• Title/Summary/Keyword: Antibiotic production

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Generation of Fibroblasts Lacking the Sal-like 1 Gene by Using Transcription Activator-like Effector Nuclease-mediated Homologous Recombination

  • Kim, Se Eun;Kim, Ji Woo;Kim, Yeong Ji;Kwon, Deug-Nam;Kim, Jin-Hoi;Kang, Man-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.564-570
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    • 2016
  • The Sal-like 1 gene (Sall1) is essential for kidney development, and mutations in this gene result in abnormalities in the kidneys. Mice lacking Sall1 show agenesis or severe dysgenesis of the kidneys. In a recent study, blastocyst complementation was used to develop mice and pigs with exogenic organs. In the present study, transcription activator-like effector nuclease (TALEN)-mediated homologous recombination was used to produce Sall1-knockout porcine fibroblasts for developing knockout pigs. The vector targeting the Sall1 locus included a 5.5-kb 5' arm, 1.8-kb 3' arm, and a neomycin resistance gene as a positive selection marker. The knockout vector and TALEN were introduced into porcine fibroblasts by electroporation. Antibiotic selection was performed over 11 days by using $300{\mu}g/mL$ G418. DNA of cells from G418-resistant colonies was amplified using polymerase chain reaction (PCR) to confirm the presence of fragments corresponding to the 3' and 5' arms of Sall1. Further, mono- and bi-allelic knockout cells were isolated and analyzed using PCR-restriction fragment length polymorphism. The results of our study indicated that TALEN-mediated homologous recombination induced bi-allelic knockout of the endogenous gene.

Phytochemicals and antioxidant capacity of some tropical edible plants

  • Hong, Heeok;Lee, Jun-Hyeong;Kim, Soo-Ki
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1677-1684
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    • 2018
  • Objective: To find biological functions such as antibacterial and antioxidant activities in several tropical plants and to investigate the possibility of antibiotic substitute agents to prevent and treat diseases caused by pathogenic bacteria. Methods: Plants such as Poncirus trifoliata fruit (Makrut), Zingiber officinale Rosc (Khing), Areca catechu L. (Mak), Solanum melongena L. I (Makkhuayao), and Solanum melongena L. II (Makhurapro) were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. The free radical scavenging activities were measured using 2-diphenyl-2-picryl hydrazyl photometric assay. Antibacterial activities with a minimum inhibitory concentration (MIC) were observed by agar diffusion assay against pathogenic strains of Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, Clostridium perfringens, and Pantoea agglomerans. Results: Poncirus trifoliata fruit methanol extract showed antibacterial activities against gram-negative and gram-positive pathogens. Additionally, this showed the strongest antibacterial activity against Burkholderia sp. and Haemopilus somnus with MIC $131{\mu}g/mL$, respectively. Areca catechu L. water extract showed antibacterial activities against Burkholderia sp., Haemopilus somnus, and Haemopilus parasuis. The MIC value for Haemopilus parasuis was $105{\mu}g/mL$ in this. Antioxidant activity of Zingiber officinale Rosc n-hexane extract showed 2.23 mg/mL effective concentration 50% ($EC_{50}$) value was the highest activity among tropical plants extracts. Total polyphenol content in Zingiber officinale Rosc methanol extract was $48.4{\mu}g/mL$ and flavonoid content was $22.1{\mu}g/mL$ showed the highest values among tested plants extracts. Conclusion: Taken together, these results suggest that tropical plants used in this study may have a potential benefit as an alternative antibiotics agent through their antibacterial and antioxidant activities.

