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Exogenous DNA Transfer by Intracytoplasmic Sperm Injection in Porcine Oocytes (돼지에 있어서 난자내 정자 직접 주입에 의한 외래 유전자 도입에 관한 연구)

  • Ahn, S. Y.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.339-347
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    • 2001
  • Sperm-mediated DNA transfer has a potential to markedly simplify techniques for the generation of transgenic animals. The exogenous DNA transfer by intracytoplasmic sperm injection (ICSI) procedure has been recently introduced in the production of transgenic animals. In this study, the developmental competence and tile expression rates of transgene were investigated after injection of spermatozoon or sperm head with enhanced green fluorescent protein (EGFP) gene into the mature porcine oocytes. The porcine oocytes were injected with intact sperm, membrane-disrupted sperm or sperm head. After injection. embryos were cultured in NCSU23 medium up to the blastocyst stage, and the developmental competence and expression rates were studied. The developmental rate (67.0%) of sperm injection group was higher than that (59.7%) of sperm head injection group, and the rates of EGFP expression were also significantly different between sperm injection and sperm head injection groups (42.1 vs 20.0%) (F<0.05). In the porcine oocytes injected with sperm treated with different methods of membrane disruption, the removal of sperm membrane did not alter the developmental competence of embryos. The rate of blastocysts at 7 days after injection with intact and membrane disrupted sperm were 15.0 and 14.2%, respectively. The EGFP expression rates, 38.4% in embryos injected with frozen-thawed sperm was higher than that, 22.4% of embryos injected with the Triton X-100 treated sperm. Prior to injection, sperm were cultured in different EGFP gene concentrations from 0.Ol to 1ng/u${mu}ell$. However, no significant difference in developmental rates of embryos among different concentrations of EGFP gene were observed. The highest expression rate of EGFP gene, 37.4% was obtained from the embryos injected with spermatozoa treated with 0.1 ng/${mu}ell$ EGFP gene. These results suggested that exogenous DNA could be attached to the membrane disrupted sperm, and that these sperm could be used as a vector carrying foreign DNA into embryos.

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Comparison of Indoor CO2 Removal Capability of Five Foliage Plants by Photosynthesis (다섯가지 관엽식물의 광합성에 의한 실내 이산화탄소 제거능력 비교)

  • Park, Sin-Ae;Kim, Min-Gi;Yoo, Mung-Hwa;Oh, Myung-Min;Son, Ki-Cheol
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.864-870
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    • 2010
  • This study was conducted to determine the effects of foliage plants on reducing indoor carbon dioxide ($CO_2$). Five foliage plants such as $Hedera$ $helix$ L., $Ficus$ $benjamina$ L., $Pachira$ $aquatica$, $Chamaedorea$ $elegans$, and $Ficus$ $elastica$ were selected and cultivated in two different growth medium (peatmoss and hydroball). Each plant was placed in an airtight chamber and then treated with the combinations of two different $CO_2$ concentrations (500 or 1,000 ppm) and two different light intensities (50 or $200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$). The change of $CO_2$ concentration (ppm) in the airtight chamber during day and night was measured and then converted into the photosynthetic rate (${\mu}mol\;CO_2{\cdot}m^{-2}{\cdot}s^{-1}$). As the results, each foliage plant reduced $CO_2$ level in the airtight chamber for one hour by photosynthesis. $Pachira$ $aquatica$ and $Ficus$ $elastica$ absorbed $CO_2$ more effectively compared to the other plants. The plants exposed to higher $CO_2$ concentration (1,000 ppm) and higher light intensity ($200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) showed more effective $CO_2$ elimination rate and photosynthetic rate. The plants that have wide leaves and big leaf areas such as $Pachira$ $aquatica$, $Hedera$ $helix$ L.,and $Ficus$ $elastica$ showed higher photosynthetic rate than the other plants that have smaller leaves. Released $CO_2$ concentration by respiration of the plants during the night was very low compared to the absorbed $CO_2$ concentration by photosynthesis during the day. There was no significant difference between peatmoss and hydroball medium on reducing $CO_2$ concentration and increasing photosynthetic rate. In conclusion, this study suggested that foliage plants can effectively eliminate indoor $CO_2$. Optimum environmental control in relation to photosyntheis and usage of right indoor foliage plants having lots of leaves and showing active photosynthesis even under low light intensity like indoor light condition would be required to increase the elimination capacity of indoor $CO_2$.

