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In Vitro Evaluation of Swine-Derived Lactobacillus reuteri: Probiotic Properties and Effects on Intestinal Porcine Epithelial Cells Challenged with Enterotoxigenic Escherichia coli K88

  • Wan, Zhilin;Wang, Li;Chen, Zhuang;Ma, Xianyong;Yang, Xuefen;Zhang, Jian;Jiang, Zongyong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1018-1025
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    • 2016
  • Probiotics are considered as the best effective alternatives to antibiotics. The aim of this study was to characterize the probiotic potential of lactobacilli for use in swine farming by using in vitro evaluation methods. A total of 106 lactic acid bacterial isolates, originating from porcine feces, were first screened for the capacity to survive stresses considered important for putative probiotic strains. Sixteen isolates showed notable acid and bile resistance, antibacterial activity, and adherence to intestinal porcine epithelial cells (IPEC-1). One isolate, LR1, identified as Lactobacillus reuteri, was selected for extensive study of its probiotic and functional properties in IPEC-1 cell models. L. reuteri LR1 exhibited good adhesion to IPEC-1 cells and could inhibit the adhesion of enterotoxigenic Escherichia coli (ETEC) to IPEC-1 cells. L. reuteri LR1 could also modulate transcript and protein expression of cytokines involved in inflammation in IPEC-1 cells; the Lactobacillus strain inhibited the ETEC-induced expression of proinflammatory transcripts (IL-6 and TNF-α) and protein (IL-6), and increased the level of anti-inflammatory cytokine (IL-10). Measurement of the permeation of FD-4 showed that L. reuteri LR1 could maintain barrier integrity in monolayer IPEC-1 cells exposed to ETEC. Immunolocalization experiments showed L. reuteri LR1 could also prevent ETEC-induced tight junction ZO-1 disruption. Together, these results indicate that L. reuteri LR1 exhibits desirable probiotic properties and could be a potential probiotic for use in swine production.

Studies on the Cellulase Producing Microorganisms(Part I) -Isolation of the Cellulase Producing Molds and their Cultural Conditions- (Cellulase 생성균(生成菌)에 관(關)한 연구(硏究) (제1보(第1報)) -유용균(有用菌)의 분리(分離) 및 그 배양조건(培養條件)에 대(對)하여-)

  • Kim, Chan-Jo;Choi, Woo-Young
    • Applied Biological Chemistry
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    • v.11
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    • pp.83-88
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    • 1969
  • 94 Cellulase producing strains were isoated from soils, composts, rotten woods and straws, and gastric contents and feces of herbivorous animals in various places. Among them, the strain MC-9, MC-10, MC-53 and MC-61 were found to be highly active in the degradation of carboxy methyl cellulose. Their cultural conditions adequate for the cellulase formation and effects of inorganic salts and various organic substances added to the wheat bran media were investigated. The results obtained are as follows; 1. Optimum conditions for the cellulase formation were MC-9: pH 5.5, temp. $35^{\circ}C$, incubation time 5 days, MC-10: pH 5.5-6.0, temp. $30^{\circ}C$, incubation time 5 days, MC-53: pH 3.5, temp. $30^{\circ}C$, incubation time 5 days, MC-61: pH 3.5-4.0, temp. 30-$35^{\circ}C$, incubation time 5 days. 2. Their cellulase activity in their optimum conditions were MC-9: CMC-LP(liquefying power). 87.7%, CMC-SP(saccharifying power) 3.20 glucose mg./gm. of the cultures/min., MC-10: CMC-LP 82.9%, CMC-SP 2.48 glucose mg./gm. of the cultures/min., MC-53: CMC-LP 72.4%, CMC-SP 1.76 glucose mg./gm. of the cultures/min., MC-61: CMC-LP 87.1%, CMC-SP 2.08 glucose mg./gm. of the cultures/min. 3. Additions of inorganic salts to the wheat bran media were not significant for the cellulase formation, but additions of soybean film and orange-peel pomace promoted the CMC-liquefying power 3 to 5 percent in wheat bran cultures of the strains.

