In this study, the effect of hydrogen peroxide (H2O2) pre-treatment for sewage sludge prior to anaerobic digestion wasassessed using a batch test with an objective to decrease nitrogen, dissolved sulfide and siloxane in sewage sludge. Atotal of 6 sets of experiments (Blank, 20, 40, 60, 80 and 100g H2O2/kg wet sludge) were carried out, each with duplicates.To assess the effect of different dosages of H2O2 on anaerobic digestion, the treated sewage sludge was used for biochemical methane potential (BMP) test and SCODcr concentration. Due to the H2O2 pre-treatment, solubilization of SCODcr in pretreated sludge increased by 89% compared to raw sewage sludge, whereas T-N and NH3-N concentrationdecreased. Cumulative methane yields were increased for all pretreated samples due to increased sludge solubilizationthrough H2O2 pre-treatment. In addition, dissolved siloxane concnetrations were decreased for all pretreated samples. Thus,a reduction in dissolved siloxane concenrtation can decrease the siloxane generation potential of sludge during anaerobicdigestion. However, dissolved sulfide concentration remained same. Although H2O2 dosage did not show any furtherimpact on dissolved sulfide, they have significantly decreased T-N, NH3-N and dissolve siloxane concentrations beforeanaerobic digestion.

In order to accelerate the biodegradation of easily organic materials in landfilled waste before excavating a closed solid waste landfill and prevent to be dried the landfilled wastes at the same time, this study has suggested the Dual Step Biostabilization System (DSBS), which could inject air with dry fog into its body. In addition, the applicability of the DSBS was estimated by means of field test at a closed landfill. As a result of field test, the reduction of oxygen consumption rate for landfilled wastes (48%) stabilized by air with dry fog was higher than that of landfilled wastes (38%) stabilized by only air. Three lysimeter experiments were, also, performed for the landfilled wastes sampled from the closed landfill. The production of cumulative carbon dioxide for landfilled wastes stabilized by air with dry fog was estimated to be highest (1,144.8 mL). In case of lysimeter that moisture was not introduced was found to be 1,051.9 mL, while another lysimeter that moisture was introduced through horizental trenches was 1,095.8 mL. It is clear that the DSBS can accelerate the biodegradation of organic compounds. In terms of volatile solids, the reduction amount of volatile solids for air with dry fog was higher than that of the other conditions.

Pluripotent cells are categorized as either "naive" or "primed" based upon their pluripotent status. According to previous studies, embryonic stem cells and embryonic germ cells are identified as naive pluripotent stem cells and epiblast stem cells are identified as primed pluripotent stem cells. In a permissive species such as the mouse, naive and primed pluripotent stem cells can be derived from embryos without genetic manipulations. In non-permissive species such as humans and pigs, primed pluripotent cells are only established from embryos. However, previous studies have shown that the embryonic germ cells of non-permissive species share similar morphology and features with naive pluripotent cells. For these reasons porcine embryonic germ cells (pEGCs) may provide a useful cell source for comparative studies on naive pluripotent cells in non-permissive species. In this study, we attempted to establish and characterize porcine embryonic germ cells. Consequently, an embryonic germ cell line was derived from the genital ridges of a porcine dpc 30 fetus in media containing bFGF. This cell line displayed a dome-shaped colony morphology. The cell line was maintained in a stable condition over an extended time period and was able to differentiate into the three germ layers in vitro. Pluripotency markers such as OCT4, SOX2, NANOG and SSEA4 were expressed in these pEGCs. Similar with pESCs, Mek/Erk signaling pathway were activated by bFGF in the cultured pEGCs. In conclusion, we were able to successfully derive embryonic germ cells from genital ridges of a porcine fetus. Unlikely naive pluripotent cells such as mESCs, pluripotency of pEGCs were regulated by Mek/Erk signaling pathway activated by bFGF. This cell line could potentially be used as naive pluripotent cell source for comparative study with porcine embryonic stem cells and other pluripotent cell lines. As porcine pluripotent cells, pEGCs could be useful candidates for preliminary studies of human disease as well as a source for generating transgenic animals.

Pluripotent stem cells are cells that have a self-renewal capacity and the ability to differentiate into all lineages. These cells can be divided into naive- and primed-state pluripotent stem cells according to their pluripotent state. Only the naive state comprises a full pluripotency or ground state that contributes to germ-line transmission. Naive states are found in specific permissive strains or species, such as 129, C57BL/6 and BALB/C in mice. However, a number of attempts have been made to derive naive-state pluripotent stem cell lines from non-permissive species, including humans and pigs, using various exogenous factors including GSK3β and MEK inhibitors (2i), LIF, hypoxic conditions and up-regulation of Oct4 or Klf4. Therefore, in this study we investigated whether a naive pluripotent stem cell line could be derived from porcine embryonic fibroblasts (PEFs) via previously reported factors. Our mouse embryonic stem cell (mESC)-like cell lines expressed the pluripotency markers Oct4, Sox2 and Nanog and a stable mESC-like morphology for more than 50 passages. In addition, these cell lines could be sequentially reprogrammed into mESC-like induced pluripotent stem (iPS) cells from secondary or tertiary fibroblast-like cells differentiated from mESC-like iPS cells by addition of doxycycline (DOX), LIF and 2i. Our results suggest that, as a non-permissive species, porcine stem cells can be induced into mESC-like iPS cells from PEFs by various exogenous factors, including continuous transgene expression, 2i and LIF. However, further work that aims to effectively induce the activation of endogenous transcription factors is necessary to derive authentic naive-state pluripotent porcine stem cells.

