Influence of substratum on the degradation processes in diesel polluted sub-Antarctic soils (Crozet Archipelago)

Biodegradation by naturally occurring populations of microorganisms is a major mechanism for the removal of oil hydrocarbons from the environment. Therefore, follow-up of bacterial populations and chemical indices of biodegradation are important components of contaminated site assessment studies. Over a 4-year period following an accidental diesel contamination of the sub-Antarctic Crozet Archipelago (51°51′E–46°25′S), a field study was carried out in the contaminated area that is located in a transition zone between an arid fell-field (upstream) and a wet vegetated area (downstream). This study included a monitoring of heterotrophic and hydrocarbon-degrading bacterial abundance and chemical analysis of the remaining hydrocarbons. Significant higher number of heterotrophic and hydrocarbon-degrading bacterial counts revealed a rapid acclimation of sub-Antarctic microbial soil communities to the diesel fuel contamination. A chemical survey conducted during the last 2 years (2002 and 2003) showed that the total extractable hydrocarbons (TPH) content in arid fell field was reduced to ≤50% of their value while it was reduced only to ≤65% in vegetated soil. In addition, the decrease of TPH was always higher in the presence of fertilizer in the arid contaminated area, while fertilizer addition was almost inefficient in the wet contaminated one. All these results demonstrate a serious influence of the soil properties on the degradation rate. However, all chemical indices showed a significant reduction of alkanes and light aromatics in both contaminated area confirming a regular oil degradation process.     

Studies on Biodegradation of Kerosene in Soil under Different Bioremediation Strategies

The effectiveness of bioremediation is often a function of the microbial population and how they can be enriched and maintained in an environment. Strategies for inexpensive in situ bioremediation of soil contaminated with petroleum hydrocarbons include stimulation of the indigenous microorganisms by introduction of nutrients (biostimulation) and/or through inoculation of an enriched mixed microbial culture into soil (bioaugmentation). To demonstrate the potential use of bioremediation in soil contaminated with kerosene, a laboratory study with the objective of evaluating and comparing the effects of bioattenuation, biostimulation, bioaugmentation, and combined biostimulation and bioaugmentation was performed. The present study dealt with the biodegradation of kerosene in soil under different bioremediation treatment strategies: bioattenuation, biostimulation, bioaugmentation, and combined biostimulation and bioaugmentation, respectively. Each treatment strategy contained 10% (w/w) kerosene in soil as a sole source of carbon and energy. After 5 weeks of remediation, the results revealed that bioattenuation, bioaugmentation, biostimulation, and combined biostimulation and bioaugmentation exhibited 44.1%, 67.8%, 83.1%, and 87.3% kerosene degradation, respectively. Also, the total hydrocarbon-degrading bacteria (THDB) count in all the treatments increased with time up till the second week after which it decreased. The highest bacterial growth was observed for combined biostimulation and bioaugmentation treatment strategy. A first-order kinetic model equation was fitted to the biodegradation data to further evaluate the rate of biodegradation and the results showed that the specific degradation rate constant (k) value was comparatively higher for combined biostimulation and bioaugmentation treatment strategy than the values for other treatments. Therefore, value of the kinetic parameter showed that the degree of effectiveness of these bioremediation strategies in the clean up of soil contaminated with kerosene is in the following order: bioattenuation < bioaugmentation < biostimulation < combined biostimulation and bioaugmentation. Conclusively, the present work has defined combined biostimulation and bioaugmentation treatment strategy requirements for kerosene oil degradation and thus opened an avenue for its remediation from contaminated soil.     

Potential for biodegradation of hydrocarbons in soil from the Ross Dependency, Antarctica

Oil spills occur in the Antarctic when fuel oils such as JP8 jet fuel are moved or stored. Hydrocarbons, both n-alkanes and aromatic compounds, have been detected in oil-contaminated soils of the Ross Dependency. In such areas hydrocarbon-degrading microbes, if naturally occurring, could be used for clean-up. Soil samples from oil-impacted and control sites were analysed for hydrocarbon-degrading microbes and for a range of parameters known to limit biodegradative activity. Soils were analysed for water content, pH, concentrations of nutrients (N and P) and electrical conductivity. Numbers of culturable heterotrophic bacteria and hydrocarbon degraders were greater in some of the oil-contaminated samples. Mineralisation studies with ¹⁴C-radiolabelled hexadecane and naphthalene demonstrated that nitrogen amendments significantly enhanced hydrocarbon mineralisation rates in an oil-impacted soil. Received: 30 June 1997 / Received revision: 15 September 1997 / Accepted: 19 September 1997     

