Microscale analytical methods for the quantitative detection of PCBs and PAHs in small tissue masses

Microscale methods (MM) were evaluated and compared to traditional methods (TM) for measuring polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) in spiked and standard reference fish and mussel tissues. MMs are advantageous because they use small tissue masses (ca. 100 mg), and maintain sensitivity through reducing final extract volume (traditionally 1 ml) by an order of magnitude or more (40 μl—PCBs; 100 μl—PAHs). Procedural losses occurred in the MMs’ combined cleanup/primary evaporation step (19% PAHs; 6% PCBs), and the final extract concentration (14% PAHs; 22% PCBs). The PAH MM performed comparably to the TM. Although most PCBs had recoveries >50%, the PCB MM generally yielded lower recoveries than the TM. Average method detection limits were 0.6 μg/kg (TM) and 1.0 μg/kg (MM) for PCBs and 25.7 μg/kg (TM) and 27.7 μg/kg (MM) for PAHs. MMs described for PCB and PAH tissue samples are potentially viable alternatives to TMs, and could lead to cost savings in bioaccumulation/toxicity tests.     

Manure management effects on grass production, nutritive content, and soil nitrogen for a grass silage-based dairy farm

Legislation in the United States has recently focused on improving water quality by establishing management practices that limit the quantities of nutrients entering the water supply. Timely application and quantification of the amount of manure applied throughout the grass-growing season can reduce the loss of nutrients into ground or surface water while improving the quality and quantity of grass harvested. During the 2001 and 2002 growing seasons, we measured the effects of different manure application rates on grass yields, grass nutritive value, and soil chemistry on a dairy farm. On-farm estimates of manure N were combined with yield estimates and forage quality measures to evaluate the effects of varying levels of manure application. Yield estimates, N content of grass, and the amount of N in soil and manure were monitored at each cutting for plots amended at different manure application rates. There are three major outcomes of this evaluation: (i) new grass seedings were at higher risk of elevated levels of nitrate N in forage; (ii) increased forage nitrate N at harvest was associated with malfermented silage and increased levels of ammonia N, which resulted in less efficient use of metabolizable protein for milk production; and (iii) increased understanding of N cycling between manure, soil, and plant provided an opportunity to reduce purchased fertilizer.     

ENVIRONMENTAL IMPACTS OF APPLYING MANURE,FERTILIZER, AND SEWAGE BIOSOLIDS ON A DAIRY FARM1

ABSTRACT: Farms that once spread only manures are now also applying sewage biosolids (sludge) and/or other wastes such as those from food processing. The objective of this study was to monitor environmental impacts at a dairy farm applying these materials. Fields were selected representing recent waste applications of manure (M1, M2), sewage biosolids (B1, B2), or fertilizer only control (F1, F2), although most fields had historical biosolids applications. Fields representing each treatment were not experimental replicates because of varying applications and soil characteristics. Septage and food processing wastes were also applied. Soil percolates were collected with wick lysimeters. Runoff was sampled at seven stream sites. Test field soils and alfalfa (Medicago sativa) were analyzed for trace elements. Cumulative trace metal loadings were low, at most only 1 percent of USEPA Part 503 limits. Surface soil enrichment was most evident for Mo, P, and S. Alfalfa tissue showed no trends of concern. The B2 site had the greatest percolate concentrations for 6 of 13 elements. Percolate Cu was somewhat elevated at Sites M1, M2, B2, and Fl. Percolate sodium was elevated on all M and B fields and sulfur was greatest at M2, B1, and B2. Soluble orthophosphate correlated with stream discharge during intensive monitoring of Stream Sites S1 (fertilizer) and S2 (biosolids). Peaks in S2 streamwater Mo lagged large runoff events by five days. Total streamwater export of Cu, Na, Mo, and soluble P were greater from the S2 biosolids subwatershed than from the S1 fertilizer subwatershed. Percolate concentrations exceeded corresponding streamwater concentrations in most cases.     

