The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.
Jung, Boknam;Park, Jungwook;Kim, Namgyu;Li, Taiying;Kim, Soyeon;Bartley, Laura E.;Kim, Jinnyun;Kim, Inyoung;Kang, Yoonhee;Yun, Ki-Hoon;Choi, Younghae;Lee, Hyun-Hee;Lee, Kwang Sik;Kim, Bo Yeon;Shon, Jong Cheol;Kim, Won Cheol;Liu, Kwang-Hyeon;Yoon, Dahye;Kim, Suhkman;Ji, Sungyeon;Seo, Young Su;Lee, Jungkwan
한국균학회소식:학술대회논문집
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2018.05a
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pp.33-33
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2018
Air-borne plant pathogenic fungus Fusarium graminearum and seed-borne plant pathogenic bacterium Burkholderia glumae are cause similar disease symptoms in rice heads. Here we showed that two pathogens frequently co-isolated in rice heads and F. graminearum is resistant to toxoflavin produced by B. glumae while other fungal genera are sensitive to the toxin. We have tried to clarify the resistant mechanism of F. graminearum against toxoflavin and the ecological reason of co-existence of the two pathogens in rice. We found that F. graminearum carries resistance to toxoflavin as accumulating lipid in fungal cells. Co-cultivation of two pathogens resulted in increased conidia and enhanced chemical attraction and attachment of the bacterial cells to the fungal conidia. Bacteria physically attached to fungal conidia, which protected bacterium cells from UV light and allowed disease dispersal. Chemotaxis analysis showed that bacterial cells moved toward the fungal exudation compared to a control. Even enhanced the production of phytotoxic trichothecene by the fungal under presence of toxoflavin and disease severity on rice heads was significantly increased by co-inoculation rather than single inoculation. This study suggested that the undisclosed potentiality of air-born infection of bacteria using the fungal spores for survival and dispersal.
The use of antibiotics in aquaculture raises environmental and food safety concerns because chronic exposure of an aquatic ecosystem to antibiotics can result in the spread of antibiotic resistance, bioaccumulation of antibiotics in the organisms, and transfer of antibiotics to humans. In an attempt to overcome these problems, high-concentration red clay was applied as an alternative antibiotic against the following common fish pathogens: Aeromonas salmonicida, Vibrio alginolyticus, and Streptococcus equinus. The growth of A. salmonicida and V. alginolyticus was retarded by red clay, whereas that of S. equinus was promoted. Phase contrast and scanning electron microscopy analyses confirmed the attachment of red clay on cell surfaces, resulting in rapid gravitational removal and cell surface damage in both A. salmonicida and V. alginolyticus, but not in S. equinus. Different cell wall properties of grampositive species may explain the unharmed cell surface of S. equinus. Significant levels of oxidative stress were generated in only the former two species, whereas significant changes in membrane permeability were found only in S. equinus, probably because of its physiological adaptation. The bacterial communities in water samples from Oncorhynchus mykiss aquacultures supplemented with red clay showed similar structure and diversity as those from oxytetracycline-treated water. Taken together, the antibiotic effects of high concentrations of red clay in aquaculture can be attributed to gravitational removal, cell surface damage, and oxidative stress production, and suggest that red clay may be used as an alternative for antibiotics in aquaculture.
