• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.414-419
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• 2018

Escherichia coli O157:H7 is one of the most common foodborne pathogens responsible for outbreaks of hemorrhagic colitis, which can lead to the life-threatening hemolytic-uremic syndrome. In this study, we identified phytochemicals that specifically inhibit the expression of LEE operon in E. coli O157:H7. Among phytochemicals, diosmin decreased the adherence of E. coli O157:H7 towards Caco-2 cells in vitro (p<0.01) and its biofilm formation activity (p<0.05). Quantitative RT-PCR analysis revealed that the transcripts of Ler-regulated genes and genes related to curli production were significantly reduced in the presence of diosmin. However, diosmin does not affect bacterial viability, indicating that the resistance rate to diosmin was remarkably low. Overall, these results provide significant insights into the development of a novel anti-infective agent that is different from conventional antibiotics.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.222-228
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• 2001

Vibrio metschnikovii strain RH530 is a non-pathogenic, industrially-important alkaline protease producer which has been isolated from wastewater. In this paper, we report on the transformation of this strain by using the method of electroporation. A field strength of $7.5\;kVcm^{-1}$ and $25\;{\mu}F$, and using a 0.2-cm cuvette, appeared to be the optimal conditions for electroporation of the cells with the recombinant pSBCm plasmid carrying the vapK alkaline protease gene and the ColE1 replicon. Cells were subjected to osmotic shock in order to remove extracelluar DNase, and adding 200 mM of sucrose to electroporation buffer cells showed an increased transformation efficiency. Maximum efficiency of transformation was obtained at an early exponential growth phase. Using all of the conditions mentioned above, we routinely obtained a transformation efficiency of more than $10^4{({\mu}g\;plasmid\;DNA)}^{-1}$. The stability of the plasmid pSBCm in V. metschnikovii RH530 was 25% after 18h of growth (27 generations) in the medium without antibiotic selection. The insertion of the par locus to the pSBCm increased the stability of the plasmid up to 42% without selective pressure. The increase in plasmid stability was accompanied by the increase in the productivity of alkaline protease in the recombinant V. metschnikovii strain RH530. Determining optimal conditions for the transformation of the industrially-important, nonpathogenic Vibrio strain, and the improvement of plasmid stability by introducing the par locus into the high copy number plasmid vector, will allow the development of procedures involved in the genetic manipulation of this strain, particularly for its use in the production of industrial enzymes such as alkaline protease.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1057-1065
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• 2010

Two experiments were conducted to compare the effects of feeding a newly-developed rare earth mineral-yeast product, zinc oxide (ZnO) or antibiotics on the performance, nutrient digestibility and serum parameters of weanling pigs. In experiment 1, 150 crossbred barrows (24 d old and 6.28 kg BW) were fed one of five dietary treatments consisting of an unsupplemented basal diet or the basal diet supplemented with antibiotics (33 ppm tiamulin and 100 ppm chlortetracycline), ZnO (1,500 or 2,500 ppm) or 0.1% peptide-bound rare earth mineral-yeast. In experiment 2, 576 crossbred barrows (28 d old and 7.20 kg BW) were fed the same diets as those used in experiment 1 modified only by the addition of 1.0% Celite 545 to all diets as a digestibility marker. However, the negative control was not included. In experiment 1, weight gain was significantly lower (p<0.05) for pigs fed the negative control than for pigs fed diets supplemented with antibiotics, ZnO, or rare earth mineral-yeast. Pig performance did not differ between pigs fed the four supplemented diets. In experiment 2, there were no differences in performance between pigs fed diets supplemented with antibiotic, ZnO or rare earth mineral-yeast. The digestibility of dry matter, crude protein, calcium, phosphorus and energy were significantly (p<0.01) higher on the rare earth mineral-yeast diet than on diets supplemented with ZnO. In addition, pigs fed the diet supplemented with rare earth mineral-yeast had significantly (p<0.05) higher digestibility of histidine, lysine, threonine and valine than pigs fed the ZnO supplemented diets. Digestibility coefficients for pigs fed antibiotics tended to be intermediate to those of pigs fed rare earth mineralyeast or ZnO. In conclusion, the performance of pigs fed rare earth mineral-yeast was basically equal to that of pigs fed antibiotics or ZnO indicating that rare earth mineral-yeast can be successfully used as a growth promoter in diets fed to nursery pigs. The effects of rare earth mineral-yeast appeared to be mediated through improvements in nutrient digestibility.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.153-160
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• 2010

