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In an extensive, multiyear study of antibiotic resistance from wastewater oxidation ponds, five mobile home park wastewater oxidation ponds in Clarke and Oconee counties were shown to be discharging high numbers of antibiotic-resistant bacteria into the waterways of North Georgia. This effluent contributed to higher nitrogen, phosphorus, and fecal coliform levels in creeks downstream from the ponds. A survey of residents revealed that many people did not complete their antibiotic prescriptions, and the majority flushed leftover antibiotic medications down the toilet. In the pond discharges, resistance was found to eighteen antibiotics: amikacin, amoxicillin/clavulanic acid, ampicillin, apramycin, cefoxitin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, imipenem, kanamycin, naladixic acid, streptomycin, sulphamethoxazole, trimethoprim/sulphamethoxazole, and tetracycline. The discharged bacteria contained both integrons and plasmids, the latter being transferable to a laboratory strain of Escherichia coil (E. coli). A turtle was found living at a pond discharge site with multiply-antibiotic-resistant bacteria in its feces. Last year, RNA fingerprinting conclusively documented the survival of three multiply-resistant important pathogenic bacteria. Ceftriaxone-resistant Stenotrophomonas maltophilia and Pseudomonas aerogenosa and a ciprofloxacin-resistant E. coli were traced through oxidation pond stages and into the discharge, thus documenting that the pathogens survived the treatment process. In addition, a potential pathogen, a serotype group D Salmonella spp., was found in the discharge. In this study, tetracycline-resistance genes C and G were detected in the first and second stages of the oxidation pond and the discharge went directly into the environment. These genes are generally found in intestinal bacteria, so it can be inferred that they are from a human source. Antimicrobial residue from the beta-lactam family of antibiotics was found in all oxidation pond stages and in the creek above the pond. Tetracycline residue was found in the first and second stages of the pond. In addition to the antibiotics, genes coding for antibiotic resistance and the antibiotics themselves were documented to survive oxidation pond treatment. Tetracycline-resistant genes were identified in the oxidation pond stages and in the discharge going into the environment. A model was also developed to study oxidation pond function in the laboratory. A biofilm was created using a highly antibiotic-resistant Salmonella typhimurium 3/97, and pond water was added. The biofilm was processed via a rotating disk bioreactor specifically designed to study biofilms in nature, but with conditions that were more favorable to bacterial inhibition than those in nature. Cultures revealed that, under these optimal conditions, S. typhimurium 3/97 was still present in this in vitro system. Thus, the competitive inhibition process that helps to remove bacteria in oxidation ponds did not effectively remove an important bacterium, S. typhimurium 3/97, in this mock oxidation pond. The bioreactor model developed in this study can be used to further investigate discharges from oxidation ponds. From this data, it is apparent that the problem is two-fold. A cost-effective technique must be developed that inactivates antibiotic-resistant bacteria in oxidation pond discharges and also removes the antibiotics. A public awareness campaign was initiated by the author to encourage proper use and disposal of antibiotics, as flushing them is a common practice in the United States.
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PMID:Antibiotic resistance from wastewater oxidation ponds. 1638 Nov 46

Eutrophication is a serious water quality problem in estuaries receiving increasing anthropogenic nutrient loads. Managers undertaking nutrient-reduction strategies aimed at controlling estuarine eutrophication are faced with the challenge that upstream freshwater segments often are phosphorus (P)-limited, whereas more saline downstream segments are nitrogen (N)-limited. Management also must consider climatic (hydrologic) variability, which affects nutrient delivery and processing. The interactive effects of selective nutrient input reductions and climatic perturbations were examined in the Neuse River Estuary (NRE), North Carolina, a shallow estuary with more than a 30-year history of accelerated nutrient loading and water quality decline. The NRE also has experienced a recent increase in Atlantic hurricanes and record flooding, which has affected hydrology and nutrient loadings. The authors examined the water quality consequences of selective nutrient (P but not N) reductions in the 1980s, followed by N reductions in the 1990s and an increase in hurricane frequency since the mid-1990s. Selective P reductions decreased upstream phytoplankton blooms, but increased downstream phytoplankton biomass. Storms modified these trends. In particular, upstream annual N and P concentrations have decreased during the elevated hurricane period. Increased flushing and scouring from storms and flooding appear to have enhanced nutrient retention capabilities of the NRE watershed. From a management perspective, one cannot rely on largely unpredictable changes in storm frequency and intensity to negate anthropogenic nutrient enrichment and eutrophication. To control eutrophication along the hydrologically variable freshwater-marine continuum, N and P reductions should be applied adaptively to reflect point-source-dominated drought and non-point-source-dominated flood conditions.
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PMID:Assessing the effects of nutrient management in an estuary experiencing climatic change: the Neuse River Estuary, North Carolina. 1645 30

