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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Management of Critical Control Points to Improve Microbiological Quality of Potentially Hazardous Foods Prepared by Restaurant Operations (외식업체에서 제공하는 잠재적 위험 식품의 미생물적 품질향상을 위한 중점관리점 관리방안)

  • Chun, Hae-Yeon;Choi, Jung-Hwa;Kwak, Tong-Kyung
    • Korean journal of food and cookery science
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    • v.30 no.6
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    • pp.774-784
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    • 2014
  • The purpose of this study was to present management guidelines for critical control points by analyzing microbiological hazardous elements through screening Potentially Hazardous Foods (PHF) menus in an effort improve the microbiological quality of foods prepared by restaurant operations. Steamed spinach with seasoning left at room temperature presents a range of risk temperatures which microorganisms could flourish, and it exceeded all microbiological safety limits in our study. On the other hand, steamed spinach with seasoning stored in a refrigerator had Aerobic Plate Counts of $2.86{\pm}0.5{\log}\;CFU/g$ and all other microbiological tests showed that their levels were below the limit. The standard plate counts of raw materials of lettuce and tomato were $4.66{\pm}0.4{\log}\;CFU/g$ and $3.08{\pm}0.4{\log}\;CFU/g$, respectively. Upon washing, the standard plate counts were $3.12{\pm}0.6{\log}\;CFU/g$ and $2.10{\pm}0.3{\log}\;CFU/g$, respectively, but upon washing after chlorination, those were $2.23{\pm}0.3{\log}\;CFU/g$ and $0.72{\pm}0.7{\log}\;CFU/g$, respectively. The standard plate counts of baby greens, radicchio and leek were $6.02{\pm}0.5{\log}\;CFU/g$, $5.76{\pm}0.1{\log}\;CFU/g$ and $6.83{\pm}0.5{\log}\;CFU/g$, respectively. After 5 minutes of chlorination, the standard plate counts were $4.10{\pm}0.6{\log}\;CFU/g$, $5.14{\pm}0.1{\log}\;CFU/g$ and $5.30{\pm}0.3{\log}\;CFU/g$, respectively. After 10 minutes of chlorination treatment, the standard plate counts were $2.58{\pm}0.3{\log}\;CFU/g$, $4.27{\pm}0.6{\log}\;CFU/g$, and $4.18{\pm}0.5{\log}\;CFU/g$, respectively. The microbial levels decreased as the time of chlorination increased. This study showed that the microbiological quality of foods was improved with the proper practices of time-temperature control, sanitization control, seasoning control, and personal and surface sanitization control. It also presents management guidelines for the control of potentially hazardous foods at the critical control points in the process of restaurant operations.

Development of Calcification-resistant Bovine Pericardium with PEO-$SO_3(II)$ -An implantation study of bovine pericardium at artery and peritoneum- (PEO-$SO_3$를 이용한 항석회화 조직첨포의 개발(II) -동맥과 복막 이식 실험연구-)

  • 김형묵;백만종;김광택;이인성;김학제;이원규;박기동
    • Journal of Chest Surgery
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    • v.31 no.11
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    • pp.1023-1030
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    • 1998
  • Background: Calcific degeneration limits durabilities of the bioprosthetic tissues implanted in the human body. The direct coupling sulphonated polyethyleneoxide(PEO-SO3) to the bioprosthetic tissues after glutaraldehyde(GA) fixation and the removal of residual aldehyde groups from the tissues can augment the effect of calcification-resistance. Materials and methods: To study the anti-calcification effect by PEO-SO3 modification and the removal of the residual aldehyde groups of tissues, surface modified bovine pericardia(BP-PEO-SO3) were preserved in aseptic saline to wash out GA(saline group) and 0.65% GA solution(GA group). And then above two groups and PERIGUARD (Bio-vascular. Co.) (product group) were evaluated with respects to calcium contents and microscopic findings using in vivo implantation models at carotid and femoral artery and peritoneum of 8 adult dogs. Results: In the tissues retrieved from carotid artery, calcium content was significantly decreased in saline group than in other two groups(saline; 2.89±0.31 vs. GA; 6.14±1.08 vs. product; 22.82±5.00 mg/g of dried tissue; p<0.05). In the tissues retrieved from femoral artery and peritoneum, calcium amount was also decreased in saline group than in other two groups, but not reached the significant difference between groups. On the other hand, the pathologic findings of pericardial tissues showed marked destructuction in GA group compared to the other two groups. Conclusions: In this study, covalently PEO-SO3 bound to bovine pericardium decreased calcifications and the anti-calcification effect of BP-PEO-SO3 could be augmented by the washing out the residual aldehyde groups using saline after GA fixation. Conclusively, the PEO-SO3 modified bovine pericardium is highly resistant to calcification and can be useful for the development of calcification-resistant cardiovascular patches and valves.

