• Title/Summary/Keyword: Bacterial challenge

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Characterization of Mitochondrial Heat Shock Protein 75 (mtHSP75) of the Big-belly Seahorse Hippocampus abdominalis (빅벨리해마(Hippocampus abdominalis)에서의 Mitochondrial Heat Shock Protein 75 유전자의 특징과 발현 분석)

  • Ko, Jiyeon;Qiang, Wan;Lee, Sukkyoung;Bathige, S.D.N.K.;Oh, Minyoung;Lee, Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.3
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    • pp.354-361
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    • 2015
  • Mitochondrial heat shock protein 75 (mtHSP75) is a member of the HSP90 family and plays essential roles in refolding proteins of the mitochondrial matrix. Mitochondria provide energy in the form of ATP and generate reactive oxygen species (ROS). Heat shock proteins (HSPs) are activated in response to stress, and protect cells. In this study, we characterized the mtHSP75 of the big-belly seahorse Hippocampus abdominalis. The protein (BsmtHSP75) is encoded by an open reading frame (ORF) of 2,157 nucleotides, has 719 amino acids (aa), and is of molecular mass 82 kDa. BsmtHSP75 has two functional domains, a histidine kinase-like ATPase (HATPase_c) domain (123-276 aa) and an HSP90 family domain (302-718 aa). BsmtHSP75 was expressed in all tested tissues of healthy seahorses. The ovary contained the highest transcription level, followed (in order) by the blood, brain, and muscle. Pouch tissue showed the lowest expression level. The expression of BsmtHSP75 was significantly (P<0.05) up-regulated on viral or bacterial challenge, suggesting that BsmtHSP75 plays a role in the immune defense against bacterial and viral pathogens.

New typhoid vaccine using sponge-like reduced protocol: development and evaluation

  • Rehab Bahy;Asmaa Gaber;Hamdallah Zedan;Mona Mabrook
    • Clinical and Experimental Vaccine Research
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    • v.12 no.1
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    • pp.70-76
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    • 2023
  • Purpose: Typhoid remains a major health problem, especially in the developing world. Furthermore, the emergence of multidrug-resistant and extensively drug-resistant strains of Salmonella typhi added a sense of urgency to develop more effective typhoid vaccines, one of which is bacterial ghosts (BGs), prepared by both genetic and chemical means. The chemical method includes incubation with numerous agents for a short time at their minimum inhibitory or minimum growth concentrations. This study included the preparation of BGs by a sponge-like reduced protocol (SLRP). Materials and Methods: Critical concentrations of sodium dodecyl sulfate, NaOH, and H2O2 were used. Moreover, high-quality BGs were visualized by scanning electron microscope (SEM). Subculturing was used to confirm the absence of vital cells. Besides, the concentrations of the released DNA and protein were estimated spectrophotometrically. In addition, the integrity of cells was proved by visualizing Gram-stained cells using a light microscope. Furthermore, a comparison between the immunogenicity and safety of the prepared vaccine and the available whole-cell killed vaccine was established. Results: Improved preparation of high-quality BGs of S. typhi, visualized by SEM, revealed punctured cells with intact outer shells. Moreover, the absence of vital cells was confirmed by subculturing. At the same time, the release of respective amounts of proteins and DNA is another evidence of BGs' production. Additionally, the challenge test provided evidence that the prepared BGs are immunogenic and have the same efficacy as the whole cell vaccine. Conclusion: The SLRP provided a simple, economical, and feasible method for BGs preparation.

Efficacy of a vaccine against Streptococcus parauberis infection in starry flounder Platichthys stellatus Pallas

  • Lee, Deok-Chan;Lee, Jae-Il;Kim, Do-Hyung;Cho, Mi-Young;Kim, Jin-Woo
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.189-195
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    • 2011
  • Starry flounder, which are recently increasingly cultured in Korea, are known to highly vulnerable to Streptococcus parauberis infection. Five groups of starry flounder (n=30 for each group) were vaccinated with S. parauberis formalin-killed whole cells by intraperitoneal injection at a final concentration of 0, 0.01, 0.1, 1 and 10 mg $fish^{-1}$. Specific antibody production of 1 and 10 mg $fish^{-1}$ administered groups significantly increased at four weeks post immunization. All vaccinated groups showed higher survival rates than a control group when five groups of fish were challenged with S. parauberis at a dose of $1.14{\times}10^4$ cfu $fish^{-1}$ and $1.14{\times}10^2$ cfu $fish^{-1}$, respectively. In particular, 0.1 or higher concentrations of formalin killed bacterial cells are able to confer the fish high protection against S. parauberis infection.

