• Title/Summary/Keyword: feed additives

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Purification and Characterization of a Novel Alkaline Protease from Bacillus horikoshii

  • Joo, Han-Seung;Choi, Jang-Won
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.58-68
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    • 2012
  • An investigation was conducted on the enhancement of production and purification of an oxidant and SDS-stable alkaline protease (BHAP) secreted by an alkalophilic Bacillus horikoshii, which was screened from the body fluid of a unique Korean polychaeta (Periserrula leucophryna) living in the tidal mud flats of Kwangwha Island in the Korean West Sea. A prominent effect on BHAP production was obtained by adding 2% maltose, 1% sodium citrate, 0.8% NaCl, and 0.6% sodium carbonate to the culturing medium. The optimal medium for BHAP production contained (g/l) SBM, 15; casein, 10; $K_2HPO_4$, 2; $KH_2PO_4$, 2; maltose, 20; sodium citrate, 10; $MgSO_4$, 0.06; NaCl, 8; and $Na_2CO_3$, 6. A protease yield of approximately 56,000 U/ml was achieved using the optimized medium, which is an increase of approximately 5.5-fold compared with the previous optimization (10,050 U/ml). The BHAP was homogenously purified 34-fold with an overall recovery of 34% and a specific activity of 223,090 U/mg protein using adsorption with Diaion HPA75, hydrophobic interaction chromatography (HIC) on Phenyl-Sepharose, and ion-exchange chromatography on a DEAE- and CM-Sepharose column. The purified BHAP was determined a homogeneous by SDS-PAGE, with an apparent molecular mass of 28 kDa, and it showed extreme stability towards organic solvents, SDS, and oxidizing agents. The $K_m$ and $k_{cat}$ values were 78.7 ${\mu}M$ and $217.4s^{-1}$ for N-succinyl-Ala-Ala-Pro-Phe-pNA at $37^{\circ}C$ and pH 9, respectively. The inhibition profile exhibited by PMSF suggested that the protease from B. horikoshii belongs to the family of serine proteases. The BHAP, which showed high stability against SDS and $H_2O_2$, has significance for industrial application, such as additives in detergent and feed industries.

Experimental studies of homogeneity and stability honeybee venom using ultra-high performance liquid chromatography (주사제 사용을 위한 봉독의 균질성 및 안정성 평가)

  • Han, Sang Mi;Kim, Se Gun;Hong, In Phyo;Woo, Soon Ok;Jang, Hye Ri;Lee, Kyung Woo
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.81-86
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    • 2016
  • Honeybee venom (BV) from Apis mellifera L. has been used as natural antimicrobial compounds in pigs, cows, dairy cattle and chicken farms in Korea. The purpose of this study was conducted to confirm homogeneity and stability of BV dissolved with distilled water or saline solution. Melittin was analyzed with ultra-high performance liquid chromatography (UPLC) for BV to secure the validation of analysis. BV at concentration of 1 mg/mL was dissolved with distilled water or saline solution at room temperature. Homogeneity of BV dissolved with distilled water or saline solution at upper, milddle, and lower layers all satisfied the accuracy and precision criteria. Stability of BV dissolved with distilled water or saline solution for 7 days all satisfied the criterion both light and dark storage condition. BV has satisfied with homogeneity and stability in distilled water or saline solution at room temperature under light or dark condition. The results of this study suggest that BV has a possibility as the substitute of natural antimicrobial agents for the animal drugs and feed additives.

In vitro selection of lactic acid bacteria for probiotic use in pigs (양돈용 생균제 개발을 위한 유산균주 선발)