Griseofulvin from Xylaria sp. Strain F0010, an Endophytic Fungus of Abies holophylla and its Antifungal Activity Against Plant Pathogenic Fungi

  • PARK, JOONG-HYEOP;CHOI, GYUNG-JA;LEE, SEON-WOO;LEE, HYANG-BURM;KIM, KYOUNG-MO;JUNG, HACK-SUNG;JANG, KYOUNG-SOO;CHO, KWANG-YUN;KIM, JIN-CHEOL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.112-117
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    • 2005
  • Abstract Griseofulvin has been used as an antifungal antibiotic for the treatment of mycotic diseases of humans and veterinary animals. The purpose of this work was to identify a griseofulvin-producing endophytic fungus from Abies holophylla and evaluate its in vivo antifungal activity against plant pathogenic fungi. Based on nuclear ribosomal ITS1-5.8SITS2 sequence analysis, the fungus was identified and labeled as Xylaria sp. F0010. Two antifungal substances were purified from liquid cultures of Xylaria sp. F0010, and their chemical identities were determined to be griseofulvin and dechlorogriseofulvin through mass and NMR spectral analyses. Compared to dechlorogriseofulvin, griseofulvin showed high in vivo and in vitro antifungal activity, and effectively controlled the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), wheat leaf rust (Puccinia recondita), and barley powdery mildew (Blumeria graminis f. sp. hordei), at doses of 50 to 150 ${\mu}$g/ml, depending on the disease. This is the first report on the production of griseofulvin and dechlorogriseofulvin by Xylaria species.

Fructans from Renga Renga Lily (Arthropodium cirratum) Extract and Frutafit as Prebiotics for Broilers: Their Effects on Growth Performance and Nutrient Digestibility

  • Vidanarachchi, J.K.;Iji, P.A.;Mikkelsen, L.L.;Choct, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.5
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    • pp.580-587
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    • 2010
  • An experiment was conducted to evaluate the effect of dietary water-soluble carbohydrate extract from Renga renga lily (Arthropodium cirratum) and a commercial product, $Frutafit^{{\circledR}}$ (both fructans) on the performance, organ weights, ileal digestibility and gut morphology of male Cobb broiler chickens. There were six treatment groups: a negative control with no supplements, a positive control supplemented with 45 ppm Zn-bacitracin, and four test diets each supplemented with Renga renga lily extract or Frutafit at 5 or 10 g/kg diet. Supplementation with low levels of Renga renga lily extract and Frutafit in the diet did not affect productive parameters, whereas the inclusion of a high level of Frutafit had a negative effect on BWG and FI compared with birds fed the negative control diet. The addition of an antibiotic to the diet significantly improved (p<0.05) the BWG and FCR of broilers. Apparent ileal digestibility of dry matter, starch, protein and fat was not affected (p>0.05) by supplementation with both levels of lily extract and the low level of Frutafit. The apparent ileal digestibility of dry matter, protein and fat was decreased (p<0.05) by the high level of Frutafit. The apparent metabolisable energy (AME) of the diets fed the high level of Frutafit was approximately 0.2 MJ/kg DM lower than that of the negative control group. The addition of Zn-bacitracin increased (p<0.05) the apparent ileal digestibility of fat. The relative weight of the liver was higher (p<0.05) in broilers supplemented with the high level of Frutafit than for negative control birds at 14 and 35 d of age. Feeding Renga renga lily extract or Frutafit had no effect on the gut morphology of birds on d 14 and 35. It can be concluded that dietary inclusion of fructans from the two sources used in this study affected broiler performance differently and in a dose-dependent manner.

Effects of Dietary Supplementation with Blended Essential Oils on Growth Performance, Nutrient Digestibility, Blood Profiles and Fecal Characteristics in Weanling Pigs