The present study was carried out for the isolation of bacteriocin-producing enterococci from indigenous sources. Gram-positive enterococci are known for having the ability to produce enterocins with good antimicrobial potential. A total of 34 strains were isolated from processed dairy products of Pakistan and seven out of them were found to be member of genus Enterococcus on selective enumeration. Biochemical and molecular characterization revealed that four of these isolates (IJ-03, IJ-07, IJ-11, and IJ-12) were Enterococcus faecalis and three (IJ-06, IJ-21, and IJ-31) were Enterococcus faecium. Local processed cheese was the source of all enterococcal isolates, except E. faecium IJ-21 and IJ-31, which were isolated from indigenous yoghurt and butter samples, respectively. Bacterial isolates were sensitive to commonly used antibiotics except methicillin and kanamycin. They also lacked critical virulence determinants, mainly cytolysin (cyl), gelatinase (gel), enterococcal surface protein (esp), and vancomycin resistance (vanA and vanB). Polymerase chain reaction amplification identified that enterocin A and P genes were present in the genome of E. faecium IJ-06 and IJ-21, whereas the E. faecium IJ-31 genome showed only enterocin P genes. No amplification was observed for genes that corresponded with the enterocins 31, AS-48, L50A, and L50B, and ent 1071A and 1071B. There were no signals of amplification found for E. faecalis IJ-11, indicating that the antimicrobial activity was because of an enterocin different from those checked by PCR. Hence, the indigenous bacterial isolates have great potential for bacteriocin production and they had antibacterial activity against a variety of closely related species.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.54-61
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• 2020

Saccharomyces boulardii is the only probiotic yeast with US Food and Drug Administration approval. It is routinely used to prevent or treat acute diarrhea and other gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. The formation of reactive oxygen species (ROS), specifically H₂O₂ during normal aerobic metabolism, contributes to programmed cell death and represents a risk to the viability of the probiotic microbe. Moreover, a loss of viability reduces the efficacy of the probiotic treatment. Therefore, inhibiting the accumulation of ROS in the oxidant environment could improve the viability of the probiotic yeast and lead to more efficacious treatment. Here, we provide evidence that supplementation with a non-reducing disaccharide, namely trehalose, enhanced the viability of S. boulardii exposed to an oxidative environment by preventing metacaspase YCA1-mediated programmed cell death through inhibition of intracellular ROS production. Our results suggest that supplementation with S. boulardii together with trehalose could increase the viability of the organism, and thus improve its effectiveness as a probiotic and as a treatment for acute diarrhea and other gastrointestinal disorders.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1604-1608
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• 2000

Penicillin antibiotics such as penicillin G, ampicillin and amoxicillin have been widely used in the pig industry to control salmonellosis, bacterial pneumonia, and urinary tract infections. Extensive use of antibiotics in veterinary clinics has resulted in tissue residues and bacterial resistance. To prevent unwanted drug residues entering the human food chain, extensive control measures have been established by both government authorities and industries. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing. In this study, we established a rapid prediction test for the detection of pigs with unacceptable tissue residues of penicillins. The recommended therapeutic doses of three penicillins, penillin G (withdrawal time, 7 days), ampicillin (withdrawal time, 7 days) and amoxicillin (withdrawal time, 14 days), were administered to three groups of 20 pigs each. Blood was sampled before drug administration and during the withdrawal period. The concentration of penicillins in plasma, determined by a semi-quantitative ELISA, were compared to that of internal standard, 4 ppb, which corresponded to the Maximum Residue Limit in milk. The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether drug residues in pig tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered residue positive, and larger than 1 negative. All 60 plasma samples from pigs were negative to three penicillins at pretreatment. Penicillin G could be detected in the plasma of the treated pigs until day 4 post-treatment and ampicillin until day 2, whereas amoxicillin could be detected until day 10 of its withdrawal period. The present study showed that the semi-quantitative ELISA could be easily adapted to detect residues of penicillin antibiotics (penicillin G, ampicillin and amoxicillin) in live pigs.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.271-276
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• 1999