In order to investigate the feasibility of biological treatment of hypersaline wastewater produced from toilet flushing with seawater at low temperature, pilot-scale studies were established with plug-flow activated sludge process at low temperature (5-9 degrees C) based on bench-scale experiments. The critical salinity concentration of 30 g/L, which resulted from the cooperation results of the non-halophilic bacteria and the halophilic bacteria, was drawn in bench-scale experiments. Pilot-scale studies showed that high COD removal efficiency, higher than 80%, was obtained at low temperature when 30 percent seawater was introduced. The salinity improved the settleability of activated sludge, and average sludge value dropped down from 38% to 22.5% after adding seawater. Seawater salinity had a strong negative effect on notronomonas and nitrobacter growth, but much more on the nitrobacter. The nitrification action was mainly accomplished by nitrosomonas. Bench-scale experiments using two SBRs were carried out for further investigation under different conditions of salinities, ammonia loadings and temperatures. Biological nitrogen removal via nitrite pathway from wastewater containing 30 percent seawater was achieved, but the ammonia removal efficiency was strongly related not only to the influent ammonia loading at different salinities but also to temperature. When the ratio of seawater to wastewater was 30 percent, and the ammonia loading was below the critical value of 0.15 kgNH4+-N/(kgMLSS.d), the ammonia removal efficiency via nitrite pathway was above 90%. The critical level of ammonia loading was 0.15, 0.08 and 0.03 kgNH4+-N/(kgMLSS.d) respectively at the different temperature 30 degrees C, 25 degrees C and 20 degrees C when the influent ammonia concentration was 60-80 mg/L and pH was 7.5-8.0.
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PMID:Pilot-scale studies on biological treatment of hypersaline wastewater at low temperature. 1645 84

Disposal of treated wastewater for more than 60 years onto infiltration beds on Cape Cod, Massachusetts produced a groundwater contaminant plume greater than 6 km long in a surficial sand and gravel aquifer. In December 1995 the wastewater disposal ceased. A long-term, continuous study was conducted to characterize the post-cessation attenuation of the plume from the source to 0.6 km downgradient. Concentrations and total pools of mobile constituents, such as boron and nitrate, steadily decreased within 1-4 years along the transect. Dissolved organic carbon loads also decreased, but to a lesser extent, particularly downgradient of the infiltration beds. After 4 years, concentrations and pools of carbon and nitrogen in groundwater were relatively constant with time and distance, but substantially elevated above background. The contaminant plume core remained anoxic for the entire 10-year study period; temporal patterns of integrated oxygen deficit decreased slowly at all sites. In 2004, substantial amounts of total dissolved carbon (7 mol C m(-2)) and fixed (dissolved plus sorbed) inorganic nitrogen (0.5 mol N m(-2)) were still present in a 28-m vertical interval at the disposal site. Sorbed constituents have contributed substantially to the dissolved carbon and nitrogen pools and are responsible for the long-term persistence of the contaminant plume. Natural aquifer restoration at the discharge location will take at least several decades, even though groundwater flow rates and the potential for contaminant flushing are relatively high.
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PMID:Long-term natural attenuation of carbon and nitrogen within a groundwater plume after removal of the treated wastewater source. 1657 69

A small-scale membrane plant for treating the domestic wastewater of a four-person household is presented. The membrane bioreactor has been in operation for 6 months and achieves elimination rates of 90, 95 and 80% for total organic carbon, chemical oxygen demand and total nitrogen, respectively. Only a small amount sludge is produced. The permeate is reused for flushing toilets and has a yellowish colour. After investigations of the effluent quality, decolourisation of the permeate, energy efficiency and control strategies in the first year, urine will be treated separately in an automated precipitation reactor where struvite is produced to improve the overall phosphate removal of the plant.
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PMID:Domestic wastewater treatment with a small-scale membrane bioreactor. 1660 19