Influence of nutrients addition and bioaugmentation on the hydrocarbon biodegradation of a chronically contaminated Antarctic soil

Complexity involved in the transport of soils and the restrictive legislation for the area makes on-site bioremediation the strategy of choice to reduce hydrocarbons contamination in Antarctica. The effect of biostimulation (with N and P) and bioaugmentation (with two bacterial consortia and a mix of bacterial strains) was analysed by using microcosms set up on metal trays containing 2·5 kg of contaminated soil from Marambio Station. At the end of the assay (45 days), all biostimulated systems showed significant increases in total heterotrophic aerobic and hydrocarbon-degrading bacterial counts. However, no differences were detected between bioaugmented and nonbioaugmented systems, except for J13 system which seemed to exert a negative effect on the natural bacterial flora. Hydrocarbons removal efficiencies agreed with changes in bacterial counts reaching 86 and 81% in M10 (bioaugmented) and CC (biostimulated only) systems. Results confirmed the feasibility of the application of bioremediation strategies to reduce hydrocarbon contamination in Antarctic soils and showed that, when soils are chronically contaminated, biostimulation is the best option. Bioaugmentation with hydrocarbon-degrading bacteria at numbers comparable to the total heterotrophic aerobic counts showed by the natural microflora did not improve the process and showed that they would turn the procedure unnecessarily more complex.     

Aromatic hydrocarbon-degrading bacteria from soil near Scott Base, Antarctica

Hydrocarbons persist in Antarctic soils when fuel oils such as JP8 jet fuel are spilled. For clean-up of hydrocarbon-contaminated soils in Antarctica, bioremediation has been proposed using hydrocarbon-degrading microbes indigenous to Antarctic soils. A number of alkane-degrading bacteria have been isolated previously from Antarctic soils. In this paper we describe the direct isolation of aromatic hydrocarbon-degrading bacteria from oil-contaminated Antarctic soil. Isolates that grew on JP8 jet fuel were characterised for their ability to degrade aromatic and aliphatic hydrocarbons and for growth at a range of temperatures. All isolates were gram-negative, oxidase-positive, rod-shaped bacteria. Representative strains were identified using 16S rDNA sequence analysis as either Sphingomonas spp. or Pseudomonas spp. Aromatic-degrading bacteria from Antarctic soils were psychrotolerant and appear similar to those found worldwide. Accepted: 27 September 1999     

Effect of drought on leaf gland secretion of the mangrove Avicennia germinans L.

This study assessed the effects of salinity changes over space and time upon leaf gland secretion in Avicennia germinans trees growing naturally in an area featuring markedly seasonal rainfall. Soil ? (, soil N MPa) during the wet season was -0.95ǂ.05 and -2.12ǂ.08 at low and high salinity sites, respectively. During the dry season, these values decreased to -3.24ǂ.09 at low salinity and to -5.75ǂ.06 at high salinity. Consequently, predawn and midday plant water potential were lowered during drought at both sites. The rates of secretion (mmol m^-2 h^-1 ) increased during drought from 0.91ǂ.12 during the wet season to 1.93ǂ.12 at low salinity, and from 1.69ǂ.12 during the wet season to 2.81ǂ.15 at high salinity. Conversely, stomatal conductance (g_s) was lowered by both salinity and drought. As xylem osmolality increased during drought, secretion tended to rise exponentially, and g_s decayed hyperbolically. Thus, a trade-off is obtained between enhancement in salt secretion and control of water loss suggested by g_s.     

