The fate of the recombinant DNA in corn during composting

In order to make regulations that safeguard food and the environment, an understanding of the fate oftransgenes from genetically modified (GM) plants is of crucial importance. A compost experiment including mature transgenic corn plants and seeds of event Bt 176 (Zea mays L.) was conducted to trace the fate of the transgene cryIA(b) during the period of composting. In bin 1, shredded corn plants including seeds were composted above a layer of cow manure and samples from the corn layer were collected at intervals during a 12-month period. The samples were tested for the transgene persistence and microbial counts and also the compost was monitored for temperature. In bin 2, piles of corn seeds, surrounded by sheep manure and straw, were composted for 12 months. A method combining nested polymerase chain reaction (PCR) and southern hybridization was developed for detection of the transgene in compost. The detection sensitivity was 200 copies of the transgene per gram of dry composted corn material. Composting commenced on day 0, and the transgene was detected in specimens from bin 1 on days 0 and 7 but not on day 14 or thereafter. The transgene in corn seeds was not detectable after 12 months of composting in bin 2. Temperatures in both bins rose to about 50 degrees C within 2 weeks and remained above that temperature for about 3 months, even when the ambient temperature dropped below -20 degrees C. Extracts from compost were inoculated onto culture plates and then were incubated at 23 to 55 degrees C. Within the first 2 weeks of composting in bin 1, the counts of bacteria incubated at 55 degrees C increased from 3.5 to 7.5 log10, whereas those incubated at 23 degrees C remained at about 7.5 log10. The counts of fungi incubated at 45 degrees C increased slightly from 2.5 to 3.1 log10, but those incubated at 23 degrees C decreased from 6.3 to 3.0 log10. The rapid degradation of the transgene during composting of Bt corn plants suggested that the composting process could be used for safe disposal of transgenic plant wastes.     

The Fate of the Recombinant DNA in Corn During Composting

Abstract

In order to make regulations that safeguard food and the environment, an understanding of the fate of transgenes from genetically modified (GM) plants is of crucial importance. A compost experiment including mature transgenic corn plants and seeds of event Bt 176 (Zea mays L.) was conducted to trace the fate of the transgene cryIA(b) during the period of composting. In bin 1, shredded corn plants including seeds were composted above a layer of cow manure and samples from the corn layer were collected at intervals during a 12-month period. The samples were tested for the transgene persistence and microbial counts and also the compost was monitored for temperature. In bin 2, piles of corn seeds, surrounded by sheep manure and straw, were composted for 12 months. A method combining nested polymerase chain reaction (PCR) and southern hybridization was developed for detection of the transgene in compost. The detection sensitivity was 200 copies of the transgene per gram of dry composted corn material. Composting commenced on day 0, and the transgene was detected in specimens from bin 1 on days 0 and 7 but not on day 14 or thereafter. The transgene in corn seeds was not detectable after 12 months of composting in bin 2. Temperatures in both bins rose to about 50°C within 2 weeks and remained above that temperature for about 3 months, even when the ambient temperature dropped below ?20°C. Extracts from compost were inoculated onto culture plates and then were incubated at 23 to 55°C. Within the first 2 weeks of composting in bin 1, the counts of bacteria incubated at 55°C increased from 3.5 to 7.5 log ₁₀, whereas those incubated at 23°C remained at about 7.5 log ₁₀. The counts of fungi incubated at 45°C increased slightly from 2.5 to 3.1 log₁₀, but those incubated at 23°C decreased from 6.3 to 3.0 log ₁₀. The rapid degradation of the transgene during composting of Bt corn plants suggested that the composting process could be used for safe disposal of transgenic plant wastes.     