Simultaneous monitoring of atmospheric methane and speciated non-methane hydrocarbon concentrations using Peltier effect sub-ambient pre-concentration and gas chromatography

Sub-ambient trapping, used to pre-concentrate atmospheric samples for non-methane hydrocarbon (NMHC) analysis by gas chromatography, can also be used to measure ambient methane concentrations. Above a sample volume of 40 ml, a dynamic equilibrium is established between ambient and trapped methane allowing for simultaneous quantitative determinations of methane and NMHC. The temperature stability of the trap is critical for quantitative methane analysis and this can be achieved by Peltier effect cooling. Simultaneous measurements of methane and NMHC reduce the equipment required for field trips and can ease the interpretation and modelling of atmospheric data. The feasibility for deployment of the system in remote locations was demonstrated by running the apparatus virtually unattended for a 5-day period. The correlations between the concentrations of methane, ethane and ethene measured during this period are discussed.     

A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana

Two spectrophotometric assays for protein commonly used in marine research (Coomassie stain, Bradford; alkaline copper, Lowry) and a more recent assay which has not been applied in this field (bicinchoninic acid, Smith) were compared for homogenates of the marine diatom Thalassiosira pseudonona using bovine serum albumin (BSA) as a standard. When homogenates were prepared by precipitating protein with trichloroacetic acid (TCA) and redissolving in 1 N NaOH, the protein content estimated by the Lowry and Smith assays agreed closely, but was consistently 20% higher than that indicated by the Bradford assay. To determine if this difference was due to the choice of a protein standard, protein from T. pseudonana was purified and compared to BSA, bovine gamma-globulin (BGG), and casein. The reactivity of the purified protein (expressed as the slope of the absorbance vs protein concentration curve) did not differ between cultures grown at high or low irradiance. For the Smith and Bradford assays the reactivity of BSA was not significantly different from algal protein, but for the Lowry assay, algal protein was significantly higher in reactivity than BSA. BGG was not significantly different in reactivity from algal protein for the Lowry and Smith assays, but BGG gave significantly lower absorbances than algal protein in the Bradford assay. These results suggest that BSA is a suitable standard for algal protein in the Bradford assays, while BGG is preferable for the Lowry assay. Either protein standard could be used for the Smith assay. Differences in purified algal protein reactivity compared to BSA could not account for the differences among the assays, nor could interference by chlorophyll a. Precipitating protein with TCA prior to analyses gave lower protein than direct analyses of homogenates for the Lowry and Smith assays, but no differences were found for the Bradford assay. As a result, the Lowry and Smith assays indicated up to 60% greater protein than the Bradford if TCA precipitation was not performed. This may be due to removal of free amino acids and small peptides which are less reactive in the Bradford assay. The 20% higher protein found in the Lowry or Smith vs Bradford assays may be due to different assay sensitivity to small peptides or other compounds which are precipitated along with proteins by TCA. Although the Smith assay is substantially simpler to perform than the Lowry, there appear to be no quantitative differences in the results. It remains unclear which spectrophotometric assay is most accurate, but the Bradford assay is faster and simpler, and is less likely to be affected by non-protein compounds found in marine phytoplankton.     

The influence of irradiance on the biochemical composition of three phytoplankton species and their nutritional value for larvae of the Pacific Oyster (Crassostrea gigas)

Three species of phytoplankton grown at high (HL) or low light (LL) were fed as saturating rations to laboratory-reared larval Crassostrea gigas. Larval C. gigas fed diets of HL grown Chaetoceros gracilis and HL grown Isochrysis aff. galbana grew faster than those fed LL grown cells of the same phytoplankton species. Faster growth of C. gigas larvae was consistently associated with increases in the percent composition of short chain saturated fatty acids (FA) 14:0+16:0 in the HL grown cells. There were no consistent and significant differences between HL and LL grown phytoplankton cells in their content of carbon, nitrogen, protein, lipid or carbohydrate. Intraspecific increases in percent composition of essential fatty acids (EFAs), 20:53 and 22:63, in the phytoplankton were not associated with improvements in the growth or survival of the oyster larvae. Oyster larvae fed diets of Phaeodactylum tricornutum with a relatively high proportion of EFAs grew more slowly than those fed C. gracilis. In this experiment the proportion of dietary EFA 20:53 was negatively correlated with oyster growth rates. The faster growing oyster larvae contained relatively more of the FAs 14:0+16:0 which may be useful as measures of larval oyster condition. After a diet of one phytoplankton species for ca. 10 d, oyster larvae acquired distinctive FA profiles resembling that of their phytoplankton prey.     

