Heavy metals in surface sediments from Minjiang Esturary—Mazu and Xiamen—Jinmen sea areas

The concentrations of Cu,Pb, Zn and Cd in surface sediments from Minjang Estuary-Mazu and Ziamen-Jinmen sea areas of China were investigated during 1995-1996.The concentration ranges of Cu,Pb ,Zn and Cd in the Minjiang Estuary-Mazu sea area were 16.4-37.2,33.4-69.6,92.1-128,0.087-0.036mg/kg(dry wt.),respectively; those for Xiamen-Jinmen sea area were 11.0-24.5,36.0-80.3,77.5-161,0.135-0.285mg/kg(dry wt.), respectively.The concentration and distributions of Cu,Pb,Zn and Cd in surface sediments of Minjiang Estuary-Mazu and Xiamen-Jinmen sea areas were analyzed and evaluated.The results showed that the values of Zn and Pb exceeded those of sediment quality criteria.The average concentration of Cu in Minjiang Estuary-Mazu sea area was higher than that in Xiamen-Jinmen sea area.The obviously higher concentrations of Cu,Pb, Zn and Cd were found at outlets of Minjiang and Jiulong River.From the estuaries to open sea, the concentrations of Cu, Pb,Zn and Cd had a decreasing trend, and then elevated in the sea areas near Mazu and Jinmen, suggesting that Cu,Pb,Zn and Cd in Minjiang Estuary-Mazu and Xiamen-Jinmen sea areas may come from both the mainland and Taiwan of China.     

Distribution of persistent organochlorine pesticides in tissue/organ of silver carp (Hypophthalmichthys molitrix) from Guanting Reservoir, China

The concentration of organochlorine pesticides（OCPs） in tissues and organs of silver carp（Hypophthalmichthys mofitrix） from Guanting Reservoir were investigated to evaluate the pollution potential and distribution of OCPs. A total of 16 OCPs were measured and the concentrations were in the range of 1.61-69.01 ng/g wet weight（ww） for total OCPs, 0.16- 0.75 ng/g ww for HCB. 0.75 -26.80 ng/g ww for SHCH（sum of α-,β-, γ- and δ-HCH） and 0.68-35.94 ng/g ww for SDDT（ sum of p, p＇-DDE, p, p＇-DDD, o, p＇-DDT and p, p＇- DDT）. The mean concentrations of total OCPs, HCB, ∑HCH, and ∑DDT were 18.04, 0.96, 7.14 and 9.28 ng/g ww, respectively. Among the organochlorine pesticides, β-HCH and p, p＇-DDE were the most dominant compounds in tissue and organ with the average concentrations of 4.42 and 8.14 ng/g, respectively. The results obtained in this study show that the levels of 16 OCP residues found in silver carps are low and pose no threat to human health and wildlife fed upon them on the basis of existing related quality guidelines. However, recent input of lindane and DDT might still exist in the area investigated and further investigation should be carried on.     

Studies on metabolism of 2-naphthylamine and its activation mechanism

2-naphthylamine was incubated with induced rat liver microsome S9 preparation and the metabolites were separated through HPLC. The following products were identified: 2-amino-5-naphthol, 2-amino-6-naphthol, 2-amino-7-naphthol and 2-amino-8-naphthol. The yields of these four metabolites are varying in quantity, and the relative contents of 2-amino-8-, -5-, -6- and -7-naphthol are 52.6%, 28.5%, 14.0% and 4.9% respectively. These results are consistent with the quantitative HMO calculation and inference based upon Di-region theory, i.e., the metabolisms of aryl amines on extra-ring (assigned the ring without the substituent of amino group) are through the epoxidation and then NIH shift, but are not the direct hydroxylation in the formation of phenols. It is shown that both the amino group and the carbon atoms on the extra-ring play duality roles of activation and detoxification in metabolism.     

