• Title/Summary/Keyword: substrate

Search Result 15,734, Processing Time 0.042 seconds

Rumen Fermentation was Changed by Feed Inoculation Method in In Vitro (사료 접종 방법에 의한 in vitro 반추위 발효 상성 변화)

  • Yoo, Dae-Kyum;Moon, Joon-Beom;Kim, Han-Been;Yang, Sung-Jae;Park, Joong-Kook;Lee, Se-Young;Seo, Ja-Kyeom
    • ANNALS OF ANIMAL RESOURCE SCIENCES
    • /
    • v.30 no.3
    • /
    • pp.111-120
    • /
    • 2019
  • The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

Analysis of the Effect of the Etching Process and Ion Injection Process in the Unit Process for the Development of High Voltage Power Semiconductor Devices (고전압 전력반도체 소자 개발을 위한 단위공정에서 식각공정과 이온주입공정의 영향 분석)

  • Gyu Cheol Choi;KyungBeom Kim;Bonghwan Kim;Jong Min Kim;SangMok Chang
    • Clean Technology
    • /
    • v.29 no.4
    • /
    • pp.255-261
    • /
    • 2023
  • Power semiconductors are semiconductors used for power conversion, transformation, distribution, and control. Recently, the global demand for high-voltage power semiconductors is increasing across various industrial fields, and optimization research on high-voltage IGBT components is urgently needed in these industries. For high-voltage IGBT development, setting the resistance value of the wafer and optimizing key unit processes are major variables in the electrical characteristics of the finished chip. Furthermore, the securing process and optimization of the technology to support high breakdown voltage is also important. Etching is a process of transferring the pattern of the mask circuit in the photolithography process to the wafer and removing unnecessary parts at the bottom of the photoresist film. Ion implantation is a process of injecting impurities along with thermal diffusion technology into the wafer substrate during the semiconductor manufacturing process. This process helps achieve a certain conductivity. In this study, dry etching and wet etching were controlled during field ring etching, which is an important process for forming a ring structure that supports the 3.3 kV breakdown voltage of IGBT, in order to analyze four conditions and form a stable body junction depth to secure the breakdown voltage. The field ring ion implantation process was optimized based on the TEG design by dividing it into four conditions. The wet etching 1-step method was advantageous in terms of process and work efficiency, and the ring pattern ion implantation conditions showed a doping concentration of 9.0E13 and an energy of 120 keV. The p-ion implantation conditions were optimized at a doping concentration of 6.5E13 and an energy of 80 keV, and the p+ ion implantation conditions were optimized at a doping concentration of 3.0E15 and an energy of 160 keV.

The synthesis of dextran from rice hydrolysates using Gluconobacter oxydans KACC 19357 bioconversion (Gluconobacter oxydans 생물전환을 통한 쌀 가수분해물 유래 dextran 합성)

  • Seung-Min Baek;Hyun Ji Lee;Legesse Shiferaw Chewaka;Chan Soon Park;Bo-Ram Park
    • Food Science and Preservation
    • /
    • v.31 no.1
    • /
    • pp.149-160
    • /
    • 2024
  • Dextran is a glucose homo-polysaccharide with a predominantly α-1,6 glycosidic linkage of microbial source and is known to be produced primarily by lactic acid bacteria. However, it can also be obtained through the dextran dextrinase of acetic acid bacteria (Gluconobacter oxydans). The dextrin-based dextran was obtained from rice starch using G. oxydans fermentation of rice hydrolysate, and its properties were studied. Both dextrin- and rice hydrolysate-added media maintained the OD value of 6 after 20 h of incubation with acetic acid bacteria, and the gel permeation chromatography (GPC) analysis of the supernatant after 72 h of incubation confirmed that a polymeric material with DP of 480 and 405, which was different from the composition of the substrate in the medium, was produced. The glucose linkage pattern of the polysaccharide was confirmed using the proton nuclear magnetic resonance (1H-NMR) and the increased α-1,4:α-1,6 bond ratio from 0.23 and 0.13 to 1:2.37 and 1:4.4, respectively, indicating that the main bonds were converted to α-1,6 bonds. The treatment of dextrin with a rat-derived alpha-glucosidase digestive enzyme resulted in a slow release of glucose, suggesting that rice hydrolysate can be converted to dextran using acetic acid bacteria with glycosyltransferase activity to produce high-value bio-materials with slowly digestible properties.

