Shukho Kim

Salmonella serovars from foodborne and waterborne diseases in Korea, 1998-2007: total isolates decreasing versus rare serovars emerging.

Salmonella enterica has been one of the most widespread foodborne pathogens in Korea. Between 1998 and 2007, a total of 9,472 Salmonella isolates were identified from foodborne and waterborne illness patients. During that time, Korea was transitioning into a developed country in industry as well as in its hygiene system. Although the isolation number of total Salmonella including serovar Typhi has decreased since 1999, the isolation of rare Salmonella serovars has emerged. Three most prevalent serovars during 1998-2007 were S. enterica Typhi, S. enterica Enteritidis, and S. enterica Typhimurium. There were remarkable outbreaks caused by rare serovars such as S. enterica Othmarschen, S. enterica London and S. enterica Paratyphi A, and overseas traveler-associated infections caused by S. enterica Weltevreden and S. enterica Anatum. Salmonella serovars from overseas travelers made a diverse Salmonella serovar pool in Korea. This study is the first review of the status of the human Salmonella infection trend in a developing country during 1998-2007. Newly emerging rare Salmonella serovars should be traced and investigated to control new type pathogens in the developed world.

Pseudomonas aeruginosa Bacteriophage PA1Ø Requires Type IV Pili for Infection and Shows Broad Bactericidal and Biofilm Removal Activities

ABSTRACT We isolated a new lytic Pseudomonas aeruginosa phage that requires type IV pili for infection. PA1Ø has a broad bactericidal spectrum, covering Gram-positive and Gram-negative bacteria, and can eradicate biofilm cells. PA1Ø may be developed as a therapeutic agent for biofilm-related mixed infections with P. aeruginosa and Staphylococcus aureus.

Complete Genome Sequence of Multidrug-Resistant Acinetobacter baumannii Strain 1656-2, Which Forms Sturdy Biofilm

Acinetobacter baumannii is a Gram-negative bacterium causing nosocomial infections worldwide. To gain quick insight into the molecular basis of biofilm formation in A. baumannii, we determined the complete genome sequence of A. baumannii strain 1656-2, which forms sturdy biofilm and is resistant to multiple drugs.

Complete Genome Sequence of Salmonella Bacteriophage SS3e

ABSTRACT A Salmonella lytic bacteriophage, SS3e, was isolated, and its genome was sequenced completely. This phage is able to lyse not only various Salmonella serovars but also Escherichia coli, Shigella sonnei, Enterobacter cloacae, and Serratia marcescens, indicating a broad host specificity. Genomic sequence analysis of SS3e revealed a linear double-stranded DNA sequence of 40,793 bp harboring 58 open reading frames, which is highly similar to Salmonella phages SETP13 and MB78.

σB‐dependent protein induction in Listeria monocytogenes during vancomycin stress

Listeria monocytogenes is a food-borne pathogen that can survive under a wide range of environmental and energy stress conditions. The general stress response controlled by sigma(B) largely contributes to stress resistance in L. monocytogenes. Moreover, the bacterial cell wall is the first defense against cellular stress and as such is the target of numerous antibiotics. We therefore hypothesize that sigma(B) contributes to monitoring the integrity of cell walls. We evaluated sigma(B) activity in wild type and DeltasigB mutant L. monocytogenes containing reporter fusions (sigma(B)-dependent opuCA promoter and a lacZ reporter gene) during the early exponential growth phase by measuring the specific activity of beta-galactosidase after vancomycin (2 microg mL(-1) final concentration) stress. sigma(B) activity is significantly induced only in the wild-type strain by addition of vancomycin. In addition, we identified sigma(B)-dependent vancomycin-inducible proteins using LC-ESI-MS/MS analysis. Two independent proteomic analyses confirmed the minimum twofold upregulation of 18 vancomycin-inducible sigma(B)-dependent stress response proteins in the wild-type strain compared with the DeltasigB mutant. The functions of these proteins are associated with cell wall biogenesis, intracellular transport, general stress response, cell metabolism and virulence. These results suggest that the sigma(B) protein may contribute to the monitoring of cell wall integrity.