Diurnal variation of number concentration and size distribution of ultrafine particles in the urban atmosphere of Beijing in winter

Number concentration and distribution of airborne particles in the size range 5.6 to 560 um diameter were measured in Beijing for a 15-d period in winter 2005.Daily average number concentrations of nucleation mode(5.6-20 nm),Aitken mode(20-100 nm), and accumulation mode(100-560 nm)particles,and total particles were 17500,32000,4000,and 53500 cm~(-3),respectively.Average particle size distribution was monomodal with a mode diameter of about 40 um at night and bimodal with mode diameters of about 10 and about 40 nm during the daytime.New particle formation events,which were connected to diurnal variation of nucleation mode particles,were observed in more than half of the observation days.The events often started around 10:00-11:00 Chinese Standard Time (CST)and ended up after 3-4 h.Concentrations of Aitken and accumulation mode particles increased from midnight and reached their maxima at about 10:00 CST,and then decreased and became the lowest in the afternoon.Analysis of diurnal cycles in traffic volume and meteorological parameters revealed that the accumulation of the particles in Aitken and accumulation modes in the morning was influenced by formation of an inversion and increase in vehicle emission,and dispersion of such particles in the afternoon was associated with more effective vertical mixing and higher wind speed.     

Field released transgenic papaya effect on soil microbial communities and enzyme activities

Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences （P〈0.05） were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences （P〈0.05） in the total number of colony forming units （CFUs） of bacteria, actinomycetes and fungi between soils amended with RP-transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher （P〈0.05） CFUs of bacteria, actinomycetes and fungi resistant to kanamycin （Km） were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Km＇ （kanamycin resistant） bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Km＇ bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase （5.4-5.9x）, polyphenol oxidase （0.7-1.4x）, invertase （0.5-0.79x）, cellulase （0.23-0.35x） and phosphodiesterase （0.16-0.2x）. The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities.     

Effect of the air pollution on atmospheric visibility in Beijing-Tianjin area

Simultaneous measurements of air pollutant concentrations and atmospheric visibility were made in Beijing and Tianjin areas in 1983-1985. The relationship between air pollution and visibility was studied. It was found that atmospheric particulates have the most high contribution to visibility decreasing. The percentages of contributions of sulfates and soot are 52-58% and 22-29% respectively. According to the results, we suggest that the emission of SO2 and particulates must be controlled in order to improve the atmospheric visibility.     

Residue and persistence of insecticide l-(2-chlorobenzoyl)-3-(4-chlorophenyl)urea in C hinese cabbage and soils in the Northern China

The residue and persistence of the insect growth regulator 1-(2-(chlorobenzoyl)-3-(4-chlorophenyl)urga(CCU)in Chinese cabbage and soil in the field were studied in Northern China during 1988-1989．The degradation of CCU in soils was also studied under laboratory conditions．The CCU was found to be unstable in Chmese cabbage and soil in field．The half-life ofCCU was 13.2-14.0 days in Chinese cabbage and 8.8-27.0 days in soil,respectively．Three ppm as the maximum residue level for CCU in Chinese cabbage and 21 days as the preharvest interval for the vegetable treated with CCU were recommendced．     

Occurrence and infection risk of waterborne pathogens in Wanzhou watershed of the Three Gorges Reservoir, China

The Three Gorges Reservoir (TGR), formed by China’s Yangtze Three Gorges Project, is the largest lake in the world, but there is too little information available about fecal contamination and waterborne pathogen impacts on this aquatic ecosystem. During two successive 1-year study periods (July 2009 to July 2011), the water quality in Wanzhou watershed of the TGR was tested with regard to the presence of fecal indicators and pathogens. According to Chinese and World Health Organization water quality standards, water quality in the mainstream was good but poor in backwater areas. Salmonella, Enterohemorrhagic Escherichia coli (EHEC), Giardia and Cryptosporidium were detected in the watershed. Prevalence and concentrations of the pathogens in the mainstream were lower than those in backwater areas. The estimated risk of infection with Salmonella, EHEC, Cryptosporidium, and Giardia per exposure event ranged from 2.9 × 10 -7 to 1.68 × 10-5 , 7.04 × 10-10 to 2.36 × 10-7 , 5.39 × 10-6 to 1.25 × 10-4 and 0 to 1.2 × 10-3 , respectively, for occupational divers and recreational swimmers exposed to the waters. The estimated risk of infection at exposure to the 95% upper confidence limit concentrations of Salmonella, Cryptosporidium and Giardia may be up to 2.62 × 10-5 , 2.55 × 10-4 and 2.86 × 10-3 , respectively. This study provides useful information for the residents, health care workers and managers to improve the safety of surface water and reduce the risk of fecal contamination in the TGR.     

