Enteropathogenic Escherichia coli (EPEC) have developed survival strategies to evade host defense systems. The intracellular level of guanosine tetraphosphate (ppGpp) controlled by RelA and SpoT can mediate immune evasion of EPEC. However, the impact of ppGpp-defective EPEC infection on phagocytes remains unknown. In this study, we report that disrupting relA and spoT of EPEC E2348/69 strain promotes its phagocytosis in porcine macrophages. Our experimental analysis showed that both uptake and killing of an E2348/69 ΔrelAΔspoT mutant by macrophages were increased compared to those of wildtype strain. These results suggest that ppGpp plays an essential role in evading phagocytosis during EPEC pathogenesis.

We investigated the epidemiological characteristics of the antimicrobial resistant Enterococcus isolates from the four major rivers of Korea in 2012. A total 316 surface water samples were collected from three distinct sites (nearby livestock farms, tributaries, and major rivers) at two different seasons (dry season: n = 76, wet season: n = 240). A total 654 bacterial cells were isolated from samples and their genus distribution were determined. We found that Gram-negative bacteria including various genera were prevalent (n = 522, 79.8%), and Enterococcus was the most common genus of Gram-positive bacteria (n = 119, 18.2%). The isolation rate of Gram-negative bacteria was higher in wet season, whereas that of Enterococcus isolates was higher in dry season. The prevalence of Enterococcus isolates was also higher nearby livestock farms than on tributaries and main rivers. Since Enterococcus isolate is a key indicator for animal fecal contamination, the following experiments focused on this microorganism. As compared to a previous report in 2006, the resistance rates in E. faecium to erythromycin (40.0% to 69.9%) and chloramphenicol (0% to 16.4%) were increased, whereas those to penicillin (56.0% to 4.1%) and teicoplanin (36.0% to 0%) were decreased. We also found that antimicrobial-resistant (AMR) E. faecium isolates from rivers and livestock samples shared similar pulsed-filed gel electrophoresis (PFGE) profiles, validating the transmission of AMR Enterococcus isolates from livestock to river. Taken together, this study provides us with detailed information about bacterial contamination status in four major rivers, and highlights the changes in AMR pattern of Enterococcus isolates, which are expected to have originated from livestock.

Enteropathogenic Escherichia coli (EPEC) is one of the etiological agents that causes diarrhea in weaning pigs. In this study, we report that mutating both relA and spoT genes in EPEC E2348/69 can promote bacterial clearance in porcine gastrointestinal tract (GIT). Our experimental analyses showed that an E2348/69 ΔrelAΔspoT mutant strain was not detected in porcine feces after 1 day post-infection (dpi), whereas its parental strain was continuously detected in porcine feces within 10 dpi. Histologic assessment revealed that the mutant strain was unable to induce moderate pathologic lesions in porcine GIT when compared to those with the wildtype strain. Taken together, our data suggest that the relA and spoT genes in EPEC play an important role in bacterial survival and pathogenesis in porcine GIT.

Salmonella is one of the most important bacterial pathogens responsible for many zoonotic food-related infectious diseases. Quantitative detection of the foodborne Salmonella contamination in various food sources is therefore critical for preventing the related disease outbreaks. In this study, we developed and evaluated a reliable real-time polymerase chain reaction (RT-PCR) assay to detect the Salmonella contamination quantitatively. The experimental results showed that our invA gene-specific quantitative RT-PCR (qRT-PCR) assay provides a strong correlation between the Cq values and the direct plate counts of Salmonella species in the artificially formulated samples. Further study may be necessary to identify more accurate correlation and equation that can apply to Salmonella spp.

In order to deal with various environmental conditions, most living organisms adapt and respond to environmental cues through nucleotide-based second-messenger signaling. Such signals regulate various endogenous factors required for environmental adaptation. In bacteria, there are five kinds of nucleotide-based second messengers, one of which is cyclic di-guanosine monophosphate (c-di-GMP). The molecule is known to regulate many cellular functions including growth, motility, biofilm formation, and virulence. Various environmental cues cause changes in the intracellular concentration of c-di-GMP, depending on the activity of specific c-di-GMP synthases and hydrolases in cells. In this review, we provide insights into nucleotide signaling in bacteria, emphasizing its impact on basic metabolism, its association with other signaling mechanisms, and its role in regulating the virulence of a wide range of bacteria. Moreover, we discuss recent studies suggesting a role for the implicated signaling molecules in bacterial persistence and antibiotic resistance.

