Journal of Microbiology

Journal Article

Environmental Adaptation of Psychrophilic Bacteria Subtercola spp. Isolated from Various Cryospheric Habitats: Hanbyul Lee , Yong-Joon Cho , Ahnna Cho , Ok-Sun Kim; J. Microbiol. 2023;61(7):663-672. Published online August 24, 2023; DOI: https://doi.org/10.1007/s12275-023-00068-y

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Abstract PDF: Subtercola boreus K300T is a novel psychrophilic strain that was isolated from permanently cold groundwater in Finland and has also been found in several places in Antarctica including lake, soil, and rocks. We performed genomic and transcriptomic analyses of 5 strains from Antarctica and a type strain to understand their adaptation to different environments. Interestingly, the isolates from rocks showed a low growth rate and smaller genome size than strains from the other isolation sources (lake, soil, and groundwater). Based on these habitat-dependent characteristics, the strains could be classified into two ecotypes, which showed differences in energy production, signal transduction, and transcription in the clusters of orthologous groups of proteins (COGs) functional category. In addition, expression pattern changes revealed differences in metabolic processes, including uric acid metabolism, DNA repair, major facilitator superfamily (MFS) transporters, and xylose degradation, depending on the nutritional status of their habitats. These findings provide crucial insights into the environmental adaptation of bacteria, highlighting genetic diversity and regulatory mechanisms that enable them to thrive in the cryosphere.

Review

[MINIREVIEW]Gain and loss of antibiotic resistant genes in multidrug resistant bacteria: One Health perspective: Misung Kim , Jaeeun Park , Mingyeong Kang , Jihye Yang , Woojun Park; J. Microbiol. 2021;59(6):535-545. Published online April 20, 2021; DOI: https://doi.org/10.1007/s12275-021-1085-9

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The emergence of multidrug resistance (MDR) has become a global health threat due to the increasing unnecessary use of antibiotics. Multidrug resistant bacteria occur mainly by accumulating resistance genes on mobile genetic elements (MGEs), made possible by horizontal gene transfer (HGT). Humans and animal guts along with natural and engineered environments such as wastewater treatment plants and manured soils have proven to be the major reservoirs and hotspots of spreading antibiotic resistance genes (ARGs). As those environments support the dissemination of MGEs through the complex interactions that take place at the human-animalenvironment interfaces, a growing One Health challenge is for multiple sectors to communicate and work together to prevent the emergence and spread of MDR bacteria. However, maintenance of ARGs in a bacterial chromosome and/or plasmids in the environments might place energy burdens on bacterial fitness in the absence of antibiotics, and those unnecessary ARGs could eventually be lost. This review highlights and summarizes the current investigations into the gain and loss of ARG genes in MDR bacteria among human-animal- environment interfaces. We also suggest alternative treatments such as combinatory therapies or sequential use of different classes of antibiotics/adjuvants, treatment with enzymeinhibitors, and phage therapy with antibiotics to solve the MDR problem from the perspective of One Health issues.

Citations

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Journal Articles

Anti-inflammatory and anti-oxidative effect of Korean propolis on Helicobacter pylori-induced gastric damage in vitro: Moon-Young Song , Da-Young Lee , Eun-Hee Kim; J. Microbiol. 2020;58(10):878-885. Published online September 2, 2020; DOI: https://doi.org/10.1007/s12275-020-0277-z

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Abstract PDF

Helicobacter pylori, present in the stomach lining, is a Gramnegative bacterium that causes various gastrointestinal diseases, including gastritis and peptic ulcers. Propolis is a natural resinous substance collected from a variety of plants, and contains several natural bioactive substances. The aim of this study was to investigate the anti-inflammatory and antioxidative effects of Korean propolis on H. pylori-induced damage in the human adenocarcinoma gastric cell line. The propolis used in this study was obtained from the Korea Beekeeping Association in South Korea. The expression of pro-inflammatory interleukins (ILs), such as IL-8, IL-12, IL-1β, tumor necrosis factor alpha, cyclooxygenase-2, and inducible nitric oxide synthase, which was increased after H. pylori infection, significantly decreased in a dose-dependent manner upon pretreatment with Korean propolis, because of the suppression of mitogen-activated protein kinases and nuclear factor κB pathway. The anti-oxidative activity of propolis was assessed using the 2,2-diphenyl-1-picrylhydrazyl hydrate free radical assay. Korean propolis showed significant anti-oxidative effects via reactive oxygen species scavenging. In addition, pretreatment with Korean propolis upregulated the expression of anti-oxidant enzymes through Nrf2 signaling activation. These findings indicate that the use of Korean propolis, which has anti-inflammatory and anti-oxidative effects, can be promising for the prevention of H. pylori-induced gastric damage.