Several Streptomyces strains were tested for potent antifungal agents active against phytopathogenic fungi. Among the tested, S. hygroscopicus MJM1004 showed a potent antifungal activity when assayed using Candida albicans as indicator organism. With the strain of MJM1004, fermentation medium for the production of an antifungal agent was developed with varying carbon sources, nitrogen sources, and mineral elements, which resulted in the highest productivity in the medium containing 2% soybean meal, 1% glucose, 2% starch, 0.3% $CaCO_3$, 0.05% $MgSO_4{\cdot}7H_2O$, 0.05% $K_2HPO_4$. The active compound showed a broad spectrum of antifungal activity against several plant pathogenic fungi. The antifungal compound was purified and showed the physicochemical characteristics similar to azalomycin F complex in NMR and MS analysis.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.325-332
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• 1998

In vitro phosphorylation experiments with a cell extract of Streptomyces coelicolor A3(2) M130 in the presence of ${\gamma}-[^32P]$]ATP revealed the presence of multiple phosphorylated proteins, including the AfsR/AfsK kinases which control the biosynthesis of A-factor, actinorhodin, and undecylprodigiosin. Phosphorylation of AfsR by a cell extract as an AfsK source was significantly inhibited by Ser/Thr protein kinase inhibitors, staurosporine and K-252a, at concentrations giving 50% inhibition ($IC_50$) of $1{\mu}M\;and\;0.1{\mu}M$, respectively. Further in vitro experiments with the cell extracts showed that phosphorylation of multiple proteins was inhibited by various protein kinase inhibitors with different inhibitory profiles. Manganese and calcium ions in the reaction mixture also modulate phosphorylation of multiple proteins. Manganese at 10 mM greatly enhanced the phosphorylation and partially circumvented the inhibition caused by staurosporine and K-252a. A calcium-activated protein kinase(s) was little affected by these inhibitors. Herbimycin and radicicol, which are known as tyrosine kinase inhibitors, did not show any significant inhibition of AfsR phosphorylation. Consistent with the in vitro effect of the kinase inhibitors, they inhibited aerial mycelium formation and pigmented antibiotic production on solid media. On the contrary, when assayed in liquid culture, the amount of actinorhodin produced was increased by staurosporine and K-252a and greatly decreased by manganese. All of these data clearly show that the genus Streptomyces possesses several protein kinases of eukaryotic types which are involved in the regulatory network for morphogenesis and secondary metabolism.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.281-286
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• 2010

A Pseudomonas fluorescens strain was isolated and found to show antagonistic activity against phytopathogenic fungi and to possess a gene responsible for production of antibiotic 2,4-diacetylphloroglucinol. For the extension of biocontrol range, a gene for an Androetonus australis Hector insect toxin 1 (AaHIT1), one of the most known toxic insect-selective peptides, was designed and synthesized according to the preferred codon usage of Pseudomonas fluorescens, cloned, and transformed into the strain by pSUP106 vector, a broad-host-range plasmid. Bioassays indicated that the engineered strain was able to produce AaHIT1 with insecticidal activity, and at the same time retain the activity against plant pathogen. The experiments for nonplanted soil and rhizosphere colonization showed that, similar to the population of the wild-type strain, that of the engineered strain remained relatively constant in the first 10 days, and the subsequent 50 days, suggesting that AaHIT1 expression in the bacterial cell does not substantially impair its long-term colonization. It is first reported that a Pseudomonas fluorescens strain expressing an active scorpion neurotoxin has dual activity against phytopathogenic fungi and insects, making at attractive for agronomic applications.

• 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.