Changes in heavy metal contents in animal feeds and manures in an intensive animal production region of China

The 360 feed and manure samples were collected from 150 animal farms in Jiangsu Province, China and analyzed for heavy metals. Concentrations of Zn and Cu in animal feeds were 15.9-2041.8 and undetected-392.1 mg/kg respectively, while Hg, As, Pb, Cd, and Cr in all feeds were below 10 mg/kg. Concentrations of Cu, Zn, and Cr in animal manures were 8.4-1726, 39.5-11379, and 1.0-1602 mg/kg respectively, while As, Cd, Hg, and Pb were 〈 10 mg/kg. The concentration of Cu, Zn, As and Cr in animal feed and manure were positively correlated （p 〈 0.001）, but the Cd, Hg, and Pb were not statistically correlated between the feed and the manure. Concentrations of Cu and Zn were highest in pig feed and manure, followed by poultry and dairy feeds and manures. During 1990- 2008, Cu, Zn, As, Cr, Cd contents increased by 771%, 410%, 420%, 220%, and 63% in pig manure, 212%, 95%, 200%, 791%, and -63% in dairy manure, and 181%, 197%, 1500%, 261, and 196% in poultry manure. Most of the increases occurred from 2002 to 2008, which reflects the extensive use of feed additives after 2002. In contrast, Pb and Hg in manures continuously decreased from 1990 to 2008. The results suggest that the heavy metal contents in animal manure have been greatly increased over 18 years and the contribution of manures to soil should be considered.     

Effects of nature organic matters and hydrated metal oxides on the anaerobic degradation of lindane, p, p '-DDT and HCB in sediments

Effects of natural organic matters(NOM) and hydrated metal oxides(HMO) in sediments on the anaerobic degradation of γ-666,p, p‘-DDT and HCB were investigated by means of removing NOM and HMO in Liaohe River sediments sequentially. The results showed that the anaerobic degradation of γ-666, p, p‘ -DDT and HCB followed pseudo-first-order kinetics in different sediments. But, the extents and rates of degradation were different, even the other conditions remained the same. Anaerobic degradation rates of γ-666, p, p ‘-DDT and HCB were 0.020 d^-1, 0.009 d^-1 and 0.035 month^-1 respectively for the sediments without additional carbon resources. However, with addition ofcarbon resources, the anaerobic degradation rates of γ-666, p, p‘-DDT and HCB were 0.071 d^1, 0.054 d^-1 and 0.088 month^-1 in the original sediments respectively. After removing NOM, the rates were decreased to 0.047 d^-, 0.037 d^-1 and 0.066 month^-1 ; in thesediments removed NOM and HMO, the rates were increased to 0.067 d^-1 , 0.059 d^-1 and 0.086 month^-1 . These results indicated that NOMin the sediments accelerated the anaerobic degradation of γ-666, p, p‘-DDT and HCB ; the HMO inhibited the anaerobic degradation of γ-666,p, p ‘-DDT and HCB.     

Experimental study on MSW gasification and melting technology

In order to develop municipal solid waste (MSW) gasification and melting technology, two preliminary experiments and a principle integrated experiment were fulfilled respectively. The gasification characteristics of MSW are studied at 500-750℃ when equivalence ratio (ER) was 0.2-0.5 using a fluidized-bed gasifier. When temperature was 550-700℃ and ER was 0.2-0.4, low heat value (LHV) of syngas reaches 4000-12000 kJ/Nm3. The melting characteristics of fly ash were investigated at 1100-1460℃ using a fixed-bed furnace. It was proved that over 99.9% of dioxins could be decomposed and most heavy-metals could be solidified when temperature was 1100-1300℃. The principle integrated experiment was carried out in a fluidized-bed gasification and swirl-melting system. MSW was gasified efficiently at 550-650℃, swirl-melting furnace maintains at 1200-1300℃ stably and over 95% of fly ash could be caught by the swirl-melting furnace. The results provided much practical experience and basic data to develop MSW gasification and melting technology.     