cDNA Cloning and Developmental Expression of Hemolin in Bombyx mandarina

  • Kang Min Uk;Kim Kyung-A;Lee Jin Sung;Kim Nam Soon;Kang Seok-Woo;Nho Si-kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.2
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    • pp.101-106
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    • 2005
  • In this study, we describe the Bombyx mandarina hemolin cDNA. A sequence analysis of cDNA revealed a single open reading frame (ORF) of 1233 nucleotides. The deduced 410 amino acid sequence of B. mandarina hemolin contains 4 imunoglobulin (Ig) C-2 type domains. B. mandarina hemolin cDNA showed the highest sequence homology to known those of B. mori. The developmental profile in terms of expression level of hemolin mRNA was determined in the absence of a bacterial challenge. Hemolin mRNA was detected only in mid-gut, but not in hemocytes, fat body, testis, and silkglands. Hemolin mRNA in mid-gut was not detected until the spinning stage of the last instar larva, however, lit dramatically increased at the beginning of spinning and gradually decreased until pupal stage.

Change in Cell Size and Buoyant Density of Pseudomonas diminuta in Response to Osmotic Shocks

  • Lee, So-Hee;Cho, Yu-Ree;Choi, Yong-Jin;Kim, Chan-Wha
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.326-328
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    • 2001
  • Pseudomonas diminuta (ATCC 19146) has been typically used in the bacterial challenge test for validation of the sterilizing filtration process. Cell size is critical for determining the retention characteristics of membrane filters with pore-size of $0.2{\mu}m$. The changes of cell sizes after osmotic shocks at 150, 260, 500, and 700 mosM were measured by a particle size analyzer and the changes of their buoyant densities were analyzed with a Percoll gradient. The results indicated that there were no significant differences when cells were cultured in 260 mosM medium and osmotically shocked at 500 and 700 mosM. However, the osmotically shocked cells at 150 mosM showed a 38% increase of the cell size compared to the cells at 260 mosM. From these study, we concluded that the worst case condition for validation of a sterilizing filter would be 500 mosM, not because of changes in the cell size, but due to decrease in cell viability under those conditions.

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Isolation of Novel Hepcidin Isoforms from the Rockbream Oplegnathus fasciatus (Perciformes)

  • Lee, Sang-Yoon;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.31-42
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    • 2011
  • Three novel hepcidin isoforms were isolated and characterized from the perciform fish species Oplegnathus fasciatus. These hepcidin isoforms (designated rbhepc5, rbhepc6 and rbhepc7) were found to share a conserved, tripartite gene structure and a considerable sequence homology one another. A comparison of their mature peptide sequences with those of other perciform hepcidin orthologs indicated that these three hepcidin isoforms as well as four other isoforms previously identified in this species, appear to belong to the HAMP2 group of hepcidin genes. Analysis of the 5'-upstream sequences showed that the proximal non-coding regions of rbhepc5~7 do not possess canonical TATA signals; instead, they harbor several binding motifs for transcription factors involved in immune modulation. Reverse transcriptase-PCR analysis demonstrated that the rbhepc5~7 are expressed predominantly in the liver, and that the transcription of rbhepc5~7 is rapidly induced in the liver, but not in other tissues, by experimental challenge with any of three different bacterial species. However, transcription of rbhepc6 appeared to be negligible under both basal and stimulated conditions, as judged by the redundancy count of randomly chosen reverse transcriptase-PCR clones.

