3^rd Global Microbiologists Annual Meeting August 15-17, 2016 Portland, Oregon, USA
(5 Plenary Forums - 1 Event)

Yuan Hu has received his MD from Guangzhou Medical University (1983) and MS (1990) in Pharmaceutical Sciences from St. John’s University, New York. He is a Research Microbiologist at U.S. Food & Drug Administration. He is also a licensed Clinical Laboratory Director in New York State. He has published over 50 peer-reviewed articles, books chapter and abstracts.

Noroviruses (NoVs) are highly infectious and cause acute gastroenteritis in humans. NoVs have high levels of genetic sequence diversities, which lead to difficulties in designing robust universal primers to efficiently amplify specific viral genomes for molecular analysis. We here described the practicality of sequence-independent amplification combined with DNA microarray analysis for simultaneous detection and genotyping of human NoVs in fecal specimens. We showed that single primer isothermal linear amplification (Ribo-SPIA) of genogroup I (GI) and genogroup II (GII) NoVs could be run through the same amplification protocol without the need to design and use any virus specific primers. Related virus could be subtyped by the unique pattern of hybridization with the amplified product to the microarray. By testing 22 clinical fecal specimens obtained from acute gastroenteritis cases as blinded samples, two were GI positive and 18 were GII positive as well as 2 negative for NoVs. NoV GII positive specimen was also identified as having co-occurrence of hepatitis A virus. The study showed that there was 100% concordance for positive NoV detection at genogroup level between the results of Ribo-SPIA/microarray and the phylogenetic analysis of viral sequences of the capsid gene. This efficient broad range microarray detection method represents a valuable tool for norovirus study and outbreaks investigation.

Hee Wook Ryu has completed his PhD from KAIST and Postdoctoral studies from University of Minnesota. He is currently a Professor at Soongsil University, South Korea. He has published more than 70 papers in reputed journals.

Trametes versicolor CBR43 was isolated from Chunma Mountain, South Korea. The fungus had high activities of laccase and Mn dependent peroxidase whereas its lignin peroxidase activity was relatively low. T. versicolor CBR43 could decolorize 200 mg/L of acid dyes (red 114, blue 62, orange 7 and black 172), reactive dyes (red 120, blue 4, orange 16 and black 5) and disperse dyes (red 1, orange 3 and black). Except acid orange 7, T. versicolor CBR43 decolorized more than 90% of acid and reactive dyes within 6 days in the PDB medium. This fungus decolorized 67% of acid orange 7 within 9 days. The decolorization efficiencies for disperse dyes were 51, ~80% at 9th day. The maximum rates of dye decolorization in the cell-free culture broth of T. versicolor CBR43 were 1,410, 44.7, 41.2 and 0.19 μmol L-1 min-1 for acid blue 62, acid black 175, reactive blue 4 and acid red 114, respectively. The inhibition effect of NaCl, Zn(II) and Cd(II) on the dye decolorization has been quantitatively compared using the half maximal inhibitory concentration (IC50) indicating the concentration of an inhibitor that is required for 50% inhibition. Based on the values of IC50, the dye decolorization in the cell-free culture broth of CBR43 was most severely inhibited by Cd(II), and the inhibition effect of NaCl was relatively low. The information on dye decolorization rates and IC50 can be utilized to design for the development of a dye wastewater treatment process employing T. versicolor CBR43 and its operating factors.

Kyung-Suk Cho has completed her PhD from Tokyo Institute of Technology (TIT) and Postdoctoral studies from TIT and KAIST. In 1994, she has joined the Faculty at Ewha Womans University, Seoul, South Korea. She operates the Environmental Bioengineering Laboratory in the Department of Environmental Science and Engineering. Her expertise is in bioremediation and ecological technology using microorganisms. She has published more than 110 papers in international journals.