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Changes in Quality Characteristics of Wild Root Vegetables during Storage (전처리 근채류의 저장과정중의 품질평가)

  • Kwak, Soo-Jin;Park, Na-Yoon;Kim, Gi-Chang;Kim, Haeng-Ran;Yoon, Ki-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1158-1167
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    • 2012
  • During the peeling, cutting, and shredding of wild root vegetables, the surface is exposed to air. This results in a rapid deterioration in quality and an increase in the potential of contamination by microorganisms, both of which reduce the shelf-life of wild root vegetables in retail markets. Thus, in this study, the effects of various washing treatments on the quality of wild root vegetables, including lotus root, burdock root, and bellflower root, were investigated at 10 and $24^{\circ}C$. Lotus root, burdock root and bellflower root were washed with 0.2% acetic acid (AA), 0.2% citric acid (CA), 500 ppm acidified sodium chlorite (ASC), and tap water (TW), which was used as a control, and stored at $10^{\circ}C$ and $24^{\circ}C$. The changes in total plate counts, coliform groups, polyphenol oxidase (PPO) activity, color, pH, and exterior appearance of the samples were then evaluated. The pH and initial microbial contamination levels were reduced when the root vegetables were washed with AA, CA, and ASC. In particular, initial population levels of total plate counts and coliform groups were not detected in lotus root and burdock root that had been washed with ASC and their growth was significantly (p<0.05) inhibited during storage at 10 and $24^{\circ}C$ when compared to the control (TW). In addition, the polyphenol oxidase (PPO) activities of the root vegetables washed with AA, CA and ASC were lower than that of root vegetables washed with TW. ASC was determined to be the most effective treatment for preventing microbial growth, tissue softening, and the development of browning and an unpleasant smell. At $10^{\circ}C$, the overall qualities of the wild root vegetables were maintained longer when compared to $24^{\circ}C$.

A study on the shelf-life extension of fresh-cut onion (Allium cepa L.) (신선편의 양파(Allium cepa L.)의 유통기한 연장연구)

  • Lee, Kyoung-Hae;Kim, Dong-Ho
    • Journal of Food Hygiene and Safety
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    • v.24 no.4
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    • pp.324-331
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    • 2009
  • Peeled whole onions (PWO) were cleaned at various hypochlorous acid (HA) concentration and steeping time and packed in LDPE bag keeping at $10^{\circ}C$ for 12 days and $35^{\circ}C$ for 3 days, in order eventually to examine microbiology, surface color and sensory quality. At the early stage of storage, it was found that total bacterial counts at H-II keeping at $10^{\circ}C$ after 1 minute steeping were $2.60\;{\pm}\;0.18\;log\;CFU/g$, and those after 3 minutes steeping were $2.10\;{\pm}\;0.18\;log\;CFU/g$ which showed less than the control. The total bacterial counts at H-III were detected after 4 days. The total bacterial counts of PWO treated HA increased as steeping time became longer, HA concentration increased, and storage temperature went down. E. coli was not detected at all treatments. It was also found that during the treatment the L-value showed decreasing trend, but the parameter a- and b- value showed increasing trend. But these trends were mitigated as HA concentration increased. The result of sensory quality evaluation for the appearance showed that the sample stored with $10^{\circ}C$ gained higher evaluation than that with $10^{\circ}C$, while the control and H-III gained highest points significantly (p < 0.05) for the sample keeping at $10^{\circ}C$ after 12 days storage. The sensory odor of onion showed similar to that for the appearance, and the 8-day treatments of H-II and H-III showed no significantly difference (p < 0.05). On the basis of the results above, it is likely to be more effective to prolong the period of circulation of PWO if you use HA over 50 ppm for washing PWO and storage at $10^{\circ}C$. This study will contribute to improve safety and quality in circulation of PWO.

Change of Particle Size of Magnesium Silicate According to Reaction Conditions and Evaluation of Its Polyol Purification Ability (반응 조건에 따른 규산마그네슘의 입도 변화 및 폴리올 정제 능력평가)