Investigation on gene transfer from genetically modified corn (Zea mays L.) plants to soil bacteria

Knowledge about the prevalence and diversity of antibiotic resistance genes in soil bacteria communities is required to evaluate the possibility and ecological consequences of the transfer of these genes carried by genetically modified (GM) plants to soil bacteria. The neomycin phosphotransferase gene (nptII) conferring resistance to kanamycin and neomycin is one of the antibiotic resistance genes commonly present in GM plants. In this study, we investigated kanamycin-resistant (Km(R)) and neomycin-resistant (Nm(R)) soil bacterial populations in a 3-year field trial using a commercial GM corn (Zea mays L.) carrying the nptII gene and its near isogenic line. The results showed that a portion (2.3 - 15.6 %) of cultivable soil bacteria was naturally resistant to kanamycin or neomycin. However, no significant difference in the population level of Km(R) or Nm(R) soil bacteria was observed between the GM and non-GM corn fields. The nptII gene was not detected in any of the total 3000 Km(R) or Nm(R) isolates screened by PCR. Further, total soil bacterial cells were collected through Nycodenz gradient centrifugation and bacterial community DNA was subjected to PCR. Detection limit was about 500 cells per gram of fresh soil. Our study suggests that the nptII gene was relatively rare in the soil bacterial populations and there was no evidence of gene transfer from a GM corn plant to soil bacteria based on the data from total soil bacterial communities.     

Fate of Bacillus thuringiensis (Bt) Cry3Bb1 protein in a soil microcosm

Transgenic crops expressing insecticidal, crystalline (Cry) Bacillus thuringiensis (Bt) proteins were commercialized in the US in 1996. There is little information in the peer-reviewed literature on the environmental fate of the coleopteran-active Bt Cry3Bb1 protein expressed in event MON863 corn. The exposure characterization of Bt proteins is unique in that the fate of the protein in soil and in crop residue must be considered. To date, the significance of macrodecomposing organisms, such as earthworms, isopods, and springtails, to the dissipation of Bt proteins present in crop residue has not been assessed. In addition, the input of Bt proteins into soil through leaching from post-harvest crop residue has not been examined. Two laboratory microcosm studies were conducted to determine the fate of Bt Cry3Bb1 in decomposing MON863 corn residue and to determine whether Bt proteins can enter soil by leaching from crop residue. In addition, the importance of macrodecomposing organisms to the degradation of Bt proteins in corn residue was assessed. Laboratory microcosms containing MON863 corn leaf, root, and stalk with and without macrodecomposers were used to examine the fate of Bt Cry3Bb1 in post-harvest MON863 corn residue. A half-life of less than five days was found for Bt Cry3Bb1 protein in decomposing MON863 corn residue. There was a trend of increasing half-life of Cry3Bb1 in microcosms containing macrodecomposers, however, this trend was only significant (p<0.05) for Bt Cry3Bb1 in MON863 leaf tissue and this increase is not likely relevant for non-target exposure. The recovery of Bt Cry3Bb1 protein from soil extracts was either below the limit of quantification (9 ng g(-1) soil) or below the limit of detection (0.7 ng mL(-1)) at all time points during the study. Based on the results from this study, Bt protein leaching from post-harvest crop residue is not a significant contributor to Bt protein input into soil.     

Extractable soil lipids and microbial activity as affected by Bt and non Bt maize grown on a silty clay loam soil

Pyrolysis-gas (Py-GC) chromatography was used to characterize extractable lipids from Bt and non-Bt maize shoots and soils collected at time of harvesting. Py-GC-MS (mass spectrometry) showed that the concentrations of total alkenes identified in non-Bt shoots and soils were 47.9 and 21.3% higher than in Bt maize shoots and soils, respectively. N-alkanes identified were of similar orders of magnitude in Bt and non-Bt maize shoots, but were 28.6% higher in Bt than in non-Bt soils. Bt maize shoots contained 29.7% more n-fatty acids than non-Bt maize shoots, whereas the concentrations of n-fatty acids in Bt soils were twice as high as those in non-Bt soils. Concentrations of unsaturated fatty acids in Bt maize shoots were 22.1% higher than those in non-Bt maize shoots, while concentrations of unsaturated fatty acids were 22.5% higher in non-Bt than in Bt soils. The cumulative CO2-C evolved from soils under Bt and non-Bt crops was 30.5% lower under Bt as compared to non-Bt crops, whereas when maize shoots were added to Bt and non-Bt soils, the decrease in CO2-C evolved were 16.5 and 23.6%, respectively. Our data showed that the cultivation of Bt maize significantly increased the saturated to unsaturated lipid ratios in soils which appeared to negatively affect microbial activity.     