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Characterization of Streptomyces netropsis Showing a Nematicidal Activity against Meloidogyne incognita (Meloidogyne incognita에 살선충활성을 보이는 신규 Streptomyces netropsis의 살선충 특성 규명)

  • Jang, Ja Yeong;Choi, Yong Ho;Joo, Yoon-Jung;Kim, Hun;Choi, Gyung Ja;Jang, Kyoung Soo;Kim, Chang-Jin;Cha, Byeongjin;Park, Hae Woong;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.21 no.2
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    • pp.50-57
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    • 2015
  • Control of nematode has become difficult owing to the restricted use of effective soil fumigant, methyl bromide, and other non-fumigant nematicides. Therefore, it is urgently necessary to develop microbial nematicide to replace chemical nematicides. In this study, the 50% aqueous methanol extraction solution of fermentation broths of 2,700 actinomycete strains were tested for their nematicidal activity against second stage of juveniles (J2s) of Meloidogyne incognita. As the results, only the 50% aqueous methanol extraction solution of AN110065, at 20% equivalent to 10% fermentation broth, showed strong nematicidal activity with 78.9% of mortality 24 h after treatment and 94.1% of mortality at 72 h. The 16S rRNA gene sequencing showed that the strain sequence was 99.78% identical to Streptomyces netropsis. The extract of S. netropsis AN110065 fermentation broth was successively partitioned with ethyl acetate and butanol and then the ethyl acetate, butanol and water layers were investigated for their nematicidal activity against the M. incognita. At $1000{\mu}g/ml$, ethyl acetate layer showed the strongest activity of 83.5% of juvenile mortality 72 h after treatment. The pot experiment using the fermentation broth of AN110065 on tomato plant against M. incognita displayed that it evidently suppressed gall formation at a 10-fold diluent treatment. The tomato plants treated with the fermentation broth of S. netropsis AN110065 did not show any phytotoxicity. The results suggest that S. netropsis AN110065 has a potential to serve as microbial nematicide in organic agriculture.

Effect o( Restricted Feeding of Layer on the Egg Productivity in Summer of Korea (산란기 제한급사가 산란성적에 미치는 영향)

  • 고태송;윤정노;주명렬;오세정
    • Korean Journal of Poultry Science
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    • v.17 no.3
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    • pp.167-177
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    • 1990
  • In order to investigate an effect of the feed restriction on the laying performance, 208 White Leghorn strain layer of 36 week-old were divided to 4 groups of 52 birds each and raised for 1 week of previous feeding and for the subsequent 3 weeks of experimental restricted feeding. The egg production, daily egg mass and feed efficiency of four groups of birds fed daily 100g, 95g, 90f and 85g of a commercial diet, respectively, which were compared with those of the original 2879 birds fed l13g of diet per day as control. During 3 weeks of experimental restricted feeding, period, daily body weight nam was decreased linearly as the degree of restriction increased in birds fed 100, 95, 90 and 85g of diet. Hen day egg production, egg weight and daily egg mass was linearly related to the diet intake level. then feed intake(x, g day$^{-1}$ ) showed a positive regression equations with the henday egg production(y, % ), egg weight(y, g egg$^{-1}$ ) and egg mass(y, g bird$^{-1}$ ) as y=38.75+0.3753$\times$(r=0.503, n=15), y=48.2+0.08868$\times$(r=0.835, n= 15) and y=15.69+0.2786$\times$(r=0.597, n=15), respectively. Feed efficiency was increased to a plateau in birds fed 95g of diet. The estimated energy utilization for egg production was reached to a plateau in birds fed 95g of diet and the highst protein utilization was shown in birds fed 90g of diet anions birds fed graded levels of diet. And the feed restriction did not affect on the egg shell contents, while protein contents of egg were shown a trend to be increased and lipids and cholesterol contents of eggs was decreased according to the diet intake lowered. The results suggested that the improved feed efficiencies of birds restricted under 16% of diet(above 95g of diet) will be due to increased energy and protein utilization for egg production and feed restriction above 16% will be aboided. In the range from 113g to 95g of diet feeding, the crude profit was increased as the feed restricted in the case of egg price 600 won kg$^{-1}$ and feed price 200 won kg$^{-1}$ .

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The Effects of Supplementation of Zeolite on the Economical Efficiency of Broiler Production (Zeolite첨가가 육계생산의 경제성에 미치는 영향)

  • 문윤영;백인기
    • Korean Journal of Poultry Science
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    • v.16 no.3
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    • pp.149-156
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    • 1989
  • Two broiler experiments were carried out to determine whether supplementation of zeolite in the diet would affect broiler performance and economical efficiency. Day old male broiler chickens of Abor Acres strain were used. In the first experiment,60 birds were distributed into f groups of 10 birds each. Three groups of chickens were randomly placed on one of the following single-stage (0-6wks) experimental diets; control diet (non-supplemented) or 2% zeolite supplemented diet (supplemented to the control). In the second experiment,216 birds were distributed into 24 groups of 9 birds each. Eight groups of chickens were randomly placed on one of the 3 two-stage diet (starter;0-4wks, finisher;5-7wks);control diet ($T_1$ , non-supplemented), 2% zeolite diet ($T_2$:isocaloric and isonitrogenous as $T_1$ ) and 2% zeolite supplemented diet ($T_3$, supplemented to the control). Birds were housed in the batteries, and feeds and water were fed ad libitum. The results of Experiment 1 showed that birds fed control diet were heavier than those fed 2% zeolite supplemented diet up to 4th wks of age. However, birds fed the latter grew more at 5th and 6th wk. Overall weight gain, feed intake and feed efficiency were not significantly different. The results of Experiment 2 showed that there were no significant differences in weight gains, feed intakes and feed efficiencies among treatments during starter, finisher to overall period. The moisture levels in excreta of the birds were significantly (P<0.01) less when zeolite was included in the diets and those in excreta significantly (P < 0.01) increased as the ambient temperature increased. Economical efficiency expressed in fed cost (won) required per kg weight gin were 346.55 (T$_3$),348.34 ($T_1$ ) and 359.85 ($T_2$).

