Growth of Tomato Plants in Soil Contaminated with Kuwait Crude Oil

This laboratory study measured growth of one plant species, Lycopersicon esculentum Big Girl (tomato), that is sensitive to the presence of soil contamination, in Kuwait soil amended with crude-oil-contaminated soil. Germinated tomato seeds were placed in containers with soil containing 0, 0.12, 0.24, 0.36, 0.48, 0.60, 1.2, and 2.4% crude oil and were grown in an indoor growth chamber. Plants grew in Kuwait soil containing up to 0.36% crude oil; however, growth and fruit production were compromised at crude oil concentrations greater than 0.12% when compared with control plants. Plants did not grow in Kuwait soil amended with 0.48% crude oil or higher.     

High-temperature hydrocarbon biodegradation activities in Kuwaiti desert soil samples

Soil samples taken monthly from the Burgan South oil field of Kuwait for one year degraded crude oil, phenanthrene, and hexadecane. Bacteria were better degraders at high-temperature (55°C) than fungi, especially in the drier, hotter months. Depending on the period of sampling, bacteria degraded hydrocarbons in the range of 46–86% (crude oil), 42–100% (hexadecane) and 5–58% (phenanthrene). Fungi alone accounted for degradation by 20–81% (crude oil), 30–95% (hexadecane) and less than 55% (phenanthrene).     

Factors affecting the natural occurrence of entomopathogenic nematodes in turfgrass

We conducted a study to determine the relationship between turfgrass management intensity and natural occurrence of entomopathogenic nematodes (EPNs). We surveyed for EPNs on putting greens, fairways, and rough areas—three distinct surface types on golf courses that are managed with different intensities. We collected 159 soil samples from putting greens, contiguous fairways, and rough areas from 19 golf courses in Ohio, USA. Nematodes were recovered from soil samples using the insect baiting technique. We also analyzed the soil samples for texture, organic matter, pH, phosphorus, calcium, magnesium and potassium. We used principal components analysis and Pearson correlation to determine the relationship between nematode occurrence and the measured parameters. Surface type was the most important factor in predicting EPNs occurrence. Putting greens differed significantly from fairways and rough areas in the number of EPN-positive sites. No EPNs were recovered from putting greens but were recovered from 43% of the fairways and 57% of the rough areas. Putting greens also differed significantly from fairways and rough areas in organic matter, pH, calcium, and phosphorus. The fairways and rough areas did not however differ in number of EPN-positive sites and measured soil parameters. Presence of EPNs correlated significantly, although weakly, with sand, silt, phosphorus, organic matter, and magnesium content, but not with clay, pH, calcium, and potassium. Nematode isolates were identified as Heterorhabditis bacteriophora, Steinernema carpocapsae, and S. glaseri. This suggests that EPNs are more likely to occur in less intensively managed sites that receive fewer inputs and have relatively high sand, and moderate silt, organic matter, phosphorus, and magnesium content.     

Cultivation-dependent characterization of bacterial diversity from British Columbia forest soils subjected to disturbance

Bacteria from forest surface organic matter and mineral soil horizons were cultivated using four methods and characterized by fatty acid methyl ester (FAME) analysis. Soil samples from a British Columbia Ministry of Forests Long-Term Soil Productivity (LTSP) installation were collected during winter and summer from two disturbance treatments (whole-tree harvesting with no soil compaction (plot N) and whole-tree harvesting plus complete surface organic matter removal with heavy soil compaction (plot S)) and from an unlogged reference plot (REF). Seventy-five percent of 1795 bacterial isolates were affiliated with 42 genera representing beta- and gamma-Proteobacteria, Actinobacteria, the Bacillus/Clostridium group, and the Cytophaga-Flexibacter-Bacteroides group. Approximately half of the culture collection represented genetic diversity confined to four bacterial genera: Pseudomonas, Bacillus, Paenibacillus, and Arthrobacter. A significantly higher proportion of bacterial isolates belonging to Actinobacteria, and the member genus Arthrobacter, were isolated from plot S soil samples compared with soil samples from plots N and REF. Twenty-five percent of bacterial isolates were not conclusively identified to genus with FAME analysis. Sherlock Tracker cluster analysis and partial 16S rRNA gene sequence analysis enabled classification of a subset of these isolates.     