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The effects of deletion of cellobiohydrolase genes on carbon source-dependent growth and enzymatic lignocellulose hydrolysis in Trichoderma reesei: Meibin Ren , Yifan Wang , Guoxin Liu , Bin Zuo , Yuancheng Zhang , Yunhe Wang , Weifeng Liu , Xiangmei Liu , Yaohua Zhong; J. Microbiol. 2020;58(8):687-695. Published online June 10, 2020; DOI: https://doi.org/10.1007/s12275-020-9630-5

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Abstract PDF

The saprophytic fungus Trichoderma reesei has long been used as a model to study microbial degradation of lignocellulosic biomass. The major cellulolytic enzymes of T. reesei are the cellobiohydrolases CBH1 and CBH2, which constitute more than 70% of total proteins secreted by the fungus. However, their physiological functions and effects on enzymatic hydrolysis of cellulose substrates are not sufficiently elucidated. Here, the cellobiohydrolase-encoding genes cbh1 and cbh2 were deleted, individually or combinatively, by using an auxotrophic marker-recycling technique in T. reesei. When cultured on media with different soluble carbon sources, all three deletion strains (Δcbh1, Δcbh2, and Δcbh1Δcbh2) exhibited no dramatic variation in morphological phenotypes, but their growth rates increased apparently when cultured on soluble cellulase-inducing carbon sources. In addition, Δcbh1 showed dramatically reduced growth and Δcbh1Δcbh2 could hardly grew on microcrystalline cellulose (MCC), whereas all strains grew equally on sodium carboxymethyl cellulose (CMC-Na), suggesting that the influence of the CBHs on growth was carbon source-dependent. Moreover, five representative cellulose substrates were used to analyse the influence of the absence of CBHs on saccharification efficiency. CBH1 deficiency significantly affected the enzymatic hydrolysis rates of various cellulose substrates, where acid pre-treated corn stover (PCS) was influenced the least. CBH2 deficiency reduced the hydrolysis of MCC, PCS, and acid pre-treated and delignified corncob but improved the hydrolysis ability of filter paper. These results demonstrate the specific contributions of CBHs to the hydrolysis of different types of biomass, which could facilitate the development of tailor-made strains with highly efficient hydrolysis enzymes for certain biomass types in the biofuel industry.

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Superantigen SpeA attenuates the biofilm forming capacity of Streptococcus pyogenes: Anshu Babbar , Israel Barrantes , Dietmar H. Pieper , Andreas Itzek; J. Microbiol. 2019;57(7):626-636. Published online June 27, 2019; DOI: https://doi.org/10.1007/s12275-019-8648-z

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Beta haemolytic Group A streptococcus (GAS) or Streptococcus pyogenes are strict human pathogens responsible for mild to severe fatal invasive infections. Even with enormous number of reports exploring the role of S. pyogenes exotoxins in its pathogenesis, inadequate knowledge on the biofilm process and the potential role of exotoxins in bacterial dissemination from matured biofilms has been a hindrance in development of effective and targeted treatments. Therefore, the present study was aimed in investigating the uncharted role of these exotoxins in biofilm process. Through our study the putative role of ciaRH in the SpeA dependent ablation of biofilm formation could be speculated and thus helping in bacterial dissemination. The seed-dispersal effect of SpeA was time and concentration dependent and seen to be consistent within various streptococcal species. Transcriptome analysis of SpeA treated S. pyogenes biofilms revealed the involvement of many transcriptional regulators (ciaRH) and response genes (luxS, shr, shp, SPy_0572), hinting towards specific mechanisms underlying the dispersal effect by SpeA. This finding opens up a discussion towards understanding a new mechanism involved in the pathogenesis of Streptococcus pyogenes and might help in understanding the bacterial infections in a better way.