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Effect of Feeding Aspergillus oryzae Inoculant Food-waste Diets on Performance, $NH_3$ Emission and Fecal Microflora in Broiler Chickens (Aspergilius Oryzae 접종 남은 음식물 사료가 육계의 생산성, $NH_3$ 발생량 및 분내 미생물 성상에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.281-289
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    • 2005
  • This study investigated the effect of dried food-waste diets(FW) fermented by Aspergillus oryzae(AO), on broiler growth performance, $NH_3$, emission and fecal microflora. Three hundreds broilers, two week old Hubbard strain, were randomly allotted to 4 experiments and fed with standards early boiler diet replaced with FW and AFW. In experiment 1, eighty four broilers were distributed into 7 treatments with 4 pens at 3 birds per replicate(pen). The dietary treatments ; T1 was com-soy bean meal based broiler diet(Control), T2, T3, T4 were for basal diet replaced with dried food waste without AO(FW) at the level of 20, 40 and $60\%$, respectively and T5, T6 and T7 followed the same levels for the basal diet but using Aspergillus oryzae inoculate food-waste(AFW). For experiments 2, 3, 4, seventy two broilers were distributed into 6 treatments with 4 pens at 3 birds per replicate(pen), respectively. The dietary treatments were the com-soy bean meal based broiler diet replacement with different combinations of FW and AFW, 1:0, 3:1, 1:1, 1:3, 0:1. at level of 20, 40 and $60\%$, respectively. In Exp. 1, it tended to be decreased in weight gain, however, there were no statistical differences among treatments except FW $60\%$ level of replacement(p<0.05). Feed intake and feed efficiency was not different among treatments. Total bacterial counts were not different between the control and FW diet, but E. coli decreased as the AFW levels of replacement were increased(p<0.05). There were no differences in weight gain, feed intake and feed efficiency among treatments in Exp. 2 and weight gains were lower fur FW diet compared with the control and AFW diet in Exp. 3(p<0.05). In Exp. 4, there were no differences in feed intakes among treatments, but lower in weight gain and feed efficiency in FW diet than that the control. In experiment 3, the $NH_3$ emission was the highest among treatments in FW/AFW 1:0 diet(p<0.05). From these results, it seems that FW would be supplemented up to $20\%$ in broiler diets and AO culture extract could improve FW value as feed supplements.

Components and Antimicrobial Activity of Veiled Lady Mushroom, Dictyophora echinovolvata (흰돌기망태버섯(가칭; Dictyophora echinovolvata)의 일반성분 및 항미생물활성)

  • Cheong, Jong-Chun;Cho, Soo-Muk;Jeong, Joon-Ho;Park, Jeong-Sik;Chung, Bong-Koo;Lee, Dong-Chul
    • The Korean Journal of Mycology
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    • v.29 no.2
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    • pp.79-85
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    • 2001
  • A strain of Dictyophora echinovolvata ASI 32002 showing good fruiting body formation was selected. Analyses of chemical and nutritional components as well as antimicrobial activity of different parts of the mushroom such as mycelium, egg, and fruiting body were carried out. There were differences in the chemical compositions and the quantities depending on developmental stages of veiled lady mushroom, D. echinovolvata ASI 32002. Nitrogen, phosphate, magnesium, and calcium in inorganic chemicals were abundant in mycelium, and potassium and mineral elements were abundant in the egg and fruiting body. Mannitol and trehalose were abundant in free sugar contents. Glutamic acid and arginine in mycelium and aspartic acid and glutamic acid in egg and fruiting body were abundant in free amino acid contents. Linoleic acid, an polyunsaturated fatty acid, was abundant in all parts of the Dictyophora species, but compositions and quantities of other fatty acids varied depending on the different parts of the mushroom. It was detected that malic acid, lactic acid and acetic acid in mycelium, formic acid, acetic acid and fumaric acid in egg, and malic acid, citric acid, lactic acid, fumaric acid in fruiting body were abundant. The methanol extracts of D. echinovolvata ASI 32002 mycelium showed antifungal activity with minimal inhibition concentration (MIC) of $62{\sim}125\;{\mu}g/ml$ that was similar levels of cyclohexamide against Aspergillus awamori, Hypocrea nigricance and Trichoderma virens. The MIC of extracts from mycelium and fruiting body against Candida albicans was $250\;{\mu}g/ml$, similar to that of tetracycline. In addition to the above results, further as food additives and ingredient of cosmetics.

