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Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

Effects of Glycogen Supercompensation in Fast and Slow Muscles on Maximal Running Time of Endurance-trained Rats (단련한 흰쥐의 속근 및 지근에 Glycogen 과저장이 운동 지구력에 미치는 영향)

  • Yeoun, Dong-Soo;Hwang, Soo-Kwan;Kim, In-Sook;Ko, Seong-Kyeong;Nam, Taick-Sang;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.22 no.1
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    • pp.63-77
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    • 1988
  • This study was carried out to investigate relationships between maximal running time (MRT) and glycogen supercompensation in fast twitch white fibers (white vastus, WV), fast twitch red fibers (red vastus, RV) and slow twitch red fibers (soleus muscle, SM) of endurance-trained rats. Male rats of a Sprague-Dawley strain were divided into the trained groups and untrained groups. Untrained groups were acquired to run on the treadmill 10 minutes for 3 days and remained rest and maintained with mixed diet for 4 weeks. For last 10 days of resting period, the untrained rats were divided into 3 groups i.e. mixed diet (untrained control), high and low carbohydrate (CHO) diet groups. And each group was subdivided into 2 groups, one group was tested for the MRT and the other was sacrificed to measure the blood glucose, blood lactate, glycogen contents of liver and muscles. The experimental groups were trained on treadmill by a modified method of Constable et al. (1984) maintained with mixed diet for 4 weeks. After measurement of MRT of this group, they were also divided into high and low CHO groups and fed with these diet for 2 days and MRT of each group was measured again to see the effect of high or low CHO feeding on the MRT. Each group was maintained with the same diet for next 2 days during which some of the rats were sacrificed at given time intervals for the measurements of blood glucose and lactate, liver and the muscles glycogen. The results were summarized as follows; 1) In the untrained group, there were no significant differences between subgroups in MRT, glycogen conent of SM, RV and WV. But blood glucose concentration and glycogen content of liver of low CHO group were significantly lower than those of mixed diet group. 2) The MRT and glycogen content of SM, RV and WV of trained mixed diet group were significantly increased compared to those of untrained mixed diet group, but there was no significant difference in glycogen content of liver. 3) MRT of trained mixed, high CHO and low CHO groups were $137{\pm}9.8,\;176{\pm}9.8\;and\;129{\pm}7.3\;min$ respectively with the significant difference between them. 4) There were no differences in blood lactate concentrations between the trained high and low CHO groups immediately after maximal running and during recovery period. 5) Glycogen contents in RV and SM of trained high CHO group were significantly increased, and glycogen contents in RV, WV and liver of trained low CHO group were significantly decreased compared to those of trained mixed diet group. 6) Immediately after maximal running, the blood glucose concentrations of trained high CHO and low CHO groups were $73{\pm}4.0\;and\;67{\pm}6.9mg%$ respecitively. The blood glucose of the trained high CHO group was fully recovered within one hour by feeding. But blood glucose concentration of low CHO group was slowly recovered up to $114{\pm}4.1mg%$ after two hours of feeding and maintained. Those values were still significantly lower than that of trained mixed diet group. The synthetic rates of glycogen in liver and muscles during the recovery period followed the similar time course of the blood glucose recoveries in each group. These results suggest that an increase in MRT of trained high CHO group was attributed to the glycogen supercompensation in slow twitch muscle fibers. And a decrease in MRT of trained low CHO may be due to decreased glycogen contents of liver and muscles. The results also suggest that glycogen supercompensation was more evident in slow twitch red fibers of endurance-trained rats and blood glucose is one of the limiting factors of glycogen synthesis.

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Plant Growth Promoting Effect and Antifungal Activity of Bacillus subtilis S37-2 (Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과)

  • Kwon, Jang-Sik;Weon, Hang-Yeon;Suh, Jang-Sun;Kim, Wan-Gyu;Jang, Kab-Yeul;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.447-453
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    • 2007
  • With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

Characteristics of Probiotics Isolated from Korean Traditional Foods and Antibacterial Activity of Synbiotics (한국전통발효식품에서 분리한 Probiotics의 특징 및 Synbiotics 항균활성 효과)

  • Moon, Chae-Yun;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.552-558
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    • 2021
  • Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.

Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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Biocontrol of Damping-Off(Rhizoctonia solani) in Cucumber by Trichoderma asperellum T-5 (Trichoderma asperellum T-5를 이용한 오이 모잘록병(Rhizoctonia solani)의 생물학적 제어)

  • Ryu, Ji-Yeon;Jin, Rong-De;Kim, Yong-Woong;Lee, Hyang-Burm;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.4
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    • pp.185-194
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    • 2006
  • A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.

Production of γ-amino Butyric Acid by Lactic Acid Bacteria in Skim Milk (탈지방우유에서 가바생성 유산균 배양을 통한 가바생성 연구)

  • Cha, Jin Young;Kim, Young Rok;Beck, Bo Ram;Park, Ji Hun;Hwang, Cher Won;Do, Hyung Ki
    • Journal of Life Science
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    • v.28 no.2
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    • pp.223-228
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    • 2018
  • Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for ${\gamma}-amino$ butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to $10,754.37{\mu}g/ml$, $13,082.79{\mu}g/ml$, $12,290.19{\mu}g/ml$, and $45.07{\mu}g/ml$ respectively in 1% monosodium glutamate-supplemented methionyl-tRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of ${\gamma}-amino$ butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production ($431.42{\mu}g/ml$) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.