Palaeorunoff estimations achieved from palaeoclimatic information for the southwest part of the North China Plain: an attempt to apply a climatological approach to palaeohydrology

Palaeoclimatic scenario projecting annual temperature and annual precipitation is firstly formalized with data available and speculations for the southwest part of the North China Plain (35- 37°N, 115-119°E) during the last 25000 years. Then, with three regression equations relating annual runoff to annual precipitation and derived with data of modern hydrological and meteorological records, values of annual runoff are calculated in terms of the corresponding values of annual precipitation from this palaeoclimatic scenario for this region during this temporal interval. These results indicate that runoff is the most during 8000-3000a B.P. and the least during 25000-12000 a B. P.; runoff occurring during 12000-8000 a B.P. and during 3000-0 a B.P. is less than the one occurring during 8000-3000 a B.P. and more than the one occurring during 25000-12000 a B. P.; and the runoff occurring during 25000-12000 a B. P., 12000-8000 a B. P., and 3000-0 a B.P. is respectively 43, 46 and 66 percent of the one occurring during 8000-3000 a B.P. Values of bankfull discharge for palaeochannels of the Yellow River flowing in this region during the same interval are calculated from available estimates of slope of stream-bed of these palaeochannels with a regression equation relating bankfull discharge to slope of stream-bed and ratios of bankfull-discharge are further calculated from these values for different groups of palaeochannel formed during different time spans embraced in this interval. To conduct a cross-check, these values and ratios of bankfull-discharge are compared to the corresponding values and ratios of runoff occurring during roughly the same time spans. The same direction and similar relative magnitude of changes of the surface water occurring in this region during the last 25000 years are indicated by these comparisons.     

Particle number concentration, size distribution and chemical composition during haze and photochemical smog episodes in Shanghai

The aerosol number concentration and size distribution as well as size-resolved particle chemical composition were measured during haze and photochemical smog episodes in Shanghai in 2009. The number of haze days accounted for 43%, of which 30% was severe （visibility 〈 2 km） and moderate （2 km 〈 visibility 〈 3 km） haze, mainly distributed in winter and spring. The mean particle number concentration was about 17,000/cm3 in haze, more than 2 times that in clean days. The greatest increase of particle number concentration was in 0.5-1μm and 1-10 μm size fractions during haze events, about 17.78 times and 8.78 times those of clean days. The largest increase of particle number concentration was within 50-100 nm and 100-200 nm fractions during photochemical smog episodes, about 5.89 times and 4.29 times those of clean days. The particle volume concentration and surface concentration in haze, photochemical smog and clean days were 102, 49, 15 μm3/cm3 and 949, 649, 206 μm2/cm3, respectively. As haze events got more severe, the number concentration of particles smaller than 50 nm decreased, but the particles of 50-200 nm and 0.5-1μm increased. The diurnal variation of particle number concentration showed a bimodal pattern in haze days. All soluble ions were increased during haze events, of which NH4, SO24- and NO3 increased great/y, followed by Na＋, IC, Ca2＋ and CI-. These ions were very different in size-resolved particles during haze and photochemical smog episodes.     