The local route of antibiotic administration can accomplish higher therapeutic doses in subgingival sites than those possible by systemic therapy. This investigation assessed on the clinical and microbiological effect of 30% Minocycline loaded polycaprolactone film (Mino-strip) on rapidly progressive periodontitis. Mino-strip was applied in the periodontal pockets of 15 patients with clinically diagnosed as a rapidly progressive periodontitis. 8sites for each patient with a 5mm probing pocket depth were selected in split mouth design and were assigned into group. i.e., placebo(group 1), supragingival scaling and R/P(group 2), Mino-strip applied only(group 3), R/P and Mino-strip applied(group 4). Supragingival scaling and oral hygiene instruction were performed 1 wk before experiment. Mino-strip was applied weekly on day 0 and 7. Clinical and microbiological test were performed on day 0, 7, 14, 28, 56. In R/P and Mino-strip applied group, Gingival index, GCF volume, probing depth and loss of attachment level were significantly reduced after the first weeks following treatment. In R/P and Mino-strip applied group, the relative proportions of spirochetes and motile rods were significantly reduced and the proportions of cocci and non motile rod were correspondingly increased for eight weeks following treatment. In R/P and Mino-strip treated group, total anaerobic and aerobic bacterial count were significantly decreased for the first two weeks following treatment and streptococcus count was decreased for eight weeks following treatment. In R/P and Mino-strip applied group, P. gingivalis, P. intermedius, B. forsythus, A. actinomycetemcomitans, F. nucleatum, E. corrodens, C. rectus counts were significantly reduced after the first week following treatment. According to this study, it is appeared that 30% Minocycline-loaded polycaprolacton film was effective in the treatment on rapidly progressive periodontitis.
Lactic acid bacteria (LAB) are widely used for various food fermentation. With the recent advances in modern biotechnology, a variety of bio-products with the high economic values have been produced using microorganisms. For molecular cloning and expression studies on the gene of interest, E. coli has been widely used mainly because vector systems are fully developed. Most plasmid vectors currently used for E, coli carry antibiotic-resistant markers. As it is generally believed that the antibiotic resistance markers are potentially transferred to other bacteria, application of the plasmid vectors carrying antibiotic resistance genes as selection markers should be avoided, especially for human consump-tion. By contrast, as LAB have some desirable traits such that the they are GRAS(generally recognized as safe), able to secrete gene products out of cell, and their low protease activities, they are regarded as an ideal organism for the genetic manipulation, including cloning and expression of homologous and heterologous genes. However, the vec-tor systems established for LAB are stil insufficient to over-produce gene products, stably, limiting the use of these organisms for industrial applications. For a past decade, the two popular plasmid vectors, pAM$\beta$1 of Streptococcus faecalis and pGK12 theB. subtilis-E. coli shuttle vector derived from pWV01 of Lactococcus lactis ssp. cremoris wg 2, were most widely used to construct efficient chimeric vectors to be stably maintained in many industrial strains of LAB. Currently, non-antibiotic markers such as nisin resistance($Nis^{r}$ ) are explored for selecting recombi-nant clone. In addition, a gene encoding S-layer protein, slp/A, on bacterial cell wall was successfully recombined with the proper LAB vectors LAB vectors for excretion of the heterologous gene product from LAB Many food-grade host vec-tor systems were successfully developed, which allowed stable integration of multiple plasmid copies in the vec-mosome of LAB. More recently, an integration vector system based on the site-specific integration apparatus of temperate lactococcal bacteriophage, containing the integrase gene(int) and phage attachment site(attP), was pub-lished. In conclusion, when various vector system, which are maintain stably and expressed strongly in LAB, are developed, lost of such food products as enzymes, pharmaceuticals, bioactive food ingredients for human consump-tion would be produced at a full scale in LAB.
The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.
To understand the effects of acclimation schemes on the formation of anode biofilms, different electrical performances are characterized in this study, with the roles of suspended and attached bacteria in single-chamber microbial fuel cells (MFCs). The results show that the generation of current in single-chamber MFCs is significantly affected by the development of a biofilm matrix on the anode surface containing abundant immobilized microorganisms. The long-term operation with suspended microorganisms was demonstrated to form a dense biofilm matrix that was able to reduce the activation loss in MFCs. Also, a Pt-coated anode was not favorable for the initial or long-term bacterial attachment due to its high hydrophobicity (contact angle = $124^{\circ}$), which promotes easy detachment of the biofilm from the anode surface. Maximum power ($655.0\;mW/m^2$) was obtained at a current density of $3,358.8\;mA/m^2$ in the MFCs with longer acclimation periods. It was found that a dense biofilm was able to enhance the charge transfer rates due to the complex development of a biofilm matrix anchoring the electrochemically active microorganisms together on the anode surface. Among the major components of the extracellular polymeric substance, carbohydrates ($85.7\;mg/m^2_{anode}$) and proteins ($81.0\;mg/m^2_{anode}$) in the dense anode biofilm accounted for 17 and 19%, respectively, which are greater than those in the sparse anode biofilm.