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Physiological and Ecological Characteristics of Indigenous Soybean Rhizobia Distributed in Korea -V. Effects of Co-inoculation of R. japonicum and A. lipoferum on the Effectiveness of Symbiotic Nitrogen Fixation with Soybean (우리 나라 토착대두근류균(土着大豆根瘤菌)의 분포상태(分布狀態)와 생리(生理) 및 생태학적(生態學的) 특성(特性) -제(第)V보(報) 대두근류균(大豆根瘤菌)과 협생질소고정균(協生窒素固定菌)과의 상호접종효과(相互接種效果))

  • Ryu, Jin-Chang;Suh, Jang-Sun;Lee, Sang-Kyu;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.307-315
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    • 1988
  • This experiment was conducted to find out the effects of fertilizer-N and co-inoculation of the Rhizobium japonicum and the Azospirillum lipoferum on nodulation, $N_2$-fixation, and growth of soybean under in situ conditions. The results obtained were summarized as follows: 1. The yield of soybean dry matter was significantly greater in the R. japonicum alone, and the mixed inoculation of R. japonicum and A. lipoferum than those of un-inoculation. But inoculum applied by different strains did not significant effect on plant growth. The effects of nitrogen applied on soybean dry matter were higher in the ammonium sulfate than potassium nitrate, and decreased with increasing rates of two forms of nitrogen applied regardless of nitrogen source. 2. Acetylene redution activity was more increased in a single inoculation of R. japonicum than those of the mixed inoculation of the R. japonicum and the A. lipoferum, in cases of Danyeup cultivar, regardless of the form of combined nitrogen used. 3. Nodule mass and total nitrogenase activity per plant showed the positively significant effect in the interrelationship between dry matter of soybean and some factors related to nitrogen fixation efficiency. 4. The highest symbiotic effect in Danyeup cultivar was obtained when a single R. japonicum 84 Dy-1 strain was inoculated and fertilized with 18 mM potassium nitrate.

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Development of Natural Antioxidants and Whitening Agents for Cosmeceuticals

  • Kim, Jong-Pyung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.11a
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    • pp.79-92
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    • 2007
  • Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

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Serotype Distribution of Invasive Group B Streptococcal Diseases in Infants at Two University Hospitals in Korea (영아에서의 침습 B군 사슬알균 감염증 및 혈청형 분석: 2개 대학병원 연구)

  • Cho, Hye-Kyung;Nam, Hye Na;Cho, Hye Jung;Son, Dong Woo;Cho, Yong Kyun;Seo, Yiel-Hea;Kim, Yae-Jean;Eun, Byung Wook
    • Pediatric Infection and Vaccine
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    • v.24 no.2
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    • pp.79-86
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    • 2017
  • Purpose: This study was aimed at analyzing the serotypes of group B streptococcus (GBS) isolated from Korean infants with invasive disease and evaluating their association with disease manifestation. Methods: Data were retrospectively collected from invasive GBS infections at Gachon University Gil Medical Center from January 2006 to June 2012 and at Samsung Medical Center from April 2010 to November 2012. Serotypes were determined by slide agglutination test. Results: A total of 37 cases were identified, which included 22 full-term infants and 15 preterm infants. Fifteen cases (40.5%) were early-onset, 19 (51.4%) was late-onset, and three (8.1%) was very late-onset. Early-onset diseases among preterm infants were higher than those among full-term infants (60.0% [9/15] vs. 27.3% [6/22], P=0.17). The most common manifestation was bacteremia (70.3%), followed by meningitis and septic arthritis. Among 24 isolates retrievable for serotyping, serotype III (41.7%) was most common, followed by V (16.7%), Ia, Ib, and II (12.5%, respectively), and non-typeable (4.2%). Serotype III was more common in isolates from full-term infants (10/22) than from preterm infants (0/15), whereas serotype V was more common in isolates from preterm infants (4/15) than from full-term infants (0/22) (P=0.002). No penicillin-resistant strain was detected, and resistance to erythromycin and clindamycin were both 64.9%. Conclusions: GBS is an important pathogen in both preterm and full-term infants, and serotype distribution of GBS causing invasive diseases can differ between preterm and full-term infants. It is necessary to monitor the nationwide epidemiology of GBS diseases, including in preterm infants, in order to prepare preventive measures without underestimating early-onset diseases.