Evaluation of quick tests for phosphorus determination in dairy manures

Nutrients in animal manure are valuable inputs in agronomic crop production. Rapid and timely information about manure nutrient content are needed to minimize the risks of phosphorus (P) over-application and losses of dissolved P (DP) in runoff from fields treated with manure. We evaluated the suitability of a commercial hand-held reflectometer, a hydrometer, and an electrical conductivity (EC) meter for determining DP and total P (TP) in dairy manures. Bulk samples (n = 107) collected from farms across CT, MD, NY, PA, and VA were highly variable in total solids (TS) concentration, ranging from 11 to 213gL(-1), in suspensions' pH (6.3-9.2), and EC (6.2-53.3 dS m(-1)). Manure DP concentrations measured using the RQFlex reflectometer (RQFlex-DP(s)) were related to molybdate-reactive P (MRP(s)) concentrations as follows: RQFlex-DP(s) = 0.471 x MRP(s) + 1102 (r2 = 0.29). Inclusion of pH and squared-pH terms improved the prediction of manure DP from RQFlex results (r2 = 0.66). Excluding five outlier samples that had pH < or = 6.9 the coefficient of determination (r2) for the MRP(s) and RQFlex-DP(s) relationship was 0.83 for 95% of the samples. Manure TS were related to hydrometer specific gravity readings (r2 = 0.53) that were in turn related to TP (r2 = 0.34), but not to either RQFlex-DP or MRP. Relationships between suspensions' EC and DP or TP were non-significant. Therefore, the RQFlex method is the only viable option for on-site quick estimates of DP that can be made more robust when complemented with TS and pH measurements. The DP quick test can provide near real-time information on soluble manure nutrient content across a wide range of handling and storage conditions on dairy farms and quick estimates of potential soluble P losses in runoff following land applications of manure.     

The presence of Bacillus thuringiensis (Bt) protein in earthworms Eisenia fetida has no deleterious effects on their growth and reproduction

Earthworms Eisenia fetida, bred in substances with stover of two genetically-engineered Bacillus thuringiensis (Bt) corns (5422Bt1 (Event Bt11) and 5422CBCL (MON810)) expressing Cry1Ab and their near-isogenic non-Bt corn (5422), were used to investigate the non-target effects of Bt corn on soil-dwelling organisms. Cry1Ab concentrations in substances, casts and guts of E. fetida were also investigated by Enzyme-linked immunosorbent assay (ELISA). More than 90% individuals of E. fetida survived over a period of 30 d, irrespective of whether they received Bt corn or non-Bt corn. Compared to 5422 treatments, significantly higher relative growth rate and more number of new offspring and cocoons of E. fetida were found in 5422Bt1 and 5422CBCL treatments. These results were unlikely to be directly caused by Cry1Ab released from Bt corns but rather by differences in other factors of plants such as plant components (soluble sugar, total organic carbon, total protein and available phosphorus of Bt corns were more than 5422). ELISA results indicated immunoreactive Cry1Ab was detectable in substances, and the casts, guts of E. fetida from Bt corns treatments, of which the highest levels were detected in substances under the corresponding experimental conditions. With the increase of treated time, a strong decline was observed in Cry1Ab from substances and casts of E. fetida, whereas Cry1Ab in guts of E. fetida from 5422Bt1 treatments gradually increased and that from 5422CBCL treatments increased between 14 and 30 d. Therefore, the presence of Cry1Ab in E. fetida had no deleterious effects on their growth and reproduction.     