Purpose: The present study investigated the outcomes of a newly-developed, simple, and practical nonsurgical treatment modality suitable for most forms of intrabony defects around failing dental implants using intrasulcular delivery of chlorhexidine solution and minocycline hydrochloride (HCl). Methods: Forty-five dental implants in 20 patients diagnosed with peri-implantitis were included. At baseline and the study endpoint, the probing pocket depth (PPD), clinical attachment level (CAL), and the presence of bleeding on probing (BOP) at 6 sites around each implant were recorded. The radiographic osseous defect morphology at the mesial or distal proximal aspect of each implant was classified as 1) narrow or wide and 2) shallow or deep. For a comparative analysis of bone changes according to the defect morphology, the distance from the implant shoulder to the most coronal bone-to-implant contact point (DIB) at the mesial and distal aspects of each implant was measured at baseline and the endpoint. Patients were scheduled to visit the clinic every 2-4 weeks for intrasulcular irrigation of chlorhexidine and delivery of minocycline HCl. Results: We observed statistically significant decreases in PPD, CAL, and BOP after treatment. At the endpoint, bone levels increased in all defects, regardless of the osseous morphology of the intrabony defect. The mean DIB change in deep defects was significantly greater than that in shallow defects. Although the mean bone gain in narrow defects was greater than in wide defects, the difference was not statistically significant. Conclusions: We propose that significant and sustainable improvements in both clinical and radiographic parameters can be expected when intrabony defects around dental implants are managed through a simple nonsurgical approach involving combined intrasulcular chlorhexidine irrigation and local delivery of minocycline HCl.
An experiment was conducted to determine whether there were any apparent differences in the microbial population, colonization pattern and digestion of guinea grass in situ, between cattle and swamp buffalo. Percentage losses in dry matter (DM), nitrogen (N) and neutral detergent fibre (NDF) of guinea grass were significantly (p<0.01) higher when incubated in the rumen of buffalo than in cattle. Buffalo also showed significantly (p<0.05) faster degradation rates than cattle for each grass component (DM, N, DNF). Light microscopy and SEM examination of the incubated grass materials showed that there were no apparent differences in the pattern of bacterial and fungal invasion and colonization of the grass materials between cattle and buffalo. Attachment of bacteria and fungal zoospores on the grass fragments occurred at 15 min after rumen incubation. After 3 h of rumen incubation, dense population of bacteria was observed in the thin-walled mesophyll and parenchyma tissues, whereas root-like fungal rhizoids were observed in both thin-walled and thick-walled cells. By 6 h, eroded zones were apparent in the thin-walled tissues and in thick-walled tissues with profuse rhizoids. After 24. 48 and 72 h of rumen incubation, most thin-walled tissues were degraded leaving mostly the thick-walled tissues. The predominant bacteria were the curved rods resembling Butyrivibrio sp., the thick rods resembling Fibrobacter sp., the diplococcoids resumbling Ruminococcus sp. And spirochetes. Fungi were predominantly those with spherical or oval sporangia. Fusiform sporangia with acuminate apices which resembled Ruminomyces sp. Were of lesser occurrence. Few protozoa were found on the grass fragments at all incubation times.
An approximately 10-fold higher level of adherence of Salmonella typhimurium strain TML to Int-407 cells was observed with organisms grown in Luria broth or in high-iron containing medium than those grown in low-iron containing medium. Iron specifically enhanced adherence, while other cations such as calcium, cobalt, copper, potassium, magnesium and manganese did not. It was suggested that iron did not act as a passive ligand - probably it stimulated production of bacterial factors necessary for adherence. A similar pattern of iron modulation of adhesiveness was also seen in Salmonella mutants with single or different combinations of multiple mutations in genes encoding the mannose sensitive hemagglutinin (type 1 fimbriae), mannose resistant hemagglutinin and flagellum. The adhesiveness of an isogenic fur mutant was modulated by iron in a manner similar to the wild-type strain, suggesting that iron modulation of adherence is independent of the fur gene product.
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