Isolation and Identification of Competitive Fungi on Medium for Black Wood Ear Mushroom in Korea and In Vitro Selection of Potential Biocontrol Agents (목이버섯 배지 오염 곰팡이균의 분리, 동정 및 생물학적 방제제 선발)

  • Seoyeon Kim;Miju Jo;Sunmin An;Jiyoon Park;Jiwon Park;Sungkook Hong;Jiwoo Kim;Juhoon Cha;Yujin Roh;Da Som Kim;Mi jin Jeon;Won-Jae Chi;Sook-Young Park
    • Research in Plant Disease
    • /
    • v.30 no.1
    • /
    • pp.66-77
    • /
    • 2024
  • Black wood ear mushroom (Auricularia auricula-judae) is one of the most economically important mushrooms in China, Japan, and Korea. The cultivation of wood ear mushrooms on artificial substrates is more efficient in terms of time and cost compared with their natural growth on trees. However, if the substrate cultivation is infected by fast-growing fungi, the relatively slow-growing ear mushroom will be outcompeted, leading to economic losses. In this study, we investigated the competitive fungal isolates from substrates infected with fast-growing fungi for the cultivation of ear mushrooms in Jangheung and Sunchon, Korea. We collected 54 isolates and identified them by sequencing their internal transcribed spacer region with morphological identification. Among the isolates, the dominant isolates were Trichoderma spp. (92.6%), Penicillium spp. (5.6%), and Talaromyces sp. (1.8%). To find an appropriate eco-friendly biocontrol agent, we used five Streptomyces spp. and Benomyl, as controls against Trichoderma spp. and Penicillium spp. Among the six Streptomyces spp., Streptomyces sp. JC203-3 effectively controlled the fungi Trichoderma spp. and Penicillium spp., which pose a significant problem for the substrates of black wood ear mushrooms. This result indicated that this Streptomyces sp. JC203-3 can be used as biocontrol agents to protect against Trichoderma and Penicillium spp.

Preparation and Gas Permeation Performance of Pd-Ag-Cu Hydrogen Separation Membrane Using α-Al2O3 Support (α-Al2O3 지지체를 이용한 Pd-Ag-Cu 수소 분리막의 제조 및 기체투과 성능)

  • Sung Woo Han;Min Chang Shin;Xuelong Zhuang;Jae Yeon Hwang;Min Young Ko;Si Eun Kim;Chang Hoon Jung;Jung Hoon Park
    • Membrane Journal
    • /
    • v.34 no.1
    • /
    • pp.50-57
    • /
    • 2024
  • In this experiment, Pd-Ag-Cu membrane was manufactured using electroless plating on an α-Al2O3 support. Pd, Ag and Cu were each coated on the surface of the support through electroless plating and heat treatment was performed for 18 h at 500℃ in H2 in the middle of electroless plating to form Pd alloy. The surface of the Pd-Ag-Cu membrane was observed through Scanning Electron Microscopy (SEM), and the thickness of the Pd membrane was measured to be 7.82 ㎛ and the thickness of the Pd-Ag-Cu membrane was measured to be 3.54 ㎛. Energy dispersive X-ray spectroscopy and X-ray diffraction analysis confirmed the formation of a Pd-Ag-Cu alloy with a composition of Pd-78wt%, Ag-8.81wt% and Cu-13.19wt%. The gas permeation experiment was conducted under the conditions of 350~450℃ and 1~4 bar in H2 single gas and H2/N2 mixed gas. The maximum H2 flux of the hydrogen separation membrane measured in H2 single gas is 74.16 ml/cm2·min at 450℃ and 4 bar for the Pd membrane and 113.64 ml/cm2·min at 450℃ and 4 bar for the Pd-Ag-Cu membrane. In the case of the separation factor measured in H2/N2 mixed gas, separation factors of 2437 and 11032 were measured at 450℃ and 4 bar.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
    • /
    • 2015.05a
    • /
    • pp.45-46
    • /
    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

  • PDF

The Treatment of Night Soil using Bacillus sp. (Bacillus sp.를 이용한 분뇨처리)