Involvement of curli fimbriae in the biofilm formation of Enterobacter cloacae

In this study, we examined the biofilm forming ability, the mRNA expression of curli genes and the morphologies of curli fimbriae and biofilms in clinical isolates of Enterobacter cloacae. The csgBA operon was found in 11 (78.6%) of the 14 isolates. The ability of E. cloacae isolates to form biofilms was significantly correlated with the mRNA expression level of the csgA and csgD genes. The curli protein fimbriae appeared as tangled fibers and the curli-proficient strain formed mature biofilms. Our data suggest that the expression of the curli fimbriae play an important role in biofilm formation in E. cloacae.

Antibacterial efficacy of lytic Pseudomonas bacteriophage in normal and neutropenic mice models.

Recently, lytic bacteriophages (phages) have been focused on treating bacterial infectious diseases. We investigated the protective efficacy of a novel Pseudomonas aeruginosa phage, PA1Ø, in normal and neutropenic mice. A lethal dose of P. aeruginosa PAO1 was administered via the intraperitoneal route and a single dose of PA1Ø with different multiplicities of infection (MOI) was treated into infected mice. Immunocompetent mice infected with P. aeruginosa PAO1 were successfully protected by PA1Ø of 1 MOI, 10 MOI or 100 MOI with 80% to 100% survival rate. No viable bacteria were found in organ samples after 48 h of the phage treatment. Phage clearing patterns were different in the presence or absence of host bacteria but PA1Ø disappeared from all organs after 72 h except spleen in the presence of host bacteria. On the contrary, PA1Ø treatment could not protect neutropenic mice infected with P. aeruginosa PAO1 even though could extend their lives for a short time. In in vitro phage-neutrophil bactericidal test, a stronger bactericidal effect was observed in phage-neutrophil co-treatment than in phage single treatment without neutrophils, suggesting phage-neutrophil co-work is essential for the efficient killing of bacteria in the mouse model. In conclusion, PA1Ø can be possibly utilized in future phage therapy endeavors since it exhibited strong protective effects against virulent P. aeruginosa infection.

Complete genome sequence of Pseudomonas aeruginosa lytic bacteriophage PA1O which resembles temperate bacteriophage D3112.

ABSTRACT A novel Pseudomonas aeruginosa lytic bacteriophage (phage), PA1Ø, was isolated, and its genome was sequenced completely. This phage is able to lyse not only P. aeruginosa but also Staphylococcus aureus. Genome analysis of PA1Ø showed that it is similar to a P. aeruginosa temperate phage, D3112, with the exception of the absence of a c repressor-encoding gene, which is known to play a critical role in the maintenance of the lysogenic state of D3112 in P. aeruginosa.

A simple colorimetric method for testing antimicrobial susceptibility of biofilmed bacteria

This study introduces a simple colorimetric method which can measure the antimicrobial susceptibility of bacteria in biofilms using trimethyl tetrazolium chloride (TTC) as an indicator of viable bacteria. The new method was utilized for the evaluation of antibiotic susceptibility of Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus biofilms.

Complete Genomic Sequence of Salmonella enterica Serovar Enteritidis Phage SE2

ABSTRACT Salmonella enterica serovar Enteritidis has remained a major food-borne pathogen in humans. We isolated a virulent S. enterica serovar Enteritidis bacteriophage, SE2, which belongs to the family Siphoviridae. Phage SE2 could lyse S. enterica serovar Enteritidis PT-4, and its virulence was maintained even at ambient temperature. The genomic sequence of phage SE2 was composed of 43,221 bp with close similarity to those of Salmonella phage SETP3 and Salmonella phage SS3e. The strong and stable lytic activity of this phage might enable its use as a therapeutic or biocontrol agent against S. enterica serovar Enteritidis.