In this study, a safety evaluation was conducted to confirm if the Enterococcus faecium CKDB003 strain obtained by selection from a mixed fermentation of fruit and milk is suitable for use as a probiotic. The MIC value for the 10 antibiotics specified in the EFSA guidance was below the acceptable cut-off value. The antibiotic resistance genes aac(6')-li, eatAv, and msr(C) exist by whole genome sequencing, but are in the chromosome and not in the plasmid, thus confirming that there is no possibility of transmission to other microorganisms. It was confirmed that cytolysin (cylA, cylB, cylI, cylL-l, cylL-s, cylM, cylR1, cylR2), aggregation substance (asa1, asp1), collagen adhesion (ace), enterococcal surface protein (esp), endocarditis antigen (efaA), hyaluronidase (hyl) and gelatinase (gelE) were not present in the genome by examining the genes of factors related to virulence. Also, the biochemical analysis showed no toxic enzyme activities, and no virulence genes were detected by the PCR method. Thus, the E. faecium CKDB003 strain can be safely used as a health functional food probiotic, based on the results of the safety assessment.
Proceedings of the Korea Society of Poultry Science Conference
/
2000.11a
/
pp.63-65
/
2000
Two experiments were conducted to investigate the effects of feeding single or combined probiotics on performance and intestinal microflora of broiler chicks for five weeks. Diets based on corn and soybean meal contained 21.50, 19.0% CP and 3,100, 3,150 kcal/kg ME for starting and finishing period, respectively. Lactobacillus salvarius isolated from chicks intestine(LSC), Lactobacillus salvarius isolated from piglet(LSP) Bacillus polyfermenticus(BP) were fed with alone and mixed ones at the level of 0.21 and 0.1% in experiment 1 and 2. Three hundred eighty four chicks were randomly assigned to eight treatment with four replicates of 12 chicks each per treatment. Weight gain, feed consumption, feed conversion ratio(FCR) were weekly measured for 5 weeks. The number of intestinal microflora was examined at the end of experiment. There were no significant different weight gain of all treatment in both experiments. Feed consumption were not shown consistency. Chicks fed LSC alone showed the lowest feed conversion ratio of all treatment in experiment 1, but was not consistency in experiment 2. The number of Lactobacillus spp. tended to increase in chicks ileum fed probiotics treatments, whereas it was not consistency in cecum. The number of ileal E. coli was not decreased in BP treatment.
The aim of this study was to investigate the efficacy of enhanced physiological activities in cultures isolated from Korean fermented watery Kimchi, Dongchimi, of single lactic acid bacteria (LAB), and when these three are mixed LAB as probiotics. Using the BIOLOG system and 16S rRNA sequencing, the isolates were characterized, and identified and assigned to Leuconostoc mesenteroides DK-3, Leuconostoc dextranicum DK-6, and Lactobacillus curvatus DK-13, respectively. Growth rate and pH changes, production of organic acids as metabolites, and physiological activities of the single and mixed LAB cultures, were monitored and compared. In mixed LAB cultures after 72 h of incubation, the maximum concentrations of lactic acid and acetic acid were approximately 340.5 mM and 191.9 mM, respectively, and pH changed from 7.00 to 3.62. Mixed LAB cultures were able to eliminate 96.3% of nitrite. Activities of antioxidant and ${\beta}$-galactosidase were 60.3% and 16.8 units/mg, respectively. Significant antibacterial activity of the concentrated supernatants was demonstrated against several food-poisoning bacteria. Physiological activities obtained from the mixed LAB cultures have been shown to be considerably higher than those of single LAB cultures. In conclusion, these studies demonstrate that compared to the single cultures, all physiological activities in mixed LAB cultures are significantly enhanced.