  • 염혜경;이은숙;이병헌;이민규;정일호;김중균
    • Journal of Life Science
    • /
    • v.12 no.6
    • /
    • pp.700-707
    • /
    • 2002
  • To study the characteristics of organic and nutrient removal by Bacillus species at high COD concentration of influent, three lab-scale batch reactors(R1, R2, R3), each of which has different substrate composition, were operated. More than 95% of $NH_4^+$-N and $COD_{cr}$, concentrations were removed under an aerobic condition, and their removal efficiencies were found to be 22.6 and 90.5%(R1), 23.9 and 65.8%(R2), 30.2 and 86.4%(R3), respectively. The removal efficiency of $NH_4^+$-N was high when an enough amount of $NO_3^{-}$-N was supplied, and that of $COD_{cr}$. was low when a high concentration of initial $NO_2^{-}$-N was added. The amount of carbon utilized in denitrification was a little. In all reactors,$NO_3^{-}$-N was removed under an anoxic condition, but in the R3 reactor, 10% of $NO_3^{-}$-N could be removed even undo, an aerobic condition. The removal efficiencies of TN and TP were 41.8 and 49.5%(R1), 40.1 and 35.8%(R2), 47.0 and 57.6%(R3), respectively. Alkalinities destructed under an aerobic condition for each reactor were 4.96, 5.41 and 3.93 mg/L (as $CaCO_3$) per each gram of $NH_4^+$-N oxidized, respectively, while 3.06, 3.17 and 2.60 mg/L (as $CaCO_3$) of alkalinities were produced for each gram of ,$NO_3^{-}$-N reduced to $N_2$. The SOUR were found to be 38.5, 52.7 and 42.0 mg $O_2$/g MLSS/hr, which indicated that Bacillus sp. had a higher cell activity than activated sludge. The OLR and sludge production were estimated to be 0.69 and 0.28(Rl), 0.77 and 0.20(R2), 0.61 kg COD/$m^3$/day and 0.25 kg MLSS/kg COD(R3), respectively. From the N-balance, the highest percentage(40.9%) of nitrogen lost to $N_2$ was obtained in the R3 reactor. From all the results, the possibility of aerobic denitrification Bacillus sp. has been shown and the B3 process seemed to have two advantages: a little amount of carbon was required in denitrification and not much amount of alkalinity was destructed under an aerobic condition.

Fly Ash Application Effects on CH4 and CO2 Emission in an Incubation Experiment with a Paddy Soil (항온 배양 논토양 조건에서 비산재 처리에 따른 CH4와 CO2 방출 특성)

  • Lim, Sang-Sun;Choi, Woo-Jung;Kim, Han-Yong;Jung, Jae-Woon;Yoon, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.45 no.5
    • /
    • pp.853-860
    • /
    • 2012
  • To estimate potential use of fly ash in reducing $CH_4$ and $CO_2$ emission from soil, $CH_4$ and $CO_2$ fluxes from a paddy soil mixed with fly ash at different rate (w/w; 0, 5, and 10%) in the presence and absence of fertilizer N ($(NH_4)_2SO_4$) addition were investigated in a laboratory incubation for 60 days under changing water regime from wetting to drying via transition. The mean $CH_4$ flux during the entire incubation period ranged from 0.59 to $1.68mg\;CH_4\;m^{-2}day^{-1}$ with a lower rate in the soil treated with N fertilizer due to suppression of $CH_4$ production by $SO_4^{2-}$ that acts as an electron acceptor, leading to decreases in electron availability for methanogen. Fly ash application reduced $CH_4$ flux by 37.5 and 33.0% in soils without and with N addition, respectively, probably due to retardation of $CH_4$ diffusion through soil pores by addition of fine-textured fly ash. In addition, as fly ash has a potential for $CO_2$ removal via carbonation (formation of carbonate precipitates) that decreases $CO_2$ availability that is a substrate for $CO_2$ reduction reaction (one of $CH_4$ generation pathways) is likely to be another mechanisms of $CH_4$ flux reduction by fly ash. Meanwhile, the mean $CO_2$ flux during the entire incubation period was between 0.64 and $0.90g\;CO_2\;m^{-2}day^{-1}$, and that of N treated soil was lower than that without N addition. Because N addition is likely to increase soil respiration, it is not straightforward to explain the results. However, it may be possible that our experiment did not account for the substantial amount of $CO_2$ produced by heterotrophs that were activated by N addition in earlier period than the measurement was initiated. Fly ash application also lowered $CO_2$ flux by up to 20% in the soil mixed with fly ash at 10% through $CO_2$ removal by the carbonation. At the whole picture, fly ash application at 10% decreased global warming potential of emitted $CH_4$ and $CO_2$ by about 20%. Therefore, our results suggest that fly ash application can be a soil management practice to reduce green house gas emission from paddy soils. Further studies under field conditions with rice cultivation are necessary to verify our findings.

Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
    • /
    • v.20 no.2
    • /
    • pp.260-268
    • /
    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Development and Research into Functional Foods from Hydrolyzed Whey Protein Powder with Sialic Acid as Its Index Component - I. Repeated 90-day Oral Administration Toxicity Test using Rats Administered Hydrolyzed Whey Protein Powder containing Normal Concentration of Sialic Acid (7%) with Enzyme Separation Method - (Sialic Acid를 지표성분으로 하는 유청가수분해단백분말의 기능성식품 개발연구 - I. 효소분리로 7% Siailc Acid가 표준적으로 함유된 유청가수분해단백분말(7%)의 랫드를 이용한 90일 반복경구투여 독성시험 평가 연구 -)

  • Noh, Hye-Ji;Cho, Hyang-Hyun;Kim, Hee-Kyong
    • Journal of Dairy Science and Biotechnology
    • /
    • v.34 no.2
    • /
    • pp.99-116
    • /
    • 2016
  • We herein performed animal safety assessment in accordance with Good Laboratory Practice (GLP) regulations with the aim of developing sialic acid from glycomacropeptide (hereafter referred to as "GMP") as an index ingredient and functional component in functional foods. GMP is a type of whey protein derived from milk and a safe food, with multiple functions, such as antiviral activity. A test substance was produced containing 7% (w/w) sialic acid and mostly-hydrolyzed whey protein (hereafter referred to as "7%-GNANA") by enzymatic treatment of substrate GMP. The maximum intake test dose level was selected based on 5,000 mg/kg/day dose set for male NOEL (no-observed-effect-level) and female NOAEL (no-observed-adverse-effect-level) determined by a dose-range finding (DRF) test (GLP Center of Catholic University of Daegu, Report No. 15-NREO-001) that was previously conducted with the same test substance. To evaluate the toxicity of a repeated oral dose of the test substance in connection with the previous DRF study, 1,250, 2,500, and 5,000 mg/kg of the substance were administered by a probe into the stomachs of 6-week-old SPF Sprague-Dawley male and female rats for 90 d. Each test group consisted of 10 male and 10 female rats. To determine the toxicity index, all parameters, such as observation of common signs; measurements of body weight and food consumption; ophthalmic examination; urinalysis, electrolyte, hematological, and serum biochemical examination; measurement of organ weights during autopsy; and visual and histopathological examinations were conducted according to GLP standards. After evaluating the results based on the test toxicity assessment criteria, it was determined that NOAEL of the test substance, 7%-GNANA, was 5,000 mg/kg/day, for both male and female rats. No animal death was noted in any of the test groups, including the control group, during the study period, and there was no significant difference associated with test substance, as compared with the control group, with respect to general symptoms, body weight changes, food consumption, ophthalmic examination, urinalysis, hematological and serum biochemical examination, and electrolyte and blood coagulation tests during the administration period (P<0.05). As assessed by the effects of the test substance on organ weights, food consumption, autopsy, and histopathological safety, change in kidney weight as an indicator of male NOAEL revealed up to 20% kidney weight increase in the high-dose group (5,000 mg/kg/day) compared with the change in the control group. However, it was concluded that this effect of the test substance was minor. In the case of female rats, reduction of food consumption, increase of kidney weight, and decrease of thymus weight were observed in the high-dose group. The kidney weight increased by 10.2% (left) and 8.9% (right) in the high-dose group, with a slight dose-dependency compared with that of the control group. It was observed that the thymus weight decreased by 25.3% in the high-dose group, but it was a minor test substance-associated effect. During the autopsy, botryoid tumor was detected on the ribs of one subject in the high-dose group, but we concluded that the tumor has been caused by a naturally occurring (non-test) substance. Histopathological examination revealed lesions on the kidney, liver, spleen, and other organs in the low-dose test group. Since these lesions were considered a separate phenomenon, or naturally occurring and associated with aging, it was checked whether any target organ showed clear symptoms caused by the test substance. In conclusion, different concentrations of the test substance were fed to rats and, consequently, it was verified that only a minor effect was associated with the test substance in the high-dose (5,000 mg/kg/day) group of both male and female rats, without any other significant effects associated with the test substance. Therefore, it was concluded that NOAEL of 7%-GNANA (product name: Helicobactrol) with male and female rats as test animals was 5,000 mg/kg/day, and it thus was determined that the substance is safe for the ultimate use as an ingredient of health functional foods.