• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1481-1489
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• 2021

Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.

• Korean Journal of Veterinary Research
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• v.47 no.1
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• pp.67-75
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• 2007

Environmental mastitis has increased particularly in well-managed or low somatic cell countherds that have successfully controled contagious pathogens. Major pathogens of environmental mastitisare Escherichia coli (E. coli) and Streptococcus uberis. The present study was conducted to investigate1,865 quaters of 241 Korean dairy farms from 2001 to 2004. Prevalence of major gram-negative bacteriaisolated from mastitis milk were E. coli (22.7%) and Enterobacter spp. (16.3%) in coliforms and Pseudomoassp. (10.3%) and Serratia spp. (7.9%) in non-coliforms. The results on antibiotic susceptibility by agardifusion test against these pathogens were 86.7% in piperaciliin, 94.6% in cefepime, 85.5% in amikacin,87.7% in gentamicin and so on. In contrast, the susceptibility against ampicillin (41.9%), cephalothin (9.9%),streptomycin (39.9%) and tetracycline (46.7%) appeared to be below 50%. Gram-negative bacteria showed(96.8%). Acording to year, distribution of high $256{\sim}64{\mu}g/ml$ on cephalothin get increased, but the othersare diferent. These findings demonstrate that major gram-negative bacteria were E. coli and Enterobacterspp. isolates, and often encountered the diverse antibiotic resistant patterns.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.816-823
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• 2022

The rapid spread of superbugs leads to the escalation of infectious diseases, which threatens public health. Endolysins derived from bacteriophages are spotlighted as promising alternative antibiotics against multi-drug resistant bacteria. In this study, we isolated and characterized the novel Salmonella typhimurium phage PBST08. Bioinformatics analysis of the PBST08 genome revealed putative endolysin ST01 with a lysozyme-like domain. Since the lytic activity of the purified ST01 was minor, probably owing to the outer membrane, which blocks accessibility to peptidoglycan, antimicrobial peptide cecropin A (CecA) was fused to the N-terminus of ST01 to disrupt the outer membrane. The resulting CecA::ST01 has been shown to have increased bactericidal activity against gram-negative pathogens including Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Enterobacter cloacae and the most affected target was A. baumannii. In the presence of 0.25 µM CecA::ST01, A. baumannii ATCC 17978 strain was completely killed and CCARM 12026 strain was wiped out by 0.5 µM CecA::ST01, which is a clinical isolate of A. baumannii and resistant to multiple drugs including carbapenem. Moreover, the larvae of Galleria mellonella could be rescued up to 58% or 49% by the administration of CecA::ST01 upon infection by A. baumannii 17978 or CCARM 12026 strain. Finally, the antibacterial activity of CecA::ST01 was verified using 31 strains of five gram-negative pathogens by evaluation of minimal inhibitory concentration. Thus, the results indicate that a fusion of antimicrobial peptide to endolysin can enhance antibacterial activity and the spectrum of endolysin where multi-drug resistant gram-negative pathogens can be efficiently controlled.

• Journal of Veterinary Clinics
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• v.21 no.2
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• pp.133-139
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• 2004

