• Animal Bioscience
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• v.36 no.2
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• pp.315-321
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• 2023

Objective: The study was conducted to investigate the dephosphorylation of Pseudomonas aeruginosa flagellin (PA FLA) by sweet potato purple acid phosphatase (PAP) and the effect of the enzyme on the flagellin-mediated inflammatory response in the A549 lung epithelial cell line. Methods: The activity of sweet potato PAP on PA FLA was assayed at different pH (4, 5.5, 7, and 7.5) and temperature (25℃, 37℃, and 55℃) conditions. The release of interleukin-8 (IL-8) and the activation of nuclear factor kappa- light-chain-enhancer of activated B cells (NF-κB) in A549 cells exposed to PA FLA treated with or without sweet potato PAP was measured using IL-8 and NF-κB ELISA kits, respectively. The activation of toll-like receptor 5 (TLR5) in TLR5-overexpressing HEK-293 cells exposed to PA FLA treated with or without sweet potato PAP was determined by the secreted alkaline phosphatase-based assay. Results: The dephosphorylation of PA FLA by sweet potato PAP was favorable at pH 4 and 5.5 and highest at 55℃. PA-FLA treated with the enzyme decreased IL-8 release from A549 cells to about 3.5-fold compared to intact PA FLA at 1,000 ng/mL of substrate. Moreover, PA-FLA dephosphorylated by the enzyme repressed the activation of NF-κB in the cells compared to intact PA FLA. The activation of TLR5 by PA-FLA was highest in TLR-overexpressing HEK293 cells at a substrate concentration of 5,000 ng/mL, whereas PA FLA treated with the enzyme strongly repressed the activation of TLR5. Conclusion: Sweet potato PAP has the potential to be a new alternative agent against the increased antibiotic resistance of P. aeruginosa and may be a new conceptual feed additive to control unwanted inflammatory responses caused by bacterial infections in animal husbandry.

• Korean Journal of Veterinary Service
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• v.36 no.3
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• pp.171-180
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• 2013

This study was carried out to investigate the antimicrobial resistance pattern and distribution of resistance gene in 44 Enterobacteriaceae and 21 Pseudomonas (P) aeruginosa isolated from hospitalized dogs and cats in animal hospital from 2010 to 2011 in Daegu. Among Enterobacteriaceae, Escherichia (E) coli was highly resistant to ampicillin (56.7%), followed by tetracycline (53.3%), cephalothin, streptomycine, sulfamethoxazole/trimethoprim, gentamicin and norfloxacin (40.0~43.3%). The remaining isolates of Enterobacteriaceae had high resistance to ampicillin (64.3%) and streptomycin (42.9%). Whereas, P. aeruginosa was low resistant to all antimicrobials tested (less than 15%). int I 1 gene was detected in 20 (57.1%) of 35 antimicrobial resistant Enterobacteriaceae and 2 (9.5%) of 21 P. aeruginosa., but int I 2 gene was not detected in all isolates. The eight resistance genes were found either alone or combination with other gene (s): $bla_{TEM}$, aadA, strA-strB, clmA, tetA, tetB, sul I and sul II. About 78% of integron-positive isolates were resistance to more than four antimicrobial agents. The findings suggest that class I integrons are widely distributed in E. coli among Enterobacteriaceae from dogs and cats and multi-drug resistance related to the presence of class I integrons. The prudent use of antimicrobials and continuous monitoring for companion animals are required.

• Microbiology and Biotechnology Letters
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• v.21 no.2
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• pp.132-138
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• 1993

The s-type pyocin was purified from the lysate of the mitomycin C-induced Pseudomonas aeruginosa 90-2-2205 cells by the other of ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Sephadex G-200 gel filtration. The purity was confirmed by the polyacry-lamide gel electrophoresis. The molecular weight of the purified pyocin was estimated 180, 000 by gel filtration. The pyocin was analyzed to be a complex of some polypeptides by the SDS-PAGE. The pyocin was stable by heat treatment and at pH 6-7.5 by adding 10-3% gelatin and 0.2M NaCl to the 10mM Tris-HCl buffer (pH7.5). Its killing action against the sensitive cells was assumably a single hit process.

• KSBB Journal
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• v.7 no.2
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• pp.118-125
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• 1992

Most of cosmetics are emulsion-type products which contain the sources of nutrition, i.e., vegetable oil, mineral oil and carbohyrate etc.. These additives are usually very susceptible to the contamination by microorganisms. The purpose of this study is to obtain the data necessary not only to prevent dermalopathia occurred by microbials but also to maintain the quality. In this experiment we observed the growth of P.aeruginosa in the cosmetics with or without antiseptics so as to prevent contamination. During the contamination period, the phase became unstable and creaming phenomina was happened together with some discoloration and bad smell. The pH of cosmetic was decreased from 7.6 to 6.0 and the concentration was increased from 1.443 to 1.453 in terms of refractive index during 40 days incubation. By adding antiseptics to the cosmetics, the number of P. aeruginosa from the challenge test method were decreased from $10^8$ cell/ml to $5{\times}10^3$ cell/ml. For the antibacterial effect against P. aeruginosa, p-hydroxy benzoic acid propyl ester in phosphoric acid buffer solution showed the best result.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.783-788
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• 2003