Response of Antarctic Soil Bacterial Assemblages to Contamination by Diesel Fuel and Crude Oil

Abstract The effects of diesel fuel and ``Arabian light' crude oil addition on Antarctic bacterial assemblages were studied in four contaminated soils during 1 year in the Terre Adelie land area. Monthly sampling allowed a regular survey of the bacterial changes occurring in the contaminated soils. All samples were analyzed for total bacteria, heterotrophic culturable microbiota, and hydrocarbon-utilizing microbiota. Crude oil contamination induced an initial increase of all bacterial parameters in all contaminated soils. Diesel oil contamination had a more complex effect. Hydrocarbon degrading bacterial abundance increases occurred after diesel oil addition. In contrast, general heterotrophic bacterial abundance could significantly decrease in the same conditions. In all cases the stimulatory effects of oil addition disappeared after several months of contamination. Received: 13 April 1999; Accepted: 24 February 2000; Online Publication: 29 May 2000     

Nutrient-limited biodegradation of PAH in various soil strata at a creosote contaminated site

The effects of nutrient addition on the in situ biodegradation of polycyclic aromatic hydrocarbons in creosote contaminated soil were studied in soil columns taken from various soil strata at a wood preserving plant in Norway. Three samples were used: one from the topsoil (0–0.5 m), one from an organic rich layer (2–2.5 m) and one from the sandy aquifer (4.5–5 m). The addition of inorganic nitrogen and phosphorous stimulated the degradation of polycyclic aromatic hydrocarbons (PAHs) in the top soil and the aquifer sand. These two soils, which differed strongly in contamination levels, responded similarly to nutrient addition with the corresponding degradation of 4-ring PAHs. The ratio between available nitrogen (N) and phosphorous (P) might explain the degree of degradation observed for the 4-ring PAHs. However, the degree of degradation of 3-ring PAHs did not significantly increase after nutrient addition. An increase in the respiration rate, after nutrient addition, could only be observed in the topsoil. In the aquifer sand, 4-ring PAH degradation was not accompanied by an increase in the respiration rate or the number of heterotrophic micro-organisms. PAH degradation in the organic layer did not respond to nutrient addition. This was probably due to the low availability of the contaminants for micro-organisms, as a result of sorption to the soil organic matter. Our data illustrate the need for a better understanding of the role of nutrients in the degradation of high molecular weight hydrocarbons for the successful application of bioremediation at PAH contaminated sites.     

Effects of oil spills on microbial heterotrophs in Antarctic soils

Oil spillage on the moist coastal soils of the Ross Sea region of Antarctica can impact on populations of microbial heterotrophs in these soils, as determined by viable plate counts and a most probable number technique. Elevated numbers of culturable hydrocarbon degraders, bacteria and fungi were detected in surface and subsurface soils from oil-contaminated sites, compared with nearby control sites. Culturable yeasts were not detected in soil from coastal control sites, yet reached >10⁵ organisms g^-1 dry weight in contaminated soils. The presence of hydrocarbons in soils resulted in a shift in the genera of culturable filamentous fungi. Chrysosporium dominated control soils, yet Phialophora was more abundant in oil-contaminated soils. Hydrocarbon degraders are most likely bacteria; however, fungi could play a role in degradation of hydrocarbons or their metabolites. Depleted levels of nitrate detected in some contaminated soils and decreased pH may be the result of growth of hydrocarbon degraders. Numbers and diversity of culturable microbes from Antarctic soil varied depending on whether a pristine site or a human-impacted (in this case, by fuel spills) site is studied.     

Fungal and bacterial biomass in tundra soils along an arctic transect from Taimyr Peninsula, central Siberia

Microscopic analyses of tundra soils from northern central Siberia, Taimyr Peninsula (74.5°N, 98.5°E) were performed in order to investigate spatial variation of fungal and bacterial biomass. Biomass figures of fungi and bacteria (µg C g^-1 dry wt.) were measured from 11 permafrost soil pits. Fungal biovolume of up to 3.5 mm³ g^-1 dry wt. (median 0.19 mm³ g^-1 dry wt.) and a maximum hyphal length of 393 m g^-1 dry wt. (median 21 m g^-1 dry wt.) were determined. Fungal biomass was found up to 455 µg C g^-1 dry wt. (median 24 µg C g^-1 dry wt.). The amounts generally decreased with depth but increased within organic horizons. Little fungal biomass was found in the unvegetated soils or deep horizons above the permafrost table. Bacterial counts ranged from 0.16 to 7.38*10⁹ g^-1 dry wt. and bacterial biomass ranged from 0.68 to 20.38 µg C g^-1 dry wt. (median 6.19 µg C g^-1 dry wt.) because of small cell volume (median 0.04 µm³). Microbial biomass was generally dominated by fungi as shown by the ratio of fungal to bacterial biomass, which was between 0 and 174.1 (median 4.5). Plant cover and soil organic matter content were found to be the important keys in understanding microbial ecology in arctic tundra soils.     