Hydrocarbon degradation in relation to cell-surface hydrophobicity among bacterial hydrocarbon degraders from petroleum-contaminated Kuwait desert environment

Forty six bacterial isolates able to grow on crude oil were isolated from various hydrocarbon-contaminated sites in Kuwait. The extent of crude oil degradation varied over a wide range (1–87%) among the isolates. Isolates were predominantly Gram-positive bacteria (79% of total isolates) belonging to the genera Bacillus (93%) and Paenibacillus (7%). Among the few Gram-negative isolates were from the genera Acinetobacter, Alcaligenes, Klebsiella, Burkholderia, Pseudomonas, and Williamsia. Analyses of their cell-surface hydrophobicity (CSH) by various methods equally showed a wide variation among the isolates. About 74% of isolates that degraded significant amounts of crude oil (>40% degradation) possessed high level of CSH, while 58% of all the isolates exhibited high levels of CSH. Statistical analyses showed significantly high correlation between the ability to degrade crude oil and CSH. The ability of the isolates to bind to polystyrene and salt-aggregation test as measures of CSH were more strongly correlated with hydrocarbon-degrading ability than adherence to hydrocarbons.     

A study of the enzymatic profile of soil isolates of Nocardia asteroides

In this study, using the API-ZYM system, we have reported the enzyme profile of 42 soil strains and 2 clinical strains of Nocardia asteroides isolated locally. Of the 19 enzymes tested, only 7 were demonstrable in over 90% of the soil isolates. These included alkaline phosphatase, esterase lipase, leucine arylamidase, acid phosphatase, phosphohydrolase, α-glucosidase and β-glucosidase. In addition, β-galactosidase activity was demonstrated in all the strains by the O-nitrophenyl-β-D-galactopyranoside (ONPG) test. The enzymes which were not demonstrable in >95% of the strains included valine arylamidase, cystine arylamidase, trypsin, chymotrypsin, α-galactosidase, β-glucoronidase, N-acetyl-β-glucosaminidase, α-mannosidase and α-fucosidase. With the exception of valine arylamidase, which was lacking in all but one isolate, the enzyme profiles of the soil isolates were comparable with the clinical isolates of N. asteroides reported in previous studies. The reasons for this difference in the two sets of isolates is not clear. The study reinforces the view that specific differences in the enzymatic profiles of Nocardia species could be used for their rapid identification. However, more extensive studies are needed to establish the reproducibility of this method. To the best of our knowledge, this is the first study of the enzymatic profile of soil isolates of N. asteroides originating from a single geographic region. This revised version was published online in June 2006 with corrections to the Cover Date.     

Compost Soil Piles for Treatment of Oil-Contaminated Soil

A number of diverse technological options are being considered for the remediation of soil contaminated with weathered crude oil in Kuwait. The bioremediation technique involving the use of composting soil piles was selected from among the most appropriate methods and evaluated on a pilot scale. The field test was conducted from November 1992 to September 1993 at the Burgan oil field. Soil piles were constructed from the contaminated soil after amendment with necessary soil additives. The piles were subjected to regular irrigation and turning, and a monitoring program was carried out, including monthly soil sample collection from each pile for the measurement of petroleum hydrocarbon PAHs, soil microbial counts, mineral and metal concentrations. The results obtained showed that the composting soil pile treatment resulted in the reduction of up to 59% total extractable matter of oil contamination within 8 months. This article describes the technology used and summarizes the results obtained.     

Oil bioremediation in a high and a low phosphorus soil

Phosphorus (P) content may influence bioremediation of soils contaminated with crude oil. A soil testing high in plant available P (Weswood, 194 mg P kg^?1 soil) and one testing low in plant available P (Lufkin, 2 mg P kg^?1 soil) were selected for laboratory experiments on oil biodegradation. Plant available P content was determined using acidified ammonium acetate at pH 4.2 as the soil extractant. Soils were amended with 3, 6, and 9% crude oil by weight and incubated for 120 d at 25°C. Treatments consisted of a factorial arrangement, with soil, N, P, and oil concentration as factors. Addition of P without N generally did not enhance biodegradation. Addition of N without P approximately tripled the quantity of oil degraded. Addition of P and N together did not increase biodegradation of oil more than addition of N alone when oil concentration was 3%. At 6 and 9% oil concentrations, CO₂ evolution increased for both soils by adding P and N together in comparison to adding N alone, and total petroleum hydrocarbon (TPH) bio‐degradation increased by 30% for the Weswood soil by 60 d and at least 25% for the Lufkin soil by 30 d. The quantity of plant‐available P or total P in soil was not very useful in predicting need for supplemental P. Addition of P to soil to enhance oil degradation was only beneficial for oil concentrations above 3% and the positive effect for higher concentrations was transitory.     