The objective of this research was to suggest the estimation method of air injection quantity for pre-stabilization of landfilled wastes in a sustainable landfill. A study on the determination of oxygen demand quantity of landfilled wastes, therefore, was conducted in two different experiments. Firstly, a batch test was performed in order to measure the oxygen quantity required to oxidize easily degradable organic matter under aerobic conditions. Secondly, a lysimeter experiment was carried out to assess the air injection period according to moisture content (20%, 30%, 40%, 50%) and to validate the oxygen demand quantity obtained by the batch test. This study assumed that landfilled wastes contain two different organic matters and two matters are sequentially utilized by microorganism. The first one provides the faster oxygen uptake rate that called the “easily degradable organics”. During the second phase of the aerobic decomposition, the other one provides the slower oxygen uptake rate that called the “moderately degradable organics”. Also, in this study, a modified logistic equation divided two terms (fractions of easily degradable organic and moderately degradable organic) was suggested to determine the oxygen demand quantity for easily degradable organic of landfilled solid waste. As a result, the oxygen demand quantity obtained by the batch test led to similar results compared with that of lysimeter experiment. Therefore, it showed that the modified logistic equation and batch test were appropriate for determination of oxygen demand quantity for decomposition of easily degradable organic matter. Also, air injection period for decomposition of easily degradable organic decreased with the increase of moisture content.

In this study, anaerobic digestion tests for municipal solid wastes were performed in order to estimate their methane production rates. To simulate methane production rate, the first derivatives of the sigmoidal equations were calculated. The sigmoidal equations used were Gompertz and Logistic equations. Also, diauxic growth was employed to simulate methane production rate from municipal solid wastes. The equations were fitted to simulate methane production rate by nonlinear regression with a Marquardt-Levenberg algorithm. This is a statistical analysing methodology to minimize the sum of the squares of the differences between the predicted and measured values. Data fittings obtained by using the equations were compared statistically by using the RMSE (Root mean square error) and AIC (Akaike's information criterion). The equations derived from sigmoidal equations successfully described methane production rate for municipal solid wastes. RMSE and AIC were decreased significantly for the equations considering diauxic growth, compared to the equations without diauxic growth. These results showed that the equations derived from sigmoidal equations with diauxic growth can be a useful tool to simulate methane production rate for municipal solid wastes.

In this study, anaerobic co-digestion experiments for mixtures consisting of sewage sludge with food wastewater and livestock wastewater were conducted to assess the methane yields, the volatile solids (VS) removal rates and the dynamic kinetics. An augmented simplex centroid design (ASCD) was employed to design the mixing rate of organic wastes for the anaerobic co-digestion. Also, synergistic effects on the anaerobic co-digestion were studied using models obtained by the ASCD. As a result, synergistic effects were not observed in terms of methane yield and VS removal rate. It was just showed that there was a linear relationship between the cumulative methane yield and the mixing rate of food wastewater. The results might be attributable that the sewage sludge and the livestock wastewater had very lower C/N ratio compared with food wastewater that had a C/N ratio within a range required for a correctly operating anaerobic co-digestion. Therefore, increasing mixing rates of food wastewater increased the methane yield and VS removal rate, but there was not a synergistic effect by the anaerobic co-digestion.

In this study, optimization of anaerobic co-digestion for food and livestock wastes was studied by an experimental design method. A central composite design (CCD) was applied in designing experiments. Selected two independent variables for this study were initial substrate concentration and mixing rate of livestock wastes. The ranges of experiment for initial substrate concentration and mixing rate of livestock wastes were 2~10 g-VS/L and 0~100%, respectively. Selected responses were methane yield, maximum methane production rate and volatile solids (VS) removal rate. The experimental design was analyzed using a response surface methodology (RSM). Models obtained by the RSM were analyzed by analysis of variance (ANOVA). ANOVA demonstrated that the models were highly significant. Optimal conditions obtained for the models were initial substrate concentration of 2.1 g-VS/L and mixing rate of livestock wastes of 48.8%, respectively. The measured values under the optimal conditions were well in agreement with the predicted values from the models. Thus, it showed that the CCD and RSM were appropriate for determination of an optimal mixing condition in the anaerobic co-digestion process for food and livestock wastes.

The objective of this study was to assess the affects of various solid waste landfill methods on mass balance of carbon. Four lysimeters simulated a conventional landfill (Lys-A), a landfill recirculated only fresh leachate (Lys-B), and two landfills recirculated leachate after pretreating with ASBR (Lys-C and Lys-D) were operated over 1,600 days. Lys-D was recirculated two times of pretreated leachate volume than that of Lys-C. Mass balance of carbon was calculated considering leachate and biogas production for each lysimeter. Lys-C and Lys-D showed that there was an increase of about 3 times in total amount of COD recovered as methane than Lys-A. This results might be attributable to the activated methanogenic bacteria and the high pH of pretreated leachate. In terms of mass balance of carbon, amount of carbon converted to landfill gas in Lys-B (25.20 g/kg-dry waste) was bigger than that of Lys-A (23.64 g/kg-dry waste), while carbon conversion rate to landfill gas for Lys-A and Lys-B showed 4.80% and 4.71%, respectively. It is assumed that only fresh leachate recirculation method can increase amount of carbon converted to landfill gas resulting from the biodegradation of organic carbon in recirculated leachate. However, in comparison with the conventional landfill method, this method should not accelerate hydrolysis of carbon from the wastes. Carbon conversion rate in the landfill recirculated leachate after pretreating with ASBR was increased due to accelerated anaerobic metabolism processes of the microbes. In Lys-C and Lys-D, about 5.9% of carbon was converted to landfill gas. Therefore, it could be seen that the landfill recirculated leachate after pretreating with ASBR could enhance carbon conversion to landfill gas more than the conventional landfill or the landfill recirculated only fresh leachate.