Eggshell (ES) is a by-product of table eggs with high content of calcium carbonate which can be used as a calcium source in feed. In this study, we have first illuminated the potential application of ES as a novel carrier for probiotics. The carriers used in the study include a SBM (Soybean meal), ESL (Eggshell powder with large particles), ESF (Eggshell powder with fine particles), and the complex carriers (SBM+ESL, SBM+ESF). The structure of carriers absorbed by L. plantarum was confirmed by SEM image. Among these carriers, the complex carrier SBM+ESF showed the highest viability of L. plantarum with pH 7~8 during four weeks storage at room temperature. The SBM+ESF was further tested as a carrier for various probiotic strains at $4^{\circ}C$ or $30^{\circ}C$. All the probiotic strains showed high viability at $4^{\circ}C$ storage. However, a significant reduction of Lactobacillus cells was observed at $30^{\circ}C$ storage. B. lichenifomis maintained high viability whereas B. subtilis, B. amyloliquefaciens, and S. cerevisiae showed the reduction of $2{\log}_{10}$ (CFU/g). These results suggest that if the ESF as a calcium source in feed was mixed with SBM, it can be used as an effective complex carrier for improving the viability of some probiotics including B. licheniformis.
This study was conducted to determine the effect of feed additives including probiotics, moisture and feed ingredients and the effect of fermented feed on digestibility and volatile fatty acid (VFA) level in finishing pigs. Feed was mixed with microbials including Saccharomyces, Lactobacillus, Enterococcus and Pediococcus together with different levels of probiotics, 0.25, 0.5, 1, 2, 3%. Addition of probiotics showed improved fermentation rate after 48 h incubation. To determine the optimal moisture level for fermentation, different levels of water, 30, 40, 50 and 60%, were added into the feed. Fermentation rate of feed with 40~50% moisture level was higher than that from 60% level at 60 h post-fermentation. In vitro fermentation rate of feed ingredients was analyzed by comparing VFA levels. Beet pulp and tapioca showed higher fermentation rate compare to other ingredients including canola meal or rapeseed meal. To determine the effect of administration of fermented feed In vivo, feces from finishing pigs were analyzed. Finishing pigs administrated with fermented feed showed improved digestibility and higher volatile fatty acid (VFA) level. In conclusion, results from the current study indicate that 40~50% of moisture with addition of beet pulp and tapioca in feed is optimal condition for fermentation. Furthermore, our data suggest that fermentation of feed can improve the feed quality and digestibility, thereby provide more nutrient in finishing pigs.
Kim, S.H.;Alam, M.J.;Gu, M.J.;Park, K.W.;Jeon, C.O.;Ha, Jong-K.;Cho, K.K.;Lee, S.S.
Asian-Australasian Journal of Animal Sciences
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v.25
no.2
/
pp.213-223
/
2012
In this study, two experiments were conducted to evaluate the total mixed ration with fermented feed (TMRF) and total mixed ration (TMR) by rumen in vitro fermentation and their effects on the growth performance and blood characteristics of Hanwoo steers. In experiment 1, three Hanwoo steers ($600{\pm}47$ kg), each permanently fitted with a ruminal cannula were used. In this experiment, three diets designated as T1, TMRF (18.4% fermented feed, tall fescue, mammoth wild rye forage and whole crop barley); T2, TMRF (17.7% fermented feed, rice straw and whole crop barley); and T3, TMR (rice straw, whole crop barley and probiotics, but no fermented feed), which were subjected to rumen in vitro fermentation for 48 h. The results demonstrated that DM disappearance rate gradually increased with advancing fermentation time, but T1 and T2 were higher than the T3 (p<0.05) from 3 h to 12 h, but insignificant (p>0.05) at 24 and 48 h. None of the specific VFAs were affected except for acetic and non volatile lactic acids, which were produced more in T2 than in T1 and T3 at 24 h and 48 h of incubation. A/P was lower in T1 and T2 than inT3 at 24 h (p<0.05) and 48 h (p>0.05) of incubation. These results confirmed that TMRF-related treatment shows a superior performance to that of TMR during the ruminal fermentation period. In experiment 2, the three diets in experiment 1 plus 1 more control diet (concentrates, probiotics and 2% rice straw of body weight) were fed to the 48 Hanwoo steers ($160{\pm}10$ kg) for a period of 168 d. The results demonstrated that the daily and total live weight gain and feed efficiency were higher (p<0.05) in the TMRF and TMR groups than in the control group. SGOT, SGPT and BUN (p<0.05) were reduced in TMRF relative to the control and TMR groups by 168 d which confirmed that TMRF shows better blood profiles than the TMR and control groups. Overall, these results appear to show that TMRF has better in vitro ruminal characteristics than those of TMR; growth performance and blood profiles were also found to be superior in TMRF than in the TMR and control groups. Thus, our findings suggest that TMRF-based feed supplies are favorable for Hanwoo cattle.