본 연구에서는, 표고버섯 균사체 발효 생물전환공법으로 생산된 미강생물전환소재(BPP-RB)가 가금티푸스의 주요 원인균인 S. Gallinarum에 감염된 닭 유래 대식세포주 HD- 11에 미치는 효과를 조사하였다. 그 결과, 미강생물전환소 재 추출액은 S. Gallinarum 277에 대한 직접적인 성장억제 효과를 보여주지 않았으며, 총단백질 및 분비단밸질 발현 양상에 어떠한 변화도 유도하지 못하였다. 하지만, 미강생 물전환소재 추출액은 (i) HD-11 대식세포의 탐식 능력 (phagocytic activity)을 활성화하였고, (ii) Th1-type cytokines (tumor necrosis factor-α, interleukin (IL)-1β, iNOS)과 immunosuppressive cytokine IL-10의 발현 증가를 유도하였으며, (iii) Th2-type cytokines (IL-4, IL-6)의 발현은 감 소시키는 것으로 확인되었다. 이러한 결과를 종합하면, 미강생물전환소재는 가금 농장에서 가금티푸스 및 다른 Salmonella종의 감염을 예방하기 위한 사료첨가제로서의 가능성을 가지고 있다고 사료된다.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a food-borne bacterial pathogen that causes various diseases in both humans and animals such as hemorrhagic colitis and hemolytic uremic syndrome. Because cattle are the main reservoir of this microorganism, undercooked meat and meat byproducts contaminated with EHEC O157:H7 are most commonly associated with epidemic disease outbreaks. As an enteric pathogen, EHEC O157:H7 enters the body via a fecal-oral route and must survive passage through the gastric stomach at pH 1.5 to 3.5 to establish an infection within the gastrointestinal tracts. Therefore, the ability to resist such acidic environments is important to the pathogenesis of EHEC O157:H7 during a host infection. In this review, we will discuss on the acid resistance (AR) mechanisms induced by EHEC O157:H7 when E. coli encounters acidic environments.

Turmeric is known for its ability to enhance immunity via anti-inflammatory and anti-oxidant effects. Salmonella enterica species contain a large number of pathogenic serotypes that are adapted to a broad range of vertebrate hosts. Our previous study revealed that bioprocessed polysaccharides from the liquid culture Lentinus edodes fungal mycelia containing turmeric (BPP-turmeric) is able to alter chicken macrophage responses and increases chick survival against Salmonella enterica infection. In this study, we examined the immunomodulatory effects of BPP-turmeric on the porcine macrophage 3D4/31 cell line infected with Salmonella enterica subsp. enterica serovar Choleraesuis (S. Choleraesuis) or S. Enteritidis. Our experimental analyses demonstrated that BPP-turmeric (i) does not alter phagocytic and killing activity of 3D4/31 against either Salmonella serotypes, but that it (ii) represses mRNA transcription of interleukin (IL)-6, IL-8, and tumor necrosis factor α in response to Salmonella infection. Collectively, these results imply that BPP-turmeric has an immunomodulatory effect that represses pro-inflammatory cytokine expression in porcine macrophages, suggesting that it may protect swine from salmonellosis via controlling Salmonella-induced hyperinflammation.

Shiga toxin-producing Escherichia coli (STEC) is a major foodborne bacterial pathogen that causes many zoonotic diseases such as mild diarrhea, hemorrhagic colitis, and a life-threatening hemolytic uremic syndrome. STEC produces one or more Shiga toxins (Stxs) with or without other virulence factors including the locus of enterocyte and effacement pathogenicity island and the 60-MDa plasmid. Because cattle are the principal reservoirs of STEC, the consumption of undercooked meat and dairy products contaminated with bovine feces is the most common transmission route of STEC in human infections. To understand the epidemiology of STEC infection in South Korea, the prevalence of STEC among bovine feces, meat products, and human patients was reviewed in this study. We found that (i) the prevalence of STEC O157:H7 (hereafter referred to as O157 STEC) isolates was generally decreased, whereas non-O157 STEC isolates be increased among bovine feces and meat products from 2000 to 2012 and (ii) no severe human outbreaks occurred. Instead, 50 to 100 sporadic cases of STEC infection per year have been reported in asymptomatic human patients or patients with mild diarrhea. Key words: Shiga toxin-producing, Escherichia coli, STEC, prevalence, South Korea