Dry Weight and Nitrogen Contents in Different Parts of 'Fuyu' Persimmon as Affected by Application Timing and Methods of Supplemental Nitrogen (질소 추비 시기와 방법이 '부유' 단감나무의 건물중과 질소함량 및 수체 부위별 분포에 미치는 영향)

  • Park, Doo-Sang;Choi, Seong-Tae;Kang, Seong-Mo
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.728-734
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    • 2010
  • We studied the accumulation and partitioning of dry weight (DW) and nitrogen (N) in different parts of field-grown 'Fuyu' persimmon to elucidate that the foliar applications of supplemental N in June or September compared favorably with the traditional soil application in securing leaf area and fruit production. We also estimated the proportion of N permanently removed from the tree at the end of a growing season. Urea was applied either to leaves in June and/or September or to the soil in June and September for three consecutive years, and the trees were excavated in November for analyses. Total DW ranged from 4.2-4.8, 8.7-9.2, and 17.1-21.5 kg in a 4-, 5-, and 6-year-old tree, respectively, without statistical difference among the four treatments. Of the total DW, 3.3-10.2% was in shoots, 5.7-10.5% in leaves, 8.3-31.4% in aerial woods, 13.0-27.0% in root, and 28.0-59.3% in fruits. As the trees became more productive, DW proportion of fruits significantly affected that of the root: in 6-year-old trees, root DW accounted for only 10.6-15.8% of the tree total when fruit DW accounted for 50-60%. N contents ranged from 24.6-28.3, 48.3-53.5, and 98.3-122.6 g in a 4-, 5-, and 6-year-old trees, respectively, without statistical difference among the treatments. Of the total N, 6.2-11.5% was in shoots, 16.7-24.3% in leaves, 17.6-23.5% in aerial woods, 17.2-37.5% in roots, and 16.9-34.4% in fruits. As in DW, the increase in the proportion of N in fruits decreased in the root most significantly. Application methods for supplemental N did not affect the proportion of DW and N removed from the tree through abscising leaves and harvested fruits. Percentage of DW removal was 41 in 4- and 5-year-old trees, but it was 61 in more productive 6-year-old trees; that of N was 39, 43, and 49%, respectively. No significant changes in the contents of DW and N in field-grown trees, as well as their percentages removed from the tree at the end of the season, demonstrated that foliar application of supplemental N was as good as soil applications with much less N.

Hedgehog Oral Viral Papilloma (고슴도치 구강유두종)

  • Elfadl, Ahmed K.;Lee, Eun-Joo;Kim, Hyeong-Mi;Kim, Yong Deuk;Chung, Myung-Jin;Park, Jin-Kyu;Kim, Tae-Hwan;Kim, Shin-Yoon;Ghim, Soong-Gu;Kim, Jun Young;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.27 no.5
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    • pp.575-578
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    • 2017
  • Here we describe the histological lesion of a viral fibro-papilloma in a hedgehog. After surgical removal from maxilla, the solitary swollen mass was round to oval, yellowish with rough surface and measuring $6{\times}3{\times}2mm$ approximately. The tumor mass was submitted to the laboratory of pathology, college of veterinary medicine, Kyungpook National University for pathological diagnosis. Pathological examination of the tumor was established, the tumor was described grossly and sample was trimmed, sectioned and routinely prepared for histopathological evaluation. The tumor mass was diagnosed as viral fibro-papilloma, as the histological picture showed characteristic features of warts caused by papillomaviruses. The tumor characterized by thickening of the stratum spinosum (acanthosis) and basale, koilocytosis, intra-nuclear inclusion bodies in keratinocytes and fibrosis of submucosa. Further, viral inclusion bodies were demonstrated by Machiavello stain giving red color to the nuclei. No lymphocytes that responsible for regression of the wart could be detected, suggesting the poor possibility of spontaneous regression of the tumor. Papillomatosis is a disease of young animals, but in our case the infected Hedgehog was 5 years old, that maybe due to an impaired immune system, which is also shown by absence of lymphocytes. To the best knowledge of the author, this case presents the first report of viral fibropapillomatosis in Hedgehog.