The effects of engineered nanoparticles on survival, reproduction, and behaviour of freshwater snail, Physa acuta (Draparnaud, 1805)

Increasing uses of engineered nanoparticles (ENPs) in commercial products and industrial applications has eventually resulted to their releases into atmospheric, terrestrial, and aquatic environments. However, knowledge gaps in ENPs toxicity, fate, and behaviour currently limit our ability to quantify risk assessment of materials with nanoscale dimensions, and therefore, the extent of the resultant environmental impacts remains unknown. In the present study, we evaluated the effects of γ-alumina, α-alumina, modified TiO(2) (M-TiO(2)), and commercial TiO(2) (C-TiO(2)) ENPs on the survival, behaviour, and early life stages of the freshwater snail Physa acuta (Draparnaud). The toxicity evaluation was carried out after spiking commercial sand with ENPs concentrations of 0.005, 0.05, or 0.5 gk g(-1). Our findings suggest that increases of γ-alumina and α-alumina concentrations at sub-lethal level concentrations caused significant reduction in the embryo growth rate and embryo hatchability. In addition, these ENPs induced observable developmental deformities of the embryos. In addition, toxicity evaluations using acute 96-h and chronic 28-d tests showed exposure duration may be a significant factor in ENPs-induced toxicity. Therefore, long-term exposure of aquatic organisms to ENPs - potentially can alter certain ecological populations at different trophic levels - and may compromise the entire aquatic ecological functionality. The percentage hatchlings in test chambers containing 0.5 gk g(-1) γ-alumina and α-alumina concentration was 50% less to those observed in the controls. Our results suggest the embryonic growth and hatchability tests are useful endpoints in chronic sediment toxicity tests for determining the toxic thresholds of ENPs in sediment environment. Although no snail mortalities were observed during the static 96-h test containing sediment spiked with different concentrations of M-TiO(2), C-TiO(2), γ-alumina and α-alumina - the antioxidant enzymatic assay results indicated a significant change in antioxidant levels which altered peroxidation at 0.05 or 0.5 gk g(-1)concentrations for both γ-alumina and α-alumina.     

First study of congener group patterns and concentrations of short- and medium-chain chlorinated paraffins in fish from the North and Baltic Sea

This study presents the first investigation of concentrations and congener group patterns of short- (SCCPs) and medium-chain chlorinated paraffins (MCCPs) in fish from the North and Baltic Sea. North Sea dab, cod and flounder were studied. High resolution gas chromatography (HRGC) coupled to low resolution mass spectrometry (LRMS) in the electron capture negative ionization mode (ECNI) was employed. Good linearity (R2>0.993, 7 measuring points) was achieved between 1 and 100 ng/g of CP mixtures for SCCPs and MCCPs. The limits of detection were 0.5-1 ng/microl of CP mixture for the major congener groups of SCCPs and MCCPs. A clean-up comprising fat extraction, adsorption chromatography on silicagel impregnated with concentrated sulphuric acid and adsorption chromatography on Florisil was employed to avoid interferences from other polychlorinated compounds. Recoveries of CPs in spiked samples ranged between 80% and 100%. Accuracy was controlled with spiked samples and deviated not more than 10% from the expected values. Quantification was performed with standards of an average chlorine content as close as possible to that of the samples (SCCPs: 59-62%, MCCPs: 53-58%). SCCP concentrations ranged between 19 and 286 ng/g liver wet weight (ww), MCCP concentrations were comparable with a range of 25-260 ng/gww. Congener group patterns were also determined and discussed. In samples from the Baltic Sea the SCCP congener pattern was similar to that of commercial SCCP mixtures or C13 congeners were most abundant. In samples from the North Sea a higher relative abundance of C10 congeners was observed.     

Dispersal and persistence of genetically modified oilseed rape around Japanese harbors

Background, aim, and scope

The possibility of gene transfer from genetically modified oilseed rape (OSR) to its cultivated or wild relatives is of concern since its commercial cultivation, because of its potential weediness and impact on the environment. Introgression of modified genes can affect conservation of agricultural crops, because there are many cultivars and wild Brassicaceae that may cross with genetically modified OSR (Brassica napus) in Japan. Japan imports more than 2 million tons of OSR a year from Canada and other countries. Since volunteers of GM OSR were found around harbors in 2004, a lot of feral GM OSR was discovered in Japan. To consider the way how to keep domestic Brassicaceae from GM contamination, we surveyed and analyzed the dispersal and persistence of GM OSR around Japanese harbors. We present the cause and abundance of GM OSR in Japan by this paper.