Real-world vehicle emission factors in Chinese metropolis city--Beijing

The dynamometer tests with different driving cycles and the real-world tests are presented. Results indicated the pollutants emission factors and fuel consumption factor with ECE15 EUDC driving cycle usually take the lowest value and with real wodd driving cycle occur the highest value, and different driving cycles will lead to significantly different vehicle emission factors with the same vehicle. Relative to the ECE15 EUDC driving cycle, the increasing rate of pollutant emission factors of CO, NOx and HC are - 0.42--2.99, - 0.32--0.81 and - 0.11--11 with FTP75 testing, 0.11--1.29, - 0.77--0.64 and 0.47--10.50 with Beijing 1997 testing and 0.25--1.83, 0.09--0.75 and - 0.58--1.50 with real wodd testing. Compared to the carburetor vehicles, the retrofit and MPI TWC vehicles‘ pollution emission factors decrease with different degree. The retrofit vehicle (Santana) will reduce 4.44%--58.44% CO, - 4.95%--36.79% NOx, -32.32%--33.89% HC, and -9.39%--14.29% fuel consumption, and especially that the MPI TWC vehicle will decrease CO by 82.48%--91.76%, NOx by 44.87%--92.79%, HC by 90.00%--93.89% and fuel consumption by 5.44%--10.55%. Vehicles can cause pollution at a very high rate when operated in high power modes; however, they may not often operate in these high power modes. In analyzing vehicle emissions, it describes the fraction of time that vehicles operate in various power modes. In Beijing, vehicles spend 90% of their operation in low power modes or decelerating.     

Groundwater contamination and natural attenuation capacity at a petroleum spilled facility in Korea

As a remedial option, the natural attenuation capacity of a petroleum contaminated groundwater at a military facility was examined. Hydrogeological conditions, such as high water level, permeable uppermost layer and frequent heavy rainfall, were favorable to natural attenuation at this site. The changes in the concentrations of electron acceptors and donors, as well as the relevant hydrochemical conditions, indicated the occurrence of aerobic respiration, denitrification, iron reduction, manganese reduction and sulfate reduction. The calculated BTEX expressed biodegradation capacity ranged between 20.52 and 33.67 mg/L, which appeared effective for the reduction of the contaminants levels. The contribution of each electron accepting process to the total biodegradation was in the order: denitrification > iron reduction > sulfate reduction > aerobic respiration > manganese reduction. The BTEX and benzene point attenuation rates were 0.0058-0.0064 and 0.0005-0.0032 day^-1, respectively, and the remediation time was 0.7-1.2 and 2.5-30 years, respectively. The BTEX and benzene bulk attenuation rates were 8.69 × 10^-4 and 1.05 × 10^-3 day^-1, respectively, and the remediation times for BTEX and benzene were 7.2 and 17.5 years, respectively. However, most of the natural attenuation occurring in this site can be attributed to dilution and dispersion. Consequently, the biodegradation and natural attenuation capacities were good enough to lower the contaminants levels, but their rates appeared to be insufficient to reach the remediation goal within a reasonable time frame. Therefore, some active remedial measures would be required.     

Numerical sim ulation of transport and transform ation of insecticide 1-(2-chlorobenzoyl)-3-(4-chlorophenyl) urea in unsaturated soil

A numerical simulation model of pesticide transport and transformation in unsaturated soil is presented in this paper．The model was developed under basic principles of movement of water and solute in soil under transient condition and overall consideration water flow：pesticide convection，diffusion, dispersion，sorption and degradation．The model was tested for the persistence and leaching of 1-(2-chlorobenzoyl)-3-(4-chlorophenyl)urea(CCU)using data from soil columns under laboratory conditions．The results of the test indicate that the modeling approach is simple and effective in describing movement and transformation of pesticides in soils．     

Distribution and elimination of polycyclic musks in three sewage treatment plants of Beijing, China

The two representative polycyclic musks, 1-(5,6,7,8-tetrahydro-3,5,5,6,8,8-hexamethyl-2-naphthenyl)-ethanone (AHTN) and 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[g]-2-benzopyrane (HHCB), were measured in aqueous samples and sludge samples of three sewage treatment plants (STPs) in Beijing, China using a gas chromatograph-mass spectrometry. The HHCB and AHTN concentration ranges in the influents were 1251.4-3003.8 ng/L and 111.9-286.3 ng/L, respectively. Meanwhile, the ranges of 492.8-1285.3 ng/...     