A 27 bp $tRNA^{Gly}$ region (att1) was identified as the integration site for a 12,384 bp Pfl-derived genomic island containing 15 open reading frames (ORFs) from PA0715 to PA0729 in P. aeruginosa strain PAOl. Homologous island was observed in P. aeruginosa strain PA14, but not in P. aeruginosa strain K (PAK). We isolated the Pfl island from PA14, and determined its 10,657 bp sequences containing 14 ORFs, with significant sequence variations near the borders. In contrast to the PAO1 Pfl island, the PA14 Pfl island was integrated at the 10 bp att2 site between PA1191 and PA1192. The attl site of PA14, however, was still occupied by a third genetic segment, whereas both attl and att2 sites of PAK remained unutilized. These results exemplify an extensive genomic variation of Pfl-related islands involving differential genetic organizations and differential att site utilizations.

• BMB Reports
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• v.37 no.4
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• pp.387-393
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• 2004

The L-asparaginase (E. C. 3. 5. 1. 1) enzyme was purified to homogeneity from Pseudomonas aeruginosa 50071 cells that were grown on solid-state fermentation. Different purification steps (including ammonium sulfate fractionation followed by separation on Sephadex G-100 gel filtration and CM-Sephadex C50) were applied to the crude culture filtrate to obtain a pure enzyme preparation. The enzyme was purified 106-fold and showed a final specific activity of 1900 IU/mg with a 43% yield. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed it was one peptide chain with $M_r$ of 160 kDa. A Lineweaver-Burk analysis showed a $K_m$ value of 0.147 mM and $V_{max}$ of 35.7 IU. The enzyme showed maximum activity at pH 9 when incubated at $37^{\circ}C$ for 30 min. The amino acid composition of the purified enzyme was also determined.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.29-36
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• 2011

Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.651-661
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• 2019

Targeting the pathogen viability using drugs is associated with development of drug resistance due to selective pressure. Hence, there is an increased interest in developing agents that target bacterial virulence. In this study, the inhibitory effect of ciclopirox, an antifungal agent with iron chelation potential, on the microbial virulence factors was evaluated in 26 clinical MDR Pseudomonas aeruginosa isolates collected from Alexandria Main University Hospital, a tertiary hospital in Egypt. Treatment with 9 ㎍/ml ciclopirox inhibited the hemolytic activity in 70% isolates, reduced pyocyanin production, decreased protease secretion in 46% isolates, lowered twitching and swarming motility, and decreased biofilm formation by 1.5- to 4.5-fold. The quantitative real-time PCR analysis revealed that treatment with ciclopirox downregulated the expression levels of alkaline protease (aprA) and pyocyanin (phzA1). Ciclopirox is used to treat hematological malignancies and the systemic administration of ciclopirox is reported to have adequate oral absorption with a satisfactory drug safety profile. It is important to calculate the appropriate clinical dose and therapeutic index to reposition ciclopirox from a topical antifungal agent to a promising virulence-modifying agent agent against P. aeruginosa, a problematic Gram-negative pathogen.

• Journal of fish pathology
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• v.13 no.1
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• pp.1-6
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• 2000

The antibacterial activity of marine bacterium(Pseudomonas aeruginosa JYMB1-3) was assayed against Vibrio anguillarum with the aim of evaluating the possible use for biocontrolling fish disease as probiotic strain. Inhibition test on the solid medium showed that vibrios were especially sensitive to the JYMB1-3. Edwardsiella tarda, Streptococcus sp. and Staphylococcus epidermidis were also sensitive to that strain, however the antibacterial abilities were varied to the pathogens. The vibriostatic activity of antibacterial substance produced from $10^7$ cells of the strain for 24 hours was equivalent to $2.5{\mu}g$ of chloramphenicol.

• KSBB Journal
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• v.18 no.2
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• pp.145-148
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• 2003

The hydrogen-producting bacterium was isolated from fresh water and identified as Enterbacter cloacae. The isolated was named Enterobacter cloacae YJ-1. In batch culture, The optimum cultivation temperature and pH of strain YJ-1 was 35$^{\circ}C$ and 7.5, respectively. All of the added glucose was consumed completely during fermentation even though pH was not controlled. Amount of hydrogen produced on each condition of 2% glucose, 4% sucrose and 5% fructose was 950, 1000 and 948 mL/L, respectively and resulted in increasing hydrogen production approximately 2.5-times more than controlled condition. The macimum hydrogen production was obtained when 50mM phosphate was added. was obtained when 50mM phosphate was added. In repeated0batch culture, yeast extract, but the production amount was not changed on the condition of over 0.5%, Most of the organic acides produced during the fermentation were formic and acetic acid, and propionic acid was moiety also generated.