Methane and unpleasant odors emitted from landfills have been caused global warming and complaints. The use of engineered biocovers is a promising strategy toward mitigating methane and odor emissions from smaller and/or older landfills. Since the filter bed in a biocover is a key factor to determine methane and odor removal efficiency and the characterization of the microbial dynamics in the filter bed is very important. Moreover, there is also a lack of information on microbial communities in biocovers installed on field landfills. The engineered biocover (10 m in length × 5 m in width × 1 m in depth) was constructed on a sanitary landfill site located at Jeollanam-do, South Korea. The mixture of soil, perlite and earthworm cast (26:6:2.5, volume ratio) was used as packing materials and inoculum sources for the biocover. Methane and odor removal efficiencies were evaluated and the dynamics of bacterial community structures in the biocover as a function of time were monitored by 16S rDNA pyrosequencing. Based on the dilution to threshold ratios derived by the air dilution sensory test, the removal efficiencies for complex odor were ranged from 95.0% to 98.6% during winter season from January to March. The representative odor compounds were ammonia, hydrogen sulfide, methanethiol, dimethyl sulfide, dimethyl disulfide, stylene, tolune, xylene, methyl ethyl ketone, acetic acid and n-butylic acid. Methane removal efficiencies were 21.5% to ~33.5%. Methylobacter (40%) and Flavobacterium (38%) were found as dominant bacteria. Rhodanobacter, Arenibacter, Arthrobacter, Algoriphagus, Nannocystis, Rhodobacter and Paenibacillus were detected in the biocover.

Ing-Lung Shih has obtained his PhD degree in Organic Chemistry from Brandies University of USA in 1987. He had a joint Postdoctoral appointment by Massachusetts Institute of Technology (MIT) and New England Deaconess Hospital (NEDH) from 1987 to 1989. He is currently a Distinguished Professor at Dayeh University of Taiwan. His research areas include chemical synthesis, enzymatic synthesis and microbial synthesis of bioactive materials (peptides, proteins, biopolymers, enzymes and antibiotics) for the applications in food, medicine, cosmetics and environment. He has published 62 journal papers (47 SCI papers), 69 conference papers, 29 technical reports, 2 book and 14 Patents.

The purpose of the present study is to investigate the feasibility of a tandem production of two invaluable biopolymers-levan and poly-ε-lysine (ε-PL) by microbial fermentation using sugarcane juice as a feedstock. The results showed that sugarcane juice can served as a good substrate for levan production by Bacillus subtilis strain natto. A high yield of levan production (22 g/L at 24 h; 22 g/L/d; 33.3% by sucrose consumption) was obtained when the bacteria were cultured in medium containing sugarcane juice (containing 78 g/L of sucrose) at pH 6.5, temperature 37 oC and agitation speed 175 rpm. The fermentation broth was concentrated by ultra filtration through a membrane of 5 kDa cutoff. The concentrate was harvested of levan by precipitation by addition of 4 vols. cold ethanol; the filtrate containing glucose was proven to be a suitable medium for ε-PL production by Streptomyces albulus when it was supplemented with yeast extract, (NH4)2SO4 and basal salts. 4.4 g/L of ε-PL accumulation was obtained in 72 h using two-stage fermentation with control of pH. Biorefinery of biomass into biopolymer is a sustainable process that not only solves environmental problems but also produced high value added bio-products; it is a green process that is worth of developing.

Enem Simon Ikechukwu has completed his PhD and he is a Senior Lecturer at the Department of Veterinary Public Health and Preventive Medicine, University of Abuja, Nigeria where he served as the immediate past Head of Department. He has over 20 journal publications to his credit and has attended many conferences both locally and internationally. He has served as a Reviewer to some journals.

The major natural reservoir of Shiga toxin producing Escherichia coli (E. coli) is cattle. Man gets infected by the consumption of contaminated cattle meat and meat products. Shiga toxin producing Escherichia coli O157 is a major cause of hemolytic colitis (HC) and hemolytic uremic syndrome (HUS) in humans. The cross sectional epidemiologic method was used in this study. Human samples were collected from sick hospital patients, apparently healthy high risk individuals (abattoir workers, cattle herdsmen, milk hawkers) and from members of the public. Freshly voided feces were collected from cattle in selected abattoirs and cattle herds. The samples were subjected to an enrichment culture and analyzed both bacteriologically and biochemically to confirm typical Escherichia coli which were then sub-cultured into plates of cefixine-tellurite sorbitol McConkey (CT-SMAC) agar. The non sorbitol fermenters stored in nutrient agar slants were further characterized using commercially procured latex agglutination test kits. A total of 572 human samples were tested for the presence of Shiga toxin producing E. coli and 6 (1.05%) was positive. Of the 718 fecal samples from cattle tested, 17 were positive. The antibiogram of the isolates to some commonly used antibiotics were tested. Ten isolates from cattle were tested and found to be sensitive to levofloxacin, streptomycin, chloramphenicol and ciprofloxacin but resistant to erythromycin, gentamycin, augumentin, tetracycline, cotrimaxazole and cloxacillin. Five Shiga toxin producing E. coli O157 isolated from humans were sensitive to levofloxacin and ciprofloxacin and resistant to the rest. The study indicated that both cattle and man within the same environment harbor Shiga toxin producing E. coli O157 proving that cattle play a major role as source of transmission of multi drug resistant Shiga toxin producing E. coli O157 to humans in Abuja, FCT.