Enhanced crude oil biodegradation in soil via biostimulation

Research on feasible methods for the enhancement of bioremediation in soil contaminated by crude oil is vital in oil-exporting countries such as Kuwait, where crude oil is a major pollutant and the environment is hostile to biodegradation. This study investigated the possibility of enhancing crude oil bioremediation by supplementing soil with cost-effective organic materials derived from two widespread locally grown trees, Conocarpus and Tamarix. Amendments in soils increased the counts of soil microbiota by up to 98% and enhanced their activity by up to 95.5%. The increase in the biodegradation of crude oil (75%) and high levels of alkB expression substantiated the efficiency of the proposed amendment technology for the bioremediation of hydrocarbon-contaminated sites. The identification of crude-oil-degrading bacteria revealed the dominance of the genus Microbacterium (39.6%), Sphingopyxis soli (19.3%), and Bordetella petrii (19.6%) in unamended, Conocarpus-amended, and Tamarix-amended contaminated soils, respectively. Although soil amendments favored the growth of Gram-negative bacteria and reduced bacterial diversity, the structures of bacterial communities were not significantly altered.     

Factors affecting the occurrence and distribution of entomopathogenic fungi in natural and cultivated soils

Factors affecting the occurrence and distribution of entomopathogenic fungi in 244 soil samples collected from natural and cultivated areas in Spain were studied using an integrated approach based on univariate and multivariate analyses. Entomopathogenic fungi were isolated from 175 of the 244 (71.7 %) soil samples, with only two species found, Beauveria bassiana and Metarhizium anisopliae. Of the 244 soil samples, 104 yielded B. bassiana (42.6 %), 18 yielded M. anisopliae (7.3 %), and 53 soil samples (21.7 %) harboured both fungi. Log-linear models indicated no significant effect of habitat on the occurrence of B. bassiana, but a strong association between M. anisopliae and soils from cultivated habitats, particularly field crops. Also, irrespective of habitat type, B. bassiana predominated over M. anisopliae in soils with a higher clay content, higher pH, and lower organic matter content. Logistic regression analyses showed that pH and clay content were predictive variables for the occurrence of B. bassiana, whereas organic matter content was the predictive variable for M. anisopliae. Also, latitude and longitude predicted the occurrence of these same species, but in opposite directions. Altitude was found to be predictive for the occurrence of B. bassiana. Using principal component analysis, four factors (1 to 4) accounted for 86 % of the total variance; 32.8, 22.9, 19.6 and 10.4 % of the cumulative variance explained, respectively. Factor 1 was associated with high positive weights for soil clay and silt content and high negative weights for soil sand content. Factor 2 was associated with high positive weights for soil organic matter content and high negative weights for soil pH. Factor 3 was associated with high positive weights for latitude and longitude of the sampled localities and factor 4, had high positive weights only for the altitude. Bi-plot displays representing soil samples were developed for different factor combinations and indicated that, irrespective of geographical location, absence of both fungal species was determined by alkaline sandy soils with low organic matter content, whereas heaviness of soil texture, acidity and increasing organic matter content led to progressively higher percentages of samples harbouring entomopathogenic fungi. These results could aid decision-making as to whether or not a particular cultivated or natural soil is suitable for using entomopathogenic fungi as a pest control measure and for selecting the fungal species best suited to a particular soil.     