  • Huang, Y.;Yoo, J.S.;Kim, H.J.;Wang, Y.;Chen, Y.J.;Cho, J.H.;Kim, I.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.5
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    • pp.607-613
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    • 2010
  • The influence of dietary supplementation with blended essential oil on growth performance, nutrient digestibility, blood profiles and fecal characteristics was evaluated in 125 crossed ((Duroc ${\times}$Yorkshire) ${\times}$Landrace) pigs (6.21${\pm}$0.20 kg initial body weight and 21 d average age). The pigs were allotted to the following treatments: i) NC (antibiotic free diet), ii) PC (NC diet+44 ppm tylosin), iii) T1 (NC diet+0.1% essential oil), iv) T2 (NC diet+0.1% essential oil (with 0.3% Benzoic acid)) and v) T3 (NC+22 ppm tylosin and 0.05% essential oil). Average daily gain (ADG) was improved in the T2 group on d 14 (p<0.05). In addition, nutrient digestibility was partially affected (both positively and negatively) by the treatments. Furthermore, the immune system was stimulated and the fecal pH and fecal noxious gases were improved in pigs that received the diets supplemented with essential oil (p<0.05). The appearance and score of diarrhea also tended to be lower in pigs that were subjected to the essential oil treatments. Collectively, the results of this study indicate that supplementation of the diet with blended essential oils could replace treatment with antibiotics to improve growth performance and fecal characteristics.

Characterization of an antifungal compound isolated from an antagonistic fungus Aspergillus terreus against phytopathogenic fungi (식물병원균 생육을 저해하는 Aspergillus terreus로부터 분리한 향균물질의 특성)

  • Kim, Keun-Ki;Kang, Jae-Gon;Choi, Yong-Lark;Yun, Han-Dae;Ha, Ho-Sung;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
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    • v.2 no.1
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    • pp.40-45
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    • 1998
  • An antagonistic fungus inhibiting growth of various phytopathogenic fungi was isolated from greenhouse soils and identified. Mophological features of fruiting structures on potato dextrose agar and colorless globose to ovate heavy walled hyaline cells from the vegetative mycelium grown on MY20 agar indicate that this antagonist is Aspergillus terreus. The antagonistic activity is due to the production of antifungal compounds. An antifungal compound was purified from its culture filtrate using chloroform extraction, column chromatography, and thin layer chromatography. The purified antibiotic was effective to various phytopathogenic fungi and identified as butyrolactone I. $ED_{50}$ values measured by petri-plate assay through effective dosage(ED)-probit analysis were 9.7, 13.7, 23.3, 42.6 and 102.7 ppm on Botrytis cinerea, Rhizoctonia solani, Pythium ultimum, Phytophthora capsici, and Fusarium oxysporum, respectively.

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Enhanced proline accumulation and salt stress tolerance of transgenic indica rice by over-expressing P5CSF129A gene

  • Kumar, Vinay;Shriram, Varsha;Kishor, P.B. Kavi;Jawali, Narendra;Shitole, M.G.
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.37-48
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    • 2010
  • [ ${\Delta}^1$ ]pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation ($T_1$) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

Microbiological Assay for the Detection of Chloramphenicol in Meat and Milk (식육 및 우유 중 클로람페니콜의 미생물학적 검출법에 관한 연구)

  • 손성완;조병훈;진남섭;박종명;박근식
    • Journal of Food Hygiene and Safety
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    • v.8 no.4
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    • pp.215-223
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    • 1993
  • Chloramphenicol (CAP) is a very effective broad-spectrum antibiotic which had been widely used in animal production. However, the drug is not approved in many countries for use in food-producing animals because of its potential toxicity and the possibility of residues in food products. In this study, a modified microbiological assay was developed for the sensitive detection of CAP residues in meat and milk. The method was characterized by the extraction of CAP with ethyl acetate, addition of $0.15\;\mu\textrm{g}$ oxytetracycline/ml in the phosphate buffer diluent (pH 6.0), a luteus ATCC 9341. The lowest levels of CAP detected in muscle tissues and milk were $0.025\;\mu\textrm{g}/ml\;and\;0.05\;\mu\textrm{g}/g$, respectively. Recovery rates free CAP from milk were 68.5%, from bovine muscle 65.1%, from swine muscle 63.8%, and from chicken muscle 59.4%, respectively, and the coefficients of variation were 1.8~15.1%. The results showed that the detection limits of CAP residues in animal products could significantly be improved by the modified microbiological assay than the conventional ones.