  • Byun, Jae-won;Kim, Gyung-tae;Bae, Hyoung-suk;Baek, Voung-jin;Lee, Wan-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.701-706
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    • 2000
  • This study was carried out to select the lactic acid bacteria(Lactobacillus, Streptococcus and Bifidobacterium) and yeast for probiotic use in pigs. Acid-tolerant 536 strains were isolated from the feces of 30 pigs. To select useful strains, the first screened strains were treated with strong acid solution(pH 2.5 to 3.0) for 3 hours and subsequentely treated with the anaerobic diluent solution containing 0.15% Oxgall for 3 hours. Among these strains, 151 strains showed strong tolerance to both acid and bile. Lactobacillus and Streptococcus tolerant to the acid and bile were treated with heat at $80^{\circ}C$ for 15 min, and at $70^{\circ}C$ for 5 min in Bifidobacterium and yeast. As a result of heat treatment, 38 strains were obtained as heat-tolerant strains. All of heat-tolerant strains were tested for antibiotic resistance against virginiamycin, sulfathiazole, aureomycin, neomycin, linsmycin, tiamulin and ASP250 which were used as feed additives for growth promotion in pigs. Finally, one strain each from Lactobacillus, Streptococcus, Bifidobacterium and yeast that showed resistance to acid, bile, heat and antibiotics was selected for probiotic use in pigs.

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Cloning, High-Level Expression, Purification, and Properties of a Novel Endo-${\beta}$-1,4-Mannanase from Bacillus subtilis G1 in Pichia pastoris

  • Vu, Thi Thu Hang;Quyen, Dinh Thi;Dao, Thi Tuyet;Nguyen, Sy Le Thanh
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.331-338
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    • 2012
  • A novel gene coding for an endo-${\beta}$-1,4-mannanase (manA) from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The ${\beta}$-mannanase showed an identity of 90.2-92.9% ${\leq}95%$) with the corresponding amino acid sequences from B. subtilis strains deposited in GenBank. The purified ${\beta}$-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDI-TOF mass spectrometry. The recombinant ${\beta}$-mannanase had an optimum temperature of $45^{\circ}C$ and optimum pH of 6.5. The enzyme was stable at temperatures up to $50^{\circ}C$ (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions ($Hg^{2+}$, $Pb^{2+}$, and $Co^{2+}$) substantially inhibited the recombinant ${\beta}$-mannanase. The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol, ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v) or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the ${\beta}$-mannanase expressed in P. pastoris could potentially be used as an additive in the feed for monogastric animals.

The Protein rich Food Mixtures for Korean Infants (유유아(乳幼兒) 및 성장기 아동을 위한 영양식품 개발에 관한 연구 -흰쥐 성장에 미치는 영향-)

  • Ho, Jin-Hee;Kim, Sook-He
    • Journal of Nutrition and Health
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    • v.3 no.2
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    • pp.95-99
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    • 1970
  • Korea is one of the typical rice eating countries where availability of animal protein is limited even for the growing generation. Nutritive food products for infants and children are not available commercially in large scale at the present time, although a limited amount of expensive milk products are produced. The present study deals with a pioneering attempt on a new food mixture to meet this demand. Several food mixtures, possibly produced in the country, consisting of rice, soybean, FPC, vitamin, minerals and other food additives are developed in this work Sixty female and male rats aged $30{\sim}40$ days were divided into five groups, twelve rats each. The rats had been maintained with the six different diets, as follow. Formulation of proposed infant food mixturesComposition F-R-1 F-S-2 F-F-3 F-P-4 S Rice 100% 40% 37% 46% 70% Sugar - 12 13 10 70 Casein - - - - 20 Bean - 40 37 24 - Yeast - 3 2 3 - Mineral Vitamain - 2 4 2 4.3 F.P.C - 3 4 7 - Fat - - 4 8 4 Cod liver Oil - - - - 3 The findings of this study presented in FER, PER, final organ weights, body weights growth, percentage of nitrogen retention in the body. F-P-4 group tended to remain the heaviest and F-R-1 the lightest in the body weight among three groups throughout the experimental period. In terms of FER (Feed Efficiency Ratio) and PER (Prortein E. R.) value, F-P-4 group kept the highest record throughout. F-P-4 group showed the highest value of nitrogen retention in the body. In comparision between F-P-4 groups and casein 20% group (standard) in all respects of this experiment, F-P-4 group which is the most superior among experimental group, revealed statisfically no significant inferior than that of standard group. This fact could be interpreted that most limiting a. a., methionine in the soy protein produced little inferiority of the experimental groups in the study.