Influence of Temperature and Oil Content on the Soil/Air Partition Coefficient for Hexachlorobenzene in Oil-Contaminated Rice Paddy Field Soil

The soil/air partition coefficients (K _SA ) for hexachlorobenzene (HCB) in oil-contaminated (crude oil and diesel) rice paddy field soils were measured in a solid fugacity meter at different oil concentrations over the temperature range of 5 to 30°C at 100% relative humidity. The results showed that values of K _SA increased with a decrease of temperature. As for oil content, there is a critical separate phase concentration (CSPC) above which K _SA increased with increasing of oil content. When oil content is above CSPC, oil plays a role as a separate phase that enhances the sorption capacity of the soil. At a given temperature (20°C) values of CSPC depended on the natural organic matter (NOM) contents of the soil, while for a given oil concentration they depended on the temperature. The normalized oil/air partition coefficients ) for HCB deduced from K _SA for oils and experimentally determined with crude oil/quartz sand system were similar and 0.7–7 times higher than the normalized organic/air partition coefficient ), which implied that oil was a super sorbent. The enthalpies (ΔH_SA) for crude oil and diesel were 64.9 and 55.7 kJ mol^?1, respectively.     

Short-Term Effects of South Louisiana and Kuwait Crude Oils on Glucose Utilization by Marine Bacterial Populations

Two crude oils, South Louisiana and Kuwait, were examined for their impact on glucose utilization by bacterial populations from the Gulf of Mexico. The uptake and mineralization of [U-¹⁴C]glucose was assayed after a 4- to 23-h exposure to various concentrations of added crude oil (0, 0.001, 0.01, and 0.1% [vol/vol]). The effects of oil were determined in a total of 15 sediment and 13 water samples collected from offshore, open-bay, and salt marsh environments. The utilization of glucose by bacterial populations usually was not affected by added oil; in 10 sediment and 11 water samples, oil had no significant effect on either glucose uptake or mineralization. Stimulation by oil was recorded in four sediment samples. Oil inhibition occurred in one sediment and two water samples, but only in the presence of the highest concentration of added oil, i.e., 0.1%. Our data suggest that short-term exposure to either South Louisiana or Kuwait crude oil, even at 0.1%, usually has no toxic effect on glucose utilization by marine bacterial populations.     

Mouse pathogenicity studies of Nocardia asteroides complex species and clinical correlation with human isolates

Abstract Nocardia asteroides complex organisms derived from human specimens between 1979 and 1992 were identified on the species level. Of 117 N. asteroides complex organisms, 34 (29%) were N. farcinica , 28 (24%) were N. nova , and 55 (47%) were N. asteroides sensu stricto . An analysis of the specimen sites from which the organisms were derived showed that isolates derived from blood, brain, or bone marrow were more likely to be N. farcinica than the other two species. A study of the virulence of ten strains of each species was undertaken, using a mouse model with intravenous inoculation. The 50% lethal doses (LD₅₀) for N. farcinica were significantly lower than those of the other two species. LD₅₀ values for N. nova and N. asteroides were not significantly different. The above data confirming the greater virulence of N. farcinica support the identification of species within the N. asteroides complex.     

Selection of thermotolerant bradyrhizobial strains for nodulation of soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) in semi-arid regions of Iran

Nodulation of soybeans grown in semi-arid region of southern parts of Iran is poor due to high air and soil temperatures. Here we identified thermotolerant isolates of soybean bradyrhizobia and evaluated the nitrogen fixation efficiency of the isolates under heat stress conditions in greenhouse and field experiments. The ability of fifty-six bradyrhizobial isolates to grow on solid or in liquid yeast extract mannitol medium at 38 and 41°C was evaluated. We identified 19 isolates, which were able to grow at 38°C and 10 isolates able to grow at 41°C. Greenhouse experiments were carried out at 28 and 38°C to study the nitrogen-fixing capacity of the isolates under optimal and high temperature conditions. Ten isolates had a symbiotic index of effectiveness of 80% or greater compared with nitrogen-fertilized treatments in greenhouse experiments at 28°C. Some thermotolerant isolates demonstrated good nitrogen-fixing performance at 38°C. Eight isolates were selected for use in a field trial in the natural high temperature environment of the Dezful region in Iran. Our results demonstrate that geographical origin can have a great influence on the successful selection of thermotolerant bradyrhizobia. Our thermotolerant isolates were mainly obtained from high-temperature regions, and improved shoot dry matter, nitrogen-uptake and seed yield of the plants.     