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Studies on Staphylococci from Subclinical Mastitic and Bulk Milk Samples (준임상형(準臨床型) 유방염(乳房炎) 및 집합유(集合乳)의 Staphylococcus 속균(屬菌)에 대하여)

  • Choi, Won-pil;Park, No-chan;Lee, Gang-log
    • Korean Journal of Veterinary Research
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    • v.26 no.2
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    • pp.237-243
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    • 1986
  • This paper deals with the incidence of bovine mastitis for 743 quarters and distribution of Staphylococci for the quarter and 70 bulk milk samples in the northern area of Gyeongbuk during the period from January to December 1984. Isolated Staphylococci were examined for species, subgroups, antibiotic resistance and penicillinase production. The results obtained were summarized as follows : A total of 25(73.5%) of 34 herds, 102(54.3%) of 188 cows and 208(30.3%) of 743 quarters were found to be infected with subclinical mastitis. A total of 83(83.1%) of 102 cows, 94(45.2%) of 208 mastitic quarters and 55(78.6%) of 70 bulk milk samples were isolated Staphylococci. Three hundred and eighteen strains of Staphylococci were classified into 11 species. Of these speoies, S. aureus from mastitis and S. sciuri from bulk milk were found most frequently, followed by S. epidermidis, S. simulans, S. cohnii, S. haemolyticus, S. xylosus, S. hyicus subsp. chromogenes, S. saprophyticus, S. warneri, S hyicus subsp. hyicus. Subgroups of catalase-positive and negative cocci were belonged most frequently to subgroup I, and subgroups III and III b, respectively. The method of Pelzer of al(97.8%) was more classified than that of Baird-Parker (68.5%). One hundred and sixty one strains(50.6%) of 318 Staphylococci isolates were resistance to one or more antibiotics such as ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, and tetracycline. Isolates from subclinical mastitis were more resistant to antibiotics than its from bulk milk. Of the 318 Staphylococci Isolates, 128(40.3%) gave positive reaction for the penicillinase test, all of ampicillin resistance strains produced this emzyme.

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Development of succinate producing Cellulomonas flavigena mutants with deleted succinate dehydrogenase gene

  • Lee, Heon-Hak;Jeon, Min-Ki;Yoon, Min-Ho
    • Korean Journal of Agricultural Science
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    • v.44 no.1
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    • pp.30-39
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    • 2017
  • This study was performed to produce succinic acid from biomass by developing mutants of Cellulomonas flavigena in which the succinate dehydrogenase gene (sdh) is deleted. For development of succinate producing mutants, the upstream and downstream regions of sdh gene from C. flavigena and antibiotic resistance gene (neo, bla) were inserted into pKC1139, and the recombinant plasmids were transformed into Escherichia coli ET12567/pUZ8002 which is a donor strain for conjugation. C. flavigena was conjugated with the transformed E. coli ET12567/pUZ8002 to induce the deletion of sdh in chromosome of this bacteria by double-crossover recombination. Two mutants (C. flavigena H-1 and H-2), in which sdh gene was deleted in the chromosome, were constructed and confirmed by PCR. To estimate the production of succinic acid by the two mutants when the culture broth was fermented with biomass such as CMC, xylan, locust gum, and rapeseed straw; the culture broth was analyzed by HPLC analysis. The succinic acid in the culture broth was not detected as a fermentation products of all biomass. One of the reasons for this may be the conversion of succinic acid to fumaric acid by sdh genes (Cfla_1014 - Cfla_1017 or Cfla_1916 - Cfla_1918) which remained in the chromosomal DNA of C. flavigena H-1 and H-2. The other reason could be the conversion of succinyl-CoA to other metabolites by enzymes related to the bypass pathway of TCA cycle.