Effects of Treadmill Exercise on Alpha-synuclein Mutation and Activated Neurotrophins in Nigrostriatal Region of MPTP-induced Parkinson Models (MPTP 파킨슨 모델의 트레드밀 운동이 알파시누크린 변성과 흑질선조체내 신경성장인자 활성화에 미치는 영향)

  • Park, Jae-Sung;Kim, Jeong-Hwan;Yoon, Sung-Jin
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.2
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    • pp.73-88
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    • 2009
  • Objectives : Neuronal changes that result from treadmill exercise for patients with Parkinson's disease(PD) have not been well documented, although some clinical and laboratory reports suggest that regular exercise may produce a neuroprotective effect and restore dopaminergic and motor functions. However, it is not clear if the improvements are due to neuronal alterations within the affected nigrostriatal region or result from a more general effect of exercise on affect areas and motivation. In this study, we demonstrate that motorized treadmill exercise improves the neuronal outcomes in rodent models of PD. Methods : We used a chronic mouse model of parkinsonism, which was induced by injecting male C57BL/6 mice with 10 doses(Every 12 hour) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (30 mg/kg) and probenecid (20 mg/kg) over 5 days. These mice were able to sustain an exercise training program on a motorized rodent treadmill at a speed of 18 m/min, $0^{\circ}$ of inclination, 40 min/day, 5 days/week for 4 weeks. At the end of exercise training, we extracted the brain and compared their neuronal and neurochemical changes with the control(saline and sedentary) mice groups. Synphilin protein is the substance that manifestly reacts with ${\alpha}$-synuclein. In this study, we used Synphilin as a manifest sign of recovery from neurodegeneration. We analyze the brain stems of the substantia nigra and striatum region using the western blotting technique. Results : There were no expression of synphilin in the saline-induced groups. The addition of MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) greatly accelerated synphilin expression which meant an aggregation of ${\alpha}$-synuclein. But, the MPTP-induced treadmill exercise group showed significantly lower expression than the MPTP-induced sedentary group. This means treadmill exercise has a definite effect on the decrease of ${\alpha}$-synuclein aggregation. Conclusions : In this study, our results suggest that treadmill exercise promoted the removal of the aggregation of ${\alpha}$-synuclein, resulting in protection against disease development and blocks the apoptotic process in the chronic parkinsonian mice brain with severe neurodegeneration.

THE SECOND MESIOBUCCAL CANAL OF UPPER PRIMARY MOLAR : CASE REPORT (제 2근심협측근관이 존재하는 상악유구치의 치험례)