Effect of Crude Oil and Chemical Additives on Metabolic Activity of Mixed Microbial Populations in Fresh Marsh Soils

Abstract Hydrocarbons increase abundance of hydrocarbon-degrading microorganisms, but also decrease microbial diversity. This could disrupt ecosystem dynamics by altering soil organic matter mineralization and resultant nutrient remineralization rates. Crude oil, which is known to contain toxins and reduce microbial diversity, was hypothesized to reduce gross metabolic activity of mixed microbial populations in wetland soils. Soil respiration and Eh were compared, for 6 months, among microcosms containing marsh soils that differed in soil organic matter (Panicum hemitomon Shult. or Sagittaria lancifolia L. dominated marshes), crude oil (Arabian crude, Louisiana crude, or no oil), and additives (a cleaner, a dispersant, fertilizer, or no additive). No treatment slowed activity; instead, Louisiana plus fertilizer and all Arabian treatments temporarily accelerated activity. Additional C respired from oiled microcosms exceeded C added as crude oil by 1.4 to 3.5 times. Thus, much additional C originated from soil organic matter rather than crude oil. Crude oils temporarily lowered soil Eh, which is consistent with accelerated metabolism and demand for electron acceptors. The lack of inhibition observed at the community level does not necessarily indicate an absence of toxicity. Instead, tolerant species with metabolic versatility probably maintained activity. Stimulation probably resulted from removal of micronutrient limitation, rather than removal of grazing pressure or macronutrient limitation. Regardless, accelerated soil organic matter mineralization surely accelerated nutrient remineralization. This might explain some reports of crude oil stimulating plant growth. These results are not inconsistent with theoretical and experimental conclusions regarding effects of biodiversity on ecosystem stability and productivity, nor are they inconsistent with conclusions that crude oils contain components that are toxic to microbes, vegetation, and fauna. However, these data do indicate that crude oils also contain components that temporarily stimulate metabolic activity of surviving microbes. Received: 27 April 1998; Accepted: 15 July 1998     

中国北亚热带天然次生林与杉木人工林土壤活性有机碳库的比较

为了了解北亚热带东部地区天然次生林转变成杉木(Cunninghamia lanceolata)人工林对土壤活性有机碳库的影响, 以浙江省富阳市庙山坞森林生态系统定位研究站杉木人工林和天然次生林为研究对象, 对达到成熟林状态的两种林分类型0-60 cm内各土层土壤活性有机碳含量进行了比较研究。结果表明: 1)天然次生林土壤总有机碳、易氧化有机碳、水溶性有机碳和轻组有机质含量均高于杉木人工林, 与人工杉木林相比, 增幅分别为19.0%-32.6%、0.8%-30.3%、3.8%-54.1%和6.3%-38.6%, 且在0-10和10-20 cm土层差异显著(p < 0.05) (水溶性有机碳仅在0-10 cm土层差异显著); 2)杉木人工林土壤水溶性有机碳与易氧化碳占总有机碳的比率均高于天然次生林; 3)两个林分土壤水溶性有机碳、易氧化碳和轻组有机质与总有机碳含量均呈现极显著相关, 且天然次生林土壤易氧化碳、轻组有机质与总有机碳的相关系数均大于杉木人工林; 4)土壤有机碳、水溶性有机碳、易氧化碳和轻组有机质与土壤养分(全氮、水解氮、速效磷、速效钾、速效钙和速效镁)的相关性均达到显著(p < 0.05)或极显著(p < 0.01)水平。     

Nitrogen mineralization and its controlling factors in various kinds of temperate humid-zone soils

The N mineralization capacity of 41 temperate humid-zone soils of NW Spain was measured by aerobic incubation for 15 days at 28°C and 75% of field capacity. The main soil factors affecting organic N dynamics were identified by principal components analysis. Ammonification predominated over nitrification in almost all soils. The mean net N mineralization rate was 1.63% of the organic N content, and varied according to soil parent materials as follows: soils on basic and ultrabasic rocks < soils over acid metamorphic rocks < soils developed over sediments < soils over acid igneous rocks < soils on limestone. The N mineralization capacity was lower in natural soils than in cropped soils or pastures. The accumulation of organic matter (C and N) seems to be due to poor mineralization which was caused, in decreasing order of importance, by high exchangeable H-ion levels, high Al and Fe gel contents and, to a lesser extent (though more markedly in cropped soils), by silty clay texture and exchangeable Al ions.     