Bioactive compounds were produced from carrot pomace by solid-state fermentation using Bacillus subtilis HA and Leuconostoc mesenteroides. The carrot pomace (CP) fermented by B. subtilis HA with 3% monosodium glutamate (MSG) showed higher production of various bioactive compounds, with 1.64 Pa·sn of consistency, 2.31% of mucilage content, 16.95 unit/g of fibrinolytic enzyme activity, 35.3 unit/g of proteolytic activity and 37.5 mg% of tyrosine content. The mucilage production was greatly dependent upon the concentration of MSG added. Most MSG added in CP was converted into mucilage (2.3%) including 0.83% poly-$gamma$-glutamic acid (PGA) with 1,505 kDa of molecular weight. The CP fermented secondly by Leuc. mesenteroides showed acidic pH and lower consistency. However, the fibrinolytic and proteolytic activities were increased. The secondly fermented CP showed the viable cell counts with $2.5{\time}108$ CFU/g of B. subtilis HA and $3.7{\time}109$ CFU/g of Leuc. mesenteroides, respectively. The freeze-dried fermented CP showed 2.88 Pa·sn of consistency, 24% of mucilage content and 104.9 unit/g of fibrinolytic enzyme activity, respectively. Also, the powder of fermented CP indicated viable cell counts of $8.0{\time}107$ CFU/g of B. subtilis and $4.0{\time}108$ CFU/g of Leuc. mesenteroides. Therefore, the fermented CP that was fortified with dietary fibers, fibrinolytic enzyme and probiotics could be utilized as valuable ingredients of functional foods in food or cosmetic industries.
Objective: This study aimed to develop and evaluate the effectiveness of a water-soluble microencapsulation method for probiotic strains using gum Arabic (GA) and skim milk (SKM) over a three-month storage period following processing. Methods: Four strains of Pediococcus acidilactici (BYF26, BYF20, BF9, and BF14) that were typical lactic acid bacteria (LAB) isolated from the chicken gut were mixed with different ratios of GA and SKM as coating agents before spray drying at an inlet temperature 140℃. After processing, the survivability and probiotic qualities of the strains were assessed from two weeks to three months of storage at varied temperatures, and de-encapsulation was performed to confirm the soluble properties. Finally, the antibacterial activity of the probiotics was assessed under simulated gastrointestinal conditions. Results: As shown by scanning electron microscopy, spray-drying produced a spherical, white-yellow powder. The encapsulation efficacy (percent) was greatest for a coating containing a combination of 30% gum Arabic: 30% skim milk (w/v) (GA:SKM30) compared to lower concentrations of the two ingredients (p<0.05). Coating with GA:SKM30 (w/v) significantly enhanced (p<0.05) BYF26 survival under simulated gastrointestinal conditions (pH 2.5 to 3) and maintained higher survival rates compared to non-encapsulated cells under an artificial intestinal juices condition of pH 6. De-encapsulation tests indicated that the encapsulated powder dissolved in water while keeping viable cell counts within the effective range of 106 for 6 hours. In addition, following three months storage at 4℃, microencapsulation of BYF26 in GA:SKM30 maintained both the number of viable cells (p<0.05) and the preparation's antibacterial efficacy against pathogenic bacteria, specifically strains of Salmonella. Conclusion: Our prototype water-soluble probiotic microencapsulation GA:SKM30 effectively maintains LAB characteristics and survival rates, demonstrating its potential for use in preserving probiotic strains that can be used in chickens and potentially in other livestock.
