• Asian Pacific Journal of Cancer Prevention
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• v.15 no.17
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• pp.7229-7234
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• 2014

Trichoderma spp. are known as a rich source of secondary metabolites with biological activity belonging to a variety of classes of chemical compounds. These fungi also are well known for their ability to produce a wide range of antibiotic substances and to parasitize other fungi. In search for new substances, which might act as anticancer agents, the overall objective of this study was to investigate the cytotoxic effects of Trichoderma harzianum and Trichoderma asperellum cultural filtrates against human cervical and breast cancer cell lines (HeLa and MCF-7 cells respectively). To achieve this objective, cells were exposed to 20, 40, 60, 80 and 100 mg/ml of both T. harzianum cultural filtrate (ThCF) and T. asperellum cultural filtrate (TaCF) for 24h, then the cell viability and the cytotoxic responses were assessed by using trypan blue and 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assays. Morphological changes in cells were investigated by phase contrast inverted microscopy. The results showed that ThCF and TaCF significantly reduce the cell viability, have cytotoxic effects and alter the cellular morphology of HeLa and MCF-7 cells in a concentration dependent manner. A concentration of 80 and 100mg/ml of ThCF resulted in a sharp decline in the cell viability percent of HeLa and MCF-7 respectively (25.2%, 26.5%) which was recorded by trypan blue assay. The half-maximal inhibitory concentrations ($IC_{50}$) of ThCF and TaCF in HeLa and MCF-7 were recorded as 16.6, 12.0, 19.6 and 0.70mg/ml respectively by MTT assay. These results revealed that ThCF and TaCF have a substantial ability to reduce the viability and proliferation of human cervical and breast cancer cells.

• Journal of Life Science
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• v.24 no.7
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• pp.757-761
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• 2014

Olive flounder (Paralichthys olivaceus) is an important aquaculture fish species in Jeju Island, South Korea. Due to the intensification of flounder fish farming, huge amounts of chemical antibiotics are used against several fish diseases. This has many harmful side effects on fish, as well as human consumers. Hence, an alternative to chemical antibiotic agents is needed for disease control. In this study, three strains of rhizobacteria (BRH433-2, TRH415-2, and THJ609-3) were isolated from the rhizosphere of plants. Assays of their antibacterial activity against fish pathogens, such as S. iniae, S. parauberis, V. anguillarum, and E. tarda, were performed with untreated broth culture (without cell separation), supernatant, and precipitated pellets separated by centrifugation. Among these, the cell suspension prepared from the precipitated pellet showed significant antimicrobial activity when compared with that of the untreated broth culture and centrifugal supernatant. These results indicate that the three isolated rhizobacterial strains exhibit antibacterial activity. Analysis of the 16S rDNA sequences of the BRH-433-2, THJ609-3, and TRH415-2 strains showed the highest similarity to Burkholderia gladioli (99.5%), Pseudomonas baetica (97.7%), and P. koreensis and P. baetica (98.4%), respectively. We suggest that the strains hold promise in disease management of fish.

• Journal of Korean Medicine Rehabilitation
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• v.25 no.2
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• pp.1-13
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• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

• Journal of Veterinary Clinics
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• v.20 no.3
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• pp.302-307
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• 2003

Although, in human medicine, strains of methicillin-resistant staphylococi have become the most important causative agents of nosocomial infections, studies on the small animals are very. limited. The aim of this study was to determine mecA gene and susceptibility to antibiotics of staphylococci strains isolated from clinically ill or healthy dogs and cats, during the period August 2002-July 2003. A total of 136 staphylococci (87 coagulase-positive and 49 coagulase-negative) were investigated for antibiotic resistance, using disk diffusion and minimum inhibitory concentration (MIC) test. The mecA gene was detected using the polymerase chain reaction. The isolates belonged to the species S. aureus (53 isolates), S. intermedius (34 isolates), S. epidermidis (26 isolates) and other coagulase-negative staphylococci (CNS, 23 isolates). Of the 136 isolates, 43 (31.6%) were mecA-positive and the frequency of the ,presence of mecA gene varied among the different species. All S. aureus strains were mecA-negative and were found to be susceptible, with an oxacillin MIC $\leq$1 $\mu\textrm{g}$/ml. Five (13.6%) isolates of 36 that exhibited oxacillin resistance on the MIC testing were found to be mecA-negative, suggesting not all mecA-positive strains may be an oxacillin resistant. However, the mecA presence of the strains was correlated with high oxacillin resistance: 71.4% (10 isolates of 14; P < 0.001) for mecA-positive S. intermedius and 72.4% (21 isolates of 29; P < 0.001) for mecA-positive CNS isolates. About 69% (94 isolates of 136) showed resistance to at least one drug, and 22.8% (31 isolates) were resistant to four or more different drug classes. Resistance (36 isolates, 71.7%) to penicillin G was a common finidng. This study suggest that the mecA-positive staphylococci are prevalent in small animals, and selection of antibiotics to treat infections caused by mecA-positive staphylococci may be very limited because of multi-drug resistance.