Materials and methods

Survey of the feral OSR was performed several times a year at different seasons when domestic OSR either grows or does not around port areas. Detection of herbicide tolerance in feral B. napus was done by test papers that cross react with the modified gene product. Two kinds of herbicide tolerance (glyphosate and glufosinate) were tested.

Results

The feral B. napus were discovered around all 13 harbors that import rapeseeds from foreign countries. Genetically modified, herbicide-tolerant OSR were frequently found in the surveyed populations. Two kinds of herbicide-tolerant OSR (glyphosate- and glufosinate-tolerant) were discovered in a natural condition 40 km from port to an oil factory where 60,000 tons of OSR seed are processed a year. The cause of voluntary growth of OSR is seed spillage during transportation by trucks from harbors to oil factories and other processing facilities. Some of the feral OSR growing along the roadsides of transport paths exhibited perennial growth spilling their seeds around the places. Alteration of the generation of feral GM OSR was discovered for the first time in Japan as a result of this study. We studied the yearly change of feral OSR abundances focusing on Yokkaichi port over the 4 years since 2004. The rate of GM OSR increases year to year, and reaches nearly 90% in 2008.

Discussion

The possibility and consequences of gene transfer from the genetically modified OSR to domestic species (B. rapa and B. juncea) were discussed in relation to impact on domestic agriculture and on environments. Evolutional meaning of the gene transfer was also discussed with respect to the gene construct of GM OSR. This study shows the importance of another pathway of modified gene transfer to non-GM relative species by seed transportation in addition to pollen transfer from commercial cultivation of genetically modified OSR.

Conclusions and recommendations

We identified unintended dispersal and persistence of GM OSR around Japanese harbors that import OSR from Canada and other countries. Both glyphosate- and glufosinate-tolerant feral B. napus were discovered. The cause of volunteer OSR is spillage of the seeds during transportation by truck to oil factory. The feral GM OSR sometimes showed perennial growth in Japanese phonological conditions which are not observed for domestic Brassicaceae. In addition, we confirmed an alteration of generations by feral GM OSR in Japan. The possibility of cross pollination and GM gene introgression to domestic varieties can occur in these environments. To improve the situation, each responsible organization, company, administration, or government should establish measures how to stop the dispersal and persistence of GM OSR in nature. Also, the GM plant developers are responsible for revising this situation.     

Elevated atmospheric ozone increases concentration of insecticidal Bacillus thuringiensis (Bt) Cry1Ac protein in Bt Brassica napus and reduces feeding of a Bt target herbivore on the non-transgenic parent

Sustained cultivation of Bacillus thuringiensis (Bt) transgenic crops requires stable transgene expression under variable abiotic conditions. We studied the interactions of Bt toxin production and chronic ozone exposure in Bt cry1Ac-transgenic oilseed rape and found that the insect resistance trait is robust under ozone elevations. Bt Cry1Ac concentrations were higher in the leaves of Bt oilseed rape grown under elevated ozone compared to control treatment, measured either per leaf fresh weight or per total soluble protein of leaves. The mean relative growth rate of a Bt target herbivore, Plutella xylostella L. larvae was negative on Bt plants in all ozone treatments. On the non-transgenic plants, larval feeding damage was reduced under elevated ozone. Our results indicate the need for monitoring fluctuations in Bt toxin concentrations to reveal the potential of ozone exposure for altering dosing of Bt proteins to target and non-target herbivores in field environments experiencing increasing ozone pollution.     