We investigated the effects of single and combined administrations of Lactobacillus species (L. plantarum, LP; L. gasseri, LG; L. casei, LC) on blood lipid metabolism and obesity in mice fed a high-fat diet (HFD). The mice were continuously supplemented with LP, LP/LG, or LP/LG/LC, along with HFD, for 12 weeks. The consumption of HFD led to significant increases in body weight, total cholesterol, and triglyceride levels compared to the normal control group. However, administration of LP, LP/LG, or LP/LG/LC to HFD-fed mice reduced body weight gain and showed a tendency to suppress the levels of total cholesterol, triglycerides, and LDL-cholesterol, while increasing HDL-cholesterol levels. The HFD group exhibited increased abdominal fat weight and larger adipocytes in the epididymal adipose tissue compared to the NC group. However, the administered probiotics led to a significant reduction in adipocyte size with decreasing tendency in abdominal fat weight compared with the HFD group. Additionally, the deposition of giant vesicular fat cells in the liver of the HFD group considerably decreased in the probiotic-administered group. Microbiome analysis revealed an imbalance in intestinal microbes in the HFD group, characterized by lower Bacteroidetes and higher Proteobacteria ratios. However, probiotic administration tended to restore the microbial distribution by controlling the abundance of Bacteroidetes and Proteobacteria, resulting in decreased Firmicutes/Bacteroidetes and Proteobacteria/Bacteroidetes ratios. These results suggest that single and combined administration of LP and other probiotics holds enormous potential in reducing obesity in HFD-fed mice as they regulate lipid metabolism, reduce adipocyte size, and restore the balance of intestinal microbes.
This experiment was conducted on 60 Hanwoo calves comprising five feed additive groups, with 12 calves in each group, to determine the effects of additives at pre- and post-weaning on growth performance and blood profile. The groups were control, antibiotic (Neomycin 110 ppm), illite (2%), fermented green tea probiotics (FGTP, 0.5%), and mixed additives (FGTP 0.25%, illite 1% and licorice 0.1%). The calves were offered experimental pellet feeds ad libitum and after one month were supplied with imported timothy hay. They moved freely within the group and suckled their mother' milk during the pre-weaning stage (birth to 3 months) and were separated from their dam during the post-weaning stage (4-5 months). During the pre-weaning stage, the highest average daily gain (ADG) was recorded in the antibiotic- and mixed additive-fed groups followed by FGTP, control and illite groups. In the post-weaning stage, significantly higher total weight gain and ADG were recorded in both the FGTP and mixed additive groups compared to the other groups (p<0.05). Feed efficiency of mixed additive- and illite-fed calves were almost similar with antibiotic-fed calves compared to the other two groups, but the ADG was lowest in illite-fed calves during the pre-weaning stage. In contrast, post-weaning calves fed FGTP and mixed additives showed better feed efficiency. The values of hematological indices, differential leukocyte count, blood proteins and immunoglobulin among the additive-fed calves were not significantly different (p>0.05), although hemoglobin and hematocrit values were lower in FGTP compared to control, but similar in mixed additive and antibiotic groups. These results indicate no detrimental effects of feed additives on the blood profile of calves at both pre- and post-weaning age. Serum albumin in post-weaning calves of all feed additive groups were similar but significantly lower (p<0.05) than in the control group. Post-weaning, IgM was significantly lower (p<0.05) in illite-fed calves compared to other treatment groups, but there was no difference at pre-weaning. Considering all factors, the mixed feed additives and FGTP can be the replacement feed formula for antibiotic for Hanwoo beef calf production, especially when used post- weaning.
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