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Influence of Hermetia illucens Larvae-Derived Functional Feed Additives on Immune Function of Broilers (동애등에(Hermetia illucens)유충 유래 기능성 사료첨가제가 육계 면역기능에 미치는 영향)

  • Ji, Sang-Yun;Park, Kwan-Ho;Kim, Ki-Hyun;Lee, Heui-Sam;Choi, Gyu-Sung;Lim, Yeon-Ji;Yu, Ri;Choi, In-Hag;Chung, Tae-Ho
    • Journal of Environmental Science International
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    • v.27 no.12
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    • pp.1305-1307
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    • 2018
  • We investigate the effects of the immune function (HI titer) in broilers fed diets containing Hermetia illucens (H. illucens) peptide extract over a 40-day period. Twenty-four broiler chicks (Arbor Acres, 1 d old) were divided into four groups and fed different diets (control, 0.1%, 0.5%, and 1% H. illucens peptide extract). To evaluate HI titer, all broilers were vaccinated with H9H2 vaccine subcutaneously on the lateral thorax, according to the manufacturer's recommendations. Similar HI titer was observed with 1% H. illucens peptide extract treatment compared to the control after 40 days (p>0.01). Groups fed 0.5% H. illucens peptide extract demonstrated the most effective immune effects (p<0.01), followed by groups fed 0.1% H. illucens peptide extract. In conclusion, using 0.1% or 0.5% H. illucens peptide extract before or after vaccination improved HI titer immune function in broilers.

Dietary turmeric (Curcuma longa L.) supplementation improves growth performance, short-chain fatty acid production, and modulates bacterial composition of weaned piglets

  • Recharla, Neeraja;Balasubramanian, Balamuralikrishnan;Song, Minho;Puligundla, Pradeep;Kim, Soo-ki;Jeong, Jin Young;Park, Sungkwon
    • Journal of Animal Science and Technology
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    • v.63 no.3
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    • pp.575-592
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    • 2021
  • In livestock nutrition, natural feed additives are gaining increased attention as alternatives to antibiotic growth promoters to improve animal performance. This study investigated the effects of dietary turmeric supplementation on the growth performance and gut health of weaned piglets. A total of 48 weaned piglets (Duroc × [Landrace × Yorkshire]) were used in a 6-week feeding trial. All piglets were allotted to two dietary treatments: corn-soybean meal basal diet without turmeric (control) and with 1% weight per weight (w/w) turmeric powder (turmeric). The results showed that dietary inclusion of turmeric with the basal diet improved final body weight and total average daily gain (p < 0.05). The concentrations of short-chain fatty acids in the fecal samples, including acetic, butyric, and propionic acids, were higher in the turmeric group (p < 0.05). The villus height-to-crypt depth ratio was higher in the ileum of turmeric-fed piglets (p = 0.04). The 16S rRNA gene sequencing of fecal microbiota indicated that, at the phylum level, Firmicutes and Bacteroidetes were the most predominant taxa in all fecal samples. Bacteroidetes were significantly decreased in the turmeric group compared to the control group (p = 0.021). At the genus level, turmeric showed a decreased abundance of Prevotella (p = 0.021) and an increasing trend of Lactobacillus (p = 0.083). Among the total detected species, nine bacterial species showed significant differences between the two groups. The results of this study indicated that turmeric altered the gut microbiota and shortchain fatty acid production. This suggests that turmeric could be used as a potential alternative growth promoter for piglets.

Effect of dietary supplementation of garlic powder and phenyl acetic acid on productive performance, blood haematology, immunity and antioxidant status of broiler chickens

