• Title/Summary/Keyword: Screening of resistant strains

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A Rapid and Efficient Screening Method for Antibacterial Compound-Producing Bacteria

  • Hettiarachchi, Sachithra Amarin;Lee, Su-Jin;Lee, Youngdeuk;Kwon, Young-Kyung;Zoysa, Mahanama De;Moon, Song;Jo, Eunyoung;Kim, Taeho;Kang, Do-Hyung;Heo, Soo-Jin;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1441-1448
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    • 2017
  • Antibacterial compounds are widely used in the treatment of human and animal diseases. The overuse of antibiotics has led to a rapid rise in the prevalence of drug-resistant bacteria, making the development of new antibacterial compounds essential. This study focused on developing a fast and easy method for identifying marine bacteria that produce antibiotic compounds. Eight randomly selected marine target bacterial species (Agrococcus terreus, Bacillus algicola, Mesoflavibacter zeaxanthinifaciens, Pseudoalteromonas flavipulchra, P. peptidolytica, P. piscicida, P. rubra, and Zunongwangia atlantica) were tested for production of antibacterial compounds against four strains of test bacteria (B. cereus, B. subtilis, Halomonas smyrnensis, and Vibrio alginolyticus). Colony picking was used as the primary screening method. Clear zones were observed around colonies of P. flavipulchra, P. peptidolytica, P. piscicida, and P. rubra tested against B. cereus, B. subtilis, and H. smyrnensis. The efficiency of colony scraping and broth culture methods for antimicrobial compound extraction was also compared using a disk diffusion assay. P. peptidolytica, P. piscicida, and P. rubra showed antagonistic activity against H. smyrnensis, B. cereus, and B. subtilis, respectively, only in the colony scraping method. Our results show that colony picking and colony scraping are effective, quick, and easy methods of screening for antibacterial compound-producing bacteria.

Microbial Inhibition by an Isolate of Pediococcus from Kimchi (김치에서 분리한 Pediococcus 의 미생물 생육 저해)

  • Park, Yun-Hee;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.29 no.2
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    • pp.207-211
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    • 1986
  • Lactic acid bacteria isolated from Kimchi were tested for inhibitory activity against E. coli, Streptococcus faecalis and Lactobacillus bulgaricus. The inhibitory strains appeared most frequently around the middle stage of fermentation during two weeks of observation, and the majority of them were identified as Pediococcus cerevisiae. The agent(s) responsible for the inhibitory activity of P. cerevisiae was inactivated by heat treatment at moderate temperature, but resistant to proteolytic enzymes. The production of the inhibitory compound(s) decreased when the pH of the medium was maintained at about 7 with phosphate buffer.

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Screening of RAPD Markers for Fluoride Resistance in Bombyx mori L.

  • Chen, Keping;Yao, Qin;Li, Muwang;Wang, ong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.1
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    • pp.11-14
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    • 2003
  • NF733xin, the near allele line was obtained by means of crossing and backcrossing the silkworm race T6, which contained fluoride resistance major gene, to race 733xin, which was highly susceptible to fluoride toxicity. Two hundred RAPD random primers were used in the RAPD analysis of these 3 strains. Two molecular markers, OPB-08850 and OPB-10917, were obtained. OPB-10917 was used to detect the backcross generations. It was found that all the fluoride resistant individuals in each backcross generation had the same special band. These results proved that this marker was reliable.

Screening of Indigenous Strains of Lactic Acid Bacteria for Development of a Probiotic for Poultry

  • Karimi Torshizi, M.A.;Rahimi, Sh.;Mojgani, N.;Esmaeilkhanian, S.;Grimes, J.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1495-1500
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    • 2008
  • In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.

Screening of Bacteriocin-producing Enterococcus faecalis Strains for Antagonistic Activities against Clostridium perfringens

  • Han, Sun-Kyung;Shin, Myeong-Su;Park, Ho-Eun;Kim, So-Young;Lee, Wan-Kyu
    • Food Science of Animal Resources
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    • v.34 no.5
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    • pp.614-621
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    • 2014
  • This study was conducted to isolate and characterize bacteriocin-producing bacteria against Clostridium perfringens (C. perfringens) from domestic animals to determine their usefulness as probiotics. Bacteriocin-producing bacteria were isolated from pig feces by the spot-on-lawn method. A total of 1,370 bacterial stains were isolated, and six were tentatively selected after identifying the inhibitory activity against the pathogenic indicator C. perfringens KCTC 3269 and KCTC 5100. The selected strains were identified as Enterococcus faecalis (E. faecalis) by 16s rRNA sequencing. Most of the isolated bacterial strains were resistant to 0.5% bile salts for 48 h and remained viable after 2 h at pH 3.0. Some E. faecalis also showed strong inhibitory activity against Listeria monocytogenes KCTC 3569, KCTC 3586 and KCTC 3710. In the present study, we finally selected E. faecalis AP 216 and AP 45 strain based on probiotic selection criteria such as antimicrobial activity against C. perfringens and tolerance to acid and bile salts. The bacteriocins of E. faecalis AP 216 and AP 45 strains were highly thermostable, showing anticlostridial activities even after incubation at $121^{\circ}C$ for 15 min. These bacteriocin-producing bacteria and/or bacteriocins could be used in feed manufacturing as probiotics as an alternative to antibiotics in the livestock industry.

Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis Using the Quantamatrix Multiplexed Assay Platform System

  • Wang, Hye-young;Uh, Young;Kim, Seoyong;Cho, Eunjin;Lee, Jong Seok;Lee, Hyeyoung
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.569-577
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    • 2018
  • Background: The increasing prevalence of drug-resistant tuberculosis (TB) infection represents a global public health emergency. We evaluated the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform [QMAP], QuantaMatrix, Seoul, Korea) to rapidly identify the Mycobacterium tuberculosis complex (MTBC) and detect rifampicin (RIF) and isoniazid (INH) resistance-associated mutations. Methods: A total of 200 clinical isolates from respiratory samples were used. Phenotypic anti-TB drug susceptibility testing (DST) results were compared with those of the QMAP system, reverse blot hybridization (REBA) MTB-MDR assay, and gene sequencing analysis. Results: Compared with the phenotypic DST results, the sensitivity and specificity of the QMAP system were 96.4% (106/110; 95% confidence interval [CI] 0.9072-0.9888) and 80.0% (72/90; 95% CI 0.7052-0.8705), respectively, for RIF resistance and 75.0% (108/144; 95% CI 0.6731-0.8139) and 96.4% (54/56; 95% CI 0.8718-0.9972), respectively, for INH resistance. The agreement rates between the QMAP system and REBA MTB-MDR assay for RIF and INH resistance detection were 97.6% (121/124; 95% CI 0.9282-0.9949) and 99.1% (109/110; 95% CI 0.9453-1.0000), respectively. Comparison between the QMAP system and gene sequencing analysis showed an overall agreement of 100% for RIF resistance (110/110; 95% CI 0.9711-1.0000) and INH resistance (124/124; 95% CI 0.9743-1.0000). Conclusions: The QMAP system may serve as a useful screening method for identifying and accurately discriminating MTBC from non-tuberculous mycobacteria, as well as determining RIF- and INH-resistant MTB strains.

Selection of indigenous starter culture for safety and its effect on reduction of biogenic amine content in Moo som

  • Tangwatcharin, Pussadee;Nithisantawakhup, Jiraroj;Sorapukdee, Supaluk
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1580-1590
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    • 2019
  • Objective: The aims of this study were to select one strain of Lactobacillus plantarum (L. plantarum) for a potential indigenous safe starter culture with low level antibiotic resistant and low biogenic amine production and evaluate its effect on biogenic amines reduction in Moo som. Methods: Three strains of indigenous L. plantarum starter culture (KL101, KL102, and KL103) were selected based on their safety including antibiotic resistance and decarboxylase activity, and fermentation property as compared with a commercial starter culture (L. plantarum TISIR543). Subsequently, the effect of the selected indigenous safe starter culture on biogenic amines formation during Moo som fermentation was studied. Results: KL102 and TISIR 543 were susceptible to penicillin G, tetracycline, chloramphenicol, erythromycin, gentamycin, streptomycin, vancomycin, ciprofloxacin and trimethoprim (MIC90 ranging from 0.25 to $4{\mu}g/mL$). All strains were negative amino acid-decarboxylase for lysis of biogenic amines in screening medium. For fermentation in Moo som broth, a relatively high maximum growth rate of KL102 and TISIR543 resulted in a generation time than in the other strains (p<0.05). These strain counts were constant during the end of fermentation. Similarly, KL102 or TISIR543 addition supported increases of lactic acid bacterial count and total acidity in Moo som fermentation. For biogenic amine reduction, tyramine, putrescine, histamine and spermine contents in Moo som decreased significantly by the addition KL102 during 1 d of fermentation (p<0.05). In final product, histamine, spermine and tryptamine contents in Moo som inoculated with KL102 were lower amount those with TISIR543 (p<0.05). Conclusion: KL102 was a suitable starter culture to reduce the biogenic amine formation in Moo som.