Woo Young Yang has completed his graduate studies from Soongsil University and he is currently a Master’s student at Soongsil University, South Korea.

As one of the methods for treating a livestock carcasses infected with foot and mouth disease, the carcasses composting using thermophilic bacteria was studied to decompose rapidly the carcasses and to minimize odor emissions. Twenty eight species of thermophilic (40~60 oC) and alkaliphilic (pH 8~10) bacteria were isolated and assessed their ability to decompose protein and lipid. Among them, five thermophilic and alkaliphilic bacteria (Bacillus sp. AL2, Bacillus sp. APJ4, Bacillus sp. OP3, Geobacillus thermodenitrificans OP4 and Aneurinibacillus thermoaerophilus OP11), which generate less odor in the decomposing process, were selected and applied to composting pig carcasses. The pilot-scale composting cells (2.5×2.5×2.0 m) were constructed and fully fermented pig manure compost was used as a feedstock for the composting carcasses. Six pig carcasses of 90 kg were buried in each cell. Compared with the control cell which not inoculated with the decomposing bacteria, the temperature of the test cell applying the thermophilic bacteria was kept higher by 40 to ~54 oC, the pH of the compost around the burial carcasses was 8.0 to ~9.5. The odors from the decomposition were less released from the cell and the buried carcasses were completely decomposed and remained clean bones after 1 year. It suggests that the use of thermophilic bacteria can quickly decompose the carcass and reduce the generation of odor.

Ching Yang is currently a Veterinary Student from National Taiwan University who has strong interest in veterinary pathology. She is actively involved in research programs in Taiwan and abroad and she has also presented her work in both international and national conferences.

Vero cell line has been routinely used for isolation and propagation of porcine epidemic diarrhea virus (PEDV) and is the most studied cells in the literature. However, isolation of PEDV in Vero cells has been proven difficult. Besides, the entry mechanism for PEDV in Vero cells is also known to be the cellular receptor (porcine aminopeptidase; pAPN; CD13)-independent suggesting that Vero cells might not be a suitable cell line for studying the interaction of PEDV with its receptor or the viral entry pathway. In the present study, a HEK 293 cell line stably expressing pAPN (HEK-pAPN) has been established. The susceptibility of the HEK-pAPN cell line to PEDV infection has been compared with Vero cells, HEK293 cells and PK-15 cells. Interestingly, similar to PEDV-infected Vero cells, cytopathic effects characterized by cell fusion, formation of multinuclear syncytial cells and cell death were observed in HEK 293 and HEK-pAPN cells after 48 hours of inoculation, but not in the PK-15 cells. Furthermore, both HEK 293 and HEK-pAPN cell lines permissive to PEDV infection confirmed by immunocytochemistry (ICC) staining, one-step growth curve and real-time PCR have found to express the APN protein. In conclusions, we have demonstrated that HEK293 cells carrying APN molecules are permissive to PEDV. The HEK293 cell line might serve as an alternative tool that could be used to study PEDV pathogenesis and entry mechanisms. Furthermore, the identification of the human origin HEK293 cell line permissive to PEDV raises the possibility of interspecies transmission of the virus is of concern.

Ruicheng Yang is currently pursuing PhD in Preventive Veterinary Medicine at Huazhong Agricultural University, China. His main research direction is bacterial meningitis, including the molecular mechanism study on the interaction between microbes and the host and the regulative mechanisms of the tight junctions by pathogens.