Effects of elevated ozone on growth and yield of field-grown rice in Yangtze River Delta, China

With rapid industrialization and urbanization in the Yangtze Delta, China, the tropospheric ozone concentration has increased to levels that induce crop yield loss. Rice, a widely grown crop in China, was investigated in field-established, open-top chambers. Four treatments were used： charcoal-filtered air （CF）, non-charcoal-filtered air （NF）, and charcoal-filtered air with two levels of additional ozone （O3-1 and O3-2）. The AOT40s （accumulated hourly mean ozone concentration above 40 ppbv） were 0, 0.91, 23.24, and 39.28 ppmv.h for treatment of CE NF, O3-1, and O3-2, respectively. The rice height and biomass were reduced in the elevated ozone concentration. Less organic matter partitioning to roots under the elevated ozone significantly decreased rice root activity. The yield loss was 14.3% and 20.2% under O3-1 and O3-2 exposure, respectively. This was largely caused by a reduction in grain weight per panicle.     

Distribution of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in surface water and sediments from Baiyangdian Lake in North China

Persistent organochlorine compounds, including hexachlorocyclohexanes (HCHs), dichlorodiphenyltrichloroethanes (DDTs) and polychlorinated biphenyls (PCBs) were analyzed in surface water and sediments from Baiyangdian Lake, North China. Total concentrations of HCHs, DDTs and PCBs in surface water were in the range of 3.13-10.60, 4.05-20.59 and 19.46-131.62 ng/L, respectively, and total concentrations of HCHs, DDTs and PCBs in sediments were 1.75-5.70, 0.91-6.48 and 5.96-29.61 ng/g dry weight, respectively. Among the groups of HCHs (sum of α-HCH, β-HCH, γ-HCH and δ-HCH) and DDTs (sum of DDT, DDD and DDE), the predominance of β-HCH, DDE and DDD in water and sediment samples was clearly observed. This observation suggested that β-HCH was resistant to biodegradation and the DDTs had been transformed to its metabolites, DDE and DDD. For PCBs, penta-, hexa- and hepta-chlorinated congeners were the most abundant compounds in the both phases. Furthermore, the partitioning of chlorinated compounds between sediment and water was investigated to understand their transport and fates in aquatic ecosystems. The results indicated that average logs of organic carbon-normalized sediment-water partition coefficients (logK_oc') for OCPs varied between 3.20 and 5.53, and for PCBs, logK_oc' values ranged from 3.19 to 5.57. The observed logK_oc' was lower than their equilibrium logK_oc predicted from linear model, which may be attributed to the solubility enhancement effect of colloidal matter in water phase and the disequilibrium between sediment and water.     

Uptake and accumulation of phenanthrene and pyrene in spiked soils by Ryegrass(Lolium perenne L.)

Phytoremediation has long been recognized as a cost-effective method for the removal of polycyclic aromatic hydrocarbons （PAHs） from soil. A study was conducted to investigate the uptake and accumulation of PAHs in root and shoot of Lolium perenne L. Pot experiments were conducted with series of concentrations of 3.31-378.37 mg/kg for phenanthrene and those of 4.22-365.38 mg/kg for pyrene in a greenhouse. The results showed that both ryegrass roots and shoots did take up PAHs from spiked soils, and generally increased with increasing concentrations of PAH in soil. Bioconcentration factors（BCFs） of phenanthrene by shoots and roots were 0.24- 4.25 and 0.17-2.12 for the same treatment. BCFs of pyrene by shoots were 0.20-1.5, except for 4.06 in 4.32 mg/kg treatment, much lower than BCFs of pyrene by roots （0.58-2.28）. BCFs of phenanthrene and pyrene tended to decrease with increasing concentrations of phenanthrene and pyrene in soil. Direct uptake and accumulation of these compounds by Lolium perenne L. was very low compared with the other loss pathways, which meant that plant-promoted microbial biodegradation might be the main contribution to plant-enhanced removal of phenanthrene and pyrene in soil. However, the presence of Lolium perenne L. significantly enhanced the removal of phenanthrene and pyrene in spiked soil. At the end of 60 d experiment, the extractable concentrations of phenanthrene and pyrene were lower in planted soil than in non-planted soil, about 83.24%-91.98% of phenanthrene and 68.53%-84.10% of pyrene were removed from soils, respectively. The results indicated that the removal of PAHs in contaminated soils was a feasible approach by using Lolium perenne L.