Bacterial pathogens were isolated from 36 dogs with respiratory signs, that were submitted to Veterinary Clinics in Jeju, including Veterinary Medical Teaching Hospital in Cheju National University. Of 36 isolates, 16 (44.4%) bacterial pathogens were Gram-positive and 20 (55.6%) were Gram-negative. Gram-positive bacteria identified with API Staph were 12 S. intermedius (33.3%), 2 S. aureus (5.6%), 1 S. haemolyticum (2.8%), and 1 S. xylosus (2.8%). Gram-negative organisms identified with API 20E or API NE included 8 Bordetella bronchiseptica (22.2%), 6 Escherichia coli (16.7%), 4 Pasteurella spp. (11.1%), 1 Enterobacter intermedius (2.8%), and 1 Oligella ureolytica (2.8%). Both Staphylococcus spp. isolates and Gram-negative pathogens were resistant to one or more antibiotics, including ampicillin (AM), amoxicillin/clavulanic acid (AMC), chloramphenicol (C), cefazolin (CZ), erythromycin (E), gentamicin (GM), kanamycin (K), lincomycin (L), oxacillin (OX), trimethoprim/sulfamethoxazole (SXT), and tetracycline (TE). All Staphylococcus spp. were susceptible to AMC, OX and VA, while many isolates were highly resistant to L (87.5%), E (68.8%), P (62.5%), and AM (56.3%). Antibiotic-resistant patterns of staphylococcal isolates were shown ranges from single to 9-resistant patterns. Resistant rates to antibiotics of Gram-negative bacteria were usually higher than those of Staphylococcus spp. in this study. Most Gram-negative bacteria were highly resistant to L (90.0%), AM (85.0%), E (85.0%), P (85.0%), OX (80.0%), and CZ (75.0%). B. bronchiseptica isolates showed 5 to 8 antibiotics-resistant patterns and Pasteurella spp., 2 to 8-resistant patterns. In particular, all 6 E. coli isolates were resistant to more than 9 different kinds of antibiotics, including one strain resistant to all antibiotics tested.

• Journal of fish pathology
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• v.5 no.2
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• pp.87-92
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• 1992

Lysozymes were isolated and purified from various organs of cultured flounder by using chitin-coated cellulose column chromatography. The molecular weights of them were compared with each other in 15% SDS-PAGE gels. The result showed that all lysozymes isolated from various organs of flounder had the same molecular weight of about 14000. To clarify the role of lysoryme as a body defence, the antibacterial activities of flounder lysozyme on seven bacterial pathogens, five Gram-negative and two Gram-positive species, were investigated. The lysozyme had substancial antibacterial activity on four strains, two Gram-negative and two Gram-positive species. These suggest that flounder lysozyme plays a role in body defence against both Gram-negative and Gram-positive bacterial pathogens.

• Clinical and Experimental Pediatrics
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• v.56 no.8
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• pp.332-337
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• 2013

Purpose: To identify the frequency of bacterial isolates in early-onset neonatal sepsis (EONS) and their antimicrobial resistance pattern. Methods: A retrospective study of EONS was conducted at the Beni Suef University Hospital from September 2008 to September 2012. A case of EONS was defined as an infant who had clinical signs of infection or who was born to a mother with risk factors for infection, and in whom blood culture obtained within 72 hours of life grew a bacterial pathogen. Results: Of 673 neonates screened, there were 138 positive blood cultures (20.5%) (confirmed EONS). Of the recovered isolates, 86.2% were gram-negative pathogens. Klebsiella pneumoniae (42.8%), Enterobacter cloacae (22.5%), and Escherichia coli (13.8%) were the commonest isolated organisms. The most common gram-positive microorganism was Staphylococcus aureus accounting for only 12 isolates (8.7%). All Klebsiella isolates and 93% of Enterobacter isolates were resistant to ampicillin. Gram-negative pathogens had the maximum overall sensitivity to imipenem, cefepime, and ciprofloxacin; whereas, gram-positive isolates were most susceptible to vancomycin, imipenem, and piperacillin. Conclusion: K. pneumoniae was the predominant causative bacteria of EONS followed by E. cloacae and E. coli. There was a high resistance to ampicillin. Imipenem had the maximum overall activity against the causative bacteria. Continuous surveillance is needed to monitor the changing epidemiology of pathogens and antibiotic sensitivity.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.1-10
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• 2019

Gram-negative pathogens, such as Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii, pose a serious threat to public health worldwide, due to high rates of antibiotic resistance and the lack of development of novel antimicrobial agents targeting Gram-negative bacteria. The outer membrane (OM) of Gram-negative bacteria is a unique architecture that acts as a potent permeability barrier against toxic molecules, such as antibiotics. The OM is composed of phospholipids, lipopolysaccharide (LPS), outer membrane ${\beta}-barrel$ proteins (OMP), and lipoproteins. These components are synthesized in the cytoplasm or in the inner membrane, and are then selectively transported to the OM by the specific transport machines, including the Lol, BAM, and Lpt pathways. In this review, we summarize recent studies on the assembly systems of OM components and analyze studies for the development of inhibitors that target these systems. These analyses show that OM assembly machines have the potential to be a novel attractive drug target of Gram-negative bacteria.