Therapeutic Effects of Bacteriophages Against Salmonella gallinarum Infection in Chickens

  • Hong, Sung Sik;Jeong, Jipseol;Lee, Jinju;Kim, Suk;Min, Wongi;Myung, Heejoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1478-1483
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    • 2013
  • In this study the isolation and characterization of three bacteriophages (ST4, L13, and SG3) infecting Salmonella gallinarum were carried out. They were further tested for their in vivo efficacy in phage therapy. All three phages belong to the Siphoviridae family with isometric heads and non-contractile tails. They have a broad host range among serovars of Salmonella enterica. The burst sizes were observed to be 1670, 80, and 28 for ST4, L13, and SG3, respectively. The in vivo efficacy of the phages was tested in chickens. Layer chickens were challenged with S. gallinarum, whereas contact chickens were cohabited without direct challenge. Each bacteriophage was orally inoculated in the form of feed additives. Mortality was observed and S. gallinarum was periodically re-isolated from the livers, spleens, and cecums of the chickens. Bacterial re-isolation from the organs and mortality decreased significantly in both challenged and contact chickens treated with the bacteriophages compared with untreated chickens serving as the control. The three bacteriophages may be effective alternatives to antibiotics for the control of fowl typhoid disease in chickens.

The Role of Nitric Oxide in Mycobacterial Infections

  • Yang, Chul-Su;Yuk, Jae-Min;Jo, Eun-Kyeong
    • IMMUNE NETWORK
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    • v.9 no.2
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    • pp.46-52
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    • 2009
  • Although tuberculosis poses a significant health threat to the global population, it is a challenge to develop new and effective therapeutic strategies. Nitric oxide (NO) and inducible NO synthase (iNOS) are important in innate immune responses to various intracellular bacterial infections, including mycobacterial infections. It is generally recognized that reactive nitrogen intermediates play an effective role in host defense mechanisms against tuberculosis. In a murine model of tuberculosis, NO plays a crucial role in antimycobacterial activity; however, it is controversial whether NO is critically involved in host defense against Mycobacterium tuberculosis in humans. Here, we review the roles of NO in host defense against murine and human tuberculosis. We also discuss the specific roles of NO in the central nervous system and lung epithelial cells during mycobacterial infection. A greater understanding of these defense mechanisms in human tuberculosis will aid in the development of new strategies for the treatment of disease.

Protective effect of bacteriophages against Salmonella Typhimurium infection in weaned piglets (이유자돈에서 Salmonella Typhimurium 감염에 대한 박테리오파지의 방어 효능)

  • Kim, Sung-Jae;Kim, Jae-Hoon;Jun, Soo-Yeon;Paik, Hyoung Rok;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.35-43
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    • 2014
  • Salmonellosis has caused heavy losses in swine industry and implications for public health. Recently, the urgent problem of antibiotic resistance due to multidrug-resistant Salmonella spp. has been on the rise. The use of host-specific bateriophages as a biocontrol is one possible alternative. In this study, clinical signs, growth performance, quantification and detection of antigen, histopathological changes of gastrointestinal tracts were analyzed comparatively in weaned piglets according to administration of bacteriophages and challenge with Salmonella (S.) Typhimurium. Piglets challenged with S. Typhimurium after administered with bacteriophages showed reduced clinical signs, higher growth performance, lower bacterial shedding, lower quantificational value of antigens in intestines, higher V/C ratio and higher the number of goblet cells in intestines than piglets administered without bacteriophage and challenged with S. Typhimurium. These results indicate that feeding contained with bacteriophages has effect to prevent infection of S. Typhimurium in weaned piglets and suggest that a use of bacteriophage can be considered a valid antibiotic alternative.

Challenging the Hypothesis of de novo Biosynthesis of Bile Acids by Marine Bacteria

  • Tueros, Felipe Gonzalo;Ellabaan, Mostafa M. Hashim;Henricsson, Marcus;Vazquez-Uribe, Ruben;Backhed, Fredrik;Sommer, Morten Otto Alexander
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.102-109
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    • 2022
  • Bile acids are essential molecules produced by vertebrates that are involved in several physiological roles, including the uptake of nutrients. Bacterial isolates capable of producing bile acids de novo have been identified and characterized. Such isolates may provide access to novel biochemical pathways suitable for the design of microbial cell factories. Here, we further characterized the ability of Maribacter dokdonensis, Dokdonia donghaensis, and Myroides pelagicus to produce bile acids. Contrary to previous reports, we did not observe de novo production of bile acids by these isolates. Instead, we found that these isolates deconjugated the amino acid moiety of bile acids present in the growth medium used in previous reports. Through genomic analysis, we identified putative bile salt hydrolases, which could be responsible for the different bile acid modifications observed. Our results challenge the hypothesis of de novo microbial bile acid production, while further demonstrating the diverse capacity of bacteria to modify bile acids.