• Journal of Life Science
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• v.19 no.11
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• pp.1672-1678
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• 2009

The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

• Journal of Life Science
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• v.24 no.8
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• pp.880-886
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• 2014

In this study, we isolate and identify bacteria from seawater collected from Jeju coast, to evaluate the antimicrobial activity against the fish pathogenic bacteria. 14 bacterial strains were isolated and identified using physiological, biochemical and molecular tools. Antibacterial activity of all the 14 isolates were screened against four major fish pathogens namely, two Gram-positive: Streptococcus iniae, Streptococcus parauberis and two Gram-negative: Vibrio anguillarum, Edwardsiella tarda. Results revealed that among the 14 isolates, MK-11 was found to have antibacterial activity against S. iniae, S. parauberis, V. anguillarum Particularly, S. iniae was susceptibility with the MIC value of $250{\mu}g/ml$. The biochemical and physio-chemical results reveal that MK-11 had the sugar-alcohol disassemble ability of the D-sorbitol and D-mannitol. Also the utilization of the yeast extract, sorbitol and di-potassium phosphate were noted to be high. The optimum culture condition such as pH and temperature was recorded as pH 6.0, $25^{\circ}C$ and along with 1% NaCl which differs from the previous reports particularly in nutrient resolutions. As results of the analysis of 16S rDNA sequences, MK-11 show the high similarity with Paenibacillus polymyxa, P. jamilae, P. brasilensis 99.78%, 99.43%, 99.39%, repectively. Hence, in the present study, the isolated Paemibacillus sp. MK-11 from Jeju seawater possesses the antibacterial activity against fish pathogens and it could be used as a new antibiotic agents against the gram positive fish pathogens.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.305-310
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• 2008

Staphylococcus aureus is one of the most significant pathogens and a causative agents of nosocomial infections. The emergence of methicillin resistant S. aureus (MRSA), in particular, has become a major clinical and epidemiological problems worldwide. In this study, we analyzed the toxin genes and investigated molecular epidemiological characteristics of S. aureus isolated from stools of diarrheal patients at the hospitals in Incheon. Of the 609 strains from 2,281 specimens, 173 strains retained enterotoxin; 68 isolates (39.30%), 100 isolates (57.80%) were classified to A and C type, respectively. In the antibiotic susceptibility, all of enterotoxin positive isolates were resistant to oxacillin. Eighty eight strains (50.86%) of 173 MRSA isolate possessed tsst gene, but eta and eth genes were not detected at all. In the detection of MRSA associated genes by PCR method, mecA genes were detected in 167 strains (96.53%). From the result of PFGE analysis, we classified tsst-positive MRSA to 10 types and 24 subtypes. Type A, H and F were the major strains comprised of 57.95% (51 strains), 10.22% (9 strains) and 9.09% (8 strains) respectively.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.564-570
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• 2003