• Journal of Veterinary Clinics
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• v.28 no.4
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• pp.348-351
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• 2011

The purpose of this study was to determine the distribution of gram-negative bacteria isolated from companion animals with sepsis, and to investigate the antimicrobial susceptibility patterns of the isolates. Bacterial pathogens were isolated from specimens of dogs and cats submitted to National Veterinary Research and Quarantine Service between 2008 and 2009. A total of 44 gram-negative pathogens were isolated from necropsied organs. The most common isolates were E. coli (n = 33), K. pneumoniae (n = 4) and B. bronchiseptica (n = 4). Most of gram-negative isolates were susceptible to ceftiofur (68.2%), colistin (84.1%), florfenicol (84.1%) and spectinomycin (61.4%). Most of those were resistant to ampicillin (77.3%), erythromycin (86.4%), flumequine (65.9%), lincomycin (97.7%), oxytetracycline (61.4%), penicillin (100%), streptomycin (63.6%), spiramycin (97.7%), sulfamethoxazole (90.9%), tylosin (97.7%) and tiamulin (100%). In conclusion, colistin and florfenicol could be useful against sepsis due to gram-negative bacteria.

• Journal of Veterinary Clinics
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• v.14 no.2
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• pp.215-222
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• 1997

Enrofloxacin-colistin combination, widely used in Gram negative infections in veterinary sector, was investigated in terms of MIC and initial killing rate using E coli k 88ab, Salmonella typhimurium, Pasteurella multocida type A, Bordetella bronchiseptica and Staphylococcus aureus as test organisms. On the basis of MICs of enrofloxacin-colistin combination against the above bacteria, killing rates of the combination of enrofloxacin and colistin at the ratio of 5:0, 4:1, 3:2, 1:1, 2:3, 1:4 and 0:5, indicated high and rapid antibacterial acitivities against all but Staphylococcus aureus R-209, with the number of bacteria reducing to less than one percent within two hours. At the MIC of enrofloxacin or colistin, both antibacterials showed the highest killing rates during 2-4 hours against Gram negatives such as E coli K88ab,Pasteurella multocida type A and Bodetella bronchiseptica but allowed the regrowth of the same pathogens thereafter. On the while, the combination of two antibacterials at a fourth MIC resulted in high killing rate without bacterial regrowth during 24 hours, suggesting the synergistic antivacterial effects. The combination, however, did not show favourable activity against Gram negatime S typhimurium and Gram positive S aureus ergistic antibacterial activity against Gram negatime pathogens but also colistin showed LPS-neutraization, we could suggest the combination should provide clinically positive therapeutic armarium in Gram negative infections.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.740-744
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• 2003

The antimicrobial substance produced by Lactobacillus bulgaricus was inactivated by pretense, which confirmed it as a bacteriocin and referred to 'bulgaricin HJ'. The bulgaricin HJ showed the inhibitory activity against mastitis pathogens, gram-positive and gram-negative bacteria. The optimal conditions for the production of bulgaricin HJ were at the temperature of $30^{\circ}C$ and 10 h after cultivation of L. bulgaricus. Staph. and Strep. agalactiae, common bovine mastitis pathogens, were treated with bulgaricin HJ by the agar well diffusion method and showed antimirobial activities to the bovine mastitis pathogens. The activity of the bulgaricin HJ was maintained at pH 6-7 and $100^{\circ}C$ for 60 min against the mastitis pathogens. The bulgaricin HJ was determined as class IV bacteriocin by various enzyme treatments. Colony forming units analysis with indicator strains by the treatments of bulgaricin HJ indicates that the mode of bacteriocin action was bactericidal rather than bacteriostatic.