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Pathomolecular epidemiology, antimicrobial resistance, and virulence genes of Streptococcus dysgalactiae subsp. equisimilis isolates from slaughtered pigs in India
Sagar M Patel, Monalisa Sahoo, Jigarji Chaturji Thakor, Dinesh Murali, Pradeep Kumar, Rajendra Singh, Karam Pal Singh, G Saikumar, Chandrakanta Jana, Shailesh Kumar Patel, Akash B Mote, Ravichandran Karthikeyan, Rajesh Kumar Vandre, Jitendra Kumar Biswal,
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T. M. Rumynska, A. R. Hural, Y. T. Konechnyi, R. B. Vynnytska, A. V. Lozynskyi, Y. T. Salyha, O. P. Korniychuk, R.B. Lesyk
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Comparative Analysis of Superantigen Genes in Staphylococcus xylosus and Staphylococcus aureus Isolates Collected from a Single Mammary Quarter of Cows with Mastitis: Karol Fijałkowski , Magdalena Struk , Jolanta Karakulska , Aleksandra Paszkowska , Stefania Giedrys-Kalemba , Helena Masiuk , Danuta Czernomysy-Furowicz , Paweł Nawrotek; J. Microbiol. 2014;52(5):366-372. Published online April 11, 2014; DOI: https://doi.org/10.1007/s12275-014-3436-2

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The purpose of this study was to analyze and compare genes encoding superantigens (SAgs) in Staphylococcus xylosus and Staphylococcus aureus isolates collected simultaneously from milk of the same cows with clinical mastitis. Genes encoding staphylococcal enterotoxins and enterotoxin-like proteins (sea-selu), toxic shock syndrome toxin 1 (tst-1) and exfolia-tive toxins (eta and etd) were investigated. It was found that among 30 isolates of S. xylosus, 16 (53.3%) harbored from 1 to 10 SAg genes. In total, in 16 SAg positive S. xylosus, 11 different enterotoxin genes were detected: sec, sed, seg, seh, sei, selm, seln, selo, selp, ser, selu and one etd gene encoding exfoliative toxin D. The most prevalent genes were ser, selu, and selo. Among all the positive isolates of S. xylosus, a total of 14 different SAg gene combinations were detected. One combination was repeated in 3 isolates, whereas the rest were detected only once. However, in the case of S. aureus all the 30 isolates harbored the same combination of SAg genes: seg, sei, selm, seln, selo and on the basis of PFGE analysis all belonged to the same clonal type. Also noteworthy was the observation that SAg genes detected in S. aureus have also been found in S. xylosus. The findings of this study further extend previous observations that SAg genes are present not only in S. aureus but also in coagulase-negative staphy-lococci, including S. xylosus. Therefore, taking into account that the SAg genes are encoded on mobile genetic elements it is possible that these genes can be transferred between different species of coexisting staphylococci.

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Hassan M. Al-Tameemi, Husam Al-Hraishawi, Murtakab Y. Al-Hejjaj, Noor S. Abdulah, Haider R. Alrafas, Yessar A. Dawood
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Influence of milk, milk fractions and milk proteins on the growth and viability of mastitis-causingStaphylococcus aureusstrain
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Menglu Shen, Yi Li, Linlin Zhang, Songbao Dai, Jiashun Wang, Yongqing Li, Lei Zhang, Jinhai Huang
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Sequence Analysis of the Gene Encoding H Antigen in Escherichia coli Isolated from Food in Morocco: Samira Badri , Aziz Fassouane , Ingrid Filliol , Mohammed Hassar , Nozha Cohen; J. Microbiol. 2010;48(2):184-187. Published online May 1, 2010; DOI: https://doi.org/10.1007/s12275-010-9182-1

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In order to develop other molecular method useful for typing of motile and non motile Escherichia coli strains, a total of 207 strains of E. coli (133 reference strains, 74 food strains) were characterized by analysis of sequences of their amplified flagellin-encoding (fliC) gene products. The collection of reference strains was used for database building of fliC gene sequences. Application of this identification system to 74 E. coli food isolates revealed a reproducible and clear cut classification with very good correlation to results obtained by HhaI restriction of the amplified flagellin gene. The proposed determination of fliC sequences variations should be helpful for epidemiological studies.

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Benjelloun Touimi Ghita, Laila Bennani, Sanae Berrada, Moussa Benboubker, Bahia Bennani
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Research Support, Non-U.S. Gov'ts

Comparative Analysis of Immune Responses to Mycobacterium abscessus Infection and Its Antigens in Two Murine Models: Bo-Young Jeon , Jeongyeon Kwak , Seung-Sub Lee , SangNae Cho , Chul Jae Won , Jin Man Kim , Sung Jae Shin; J. Microbiol. 2009;47(5):633-640. Published online October 24, 2009; DOI: https://doi.org/10.1007/s12275-009-0139-1