PlANT-ASSOCIATED BACTERIA AS TOOLS FOR THE PHYTOREMEDIATION OF OILY NITROGEN-POOR SOILS

The rhizospheres and phyllospheres of peas, beans, tomatoes, and squash raised in a desert sand soil mixed with 0.5% crude oil were rich in oil-utilizing bacteria and accommodated large numbers of free-living diazotrophic bacteria, with potential for hydrocarbon utilization. According to their 16S rRNA-sequences, the cultivable oil-utilizing bacteria were affiliated with the following genera, arranged in decreasing frequency: Bacillus, Ochrobactrum, Enterobacter, Rhodococcus, Arthrobacter, Pontola, Nocardia, and Pseudoxanthomonas. Diazotrophic isolates were affiliated with Rhizobium, Bacillus, Rhodococcus, Leifsonia, Cellulosimicrobium, Stenotrophomonas, Kocuria, Arthrobacter, and Brevibacillus. The crude oil–utilizing and diazotrophic isolates grew, with varying growth intensities, on individual aliphatic (C₈ to C₄₀) and aromatic hydrocarbons, as sole sources of carbon and energy. Quantitative gas liquid chromatographic measurements showed that representative bacterial isolates eliminated pure n-hexadecane, n-decosane, phenanthrene, and crude oil from the surrounding liquid media. Cultivation of oily sand–soil samples with any of the four tested crops led to enhanced oil degradation in that soil, as compared with the degradation in uncultivated oily sand–soil samples.     

Biodegradation of [(sup14)C]Benzo[a]pyrene Added in Crude Oil to Uncontaminated Soil

To investigate the possible cometabolic biodegradation of benzo[a]pyrene (BaP), crude oil spiked with [7-(sup14)C]BaP and unlabeled BaP was added to soil with no known pollution history, to give 34 g of oil and 67 mg of BaP/kg of dry soil. The oil-soil mixture was amended with mineral nutrients and incubated in an airtight container with continuous forced aeration. Total CO(inf2) and (sup14)CO(inf2) in the off-gas were trapped and quantified. Soil samples were Soxhlet extracted with dichloromethane at seven time points during the 150-day incubation period, and the extracted soil was subjected to further fractionation in order to recover reversibly and irreversibly bound radiocarbon. Radiocarbon recovery was 100% (plusmn) 3% for each time point. During the first 50 days of incubation, no (sup14)CO(inf2) was evolved, but over the next 100 days, 50% of the BaP radiocarbon was evolved as (sup14)CO(inf2). At 150 days, only 5% of the intact BaP and 23% of the crude oil remained. Of the remaining radiolabel, 20% was found in solvent-extractable metabolites and 25% was incorporated into soil organic matter. Only 1/10 of this could be solubilized by chemical hydrolysis. An abiotic control experiment exhibited binding of only 2% of the BaP, indicating the microbial nature of the BaP transformations. We report that in soil containing suitable cosubstrates, BaP can be completely degraded.     

Soil carbon stocks and changes after oil palm introduction in the Brazilian Amazon

As oil palm has been considered one of the most favorable oilseeds for biodiesel production in Brazil, it is important to understand how cultivation of this perennial crop will affect the dynamics of soil organic carbon (SOC) in the long term. The aim of this study was to evaluate the changes in soil C stocks after the conversion of forest and pasture into oil palm production in the Amazon Region. Soil samples were collected in March 2008 and September 2009 in five areas: native forest (NARF), pasture cultivated for 55 years (PAST), and oil palm cultivated for 4 (OP‐4), 8 (OP‐8) and 25 years (OP‐25), respectively. Soils were sampled in March 2008 to evaluate the spatial variability of SOC and nitrogen (N) contents in relation to the spacing between trees. In September 2009, soils were sampled to evaluate the soil C stocks in the avenues (inter rows) and frond piles, and to compare the total C stocks with natural forest and pasture system. Soil C contents were 22–38% higher in the area nearest the oil palm base (0.6 m) than the average across the inter row (0–4.5 m from the tree), indicating that the increment in soil organic matter (SOM) must have been largely derived from root material. The soil C stocks under palm frond piles were 9–26% higher than in the inter rows, due to inputs of SOM by pruned palm fronds. The soil carbon stocks in oil palm areas, after adjustments for differences in bulk density and clay content across treatments, were 35–46% lower than pasture soil C stocks, but were 0–18% higher than the native forest soil C content. The results found here may be used to improve the life cycle assessment of biodiesel derived from palm oil.