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Culture-dependent DGGE (CD DGGE) fingerprinting of the 16S rRNA gene was used to characterize mixed bacterial communities recovered on agar plates. Using R2A Agar as a growth medium, CD DGGE analysis resulted in clear banding patterns of sufficient complexity (16-32 major bands) and reproducibility to investigate differences in bacterial communities in a silt loam soil. Replicate CD DGGE profiles from plates inoculated with less-dilute samples (10−3) had a higher band count and were more similar (72-77%) than profiles from more-dilute samples (51-61%). Different culture media and incubation conditions resulted in distinct community fingerprints and increased the cumulative number of unique bands detected. When CD DGGE fingerprints were compared to profiles constructed from 16S rRNA genes obtained from culture-independent clone libraries (CB DGGE profiles) 34% of the bands were unique to the culture-dependent profiles, 32% were unique to the culture-independent profiles and 34% were found in both communities. These data demonstrate that culture-independent DGGE profiles are supplemented by the distinct bands detected in culture-dependent profiles. CD DGGE can be a useful technique to follow the dynamics of distinct culturable fractions of the soil bacterial community. 相似文献
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Coleman Jerry Hench Keith Garbutt Keith Sexstone Alan Bissonnette Gary Skousen Jeff 《Water, air, and soil pollution》2001,128(3-4):283-295
Three common Appalachian plant species (Juncus effusus L., Scirpus validus L., and Typha latifolia L.) were planted into small-scale constructed wetlands receivingprimary treated wastewater. The experimental design includedtwo wetland gravel depths (45 and 60 cm) and five plantingtreatments (each species in monoculture, an equal mixture of the three species, and controls without vegetation), with two replicates per depth × planting combination. Inflow rates (19 L day-1) and frequency (3 times day-1) were designed to simulate full-scale constructed wetlands as currently used for domestic wastewater treatmentin West Virginia. Influent wastewater and the effluent from each wetland were sampled monthly for ten physical, chemical and biological parameters, and plant demographic measurements were made. After passing through these trough wetlands, the average of all treatments showed a 70% reduction in total suspended solids (TSS) and biochemical oxygen demand (BOD), 50 to 60% reduction in nitrogen (TKN), ammonia and phosphate, anda reduction of fecal coliforms by three orders of magnitude. Depth of gravel (45 or 60 cm) had little effect on wetland treatment ability, but did influence Typha and Scirpus growth patterns. Gravel alone provided significant wastewater treatment, but vegetation further improved many treatment efficiencies. Typha significantly out-performedJuncus and Scirpus both in growth and in effluent quality improvement. There was also some evidence that the species mixture out-performed species monocultures.Typhawas the superior competitor in mixtures, but a decline in Typha growth with distance from the influent pipe suggested that nutrients became limiting or toxicities may have developed. 相似文献
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Timothy B. Parkin Heinrich F. Kaspar Alan J. Sexstone James M. Tiedje 《Soil biology & biochemistry》1984,16(4):323-330
A method was developed for rapid measurement of soil denitrification under conditions where natural soil structure and aeration status is maintained. Air was continuously recirculated by means of a membrane pump through a soil core and a sample loop of a gas chromatograph equipped with an electron capture detector. Addition of acetylene to the recirculating air permitted measurement of denitrification in the soil core. Because of the rapid distribution of C2H2 and removal of N2O provided by the gas flow, denitrification rates could usually be determined in less than 2 h. By means of external 6-way and 8-way valves, four soil cores could be simultaneously analyzed on one gas Chromatograph equipped with dual detectors. Soil cores could also be stored at 4°C for later analysis without affecting the denitrification rate. The detection limit for denitrification rate measurements was 0.5 ngN g?1 soil day?1 or approximately 2.6 g N ha?1 day?1. Coefficients of variation for repeated measurements on the same soil core were usually less than 15%, but coefficients of variation for repacked or natural cores of the same soil were much higher (70–90%) Disruption of the natural soil structure by sieving increased the denitrification rate in an aggregated clay loam soil, but decreased the rate in a non-aggregated sandy soil. These results illustrate the importance of maintaining natural soil structure during denitrification measurements. The effect of pumping gas through soil was evaluated by comparing denitrification rates in soil cores where C2H2 was allowed to distribute into the soil by passive diffusion with rates obtained by pumping. Lower denitrification rates were observed in the static incubation presumably due to limited diffusion of C2H2 into or N2O out of the denitrifying sites in the soil. This diffusion limitation could be overcome in the static incubations if C2H2 was initially distributed through the soil by pumping. This gas flow method is well suited to the study of soil denitrification rates under nearly natural conditions because the indigenous substrates and anaerobic microsites are preserved, the rapidity in which denitrification rates can be measured, and the high sensitivity and relatively low analytical variability of the method. 相似文献
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Open pasture,silvopasture and sward herbage maturity effects on nutritive value and fermentation characteristics of cool‐season pasture 
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下载免费PDF全文 In Appalachian USA, silvopasture offers promise of increased farm productivity. A synchronized, temporal understanding of open pasture (OP) and silvopasture (SP) nutritive characteristics is essential for grazing system development. We examined pasture‐type nutritive‐value relationships when herbage was harvested based on morphological maturity rather than calendar date. Neutral detergent fibre and acid detergent fibre (ADF and NDF) content were greater in silvopasture, while organic matter (OM) was lower (P < 0·05). Digestibility of SP herbage dry matter (DM) and OM tended (P = 0·10) to be lower (418 vs. 471 and 437 vs. 491 g kg?1 respectively). Neutral detergent fibre digestibility was greater (P < 0·05) for OP than SP forage (538 vs. 480 g kg?1), and ADF tended to be greater (P = 0·10; 551 vs. 501). Open‐pasture forage fermentation effluent exhibited slightly higher microbial richness and Shannon diversity than SP. However, overall community composition of both bacteria and archaea did not differ between pasture types or sampling times. Pasture types show proximate analyses differences generally favourable to OP, although both have similar overall nutritive value. In addition, the SP sward exhibited a 4‐ to 6‐days delay in reaching equivalent maturity. Conversion of farm woodlots to SP would increase overall herbage production and improve pasture management flexibility. 相似文献
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Charles E. Turick Alan J. Sexstone Gary K. Bissonnette 《Water, air, and soil pollution》1988,40(3-4):449-460
A species of commercially obtained freshwater mussel (Elliptio complanata) was shown to actively filter, concentrate, and retain fecal coliform (FC) bacteria from a variety of freshwater stream environments. Fecal coliform densities in mussel viscera were maximum within 48 hr of in situ exposure. Significant FC reproduction did not occur in mussel viscera at temperatures below 30 °C. Variation in replicate FC determinations from mussels exposed in situ for 48 hr was less than that obtained directly from water samples. Numbers of FC concentrated in mussel viscera were retained for several hours after FC densities declined in the surrounding water, suggesting the utility of mussels to record recent episodes of fecal pollution. 相似文献
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