Back to Basics

ABSTRACT

Tests of the compatibility of geomembrane (GM) samples with waste were conducted using U.S. Environmental Protection Agency (EPA) Method 9090 and the Comprehensive Testing System (CTS). The CTS is a multi-axial performance test capable of simultaneous cyclic mechanical loads and chemical exposure. The test chemicals consisted of solvents, transportation-related compounds, and synthesized landfill leachate. Method 9090 testing was unable to distinguish between the effects of individual chemicals to which the GM was subjected, while the CTS was able to provide statistically-significant differences that were also traceable to chemical properties of the solvent and the GM liner. Further, the time required for changes in mechanical properties of the GM was significantly shorter than would be expected based upon diffusion of the solvent into the GM alone. The combination of chemical attack with mechanical load was found to enhance both reduction in mechanical properties and the ability of the solvent to diffuse into the GM. The CTS is a more realistic test than the existing standard test methods because of its ability to provide multi-axial loads and chemical exposure simultaneously.     

Colloidal gold probe-based immunochromatographic assay for the rapid detection of lead ions in water samples

One-step immunochromatographic assay (ICA) has been developed using colloidal gold-labeled monoclonal antibody probe for the rapid detection of lead ions in water samples. The ICA was based on the theory of competitive reactivity, and the results can be easily judged based on the presence or absence of a red colored test line with visual detection. Under optimal conditions, this method shows high detecting sensitivity with a LOD (limit of detection) of 50 ng/ml. Stability test indicates that the immunochromatographic strips are stable for 8 weeks at room temperature. During practical application, nanometer TiO₂ is used to enrich the lead ions in water samples. The ICA is successfully applied in the measurement of lead ion concentrations in local water samples, and the results are highly consistent with that of ICP-MS. Detecting lead ions with ICA can be done within 4 min and is very useful for the rapid onsite testing.     

Arsenic species and chemistry in groundwater of southeast Michigan

Groundwater samples, taken from 73 wells in 10 counties of southeast Michigan in 1997 had arsenic concentrations in the range of 0.5 to 278 microg/L the average being 29 microg/l. About 12% of these wells had arsenic concentrations that exceeded the current USEPA's maximum contaminant level of 50 microg/l. Most (53-98%) of the arsenic detected was arsenite [As(III)] and other observations supported the arsenic species distribution (low redox potential and DO). In shallow groundwater (< 15 m), arsenic concentrations are low likely due to the formation of insoluble ferrosoferric hydroxide complex. In deep groundwater (> 15 m), the concentration of arsenic is possibly controlled by reductive dissolution of arsenic-rich iron hydroxide/oxyhydroxide and dissolution of arsenic sulfide minerals.     

A simplified headspace biochemical oxygen demand test protocol based on oxygen measurements using a fiber optic probe.

Batch respirometric tests have many advantages over the conventional biochemical oxygen demand (BOD) method for analysis of wastewaters, including the use of nondiluted samples, a more rapid exertion of oxygen demand, and reduced sample preparation time. The headspace biochemical oxygen demand (HBOD) test can be used to obtain oxygen demands in 2 or 3 days that can predict 5-day biochemical oxygen demand (BOD5) results. The main disadvantage of the HBOD and other respirometric tests has been the lack of a simple and direct method to measure oxygen concentrations in the gas phase. The recent commercial production of a new type of fiber optic oxygen probe, however, provides a method to eliminate this disadvantage. This fiber optic probe, referred to here as the HBOD probe, was tested to see if it could be used in HBOD tests. Gas-phase oxygen measurements made with the HBOD probe took only a few seconds and were not significantly different from those made using a gas chromatograph (t test: n = 15, R2 = 0.9995, p < 0.001). In field tests using the HBOD probe procedure, the probe greatly reduced sample analysis time compared with previous HBOD and BOD protocols and produced more precise results than the BOD test for wastewater samples from two treatment plants (University Area Joint Authority [UAJA] Wastewater Treatment Plant in University Park, Pennsylvania, and The Pennsylvania State University [PSU] Wastewater Treatment Plant in University Park). Headspace biochemical oxygen demand measurements on UAJA primary clarifier effluent were 59.9 +/- 2.4% after 2 days (HBOD2) and 73.0 +/- 3.1% after 3 days (HBOD) of BOD, values, indicating that BOD5 values could be predicted by multiplying HBOD2 values by 1.67 +/- 0.07 or HBOD3 by 1.37 +/- 0.06. Similarly, tests using PSU wastewater samples could be used to provide BOD5 estimates by multiplying the HBOD2 by 1.24 +/- 0.04 or by multiplying the HBOD3 by 0.97 +/- 0.03. These results indicate that the HBOD fiber optic probe can be used to obtain reliable oxygen demands in batch respirometric tests such as the HBOD test.     