Bioenergetics has been defined as the biology of energy transformations and energy exchanges within and between living organisms and their environment; this field now includes the concept of bioenergetic medicine, e.g., therapeutic approaches involving biophotons. QELBY® powder is a patented quantum energy-radiating material (patent No. 10-1172018), to be precise, a biologically active silicon dioxide-containing mineral powder that radiates reductive energy in infrared wavelength. In this study, we examined possible biological effects of indirect contact with QELBY® powder on various mammalian cell lines derived from macrophagic (MØ) and nonmacrophagic cells, including Raw 264.7 (mouse-derived MØ cell line), HD11 (chicken-derived MØ cell line), and HeLa (human cervical cancer cell line). Our comparison among the cells with and without indirect contact with QELBY® powder showed that this indirect contact significantly (i) increased the mitochondrial membrane potential (up to 1.36-fold) regardless of the cell type (p < 0.05), (ii) decreased the intracellular concentration of ATP in HeLa cells but not in the MØ-derived cells (p < 0.05), and (iii) protected DNA from damage during oxidative stress according to a standard comet assay (single-cell alkaline gel electrophoresis). Taken together, these results imply that indirect contact with QELBY® powder can make cells more metabolically active by increasing the mitochondrial membrane potential and by alleviating DNA damage caused by oxidative stress.

Shiga toxins (Stxs), some of the most important virulence factors in enterohemorrhagic Escherichia coli (EHEC) O157:H7, are known to be induced and released by various environmental cues, such as DNA damage responses and stress-inducing chemicals. In order to investigate the possible effects of growth media on Stxs expression, we analyzed the growth kinetics and expression of Stxs (Stx1 and 2) in cells grown in Luria-Bertani (LB) and E. coli (EC) media, which are widely used for EHEC O157:H7. Through direct plating and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), it was revealed that, when EHEC O157:H7 was grown in EC medium, the total bacterial count was reduced significantly and the stx1 transcription was greatly increased during the stationary growth phase than that in LB. Here we report that bile salts and lactose, which are the two only components in EC medium that are absent in LB, function as negative and positive regulatory signals, respectively, for the transcription of both stx1 and stx2. Indeed, stx transcription was significantly increased (~5.7 and ~21.8 fold for stx1 and stx2, respectively; p < 0.05) in an EC medium lacking bile salts when compared to the normal EC. In contrast, EHEC O157:H7 grown in an EC medium lacking lactose did significantly decrease these transcriptions (~93.5 and ~4.3 fold for stx1 and stx2, respectively; p < 0.05). Consistently, stx transcription was drastically increased in an LB medium supplemented with lactose, implying that lactose might be an environmental trigger for the expression of Stxs.

Lacticin NK34 is a small nisin-like bacteriocin present in the supernatants of an isolate of Lactococcus lactis from jeotgal, a salted and fermented Korean food made with seafood such as shrimp, oysters, and fish. Recently, we demonstrated that a partially purified NK34 is highly effective against various Staphylococcus species in a murine infection model. In this study, the two major bacterial pathogens associated with bovine mastitis, Streptococcus aureus and S. agalactiae, were evaluated for their susceptibility to NK34 in vitro using a standard teat-dip assay as well as in vivo using mastitic cows infected with one of these pathogenic strains. The experimental analyses showed a significant decrease (up to 98 times) in the bacterial numbers between the NK34-treated and -untreated teats. Moreover, a dramatic reduction in somatic cell counts was observed at 3 days post-treatment with 10 ml of NK34 administered directly into the mastitic cows. Neither S. aureus nor S. agalactiae were recovered. Taken together, these results imply that lacticin NK34 is an alternative antimicrobial substitute for the treatment of bovine mastitis, caused especially by either S. aureus or S. agalactiae.

Mesenchymal stem cell (MSC) based cell therapy has emerged as a promising therapeutic approach for treatment of several degenerative, infectious and non-infectious diseases. Numerous studies have demonstrated the remarkable immunosuppressive and antibacterial effects of MSCs both in vitro and in vivo, in animal models and in humans. However, the antibacterial effects of MSCs rely heavily on their paracrine factors rather than direct cell-to-cell contact and the effect is specific to disease and site of infection or injury. Furthermore, recent studies have demonstrated the double-edged sword effect of MSCs in bacterial infectious diseases. Despite their inherent potential for repair of damaged tissues, immunosuppression, and alleviation of various autoimmune as well as infectious diseases, MSCs also play a critical role in promoting persistent bacterial infection and disease progression. Therapeutic administration of MSCs successfully inhibited the bacterial growth and enhances survival by improved clearance of pathogenic bacteria in sepsis and pneumonic conditions. However, due to their abnormal transformation, they assist in long lasting survival and persistent infection of Mycobacterium tuberculosis (M. tuberculosis) and may also be responsible for progression of gastric cancer. This review focuses on recent advances that have broadened our understanding of MSC based therapy for bacterial diseases and provides new insight into the possible therapeutic targets of fatal bacterial diseases.