The antibacterial mechanism of enterobacter Salmonella typhimurium was studied. The rfa (Waa) gene cluster of S. typhimurium encodes the core oligosaccharide biosynthesis of lipopolysaccharide (LPS). Among the rfa gene cluster, we recently cloned the rfaE gene, which is involved in ADP-L-glycero-D-manno-heptose biosynthesis. The rfaE mutant synthesizes heptose-deficient LPS, which consists of only lipid A and 3-deoxy-D-manno-octulosonic acid (KDO), thus making an incomplete LPS and a rough phenotype mutant. S. typhimurium deep-rough mutants with the heptose region of the inner core show a reduced growth rate, sensitivity to high temperature, and hypersensitivity to hydrophobic antibiotics such as baicalin isolated from the medicinal herb of Scutellaria baicalensis Georgi. Thus, in this study, the cloned rfaE gene was added to the S. typhimurium rfaE mutant strain SL1102 (rfaE543), which makes heptose-deficient LPS and has a deep-rough phenotype. The complementation created a smooth phenotype in the SL1102 strain. The sensitivity of SL1102 to bacteriophages was also recovered to that of wild-type strain, indicating that LPS is used as the receptor for bacteriophage infection. The permeability barrier of SL1102 to hydrophobic antibiotics such as novobiocin and baicalin was restored to that of the wild-type, suggesting that antibiotic resistance of the wild-type strain is highly correlated with their LPS. Through an agar diffusion assay, the growth-inhibition activity of baicalin was fully observed in the mutant SL1102 strain. However, only a half of the inhibitory activity was detected in the rfaE complemented SL1102 strain. Furthermore, the LPS produced by the rfaE-complemented SL1102 strain was indistinguishable from LPS biosynthesis of smooth strains.

• Korean Journal of Organic Agriculture
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• v.22 no.3
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• pp.469-481
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• 2014

IIn vitro antimicrobial and anti-inflammatory activities of nargenicin and its derivatives were investigated. Nargenicin, an unusual macrolide antibiotic with potent anti-MRSA (methicilin-resistant Staphylococcus aureus) activity, was purified from the culture broth of Nocardia sp. CS682. And variety of novel nargenicin derivatives was synthesized from nargenicin. Two compounds (4 and 5) exhibit a broad spectrum of antimicrobial activities against infectious bacteria. The antimicrobial activity of derivatives against fifteen organisms was assessed using the minimum inhibitory concentration (MIC). The MIC values were in the ranges of $0.15{\sim}80{\mu}g/mL$ (w/v) for compound 1 and 2, $5{\sim}80{\mu}g/mL$ (w/v) for compound 3, $1.25{\sim}40{\mu}g/mL$ (w/v) for compound 4, and $1.25{\sim}80{\mu}g/mL$ (w/v) for compound 5, depending on the pathogens studied. In vitro, we investigated cytotoxicity and inhibition of nitric oxide (NO) production of synthesized compounds 1-5 in Raw 264.7 cells. LPS-induced nitric oxide releases were significantly blocked by compound 3, 4 and 5 in a dose-dependent manner. At high concentrations ($5{\mu}g/mL$) compound 5 inhibited the NO production by 95%. Compound 4 inhibited the release of NO in LPS-activated Raw 264.7 cells by 75% at the concentration of $10{\mu}g/mL$. Compound 3 inhibited the release of NO in LPS-activated Raw 264.7 cells by 65% at the concentration of $100{\mu}g/mL$. On the other hand, nargenicin, compound 1 and 2 did not inhibit NO production. These results demonstrated that compound 4 and 5 displayed antimicrobial activity and blocked LPS-induced pro-inflammatory mediators such as NO in macrophages, which might be responsible for its therapeutic application.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.4
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• pp.342-348
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• 2020

Uropathogenic Escherichia coli (UPEC) is a major cause of urinary tract infections (UTIs), which is one of the most common infectious diseases in humans worldwide. Since UPEC is increasingly gaining resistance to many antimicrobial agents, antibiotic therapy of UTI has recently become a great concern. This study examined the epidemiological relationship, and antimicrobial resistance patterns of 84 UPEC isolates obtained from UTI patients in Daejeon, from March to December 2017. Molecular epidemiology was investigated by multilocus sequence typing (MLST), and an antimicrobial susceptibility test was determined using an E-test. In this study, UTI was more common in females (73.8%) than in males (26.2%), and the highest incidence of UTI was observed in the age group in their 70s. Among the 84 UPEC isolates, 59 isolates (70.2%) were multidrug-resistant (MDR), and the major sequence type was ST131 (44 isolates, 52.4%). Interestingly, the rates of MDR in non-ST131 isolates (72.5%) were higher compared to ST131 isolates (68.2%). These results indicate the possibility of the development and spread of MDR in non-ST131 isolates. Effective surveillance networks and continuous research need to be conducted globally to prevent the emergence and international spread of MDR non-ST131 isolates.