Effects of Forced Molting and Storage Time on the Egg Quality (강제환우와 보존기간이 난질에 미치는 영향)

  • Oh, Hong-Rock;Lee, Bong-Duk;Lee, Soo-Kee;Liu, Xian-De;Yu, Dong-Jo
    • Korean Journal of Agricultural Science
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    • v.33 no.1
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    • pp.17-24
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    • 2006
  • An experiment was conducted to investigate the effects of forced molting and egg storage time on the various egg qualities. A total of 240 ISA Brown layers (60 wk of age) were employed as the unmolted treatment (Control). Two hundred and forty ISA Brown layers, molted at the age of 55 wk, were used as a forced molting treatment (T1), and the same number and strain of layers, molted at the age of 70 wk, were also used as the another forced molting treatment (T2). A total of 120 eggs were sampled from each treatment, and divided into six sets, 20 eggs per set. These six sets were stored for 1, 3, 6, 9, 12, and 15 days at $18^{\circ}C$ temperature, respectively. Eggs from T1 were collected from laying hens at the age of 68 wk, which started molting at 60 wk of age and achieved 50% egg production at 63 wk of age. Eggs from T2 were collected from hens at 82 wk of age, which started molting at 70 wk of age and achieved 50% egg production at 78 wk of age. The eggshell strength of T1 was significantly (p<0.05) higher than the Control and T2, and the storing periods did not affect the eggshell strength at all. Neither the forced molting nor the storing periods did not exert any consistent effect on the egg weight, eggshell thickness, eggshell color and egg yolk color. The albumin heights of T1 and T2 were significantly (p<0.05) lower than the Control, and it was remarkably reduced gradually as the storage periods increased in all three treatments. The Haugh unit showed very similar trends as the albumin height, indicating that both albumin height and Haugh unit were very much related to each other. In conclusion, the forced molting improves the eggshell strength, but decreases the albumin height and Haugh unit. The storage of eggs also decreases the albumin height and Haugh unit regardless of molting.

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Physico-chemical Characteristics of Yogurt by Lactobacillus spp. from Pickles (젓갈에서 분리한 Lactobacillus spp.로 제조한 요구르트의 이화학적 특성)

  • Rhee, Young-Hwan;Na, Han-Ju;Lee, Yong-Kyu;Shin, Seung-Yee;Kim, Jong-Hyun
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.12-17
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    • 1997
  • Three strains of lactic acid bacteria were isolated from fish and shrimp pickles. Two strains were identified as Lactobacillus casei, and one strain as L. Pentosus, respectively. All three strains were used as starters in producing yogurts. The physico-chemical characteristics of yogurts were examined. The original pH, titratable acidity, visicosity and viable cell counts of the yogurts were $4.03{\sim}4.26,\;1.049{\sim}l.217%,\;1,772{\sim}2,232\;cps\;and\;1.4{\times}10^9{\sim}1.6{\times}10^9\;cfu/ml$, respectively. In evaluating buffer capacity, $12.50{\sim}14.06\;ml$ 1.0N HCl was consumed to titrate 100 ml of yogurt to pH value 2 units below the original pH value and $9.46{\sim}13.06\;ml$ of 1.0N NaOH was consumed to pH value 4 units above the original pH value. The ${\beta}-galactosidase$ activity reached maximum at 48 hrs, and reduced gradually during fermantation. After 2 hr incubation of yogurts at $37^{\circ}C$ under different pH conditions, ${\beta}-galactosidase$ activities of three strains were reduced to 50% at pH 3.5, but there were no remaining activities neither at pH 2.5 nor at pH 1.5. Under the same pH conditions the number of viable cells decreased to $1.9{\times}10^6{\sim}1.8{\times}10^8\;cfu/ml$ at pH 2.5 and $1.0{\times}10^3{\sim}2.4{\times}10^5$ at pH 1.5, respectively. However, no significant difference was found at pH 3.5.

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THE ANTIMICROBIAL EFFECT OF HORSERADISH (ARMORACIA RUSTICANA) ROOT EXTRACTS AGAINST OBLIGATE ANAEROBES IN ROOT CANAL (치근관 내 편성 혐기성 세균에 대한 서양산 고추냉이 뿌리 추출물의 항균효과)

  • Lee, Won-Ju;Park, Ho-Won;Shin, Il-Sik;Lee, Ju-Hyun;Seo, Hyun-Woo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.36 no.2
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    • pp.237-244
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    • 2009
  • When the symptom of periapical infection is not released by mechanical instrumentation. anti-microbial agents including antibiosis become necessary in order to remove microorganisms from the root canal. Since anti-microbial agents of natural origins are currently popular, more natural remedies are being sought out. As it turns out, it is well known isothiocyanates (ITCs) in horseradish root extract have anti-microbial activity from many studies. In this research, anti-microbial effects of horseradish root extract and chlorhexidine, a typical anti-microbial agent, were investigated and compared against two kinds of obligate anaerobes. Fusobacterium nucleatum and Prevotella nigrescens, that are often discovered in infected root canal, and Clostridium perfringens, which is resistant to antibiotics and frequently used as a control strain for antibacterial studies 1. The MIC and MBC of horseradish root extract were ranged from 87 to 470 ppm and from 156 to 625 ppm against three kinds of obligate anaerobes, respectively. Horseradish root extract showed the strongest anti-bacterial activity (MBC, 156 ppm) against F. nucleatum and also showed anti-bacterial activity against antibiotic resistant obligate anaerobes. C. perfringens. 2. The MIC and MBC of chlorhexidine were ranged from 3.12 to 6.25 ppm and 10.94 ppm against three kinds of obligate anaerobes, respectively. 3. The MIC with 87-470 ppm of horseradish root exact has the same growth inhibiting effect as the one of 3.12-6.25 ppm of chlorhexidine. Likewise, the MBC with 156-625 ppm of horseradish has the similar bactericidal effect as 10.94 ppm of chlorhexidine.

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