  • Ismail, I.E.;Alagawany, M.;Taha, A.E.;Puvaca, N.;Laudadio, V.;Tufarelli, V.
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.363-370
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    • 2021
  • Objective: The effect of garlic powder (GP) and phenyl acetic (PA) acid throughout the fattening period of broiler chickens on performance, blood parameters, immune, and antioxidant parameters as well as carcass traits was evaluated. Methods: A total of 210 day-old Cobb broiler chicks were randomly distributed into seven dietary treatments having five replications with six chicks per replicate. The first group (control) fed a basal diet without supplements, whereas the 2nd, 3rd, and 4th group were fed basal diet plus 0.25, 0.50, and 0.75 g GP/kg diet, respectively and the group 5th, 6th, and 7th were fed on the basal diet plus 0.25, 0.50, and 0.75 g PA/kg diet. Results: Broiler body weight and gain at 21 and 42 days were increased (p<0.05) with diets supplemented with GP and PA. Red blood cells and hemoglobin were improved in chickens fed diets enriched with GP. Broiler chickens received diets containing either GP or PA recorded the higher values (p<0.05) of total protein, globulin, high-density lipoprotein, immunoglobulin M (IgM), and IgG, superoxide dismutase and total antioxidant capacity; while, blood total cholesterol, low-density lipoprotein, aspartate-aminotransferase, and malondialdehyde were lowered (p<0.05) compared to control-diet. Liver and immune-related organs weight were improved (p<0.05) in broilers fed diet supplemented with GP and PA. Conclusion: Feeding of GP or PA in diet had positive effects on performance traits and immunological, antioxidant and physiological status of broilers. Thus, the use of tested feed additives as an eco-friendly alternative to antibiotics produced a positive effect on animal health.

Effect of Oral Administration of Seaweed-derived Fucoidan on the Resistance of the Rockfish Sebastes schlegelii to High Temperature and Streptococcus iniae Infection (해조류 유래 후코이단(Fucoidan) 경구 투여가 조피볼락(Sebastes schlegelii)의 고수온 내성 및 Streptococcus iniae 감염 저항성에 미치는 영향)

  • Song, Jun-Young;Choi, Yong-Jun;Lee, Nam-Sil;No, Chi-Won;Kim, Ahran;Do, Yong Hyun;Choi, Hye Sung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.5
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    • pp.567-574
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    • 2022
  • Immunostimulating effects of fucoidan administration to rockfish Sebastes schlegelii at a concentration of 20 g/kg of diet were evaluated under high water temperature condition. The oral administration of fucoidan mixed with feed at a concentration of 20 g/kg of diet for 2 weeks increased the interleukin 1β gene in the intestine and kidney of fish by 5.7 and 6.3 times, respectively. In addition, when the water temperature was gradually increased from 24 to 31.4℃ for 2 weeks, LT50 delayed by 24 h in the fucoidan treated group compared to that in the control group, and mortality also reduced. Streptococcus iniae infection at a concentration of 1.50×100 CFU/fish at 28℃ delayed LT50 by 12 h in the fucoidan-treated group. Furthermore, the overall survival rate was 0% in the control group and 20% in the fucoidan-treated group. This study confirmed the applicability of dietary additives such as fucoidan as an immune activator of rockfish under high temperature condition.

Safety Assessment Systems for Microbial Starters Derived from Fermented Foods

  • Heo, Sojeong;Kim, Tao;Na, Hong-Eun;Lee, Gawon;Park, Jung-Hyun;Park, Hee-Jung;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1219-1225
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    • 2022
  • Microorganisms involved in food fermentation not only improve the aroma and taste of the food, but also enhance its preservation. Thus, they are added as starter cultures to boost the final product quality of commercial fermented foods. Although these microorganisms originate from fermented foods and have a long history of consumption, the European Union recently applied the concept of Qualified presumption of Safety (QPS), which is a safety evaluation system for microorganisms used in food or feed in Europe. The QPS system is a species-level safety system and shares results with the European Novel Food System, a strain-level safety evaluation system. In the United States, microorganisms added to fermented foods are considered as food additives or Generally Recognized as Safe substance. In Korea, food microbe lists are presented at the species level. Moreover, the nation has established a strain-oriented evaluation system that applies temporary safety evaluation methods for food raw materials as well as new raw materials. However, when it comes to microorganisms isolated from traditional fermented foods and other fermented food products, there is no definition of the term "species," and there is a lack of an evaluation system at the species level. Therefore, such an evaluation system for microbial species used in Korean fermented foods is necessary.