  • Yoo, Jhongryul;Jeong, Hongin;Kang, Donggyun;Park, Sungho
    • Korean Chemical Engineering Research
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    • v.58 no.1
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    • pp.84-91
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    • 2020
  • The efficiency of the synthetic magnesium silicate used in basic polyols and edible oil purification is evaluated by its purification ability and filtration rate and is affected by the particle size and surface area of magnesium silicate. In this study, it was investigated the change on the particle size of magnesium silicate was influenced by the reaction temperature, injection rate, injection order (Si, Mg) and Mg/Si reaction mole ratio. The synthesized magnesium silicate was compared and analyzed for the synthesis, grinding, and refining processes. In the synthesis process, the reaction temperature and feed rate did not affect the average particle size change of magnesium silicate, while the reaction molar ratio of Mg / Si and the order of injection acted as main factors for the change of average particle size. The average particle size of magnesium silicate increased by 8.7 ㎛ from 54.4 ㎛ to 63.1 ㎛ at Mg injection when Mg molar ratio increased from 0.125 to 0.500, and increased by about 4.8 ㎛ from 47.3 ㎛ to 52.1 ㎛ at Si injection. The average particle size according to the order of injection was 59.1 ㎛ for Mg injection and 48.4 ㎛ for Si injection and the difference was shown 10.7 ㎛, therefore the filtration rate was about 2 times faster under the condition of Mg injection. That is, as the particle size increases, the filtration time is shortened and washing filtration rate can be increased to improve the productivity of magnesium silicate. The cake form of separated magnesium silicate after filtration becomes a solid through drying process and is used as powdery adsorbent through the grinding process. As the physical strength of the dried magnesium silicate increased, the average particle size of the powder increased and it was confirmed that this strength was affected by the reaction molar ratio. As the reaction molar ratio of Mg / Si increased, the physical strength of magnesium silicate decreased and the average particle size after grinding decreased by about 40% compared to the average particle size after synthesis. This reduction of strength resulted in an improvement of the refining ability due to the decrease of the average particle size and the increase of the amount of fine particle after the pulverization, but it resulted in the decrease of the purification filtration rate. While the molar ratio of Mg/Si was increased from 0.125 to 0.5 at Mg injection, the refining ability increased about 1.3 times, but the purification filtration rate decreased about 1.5 times. Therefore, in order to improve the productivity of magnesium silicate, the reaction molar ratio of Mg / Si should be increased, but in order to increase the purification filtration rate of the polyol, the reaction molar ratio should be decreased. In the synthesis parameters of magnesium silicate, the order of injection and the reaction molar ratio of Mg / Si are important factors affecting the changes in average particle size after synthesis and the changes of particle size after grinding due to the changes of compressive strength, therefore the synthetic parameter is an important thing that determines productivity and refining capacity.

Cooling Properties and Quality Changes during Storage of Citron (Citrus junos) (유자의 냉각특성 및 저장중 품질변화)

  • Jeong, Jin-Woong;Lee, Young-Chul;Kim, Jong-Hoon;Kim, Oni-Woung;Nahmgung, Bae
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1071-1077
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    • 1996
  • Quality changes in citron (Citrus junos) during storage were studied to investigate the efficiency, cooling properties and the washing and storage effects of hydrocooling method. As a result of plotting the nondimensionalized citron temperature versus cooling time, its cooling rate coefficient was shown to be $-0.012\;min^4{\sim}\;-0.017\;min^4\;(R^2=0.97{\sim}0.99)$ at center, and to be $-0.033\;min^4{\sim}\;0.075\;min^4\;(R^2=0.89{\sim}0.93)$ at surface. During storage, weight loss was more affected by storage temperature than by pretreatment condition and in reached $22{\sim}23%$ after 7 weeks at $15^{\circ}C\;and\;10{\sim}11%$ after 8 weeks at $5^{\circ}C$ in all samples. However, changes in moisture contents of hydrocooled citron were shown to be about $1{\sim}2%$ after 7 weeks while that of non-treated citron was about 3% after 1 week of storage at $5^{\circ}C$. And the change of pH, acidity and soluble solid content were not significantly different between each treatments during storage $5^{\circ}C\;and\;15^{\circ}C$. Changes in Hunter L, a, and b values of hydrocooled citron were lower than those of non-treated one as the storage time increased. The respiration rate of hydrocooled citron during storage at $15^{\circ}C$ was $103.63\;mg{\cdot}CO_2/kg{\cdot}hr$, which is about 50% of that of non-treated citron.

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The Effect of Rootzone Mix and Compaction on Nitrogen Leaching in Kentucky bluegrass (토양의 종류와 답압이 켄터키블루그래스 토양층에서 질소용탈에 미치는 영향)