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Mycobacterium abscessus has been identified as an emerging pulmonary pathogen in humans. Because little is known regarding immune responses elicited by M. abscessus or its antigens, immunological responses were studied in two murine models subjected to intravenous (high-dose or systemic infection) or pulmonary (low-dose or local infection) inoculation with M. abscessus ATCC 19977. An overall comparison between the two models showed similar patterns of bacterial survival and host immune responses. The colonization of M. abscessus was the highest at 5 days post-infection (dpi) and its elimination was positively correlated with cell-mediated immunity in both challenges. However, an inverse relationship was observed between progressive inflammation and mycobacterial colonization levels in mice infected with a high dose at 14 dpi. Regarding antigens, culture filtrate (CF) of M. abscessus strongly induced IFN-γ secretion, whereas cellular extract (CE) antigen elicited strong antibody responses. The antibody response to M. abscessus antigens in mice subjected to low-dose infection increased when the cellular immune response decreased over 14 dpi. However, the antibody response for the high-dose infection increased promptly after the infection. In comparison of cytokine expression in lung homogenates after M. abscessus infection, Th1 and Th2 cytokines increased simultaneously in the high-dose infection, whereas only cell-mediated immunity developed in the low-dose pulmonary infection. These findings not only enhance our understanding of the immune response to M. abscessus infection according to systemic or pulmonary infection, but may also aid in immunological diagnosis and vaccine development.

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Molecular Characterization and Phylogenetic Analysis of H3N2 Human Influenza A Viruses in Cheongju, South Korea: Yun Hee Baek , Jeung Hyun Park , Young Jun Song , Min-Suk Song , Philippe Noriel Q. Pascua , Yoon-Soo Hahn , Heon-Seok Han , Ok-Jun Lee , Ki-Soon Kim , Chun Kang , Young-Ki Choi; J. Microbiol. 2009;47(1):91-100. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0207-y

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To investigate the genetic characteristics of human influenza viruses circulating in Chungbuk province, we tested 510 clinical samples of nasopharyngeal suction from pediatric patients diagnosed with respiratory illness between June 2007 and June 2008. Genetic characterization of the HA genes of H3N2 isolates indicated the relative higher similarity to A/Virginia/04/07 (99.6%) rather than that of A/Wisconsin/67/2005 (98.4%), a Northern Hemisphere 2007~2008 vaccine strain, based on amino acid sequences. We found several altered amino acids at the H3 HA1 antigenic sites compared with the vaccine strain; K140I at site A, K158R at site B, and K173N (H471) or K173Q, and S262N at site E, but there was no antigenic shift among the H3N2 viruses. Interestingly, A/Cheongju/H383/08 and A/Cheongju/H407/08 isolates had single amino acid substitution at D151G on the catalytic site of the N2 NA while A/Cheongju/H412/08 and A/Cheongju/H398/07 isolates had one amino acid deletion at residue 146. Furthermore, we found that 25% (3 out of 12 isolates) of the H3N2 subtype viruses had the amino acid substitution at position 31 on the M2 protein (Aspartic acid to Asparagine) and confirmed their drug-resistance by biological assays. Taken together, the results of this study demonstrated continuous evolutions of human H3N2 viruses by antigenic drift and also highlighted the need to closely monitor antigenic drug resistance in influenza A viruses to aid in the early detection of potentially pandemic strains, as well as underscore the need for new therapeutics.

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Enhancement of Immunotherapeutic Effects of HPV16E7 on Cervical Cancer by Fusion with CTLA4 Extracellular Region: Yi Zheng , Yijuan Zhang , Jun Wan , Chaofan Shi , Laiqiang Huang; J. Microbiol. 2008;46(6):728-736. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0087-1

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Cervical cancer is caused by infection by high-risk human papillomavirus (HPV), especially HPV16. Limitations in current treatments of cervical cancers call for the development of new and improved immunotherapies. This study aims at investigating the efficacy of a novel vaccine consisting of modified HPV 16E7 fused with human cytotoxic T-lymphocyte antigen 4 (CTLA4). The regions in HPV16 E7 gene associated with its transformation and CTL-enhanced response were modified; the resultant HPV16mE7 was fused with extracellular region of CTLA4 to generate HPVm16E7-eCTLA4 fusion protein. Binding of this fusion protein to B7 molecules expressed on antigen presenting-cells (APCs) was demonstrated. C57BL/6 (H-2b) mice immunized with low dose of the fusion protein (10 μg) produced higher titer antibody and stronger specific CTL response, and expressed higher levels of IFN-γ and IL-12, compared with those immunized with HPVm16E7 only or admixture of HPVm16E7 and CTLA4, or PBS; and were protected from lethal dose tumor challenge. Tumor growth was retarded and survival prolonged in mouse models with the fusion protein treatment. Our results demonstrate that fusion of HPV16 E7 with eCTLA4 targeting APCs resulted in enhanced immunity, and that this fusion protein may be useful for improving the efficacy of immunotherapeutic treatments of cervical cancer and other HPV16 infection-associated tumors.