The aim of our study was to estimate the viability of cat epididymal sperm in short time storage at +4 degrees C and in long term storage at -196 degrees C and to assess the percentage of live sperm in fresh semen using eosin/nigrosin staining compared to the flow cytometry method. The testes with epididymides were obtained after routine castration procedure. The sperm for further research were collected after flushing the epididymides using extender consist of: Tris 2.4 g, citric acid 1.4 g, glucose 0.8 g, 0.06% (w/v) Na-benzylpenicillin, 0.1% (w/v) streptomycin sulphate and distilled water. Half of each sample was equilibrated with the dilution and loaded in 0.25 ml plastic straws. The straws were placed on a rack in liquid nitrogen vapour at -120 degrees C for 10 min, plunged in liquid nitrogen for 10 min, replaced to marked goblets and loaded into canes for long term storage in liquid nitrogen at -196 degrees C. Sixty percent of motile spermatozoa was accomplished after thawing. However, the percentage of the sperm with intact acrosomes was decreased and the share of cells with midpiece and tail defects was increased. The storage of sperm flushed from epididymides at +4 degrees C for a short time and the usage of sperm during 2-3 days after collection seems to be better than cryopreservation. In our study, normospermia was present in 72.7 +/- 8.8% of fresh semen. The most common defect was the presence of distal droplets, imperfect heads or abnormal acrosomal outline. The motility of fresh sperm flushed from epididymides achieved 77.9 +/- 6.8%. The viability of sperm amounting to 52.5 +/- 13.8% was achieved on third day of conservation in the liquid extender. The percentage of viable sperm in fresh epididymal spermatozoa was 84.9 +/- 7.8%. Compared to these results, the percentage of live cells using SYBR-14/propidium iodide staining was insignificantly lower (82.2 +/- 8%). The live, non-apoptotic cells were 79.0 +/- 7.8%. The share of live, early-apoptotic spermatozoa and late-apoptotic spermatozoa was, respectively, 2 +/- 1.4% and 1.5 +/- 0.9%. The viability of sperm estimated by eosin/nigrosin staining was confirmed by the flow cytometry method. There was no statistical differences between the staining. The usage of apoptosis detection kit revealed, that the percentage of early-apoptotic and late-apoptotic cells was insignificant.
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PMID:Preservation of tomcat (Felis catus) semen in variable temperatures. 1672 86

The objective of this study was to investigate the in vitro and in vivo developmental abilities of equine embryos cryopreserved by vitrification. Twenty-eight embryos were recovered from Native pony and Thoroughbred mares at Days 5 to 7 by nonsurgical uterine flushing (detection of ovulation=Day 0). The vitrification solution contained 40% ethylene glycol, 18% Ficoll, and 0.3 M sucrose in PBS. The embryos were placed for 1 to 2 min in vitrification solution (Group 1) or following exposure to 20% ethylene glycol in PBS for 10 to 20 min (Groups 2 and 3). Single embryos were loaded in 0.25-ml straws, cooled for 1 min in liquid nitrogen vapor and immersed in liquid nitrogen. Straws were warmed in water (20 degrees C, 20 sec), and the contents were expelled with 0.5 M sucrose in PBS. Then the sucrose was diluted in 1-step (Groups 1 and 2) or 4-steps (Group 3). Embryos (n=21) were cultured for 120 h in TCM199 supplemented with 10% fetal bovine serum at 37 degrees C in 5% CO2 in air and evaluated morphologically. Development to the hatching or hatched blastocyst stage was obtained in 0 7 , 4 7 and 4 7 embryos in Groups 1, 2 and 3, respectively. An additional 7 embryos were vitrified-warmed according to the treatment of Group 2 (4 embryos) and Group 3 (3 embryos). Five embryos were selected after in vitro culture for 4 h and were transferred nonsurgically into the uterine horn of Day-4 recipient mares. Transfer of 2 embryos (both Day-6 blastocysts: Group-2 treatment) resulted in pregnancies with a viable fetus at Day-60 of the gestation period.
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PMID:Pregnancies following transfer of equine embryos cryopreserved by vitrification. 1672 55