Evaluation of sample recovery of malodorous livestock gases from air sampling bags, solid-phase microextraction fibers, Tenax TA sorbent tubes, and sampling canisters

Odorous gases associated with livestock operations are complex mixtures of hundreds if not thousands of compounds. Research is needed to know how best to sample and analyze these compounds. The main objective of this research was to compare recoveries of a standard gas mixture of 11 odorous compounds from the Carboxen/PDMS 75-microm solid-phase microextraction fibers, polyvinyl fluoride (PVF; Tedlar), fluorinated ethylene propylene copolymer (FEP; Teflon), foil, and polyethylene terephthalate (PET; Melinex) air sampling bags, sorbent 2,b-diphenylene-oxide polymer resin (Tenax TA) tubes, and standard 6-L Stabilizer sampling canisters after sample storage for 0.5, 24, and 120 (for sorbent tubes only) hrs at room temperature. The standard gas mixture consisted of 7 volatile fatty acids (VFAs) from acetic to hexanoic, and 4 semivolatile organic compounds including p-cresol, indole, 4-ethylphenol, and 2'-aminoacetophenone with concentrations ranging from 5.1 ppb for indole to 1270 ppb for acetic acid. On average, SPME had the highest mean recovery for all 11 gases of 106.2%, and 98.3% for 0.5- and 24-hr sample storage time, respectively. This was followed by the Tenax TA sorbent tubes (94.8% and 88.3%) for 24 and 120 hr, respectively; PET bags (71.7% and 47.2%), FEP bags (75.4% and 39.4%), commercial Tedlar bags (67.6% and 22.7%), in-house-made Tedlar bags (47.3% and 37.4%), foil bags (16.4% and 4.3%), and canisters (4.2% and 0.5%), for 0.5 and 24 hr, respectively. VFAs had higher recoveries than semivolatile organic compounds for all of the bags and canisters. New FEP bags and new foil bags had the lowest and the highest amounts of chemical impurities, respectively. New commercial Tedlar bags had measurable concentrations of N,N-dimethyl acetamide and phenol. Foil bags had measurable concentrations of acetic, propionic, butyric, valeric, and hexanoic acids.     

Mycobiota and mycotoxins in bee pollen collected from different areas of Slovakia

Contamination by microscopic fungi and mycotoxins in different bee pollen samples, which were stored under three different ways of storing as freezing, drying and UV radiation, was investigated. During spring 2009, 45 samples of bee-collected pollen were gathered from beekeepers who placed their bee colonies on monocultures of sunflower, rape and poppy fields within their flying distance. Bee pollen was collected from bees’ legs by special devices placed at the entrance to hives. Samples were examined for the concentration and identification of microscopic fungi able to grow on Malt and Czapek-Dox agar and mycotoxins content [deoxynivalenol (DON), T-2 toxin (T-2), zearalenone (ZON) and total aflatoxins (AFL), fumonisins (FUM), ochratoxins (OTA)] by direct competitive enzyme-linked immunosorbent assays (ELISA). The total number of microscopic fungi in this study ranged from 2.98 ± 0.02 in frozen sunflower bee pollen to 4.06 ± 0.10 log cfu.g^?1 in sunflower bee pollen after UV radiation. In this study, 449 isolates belonging to 21 fungal species representing 9 genera were found in 45 samples of bee pollen. The total isolates were detected in frozen poppy pollen 29, rape pollen 40, sunflower pollen 80, in dried poppy pollen 12, rape pollen 36, sunflower 78, in poppy pollen after UV radiation treatment 54, rape 59 and sunflower 58. The most frequent isolates of microscopic fungi found in bee pollen samples of all prevalent species were Mucor mucedo (49 isolates), Alternaria alternata (40 isolates), Mucor hiemalis (40 isolates), Aspergillus fumigatus (33 isolates) and Cladosporium cladosporioides (31 isolates). The most frequently found isolates were detected in sunflower bee pollen frozen (80 isolates) and the lowest number of isolates was observed in poppy bee pollen dried (12 isolates). The most prevalent mycotoxin of poppy bee pollen was ZON (361.55 ± 0.26 μg.kg^?1), in rape bee pollen T-2 toxin (265.40 ± 0.18 μg.kg^?1) and in sunflower bee pollen T-2 toxin (364.72 ± 0.13 μg.kg^?1) in all cases in frozen samples.     

