probiotic treatments (P <0.05) than those of TRT3 treatment. The TRT1 and TRT 2 treatments were significantly higher (P <0.05) than the control and probiotic TRT3 treatments and prebiotic treatment treatments, respectively. The feed efficiency of TRT1 was significantly higher than that of the control (P <0.05).

In conclusion, the body weight gain and feed efficiency of TRT1 and TRT2 treated with synbiotics were significantly higher than those of the control (P <0.05), and compared with simple prebiotic or probiotic treatments It has been confirmed that the benefit of a combined form of synbiotics, rather than a single formulation in the form of prebiotics and probiotics, is more effective in improving the productivity of the weaned piglets.

Items CON TRT1 TRT2 TRT3 TRT4 SEM ² Weight, kg Early 5.35 5.34 5.30 5.31 5.33 0.02 2 parking 9.01 ^b 9.76 ^a 9.77 ^a 9.10 ^ab 9.11 ^ab 0.18 4 parking 14.66 ^b 16.99 ^a 16.87 ^a 15.38 ^ab 15.38 ^ab 0.45 2 parking ADG, g 259 ^b 316 ^a 319 ^a 268 ^ab 270 ^ab 13 ADFI, g 304 335 335 307 300 10 GF 0.851 ^b 0.943 ^a 0.952 ^a 0.870 ^b 0.899 ^ab 0.017 FCR 1.175 ^a 1.06 ^b 1.05 ^b 1.149 ^a 1.115 ^ab 0.021 4 parking ADG, g 403 ^b 516 ^a 507 ^a 449 ^ab 448 ^ab 24 ADFI, g 668 ^b 703 ^a 704 ^a 674 ^b 672 ^b 7 GF 0.603 ^b 0.735 ^a 0.719 ^ab 0.666 ^ab 0.665 ^ab 0.032 FCR 1.668 ^a 1.363 ^b 1.427 ^ab 1.513 ^ab 1.513 ^ab 0.061 Sum ADG, g 331 ^b 416 ^a 413 ^a 358 ^ab 359 ^ab 16 ADFI, g 486 519 520 491 486 9 GF 0.681 ^b 0.802 ^a 0.794 ^a 0.73 ^ab 0.737 ^ab 0.022 FCR 1.472 ^a 1.247 ^b 1.272 ^b 1.375 ^ab 1.363 ^ab 0.035 ¹ Abbreviation: CON, foundation feed; TRT1, CON + 0.05% synbiotics; TRT2, CON + 0.1% synbiotics; TRT3, CON + 0.1% probiotic; TRT4, CON + 0.05% prebiotic; ² Mean standard error Different characters in columns such as ^{a and b} show significant differences (P <0.05).

Korea Microorganism Conservation Center (overseas) KCCM11856P 20160629

Claims (5)

Pediococcus acidilactici GB-U15 (accession number: KCCM11856P), which has antibacterial activity against E. coli and Salmonella,
11. A synbiotic composition comprising the strain of claim 1 and lactulose.
A feed additive for pigs comprising a synbiotic composition according to claim 2.
The process for producing a synbiotic composition according to claim 2, wherein the strain according to claim 1 and lactulose are mixed together in a soybean meal / wheat flour / molasses mixed medium to perform solid fermentation.
The method of claim 4, wherein the initial moisture content of the fermentation is 50% or more.

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