Using high sensitivity differential scanning calorimetry (HSDSC), the phase transitions of dimyristoylphosphatidylcholine (DMPC) liposomal bilayers and their interaction with the model steroid beclomethasone dipropionate (BDP) were found to be dependent on the method of liposome manufacture. Ethanol-based proliposomes produced liposomes having no phospholipid pretransition, a main transition of high enthalpy and a low onset temperature, and a very low incorporation of the steroid (maximum 1 mol%). This was attributed to an alcohol-induced interdigitation of the bilayers, which was not apparently reversed by flushing the liposome dispersion with nitrogen in an attempt to remove ethanol. For liposomes manufactured by thin film or particulate-based proliposome methods, 1-2.5 mol% steroid was optimal for incorporation within bilayers, although the nature of the steroid interaction with the bilayers differed between the two methods. For liposomes manufactured by the thin film method, a higher steroid concentration resulted in a broadened main transition and a reduced melting cooperativity. This suggests that BDP formed separate domains within the bilayers which caused non-ideal mixing and phase separation at 5 mol% steroid. This observation was absent for liposomes generated from particulate-based proliposomes, indicating separate steroid domains were not formed and subsequent non-ideal mixing and phase separation did not occur. In addition, liposomes generated from particulate-based proliposomes showed reduced pretransition and main transition enthalpies. These differences were attributed to the employment of sucrose to manufacture the particulate-based proliposomes. This study has shown that the thermal behaviour of liposomes and their interaction with beclomethasone dipropionate were dependent on the method of liposome manufacture. Moreover, particulate-based proliposomes may provide a reasonable alternative to the conventional thin film method in producing liposomes incorporating this steroid.
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PMID:A calorimetric study of dimyristoylphosphatidylcholine phase transitions and steroid-liposome interactions for liposomes prepared by thin film and proliposome methods. 1676 1

We conducted experiments in laboratory microcosms to simulate input of stemflow nutrients and flushing of metabolites in the tree hole habitats of larval Ochlerotatus triseriatus (Say). In the first experiment, we simultaneously examined the effects of nutrient additions (nitrogen, phosphorus, glucose, or combination) and flushing (removal of one-half of water volume replaced by deionized water) on mosquito production. The combination of nutrients had the greatest positive effects on mosquito production, with nitrogen (N) likely accounting for most of the increase in adult emergence and adult mass. Dilution of the nutrient pool via simulated flushing reduced mosquito growth, suggesting that the primary effect of stemflow input was nutrient addition as opposed to dilution of any latent toxic metabolites. In a second experiment, N additions were crossed with larval presence or absence to examine effects on key microbial processes. N increased leaf decay rates, soluble carbohydrate concentrations, fungal biomass and leaf-associated carbohydrase activity, but it did not stimulate bacterial productivity. Leaf decay was enhanced and bacterial production on leaves and in the water column was depressed in the presence of larvae. We conclude that the inputs of soluble N stimulated fungal growth, which made more fungal biomass available because of both its absolute increase and via the softening of the leaf particulate matter that could allow direct ingestion by larvae.
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PMID:Indirect effects of soluble nitrogen on growth of Ochlerotatus triseriatus larvae in container habitats. 1689 24

Submerged cultures were used to identify growth-limiting nutrients by Antrodia cinnamomea strains. The mycelial biomass and EPS production by A. cinnamomea BCRC 35396 were markedly higher than other A. cinnamomea strains. A relatively high C/N ratio was favorable for both the mycelial growth (5.41 g/l) and EPS production (0.55 g/l); the optimum ratio was 40. The glucose was available utilized preferentially for mycelial growth, rather than for EPS production. Flushing the culture medium with nitrogen had a stimulating effect on both mycelial growth and EPS production. In addition, peptone, yeast extract and malt extract appeared to be important and significant component for EPS production. Phosphate ion, magnesium ion and thiamine were probably not essential for mycelial growth. By optimizing the effects of additional nutrition, the results showed that 5% (w/v) glucose, 0.8% (w/v) peptone, 0.8% (w/v) yeast extract, 0.8% (w/v) malt extract, 0.03% (w/v) KH2PO4, 0.1% (w/v) MgSO4 .7H2O and 0.1% (w/v) thiamine could lead to the maximum production of EPS (1.36 g/l).
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PMID:Factors affecting mycelial biomass and exopolysaccharide production in submerged cultivation of Antrodia cinnamomea using complex media. 1707 Oct 80


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