Escherichia coli is the most common Gram negative bacterium that possesses the ability to cause neonatal meningitis, which develops as circulating bacteria penetrate the blood-brain barrier (BBB). However, whether meningitic E. coli could induce disruption of the BBB and the underlying mechanisms are poorly understood. Our current work highlight for the first time the participation of VEGFA and Snail-1, as well as the potential mechanisms, in meningitic E. coli induced disruption of the BBB. Here, we characterized meningitis causing E. coli PCN033 and demonstrated that PCN033 invasion could increase the BBB permeability through down-regulating and remodeling the tight junction proteins (TJs). This process required the PCN033 infection induced up-regulation of VEGFA and Snail-1, which depend on the activation of TLR2-MAPK-ERK1/2 signaling cascade. Moreover, production of proinflammatory cytokines and chemokines in response to infection also promoted the up-regulation of VEGFA and Snail-1, therefore further mediating the BBB disruption. Our observations reported here directly support the involvement of VEGFA and Snail-1 in meningitic E. coli induced BBB disruption and VEGFA and Snail-1 would therefore represent the essential host targets for future prevention of clinical E. coli meningitis.

Narendra Kumar Kata is currently pursuing his Masters in Plant Sciences at Wageningen University, Netherlands

Currently, biodiesel derived from rapeseed is classified as sustainable in European Union (EU) although, it has relatively low savings on greenhouse gas (GHG) emissions. Europe’s Renewable Energy Directive (RED), which was introduced in 2009, requires that the production and use of biofuels for transport save 35% or more on GHG emissions. From 2017, savings of 50% are required. Currently, rapeseed biodiesel counts for a 35% saving of GHG emissions. Therefore, we focus in this report on the possibilities to increase savings of GHG emissions related to the production of rapeseed. Moreover, Germany is Europe’s largest producer of rapeseed. Hence, the focus of this report is on sustainable rapeseed production for biodiesel in Germany. Considering that EU uses GHG emissions to qualify biofuel feedstock, changes in the crop should lead to lower emissions. Currently, the greatest share of the GHG emissions is related to the cultivation of rapeseed. Nitrogen and fossil fuel use are the major contributors to GHG emissions of rapeseed cultivation. More efficient production could reduce GHG emissions of rapeseed biodiesel significantly. Oils optimal for the production of biodiesel in Europe contain mostly oleic acid (C18:1). Rapeseed oil, however, contains long chain fatty acids and polyunsaturated fatty acids. This can be done by changing the fatty acid synthesis pathway in rapeseed by targeting the genes that regulate the production of enzymes responsible for the elongation and desaturation of the fatty acids. Oil and protein from rapeseed are produced in the seeds. Thus, the breeding targets focus on seed production content and composition of fat and protein in the seed. Main objectives of the study are seeds with high oil content (Hadrian), seeds with high C18:1 oil content (V140OL) and seeds with high protein content (Mentor). The selection of these targets should be done by genotyping. For the high oil and high C18:1 oil content, there are QTL’s known, which can be selected by marker assisted selection. Another strategy to reduce the GHG emissions is by improving the agronomy. According to the study by Zhang et al. (2012), by changing the plant density, the seed yield can increase. By optimizing the oil composition and by increasing the oleic acid concentration to approximately 85%, the GHG emission savings are increased with 5% with increased oil content in the seed, value of the rapeseed meal, applying more nitrogen. This results in 8% more GHG emissions savings. The 50% savings can only be reached by adding up all changes. It is, however, questionable if this will work in practices.

Jacqueline Mazzotti Cavalcanti da Silva is a graduate of The Federal University of Sergipe (UFS), a public institution of higher education in Brazil. After completing her Medical School in 2014, she has begun her training as a Resident in Gynecology and Obstetrics at The Federal University of Bahia, Brazil. Her research has focused on human papillomavirus infection and human sexuality and has been published in nationally recognized scientific journals.

Aim: Knowing the prevalence of reported cases of congenital syphilis in pregnant women and in the state of Sergipe between 2007 and June 2013. Methods: Analysis of cross-sectional, descriptive and retrospective component using only secondary data from the Information System for Notifiable Diseases, Ministry of Health of Brazil. The data collected refer to the period 2007 to June 2013. It was used SPSS 17.0 for data analysis. Results: It has reported 1450 cases of syphilis in pregnant women, 47% with latent syphilis, 30% with treatment considered inadequate, 42% of partners were not treated. As for congenital syphilis cases were reported in 1371, 48.5% female, 50% of mothers received treatment, 68.9% underwent prenatal, 48% received a diagnosis at the time of delivery or curettage. Conclusion: The analysis shows that, as in nationwide; the central problem of the high prevalence of vertical transmission of syphilis is the ineffectiveness of prenatal care offered to pregnant women.