Detection of fullerenes (C60 and C70) in commercial cosmetics

Detection methods are necessary to quantify fullerenes in commercial applications to provide potential exposure levels for future risk assessments of fullerene technologies. The fullerene concentrations of five cosmetic products were evaluated using liquid chromatography with mass spectrometry to separate and specifically detect C₆₀ and C₇₀ from interfering cosmetic substances (e.g., castor oil). A cosmetic formulation was characterized with transmission electron microscopy, which confirmed that polyvinylpyrrolidone encapsulated C₆₀. Liquid-liquid extraction of fullerenes from control samples approached 100% while solid-phase and sonication in toluene extractions yielded recoveries of 27-42%. C₆₀ was detected in four commercial cosmetics ranging from 0.04 to 1.1 μg/g, and C₇₀ was qualitatively detected in two samples. A single-use quantity of cosmetic (0.5 g) may contain up to 0.6 μg of C₆₀, demonstrating a pathway for human exposure. Steady-state modeling of fullerene adsorption to biosolids is used to discuss potential environmental releases from wastewater treatment systems.     

Detection of antibiotics in chicken eggs obtained from supermarkets in Ho Chi Minh City,Vietnam

The residual levels of antibiotics in Vietnamese eggs were monitored from 2014 to 2015. A total of 111 egg packages, distributed by 11 different companies, were collected from supermarkets in Ho Chi Minh City and the levels of 28 antibiotics were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method. Sixteen samples tested positive for antibiotics; a total of eight compounds (enrofloxacin, ciprofloxacin, norfloxacin, sulfadimethoxine, sulfamethazine, sulfamonomethoxine, tilmicosin and trimethoprim) were detected. Enrofloxacin was detected in eight samples, with two samples exhibiting concentrations exceeding 1,000 µg kg^?1. Tilmicosin was detected in three samples at a range of 49–568 µg kg^?1. We observed that two of the 11 companies frequently sold antibiotic-contaminated eggs (detection rates of 56 and 60%), suggesting that a number of companies do not regulate the use of antibiotics in egg-laying hens. Our findings indicate that livestock farmers require instruction regarding antibiotic use and that continual antibiotic monitoring is essential in Vietnam.     

Rapid enzyme immunoassays for the detection of carbaryl and methoprene in grains

In order for grain handlers and traders to reliably estimate residues of grain protectants in the field, antibody-based rapid tests were developed for carbaryl (1-naphthyl methylcarbamate) and methoprene [isopropyl (E,E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2,4-dienoate]. To complement the rapid analysis, a simple and rapid extraction technique was developed. In these tests, a pesticide-containing methanol extract of the grain sample and an enzyme-labeled component are added to precoated strips. After a brief incubation, the strips are washed and a substrate/chromogen for the enzyme is added. The color developed is stopped by acidification and the results are read either by eye or in a portable field photometer. The overall test time is under 20 minutes. For carbaryl, the test had a limit of detection of 4.5 ppb (1.1 ppm in grain), while the methoprene test had a limit of detection of 4 ppb (1 ppm in grain) based on the lower datum point, which is 15% inhibition, in the standard curves. Both assays can be used as a screening test for carbaryl and methoprene in animal feed grains.