  • Lee, Sang-Kook;Frank, Kevin W.;Crum, James R.
    • Asian Journal of Turfgrass Science
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    • v.24 no.1
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    • pp.45-49
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    • 2010
  • Research on nitrate-nitrogen ($NO_3-N$) leaching in turfgrass indicates that in most cases leaching poses minimal risk to the environment. Although there have been many studies investigating $NO_3-N$ leaching, there has been little research to investigate the effect of compaction level and rootzone mix on nitrogen (N) leaching. The research objective is to determine the effect of compaction level and rootzone mix on nitrogen leaching. The four rootzone mixes are 76.0:24.0, 80.8:19.2, 87.0:13.0 and 93.7:6.3 % (sand:soil). The four levels of compaction energies are 1.6, 3.0, 6.1, and 9.1 J $cm^{-2}$. Nitrogen was applied using urea at a rate of 147 kg $ha^{-1}$ split among three applications. Rootzone was packed into a polyvinylchloride pipe with a perforated bottom to facilitate drainage. Rootzone depth was 30 cm over a 5 cm gravel layer. Each column was sodded with Poa pratensis L. Hoagland solution designed for coolseason grasses, minus N, was used to ensure adequate nutrition in the rootzone. Turf grass quality and clipping yield were recorded from each tube at two-week intervals. The clippings were oven-dried at a temperature of $67^{\circ}C$ for 24 h and weighed. At the end of the study, root dry weight was determined by washing and oven-drying samples at $67^{\circ}C$ for 24 h. Leachate solution was collected weekly for analysis. More than 6.1 J $cm^{-2}$ of compaction energy increased possibilities of surface runoff. The compaction energy between 3.0 and 6.1 J $cm^{-2}$ produced more clipping dry weight and less N leaching than 9.1 J $cm^{-2}$.

Preparation of Mesoporous Titanium Oxides by Template Synthesis and Phase Transition of TiO2 inside Mesoporous Silica (주형합성을 통한 메조포러스 TiO2 제조 및 실리카 메조포어 내부에서의 TiO2 상전이 거동 변화)

  • Bang, Gyeong-Min;Kim, Young-Ji;Kim, Seung Han;Choi, Yerak;Lee, In Ho;Ko, Chang Hyun
    • Korean Chemical Engineering Research
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    • v.56 no.2
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    • pp.261-268
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    • 2018
  • To prepare mesoporous $TiO_2$ ($meso-TiO_2$) with anatase and rutile crystal structures, hydrothermal and template synthesis were used. $Meso-TiO_2$ with anatase structure was obtained by hydrothermal synthesis. The crystal structure of $meso-TiO_2$ by hydrothermal synthesis converted from anatase to rutile by simple heating at $600^{\circ}C$ and above. However, their mesopore structure collapsed due to phase transition. To prepare $meso-TiO_2$ with rutile structure, template synthesis method was applied using mesoporous silica KIT-6 as a template. Once we incorporated anatase $TiO_2$ inside mesopores of silica, the phase transition temperature of $TiO_2$ confined inside KIT-6 was much higher ($900^{\circ}C$) than that of free-standing $TiO_2$ ($600^{\circ}C$). The suppression of $TiO_2$ phase transition inside mesopores of KIT-6 is closely related with the interaction between $TiO_2$ surface and silica walls in KIT-6 because oxygen vacancy in $TiO_2$ is regarded as the starting point for phase transition. After removal of silica template by NaOH solution washing, $meso-TiO_2$ with mixed phase between anatase and rutile was obtained.

Investigations on the Adsorption Characteristics of $SO_2$ Gas on Fixed Bed Manganese Nodule Column (고정(固定) 흡착층(吸着層)에서 망간단괴(團塊)의 $SO_2$ 가스 흡착(吸着) 특성(特性)에 관한 연구(硏究))

  • Baek, Mi-Hwa;Kim, Dong-Su;Jung, Sun-Hee;Park, Kyoung-Ho
    • Resources Recycling
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    • v.15 no.4 s.72
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    • pp.3-12
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    • 2006
  • The feasibility for the employment of manganese nodule as an adsorbent for $SO_{2}$ gas has been investigated. The specific surface area of manganese nodule particle, which used in the experiments, was ca. $221.5m^{2}/g$ and the content of sulfur in manganese nodule was observed to significantly increase after $SO_{2}$ was adsorbed on it. The EPMA for the distilled water-washed and methanol-washed manganese nodule particle after $SO_{2}$ adsorption showed that its sulfur content was slightly decreased to 14.7% and 13.1% respectively, from 15.4% before washing. The XRD analysis of manganese nodule showed that todorokite and birnessite, which are manganese oxides, and quartz and anorthite were the major mineralogical components and weak $MnSO_{4}$ peaks were detected after $SO_{2}$ was adsorbed on manganese nodule. For an comparative investigation, limestone was also tested as an adsorbent for $SO_{2}$, however, no peaks for $CaSO_{4}$ were found by XRD analysis after the adsorption of $SO_{2}$. As the size of adsorbent increased, time for breakthrough was decreased and the adsorbed amount of $SO_{2}$ was also diminished. The $SO_{2}$ adsorption was hindered when its flow rate became high and the adsorption capacity of manganese nodule was observed to be superior to that of limestone. In addition, the mixture of manganese nodule and limestone did not show an increase in the adsorption of $SO_{2}$. Finally, as the temperature was raised, the adsorbed amount of adsorbate on manganese nodule was found to be decreased.