  • Kim, Chang-Gi;Hong, Seong-Soo;Ko, Sung-Back;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.2
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    • pp.139-145
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    • 2002
  • In a primary teeth, dental caries is rapidly advanced the pulp disease, because the primary teeth have the thinner and the weaker enamel layer and the wider pulp chamber than relatively the permanent teeth. And the pulps of primary teeth are exposed during caries removal or even they are exposed by unexpected movement of the children or by trauma. For successive pulp treatment in primary teeth, it is necessary to understand completely about multiple canal morphology, variation of root canal anatomy and specific problem related to root formation and resorption of primary teeth. In upper primary molar, canal configuration of mesial root has the most variation same as upper molar. If not canal treatment is completely, most of all endodontic treatment should be failed. In a clinical case report, upper primary molars existed persistent pain or bleeding during treatment were founded the second MB canal and were performed the endodontic treatment of theirs. As a result, the upper primary molars have no symptom and good prognosis. In the examination of extracted upper primary molar, we found that 8 of 35 teeth(22.8%) in the upper primary first molars and 22 of 33 teeth(66.6%) in the upper primary second molar had the second mesiobuccal canal. It has revealed the high prevalence of two canals in mesiobuccal roots of upper primary molars. The frequency of occurrence of the second mesiobuccal canal must be taken into consideration when endodontic treatment is planned and as a possible cause of otherwise un explained failure.

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Evaluation of Cleanness and Physical Properties of W/O Microemulsion (W/O Microemulsion 세정제의 물성 및 세정성 평가)

  • Lee, Myung Jin;Han, Ji Won;Lee, Ho Yeol;Han, Sang Won;Bae, Jae Heum;Park, Byeong Deog
    • Korean Chemical Engineering Research
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    • v.40 no.6
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    • pp.769-777
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    • 2002
  • Using four components - nonionic surfactants, water, hydrocarbon oil and an alcohol as cosurfactant, 12 types of cleaning agents were prepared, and their physical properties such as surface tension, viscosity, electroconductivity and phase stability were measured. As the formulated cleaning agents have low surface tensions(30.5-31.1 dyne/cm) and low viscosities (1.6-7.2 c.p.), they are satisfied with the general physical properties of water-in-oil(W/O) microemulsions for their industrial use. They showed a tendency that their temperature range for stable one-phase microemulsion decreased in accordance with the increase of alcohol/surfactant(A/S) ratio in the formulations. However, the temperature range of one-phase microemulsion was much more affected by hydrophilic lipophillic balance(HLB) value of the nonionic surfactant which increased its temperature range and it increased in accordance with the higher HLB value in the formulations. And the maximum content of water which can keep stable one-phase W/O microemulsion was measured at each sample. In addition, their temperature range for stable one-phase microemulsion was also measured. It was confirmed that the selection of surfactant type was very important for formulating a cleaning agent, since the W/O microemulsion system with the nonionic surfactant of the lower HLB value showed better cleaning efficacy that of the higher HLB value for abietic acid as a soil, which was used for preparing a rosin-type flux. In the formulated cleaning agents with the increase of A/S ratio in the formulations, however, there was no significant difference in cleaning efficacy. It was investigated that the differences of their cleaning efficacy was affected by the change of the condition of temperature and sonicating frequency as important factors in the industrial cleaning. That is, the higher, their cleaning temperature and the lower, their sonicating frequency, the more increased, their cleaning efficacy. Furthermore, using optical instruments like UV/Visable Spectrophotometer and FT-IR Spectrometer, their cleaning efficacy for abietic acid was measured. The removal of soil from the contaminated rinse water was measured by gravity separation method in the rinse bath. As a result, the cleaning agent system having the nonionic surfactant of HLB value 6.4 showed over 85% water-oil separation efficacy at over $25^{\circ}C$. Therefore, it was demonstrated in this work that the formulating cleaning agents were very effective for cleaning and economical in the possible introduction of water recycling system.