Soil microbial community and bacterial functional diversity at Machu Picchu, King George Island, Antarctica

The objective of this study was to evaluate the potential contribution of the soil microbial community in the vicinity of two plant covers, Sanionia uncinata and Deschampsia antarctica, at Machu Picchu Station, King George Island, Antarctica. Soil samples were collected at the study site during the southern (pole) summer period from 0-5 cm and 5-10 cm depths, for chemical and biological analyses. Soil microbial biomass reached a maximal value of 144 µg g^-1 in soil samples taken from under the S. uncinata upper layer plant. qCO₂ ranged from 167 to 239 µg CO₂^.mgC_mic^.h^-1 at the 0-5 and 5-10 cm depths, respectively. CO₂ evolution showed values of 54.3 mg^.m^-2 h^-1 beneath plant cover and 55.9 mg^.m^-2 h^-1 in the open space. CO₂ evolved by substrate induced respiration in the soil samples taken under the plant cover in the summer period, oscillated between 0.25 and 4.78 µg CO₂ g^-1 h^-1. The data obtained from this short study may provide evidence that both activity and the composition and substrate utilization of the microbial community appear to change substantially across the moisture level and sample location.     

Fecal bacterial contamination in natural water reservoirs as an indicator of seasonal infection by Opisthorchis viverrini in snail intermediate hosts

Opisthorchis viverrini, a carcinogenic liver fluke, requires Bithynia snails as the first intermediate host, which release cercariae after ingesting fluke eggs from contaminated water. Fecal bacterial contamination and O. viverrini-infected Bithynia snails were investigated in samples collected from natural water reservoirs in Ban Phai, Chonnabot and Muang Districts (Ban Lerngpeuy) in Khon Kaen Province, northeast Thailand, where there is a high incidence of cholangiocarcinoma. Water was sampled and examined six times (February, April, June, August, October and December 2006). The most probable number (MPN) index and coliform counts were utilized to evaluate fecal contamination; the cercarial shedding method was conducted for detecting infected snails. The data revealed that all water samples had a high MPN index number, and fecal coliform levels above the WHO standard. This indicated that water in these reservoirs was contaminated with feces or manure constituents. Water sampling from Ban Lerngpeuy showed full-scale bacterial contamination (> 1609 MPN index) throughout the year. This finding was correlated with the highest prevalence of O. viverrini-infected snails, which were found nearly all year round in this area. Slightly lower fecal contamination levels were detected in water samples from Chonnabot and Ban Phai, with high MPN index numbers and coliform counts from April to October. This corresponded with the higher recovery of infected snails in June and August, but with relatively lower prevalence than those found in Ban Lerngpeuy. Among the sampling sites, the people in Ban Lerngpeuy live nearer to the reservoir than do those in Ban Phai and Chonnabot. These results indicate that fecal bacterial contamination in natural water reservoirs is an important indicator of seasonal transmission of O. viverrini eggs to snail intermediate hosts. Sanitation improvement is essential and future investigations on the sources of contamination are needed.     

Colonization of contaminated soil by an introduced bacterium: effects of initial pentachlorophenol levels on the survival of Sphingomonas chlorophenolica strain RA2

The survival of a Sphingomonas species that was introduced into pentachlorophenol (PCP)-contaminated soil was monitored with two complementary methods, a respiration-based assay and a most probable number (MPN) technique. Sphingomonas chlorophenolicastrain RA2 is a PCP-mineralizing bacterium that was introduced into soil contaminated with a range of PCP concentrations (0–300 μg PCP g⁻¹ soil). The population of introduced microorganisms was followed for 170 days using a substrate-induced growth-response method and a MPN assay that specifically targets PCP-mineralizing bacteria. Varying the initial PCP concentration resulted in the emergence of three distinct patterns of survival. In soil contaminated with 300 μg PCP g⁻¹ the population of S. chlorophenolica strain RA2 immediately declined following introduction, increased by 200-fold and leveled off by the end of the 170-day incubation. In contrast, populations of S. chlorophenolica strain RA2 declined to levels below detection limits in uncontaminated soil by the end of the experiment. Intermediate PCP concentrations (10–100 μg PCP g⁻¹ soil) resulted in the establishment of S. chlorophenolica strain RA2 that slowly declined in numbers. These results indicate that Sphingomonas chlorophenolica strain RA2 is an effective colonizer of PCP-contaminated soil but will not persist in the absence of PCP. Received 14 April 1999/ Accepted in revised form 24 June 1999     