Draft Genome Analysis of Antimicrobial Streptomyces Isolated from Himalayan Lichen

  • Kim, Byeollee;Han, So-Ra;Lamichhane, Janardan;Park, Hyun;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1144-1154
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    • 2019
  • There have been several studies regarding lichen-associated bacteria obtained from diverse environments. Our screening process identified 49 bacterial species in two lichens from the Himalayas: 17 species of Actinobacteria, 19 species of Firmicutes, and 13 species of Proteobacteria. We discovered five types of strong antimicrobial agent-producing bacteria. Although some strains exhibited weak antimicrobial activity, NP088, NP131, NP132, NP134, and NP160 exhibited strong antimicrobial activity against all multidrug-resistant strains. Polyketide synthase (PKS) fingerprinting revealed results for 69 of 148 strains; these had similar genes, such as fatty acid-related PKS, adenylation domain genes, PfaA, and PksD. Although the association between antimicrobial activity and the PKS fingerprinting results is poorly resolved, NP160 had six types of PKS fingerprinting genes, as well as strong antimicrobial activity. Therefore, we sequenced the draft genome of strain NP160, and predicted its secondary metabolism using antiSMASH version 4.2. NP160 had 46 clusters and was predicted to produce similar secondary metabolites with similarities of 5-100%. Although NP160 had 100% similarity with the alkylresorcinol biosynthetic gene cluster, our results showed low similarity with existing members of this biosynthetic gene cluster, and most have not yet been revealed. In conclusion, we expect that lichen-associated bacteria from the Himalayas can produce new secondary metabolites, and we found several secondary metabolite-related biosynthetic gene clusters to support this hypothesis.

Probiotic Properties of Lactobacillus strains Isolated from Kimchi (김치로부터 분리된 Lactobacillus strains의 probiotic 특성)

  • Choi, Hye Jung;Lim, Bo Ram;Kim, Dong Wan;Kwon, Gi-Seok;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1231-1237
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    • 2014
  • The objective of this study was to evaluate the safety and functional properties of four potential probiotic strains isolated from Kimchi, traditional Korean fermented vegetables. Based on being higher tolerance to bile salts and showing higher acid resistance or hydrophobic properties, one Lactobacillus arizonensis strain (BCNU 9032) and three L. brevis strains (BCNU 9037, BCNU 9098 and BCNU 9101) were selected in the screening experiment. All strains can survived up to 99% after 3h culture in pH 2.5 and resistant to 1% bile salts. These strains also showed good antimicrobial activities against a number of food borne pathogens, especially against Escherichia coli and Shigella sonnei. The ability to lower cholesterol levels of L. arizonensis BCNU 9032 and L. brevis 9037 were demonstrated by bile salt hydrolytic activity and cholesterol assimilation tests. Moreover, L. brevis BCNU 9098 and BCNU 9101 showed higher adherence to Caco-2 cells (12.76 and 11.86%, respectively) than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. The results suggest that these strains could be used as probiotics.

Judgement of Resistant Cultivar by Screening method for Resistance of Oyster Mushroom to Trichoderma disease in vitro (푸른곰팡이균의 저항성 품종 검정방법에 의한 느타리버섯 균주의 저항성 판별)

  • Jhune, Chang-Sung;Leem, Hoon-Tae;Lee, Chan-Jung;Kong, Won-Sik;Jang, Kab-Yeul;Sung, Gi-Ho
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.170-179
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    • 2011
  • In coculturing with strains of Trichoderma and oyster mushroom, we could detect the difference in the resistance of oyster mushroom against Trichoderma with the phenomena of barrage reaction, overgrowth and lysis. We selected the isolates ASI 2183, ASI2504 and ASI 2477 as varieties that showed the resistance. The isolates ASI 2240, ASI 2479 and ASI 2181 were the best in their resistance against Trichoderma in the method using culture filtrate. In common, the isolates ASI 2479 and ASI 2240 were selected in both methods. In post-inoculation method, the isolates ASI 2479, ASI 2333 and ASI 2181 were selected and ASI 2302 was susceptible. For the same isolate of Trichoderma, the resistance varied depending on the isolates of oyster mushroom used in the experiments. Because we could detect the interactions between Trichoderma and oyster mushroom, it is possible to detect the level of the resistance that differs in the varieties. However, there were the cases of detecting the level of the resistance in repetitions with the same isolate, which may be caused by the vitality of isolates of Trichoderma and oyster mushroom. It is efficient to test the resistance with the resistant isolate of Pleurotus salmoneostramine and the susceptible isolate of ASI 2302.