Disinfection of Fusarium-infected Rice Seeds by Prochloraz and Gaseous Chlorine Dioxide

  • Jeon, Young-ah;Lee, Young-yi;Lee, Ho-sun;Sung, Jung-sook;Lee, Seokyoung
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.25-25
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    • 2014
  • Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

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Study on the Manufacturing techniques & Conservation of Iron Pot from Cheonmachong Ancient Tomb (천마총 출토 철부(鐵釜)의 제작기법 및 보존처리)

  • Lee, Seung Ryul;Shin, Yong Bi;Jung, Won Seob
    • Journal of Conservation Science
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    • v.30 no.3
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    • pp.263-275
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    • 2014
  • It's shown how to proceed the study on Manufacturing techniques & Conservation to the Iron Pot from Cheonmachong Ancient Tomb(the 155th Tomb in Hwangnam-dong). In order to investigate manufacturing techniques of the Iron Pot, some parts of the relic were gathered. After mounting, polishing and etching on the relic, analyzing the metal microstructure was conducted. Also it's conducted a SEM-EDS analysis on the nonmetallic inclusion. White iron structure was observed in the metallurgical structure inspection, SEM-EDS analysis. It seems to be dried slowly at room temperature after casting, doesn't look as particular heat treatment to improve brittleness. It is estimated that it's as the handle seam side were verified about 3cm inch wide, 1.5 thick in center of body, so 2 separate half-completed products was cast with width-type mould. The manufacturing techniques Using white cast iron structure, width-type mould are observable to the Iron Pot excavated from Sikrichong Ancient Tomb & Hwangnamdaechong grand Ancient Tomb around those were constructed the same time. It's able to recognize that it's almost identical manufacturing techniques at that time. Conservation is generically following those are survey of pretreatment, foreign material removal, stabilization, restoration and color matching in the order. cleaning & drying were added to the process as occasion demands. The strengthening treatment were difficult with artifact's volume, low concentration Paraloid NAD-10 solution was spread two or three times with a brush, surface hardening also came up with 15wt% Paraloid NAD-10 solution after the conservation was complete. There were connection & restoration for the restoration to the damage after modeling forms that it's similar to damaged parts by using the Fiber Reinforced Plastic resins(POLYCOAT FH-245, mold laminated type). Throughout this research, capitalizing on accumulations of measurements about the production technique of Iron Pot in the time of the fifth and 6th centuries is no less important than the Iron artifact's conservation for a better study in the future.

Studies on a Feasibility of Swine Farm Wastewater Treatment using Microbial Fuel Cell (미생물연료전지의 가축분뇨 처리 가능성 연구)

  • Jang, Jae-Kyung;Kim, Se-Hee;Ryou, Young-Sun;Lee, Sung-Hyoun;Kim, Jong-Gu;Kang, Young-Goo;Kim, Young-Hwa;Choi, Jung-Eun
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.461-466
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    • 2010
  • In this study the feasibility of simultaneous electricity generation and treatment of swine farm wastewater using microbial fuel cells (MFCs) was examined. Two single-chamber MFCs containing an anode filled with different ratio of graphite felt and stainless-steel cross strip was used in all tests. The proportion of stainless-steel cross strip to graphite felt in the anode of control microbial fuel cell (CMFC) was higher than that of swine microbial fuel cell (SMFC) to reduce construction costs. SMFCs produced a stable current of 18 mA by swine wastewater with chemical oxygen demand (COD) of $3.167{\pm}80\;mg/L$ after enriched. The maximum power density and current density of SMFCs were $680\;mW/m^3$ and $3,770\;mA/m^3$, respectively. In the CMFC, power density and current density was lower than that of SMFC. CODs decreased by the SMFC and CMFC from $3.167{\pm}80$ to $865{\pm}21$ and $930{\pm}14\;mg/L$, achieving 72.7% and 70.6% COD removal, respectively. The suspended solid (SS) of both fuel cells was also reduced over 99% ($4,533{\pm}67$ to $24.0{\pm}6.0\;mg/L$). The concentration of nutritive salts, ${NH_4}^+$, ${NO_3}^-$, and ${PO_4}^{3-}$, dropped by 65.4%, 57.5%, and 73.7% by the SMFC, respectively. These results were similar with those of CMFC. These results show that the microbial fuel cells using electrode with mix stainless-steel cross strip and graphite felt can treat the swine wastewater simultaneously with an electricity generation from swine wastewater.