Climatic Variation and Solute Concentration and Flux in Meltwaters Draining from an Alpine Glacier

Measurements of electrical conductivity and discharge ofmeltwaters in the Gornera, which drains from the 83%glacierised basin containing Gornergletscher, PennineAlps, Switzerland, were undertaken between May andSeptember in both 1979 and 1998. Discharge in theGornera was 43% higher in 1998, average air temperatureduring the ablation season being 2.1 °C warmer andpreceding winter precipitation 28% lower than in 1979. Mean electrical conductivity of meltwater in 1998 wasreduced by 40%. In the same 60 day period in 1998,however, solute flux was augmented by only 2% bycomparison with 1979. Year-to-year climatic variations,reflected in discharge variability, strongly affectsolute concentration in glacial meltwaters, but havelimited impact on solute flux. Climatic conditionstranslate into meltwater quality through inter-relationships between mineral reaction rates, subglacialresidence time in contact with sediment, and discharge. Annual variability in solute flux depends on the extentto which volume of flow can offset decline in soluteconcentration brought about by reducing residence time.     

Comparison of an enzyme-linked immunosorbent assay (ELISA) to gas chromatography (GC)--measurement of polychlorinated biphenyls (PCBs) in selected US fish extracts

The analysis of PCBs in fish tissues by immunoassay methods was evaluated using fish collected from a US monitoring program, the National Contaminant Biomonitoring Program of the US Department of Interior, Fish and Wildlife Service. Selected composite whole fish samples, which represented widely varying concentrations and sources of PCBs, were extracted and subjected to congener PCB analysis by gas chromatography (GC) and total PCB analysis using an ELISA (ePCBs) calibrated against technical Aroclor 1248. PCB congener patterns in these fishes were different from the patterns found in commercial Aroclors or their combinations as demonstrated by principal component analysis of normalized GC congener data. The sum of the PCB congeners measured by GC (total-PCBs) ranged from 37 to 4600 ng/g (wet weight). Concentrations of PCBs as determined by the ELISA method were positively correlated with total-PCBs and the ePCBs/total-PCBs ratios for individual samples ranged from 1 to 6. Ratios of ePCBs/total-PCBs for dilutions of Aroclors 1242, 1254, and 1260 and for matrix spikes range from 0.6 for 1242 to 2.5 for 1254 and 1260. These results suggest that higher chlorinated PCB congeners have higher affinity for the anti-PCB antibodies. Partial least squares with latent variable analysis of GC and ELISA data of selected Aroclors and fish samples also support the conclusion that ELISA derived PCB concentrations are dependent on the degree of chlorination.     

Metacommunity models predict the local-regional species richness relationship in a natural system

Many natural communities exhibit positive relationships between local and regional species richness (LSR-RSR relationships), which can be either linear or curvilinear. Previous models have shown that the form of this relationship depends on the relative rates of colonization and extinction and the sensitivity of these rates to competition. We use simple models to show that the LSR-RSR relationship also depends on the type of metacommunity structure (Levins-like or mainland-island), and our models generate a wider range of realistic forms than do most previous models. We parameterize and test our models with two independent data sets for Daphnia in rock pools on islands in Finland and Sweden. We find that the Levins-like model with competition correctly predicts the observed LSR-RSR relationship and provides the best fit to the average local species richness per island. Simulations show that our models are robust to relaxing our assumption of identical species properties. Our study is one of the first to make and successfully test quantitative predictions for how a widely studied community pattern, the LSR-RSR relationship, arises from metacommunity dynamics.