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Genetic Characterization of the Escherichia coli O66 Antigen and Functional Identification of its wzy Gene: Jiansong Cheng , Bin Liu , David A. Bastin , Weiqing Han , Lei Wang , Lu Feng; J. Microbiol. 2007;45(1):69-74.; DOI: https://doi.org/2488 [pii]

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Abstract PDF: Escherichia coli is a clonal species, and occurs as both commensal and pathogenic strains, which are normally classified on the basis of their O, H, and K antigens. The O-antigen (O-specific polysaccharide), which consists of a series of oligosaccharide (O-unit) repeats, contributes major antigenic variability to the cell surface. The O-antigen gene cluster of E. coli O66 was sequenced in this study. The genes putatively responsible for the biosynthesis of dTDP-6-deoxy-L-talose and GDP-mannose, as well as those responsible for the transfer of sugars and for O-unit processing were identified based on their homology. The function of the wzy gene was confirmed by the results of a mutation test. Genes specific for E. coli O66 were identified via PCR screening against representatives of 186 E. coli and Shigella O type strains. The comparison of intergenic sequences located between galF and the O-antigen gene cluster in a range of E. coli and Shigella showed that this region may perform an important function in the homologous recombination of the O-antigen gene clusters.

Independent regulation of antigen processing and presentation on induction of antibody responses to various bacterial antigens in C3H/He mice: Kim, Hyung Su , Jeong, Ga Jin; J. Microbiol. 1995;33(4):355-362.

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Abstract PDF: Induction of antibody production in C3H/He mice by bacterial infection is regulated through the processing exerted by antigen presenting cells. From the studies with Psudomonas aeruginosa, Salmonella typhimurium, and Micrococcus luteu, lipopolysaccharides (LPS) in Gram negative bacteria, which are known to be T-cell independent B cell mitogen, seem to be the major factor stimulating immune responses via activation of macrophages. Activation of macrophage, however, does not seem to correlate with antibody production. M. luteus was easily eliminatd by activated macrophages, while the processed antigens were immediately releasedd into culture medium before presentation. Nevertheless, antigens from Gram positive bacteria, Staphylococcus aureus and Bacillus subtilis, were very very active in chemotaxis and activation of periotoneal macrophages as well as in antien presnetation, while the very nature of the antigens is not yet clearly understood.

Efficient Generation of BLCL Expressing Foreign Antigen as Antigen-presenting Cells with Recombinant Retroviruses: Hyun-Il Cho , Soon-Young Paik , Il-Hoan Oh , Kyu-Jung Ahn , Dong-Wook Kim , Jin-Han Kang , Wan-Shik Shin , Chun-Choo Kim , Hoon Han; J. Microbiol. 2001;39(4):300-304.

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Abstract PDF: Epstein-Barr Virus (EBV)-transformed lymphoblastoid B cell lines, BLCL, which expresse antigens, are potential antigen-presenting cells (APCs) for the induction of CTL in vitro. However, transfection of BLCLs with subsequent selection by antibiotics is notoriously difficult because plating efficiencies of BLCLs are reported to be 1% or less. To generate stable transfectants of BLCLs, we produced high titers of retroviruses encoding pp65 antigen of human cytomegalovirus as foreign antigens and transduced them to BLCLs. The pp65 gene was cloned into the retroviral vector pLXSN. The recombinant retroviral vector was transfected to ecotropic packaging cell line, GP&E86, and this polyclonal recombinant retrovirus was transduced to PA317 that is amphotropic packaging cell line. The titers of cloned PA317 amphotropic retroviruses ranged from 5 to 7 X 10^6 colony forming units (CFU) per ml (CFU/ml). We performed three rounds of consecutive transductions to BLCLs in order to improve the cloning efficiencies. The expression of recombinant HCMV-pp65 antigen was more than 20% after the final transduction. The third-transduced BLCLs were easily selected in optimal concentration of G418. BLCLs expressing foreign antigens could be used as target cells for CTL assay and/or as APCs for induction of in vitro CTL responses specific for viral and tumor antigens.

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