Measurement of hydrocarbon-degrading microbial populations by a 96-well plate most-probable-number procedure

A 96-well microtiter plate most-probable-number (MPN) procedure was developed to enumerate hydrocarbondegrading microorganisms. The performance of this method, which uses number 2 fuel oil (F2) as the selective growth substrate and reduction of iodonitrotetrazolium violet (INT) to detect positive wells, was evaluated by comparison with an established 24-well microtiter plate MPN procedure (the Sheen Screen), which uses weathered North Slope crude oil as the selective substrate and detects positive wells by emulsification or dispersion of the oil. Both procedures gave similar estimates of the hydrocarbon-degrader population densities in several oil-degrading enrichment cultures and sand samples from a variety of coastal sites. Although several oils were effective substrates for the 96-well procedure, the combination of F2 with INT was best, because the color change associated with INT reduction was more easily detected in the small wells than was disruption of the crude oil slick. The method's accuracy was evaluated by comparing hydrocarbon-degrader MPNs with heterotrophic plate counts for several pure and mixed cultures. For some organisms, it seems likely that a single cell cannot initiate sufficient growth to produce a positive result. Thus, this and other hydrocarbon-degrader MPN procedures might underestimate the hydrocarbon-degrading population, even for culturable organisms.     

华西雨屏区苦竹林土壤呼吸对模拟氮沉降的响应

2007年11月至2008年11月, 对华西雨屏区苦竹(Pleioblastus amarus)人工林进行了模拟氮沉降试验, 氮沉降水平分别为对照(CK, 0 g N·m^-2·a^-1)、低氮(5 g N·m^-2·a^-1)、中氮(15 g N·m^-2·a^-1)和高氮(30 g N·m^-2·a^-1)。每月下旬, 采用红外CO₂分析法测定土壤呼吸速率, 并定量地对各处理施氮(NH₄NO₃)。结果表明: 2008年试验地氮沉降量为8.241 g·m^-2, 超出该地区氮沉降临界负荷。在生长季节, 苦竹林根呼吸占总土壤呼吸的60%左右。模拟氮沉降促进了苦竹林土壤呼吸速率, 使苦竹林土壤每年向大气释放的CO₂增加了9.4%~28.6%。在大时间尺度上(如1 a), 土壤呼吸主要受温度的影响。2008年6~10月, 土壤呼吸速率24 h平均值均表现为: 对照<低氮<中氮<高氮。氮沉降处理1 a后, 土壤微生物呼吸速率和土壤微生物生物量碳、氮增加, 并且均与氮沉降量具有相同趋势。各处理土壤呼吸速率与10 cm土壤温度、月平均气温呈极显著指数正相关关系, 利用温度单因素模型可以解释土壤呼吸速率的大部分。模拟氮沉降使得土壤呼吸Q₁₀值增大, 表明氮沉降可能增强了土壤呼吸的温度敏感性。在氮沉降持续增加和全球气候变暖的背景下, 氮沉降和温度的共同作用可能使得苦竹林向大气中排放的CO₂增加。     

Fatty acid composition of lipid-rich myctophids and mackerel icefish (<Emphasis Type="Italic">Champsocephalus gunnari</Emphasis>) – Southern Ocean food-web implications

The lipid content, fatty acid composition and calorific value of seven species of mesopelagic deep-sea fish of the family Myctophidae and the mackerel icefish, Champsocephalus gunnari, important in the diet of Southern Ocean marine predators, are presented. Fish were sampled at the Kerguelen Plateau (KP) and Macquarie Ridge (MR) in the Indian and Pacific sectors of the Southern Ocean respectively, to examine geographic variation in lipid compositon. All species of myctophid from KP and Electrona antarctica from MR were high in lipid content (6-18% wet mass), particularly Gymnoscopelus nicholsi (18%) and E. antarctica (15%). The mackerel icefish, and G. fraseri and Protomyctophum tenisoni from MR were generally lower in lipid content (3-5%) and varied significantly in fatty acid composition from KP species. KP myctophids were high in calorific content (9.3 kJ g^-1 wet mass) when compared with icefish (5.4 kJ g^-1 wet mass) and other published values for prey items of marine predators such as squid (1.7-4.5 kJ g^-1). KP myctophids were distinguished from each other and from C. gunnari and MR specimens by cluster and discriminant function analysis using six fatty acids (16:0, 18:1̩, 20:1̩, 22:1Ὃ, 20:5̣, 22:6̣). Findings presented here highlight trophic links between high-latitude fish and their prey and emphasise the importance of myctophids as a significant energy source for marine predators foraging in the Polar Frontal Zone.     

