• Title/Summary/Keyword: Cell Observation

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Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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Neuroprotective Effect of Cyclosporin A on Spinal Cord Ischemic Injury in Rabbits (토끼를 이용한 척수 허혈 손상 모델에서 Cyclosporin A의 척수 손상에 대한 보호 효과)

  • Shin Yoon-Cheol;Choe Ghee-Young;Kim Won-Gon
    • Journal of Chest Surgery
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    • v.39 no.10 s.267
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    • pp.739-748
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    • 2006
  • Background: The purpose of this study is to ascertain the neuroprotective effect of cyclosporin A on the 25-min surgical ischemia model in the spinal cords of rabbits with neuropathological correlation and histoimmunochemical analyses, Material and Method: Thirty-two New Zealand white rabbits were randomly divided into four groups: Rabbits were randomly divided into four groups: the control 12 group (n=8), the control 17 group (n=8), the cyclosporin Cs2 group (n=8), and the cyclosporin Cs7 group (n=8). The 12 group underwent a 25-min aortic cross- clamp without intervention and were sacrificed on the 2nd day postoperatively, while the 17 group underwent a 25- min of aortic cross-clamp without intervention and were sacrificed on the 7th day postoperatively. The Cs2 group received cyclosporin A (25 mg/kg) intravenously 15 min after the 25-min cross-clamp and were sacrificed on the End day postoperatively, while the Cs7 group received cyclosporin A (25 mg/kg) intravenously 15 min after the 25-min cross-clamp and were sacrificed on the 7th day postoperatively. The rabbits underwent 25-min surgical aortic cross-clamp. Neurologic functions were evaluated on the 2nd day and 7th postoperative day using Tarlov scoring system. After scoring neurologic function, all rabbits were sacrificed for histopathologic observation. Result: All rabbits survived the experimental procedure. The values of Tarlov score did not show any differences between the control and cyclosporin groups on the 2nd day. The scores of group Cs7 ($2.75{\pm}0.89$) were significantly higher than those of group 17 ($1.25{\pm}1.39$) on the 7th day (p<0,05). On the histologic exanminations, specimens of the spinal cord showed necrosis and apoptosis. The pathologic scores of group Cs7 ($1,0{\pm}0.53$) was less than those of group 17 ($2.13{\pm}1.36$, p<0.05). TUNEL staing showed apoptosis of the specimen in group 12 and Cs2 but there was no stastically significant difference between groups on the score. There were more overexpression of HSP70 and nNOS in cyclosporine group than in control group. Conclusion: We think that cyclosporin A may decrease neuronal cell death with induced upregulation of HSP70 against 25-min ischemia of the spiral cord in the rabbit.

Testicular Development and Serum Levels of Gonadal Steroids Hormone during the Annual Reproductive Cycle of the Male Koran Dark Sleeper, Odontobutis platycephala (Iwata et Jeon) (동사리, Odontobutis platycephala (Iwata et jeon) 수컷의 생식주기에 따른 정소 발달과 혈중 생식소 스테로이드의 변화)

  • 이원교;양석우
    • Journal of Aquaculture
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    • v.11 no.4
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    • pp.475-485
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    • 1998
  • To clarify annual reproductive cycle of Koran dark sleeper, odontobutis platycephala, we examined the seasonal changes of gonadosomatic index(GSI), testicular development stages and sex steroid hormones in blood from December 1995 to November 1997. Testis was podlike shape from July to October, and tadpole-like shape from November because of its expanded posterior part. GSI was 0.14~0.18 from July to September and increased to $0.43{\pm}0.04$ in October and then was not changed significantly until February. GSI was reincreased to $0.52{\pm}0.09$ from March and then was kept at similer levels until May, but fell down to $0.28{\pm}0.05$ in June. As results of histological observation, testis was divided into 3 parts(anterior, boundary, posterior) in the development progress of germ cells. In July, the testis was composed of only spermatogonia without seminiferous tubules in most fishes. In the anterior part of testis, the ferquency of spermatogenesis stage seminiferous tubules appearing in August was more than 80% from September to December. decreased gradually from January to March and drastically in April, and then disappeared in June. The frequency of spermiogenesis stage seminiferous tubules appearing in December, increased gradually from January to March and drastically to 80% in April, and reached to 90% the highest levels of the year in June. Post-spawning stage seminiferous tubules did not appear throughout the year. The frequency of spermatogonia was 100% and 65% in July and August, and less than 20% in the rest period of the year. In the boundary part, the frequency of spermatogenesis stage seminiferous tubules appearing in August increased from September and reached to 82% in November, decreased from December, adn disappeared in March. The frequency of spermiogenesis stage seminiferous tubules appearing in November was less than 18% until February, and increased to 29%~57% from March to June. The frequency of post-spawning stage seminiferous tubules appeared 12%~25% only from March to June. The frequency of spermatogonia was 100% in July, decreased to 85% in August and 10% in November, and increased gradually from December to 50% in April, and decreased again from May to June. In the posterior part, seminiferous tubules with some seminiferous tubules increased drastically 80%~85% in August and September, decreased drastically from October to November and remained below 10% until February, and disappeared after March. The frequency of spermiogenesis stage seminiferous tubules appearing in August increased sharply from October and reached to 75% in November. decreased to 15% in December and no significant changes until March, and disappeared after April. The frequency of post-spawning stage seminiferous tubules appearing very early in November increased to 82% in December and 85%~95% until June. The frequency of spermatogonia was 100% in July, decreased drastically to 15% in August, disappeared from October to Mrch, but reappeared from April and kept at less than 10% until June. The blood level of testosterone (T) increrased gradually from August was $0.61{\pm}0.09 ng/m\ell$ in November, increrased drastically to $3.99{\pm}1.22 ng/m\ell$ in December and maintained at in similar level until March, and decreased to $0.25{\pm}0.14 ng/m{\ell} ~ 0.17{\pm}0.13ng/m{\ell}$ in April and May and no significant changes until July (P<0.05). The blood level of 17, 20 -dihydroxy-4-pregnen-3-one $ng/m{\ell}$in the rest of year without significant changes(P<0.05). Taken together these results, the germ cell development of testis progressed in the order of posterior, boundary, anterior part during annual reproductive cycle in Korean dark sleeper. The testicular cycle of Korean dark sleeper was as follows. The anterior part of testis : i.e. spermatogonial proliferation period (July), early maturation period (from August to November), mid maturation period (from December to March), late maturation period (from April to May) and functional maturation period (June) were elucidated. The boundary of testis, i.e. spermatogonial proliferation period (July), early maturation period (from August to October), mid maturation period (from November to February) and the coexistence period of late maturation, functional maturation and post-spawn (from March to June) were elucidated. The posterior of testis, i.e. spermatogonial proliferation period (July), mid maturation period (from August ot September), late maturation period (October), functional maturation period (November) and post-spawn period (from December to June) were elucidated. It was showed that the changes of sex steroid hormone in blood played a important roles in the annual reproductive cycle of Korean dark sleeper.

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