Influence of long-term diesel fuel pollution on nitrite-oxidising activity and population size of nitrobacter spp in soil

Previous investigations have shown that ammonia oxidation is not inhibited by diesel fuel in a soil with a long history of contamination contrary to a non-contaminated soil. As a consequence, ammonia oxidation does not constitute a limited step in nitrification process (Appl. Environ. Microbiol. 65 (1999) 4008). Moreover, this type of soil also has had the opportunity to develop an abundant microbial population able to metabolise the diesel hydrocarbons. Whether the properties of soil with a long history of diesel fuel contamination may affect the activity of nitrite-oxidising bacteria was investigated. It was observed that re-exposure of soil to diesel fuel apparently stimulated the proliferation of nitrite-oxidising bacteria, as determined by most probable number (MPN) culture technique and MPN-polymerase chain reaction technique. The potential of nitrite-oxidising activity in soil treated with diesel fuel was about 4 times higher than in the control without addition. In the presence of diesel fuel and ammonium, the potential nitrite-oxidising activity was 40% higher than in presence of ammonium only. However, in the presence of hydrocarbon only, low proliferation of Nitrobacter was observed, probably because the heterotrophic bacteria were strongly limited by lack of nitrogen and did not produce sufficient organic metabolites that could be used by the Nitrobacter cells.     

Microbiological characterization of a coke oven contaminated site and evaluation of its potential for bioremediation

The soil microbial population of a coke oven site was investigated in order to evaluate its potential for bioremediation. The study was carried out in soil samples with distinct polynuclear aromatic hydrocarbon (PAH) contamination levels, comparing the population profiles constituted by total heterotrophic and PAH-utilizing strains. Isolation of degrading strains was performed with phenanthrene or pyrene as sole carbon sources. The ability to degrade other PAHs, such as anthracene, fluorene and fluoranthene was also investigated. The results showed a reduction of 30% in species diversity and microbial density drops one order of magnitude in contaminated samples. Furthermore, the number of PAH-utilizing colonies was higher in the contaminated area and about 20% of the isolates were able to degrade phenanthrene and pyrene, while this value decreased to 0.15% in uncontaminated samples. Three PAH-degrader strains were identified as: CDC gr. IV C-2, Aeromonas sp. and Pseudomonas vesicularis. The ability of these strains to degrade other PAHs was also investigated.     

Bioremediation of hydrocarbon-contaminated polar soils

Bioremediation is increasingly viewed as an appropriate remediation technology for hydrocarbon-contaminated polar soils. As for all soils, the successful application of bioremediation depends on appropriate biodegradative microbes and environmental conditions in situ. Laboratory studies have confirmed that hydrocarbon-degrading bacteria typically assigned to the genera Rhodococcus, Sphingomonas or Pseudomonas are present in contaminated polar soils. However, as indicated by the persistence of spilled hydrocarbons, environmental conditions in situ are suboptimal for biodegradation in polar soils. Therefore, it is likely that ex situ bioremediation will be the method of choice for ameliorating and controlling the factors limiting microbial activity, i.e. low and fluctuating soil temperatures, low levels of nutrients, and possible alkalinity and low moisture. Care must be taken when adding nutrients to the coarse-textured, low-moisture soils prevalent in continental Antarctica and the high Arctic because excess levels can inhibit hydrocarbon biodegradation by decreasing soil water potentials. Bioremediation experiments conducted on site in the Arctic indicate that land farming and biopiles may be useful approaches for bioremediation of polar soils.     

Effect of genetically modified Pseudomonas fluorescens introduced into soil contaminated with copper (II) on microbial community diversity in the soil and rhizosphere

When genetically altered strains of Pseudomonas fluorescens were introduced into soil contaminated with copper in a laboratory study, they did not affect the phenotypic diversity (eco-physiological index) of indigenous microflora in the rhizosphere and bulk soil. Soil contamination with copper also did not cause shifts in the distribution of bacterial classes within the total community of heterotrophic or Gram-negative bacteria.