Antimicrobial resistance among Pseudomonas spp. and the Bacillus cereus group isolated from Danish agricultural soil

From four Danish pig farms, bacteria of Pseudomonas spp. and the Bacillus cereus group were isolated from soil and susceptibility towards selected antimicrobials was tested. From each farm, soil samples representing soil just before and after spread of animal waste and undisturbed agricultural soil, when possible, were collected. Soil from a well-characterized Danish farm soil (H?jbakkegaard) was collected for comparison. The Pseudomonas spp. and B. cereus were chosen as representative for Gram-negative and Gram-positive indigenous soil bacteria to test the effect of spread of animal waste on selection of resistance among soil bacteria. No variations in resistance levels were observed between farms; but when the four differently treated soils were compared, resistance was seen for carbadox, chloramphenicol, nalidixan (nalidixic acid), nitrofurantoin, streptomycin and tetracycline for Pseudomonas spp., and for bacitracin, erythromycin, penicillin and streptomycin for the B. cereus group. Variations in resistance levels were observed when soil before and after spread of animal waste was compared, indicating an effect from spread of animal waste.     

Development of a rapid method for direct detection of tet(M) genes in soil from Danish farmland

A method for direct detection of antibiotic resistance genes in soil samples has been developed. The tetracycline resistance gene, tet(M), was used as a model. The method was validated on Danish farmland soil that had repeatedly been treated with pig manure slurry containing resistant bacteria. The tet(M) gene was directly detected in 10-80% of the samples from the various farmland soils and could be detected in all samples tested after selective enrichment. To validate the obtained results, the method was applied to garden soil samples where lower prevalence of resistance was found. RESULT: A detection limit of 10(2)-10(3) copies of the tet(M) gene per gram of soil (in a Bacillus cereus group bacterium) was achieved. tet(M) gene was detected in soil samples with the highest prevalence on farmland treated with pig manure slurry.     

Utilization of petroleum hydrocarbons by Pseudomonas fluorescens isolated from a petroleum-contaminated soil

A strain of Pseudomonas fluorescens, isolated from petroleum hydrocarbon-contaminated soil was examined for its ability to utilize a variety of hydrocarbon substrates. Surface tension measurements indicated the production of biosurfactant during the microbial degradation of hydrocarbon. The organism utilized both short and long chain n-alkanes. It emulsified a number of aliphatic and aromatic hydrocarbons.     

Denitrification with acrylonitrile as a substrate using pure bacteria cultures isolated from acrylonitrile-butadiene-styrene wastewater

This study attempted to isolate and identify the denitrifying bacteria that utilize acrylonitrile as a substrate from acrylonitrile-butadiene-styrene (ABS) resin wastewater. The performance of the denitrifying bacteria for treating different initial acrylonitrile concentrations was also investigated under anoxic conditions. The results showed that seven strains of denitrifying bacteria that can use acrylonitrile or acrylic acid as a substrate were isolated from the denitrification tank of a wastewater treatment plant in a ABS resin manufacturing plant and a lab-scale anoxic granular activated carbon-fluidized bed. The bacteria strains Acidovorax facilis B and Pseudomonas nautica could utilize acrylonitrile up to 279 mg/l as a substrate for denitrification. For complete nitrate removal, an adequate supply of acrylonitrile was necessary. Under the assumption that the acrylic acid would be completely removed, the removal of 1 mg/l nitrate by A. facilis B or P. nautica, about 0.64-0.74 mg/l acrylonitrile or 0.87-1 mg/l acrylic acid was needed. Because strains A. facilis B and P. nautica could utilize acrylonitrile for denitrification, they are expected to play an important role in the treatment of acrylonitrile in the wastewater treatment plant (denitrification and nitrification processes) and lab-scale granular activated carbon-fluidized bed.     

Quantifying the transfer of radionuclides to food products from domestic farm animals

Databases have been compiled to derive parameter values relevant to the transfer of radionuclides from feedstuffs to domestic animal products to provide a revision to the IAEA Handbook on transfer parameters TRS 364. Significant new data inputs have been incorporated into the databases from an extensive review of Russian language information and inclusion of data published since the early 1990s. Fractional gastrointestinal absorption in adult ruminants presented in the revised handbook are generally similar to those recommended for adult humans by the ICRP. Transfer coefficient values are presented in the handbook for a range of radionuclides to farm animal products. For most animal products, transfer coefficient values for elements additional to those in TRS 364 are provided although many data gaps remain. Transfer coefficients generally vary between species with larger species having lower values than smaller species. It has been suggested that the difference is partly due to the inclusion of dietary dry matter intake in the estimation of transfer coefficient and that whilst dietary intake increases with size nutrient concentrations do not. An alternative approach to quantifying transfer by using concentration ratios (CR), which do not consider dietary intake, has been evaluated. CR values compiled for the handbook vary considerably less between species than transfer coefficient values. The advantage of the CR approach is that values derived for one species could be applied to species for which there are no data. However, transfer coefficients will continue to be used as few studies currently report CR values or give data from which they can be estimated.     

Airborne dissemination of bacteria from sewage treatment plants

During the last 5 yr, the air of sewage treatment plants was examined bacteriologically under different climatic conditions. Methods of examination were sedimentation on nutrient agar, membrane filtration and the use of an impinger. At each sampling site wind velocity and humidity of air were measured. In addition to colony counts, the presence of fecal indicators (E. coli, Clostridia), Salmonella, non-agglutinable cholera vibrios and Pseudomonas aeruginosa was determined. The results were as follows: The formation of mist above the sewage in aeration tanks was preferably observed during the winter season. Correspondingly, the bacteriological results were less favourable in winter-time; trickling filters are also capable of spreading bacteria into ambient air; E. coli could be found up to a distance of 300 m downwind from the aeration tanks, Salmonella up to a distance of 10 m; higher wind velocity resulted in a higher number of positive findings, sunshine diminished it, covering of the aeration tanks clearly resulted in a lower number of positive findings, parallel examinations performed at other sampling sites far from sewage treatment plants, such as underground stations, shops, or sports facilities revealed that the bacterial level at such sites was much lower than in the air of sewage treatment plants. Quantitative studies (Salmonella was taken as an example) revealed that the dissemination by air was low in relation to the Salmonella content of the sewage itself, so that the infectious dose was hardly attained by direct ingestion. Nevertheless, hygienic measures, such as hand washing, use of protecting clothes and avoidance of contact with foods in the open air should be performed in sewage treatment plants so that the infectious dose also cannot be indirectly reached. Beyond this, the possibility cannot be ruled out that conditions are quite different for other pathogens having much lower infectious doses.     

Aerobic spore-forming bacteria from the region of salt mining

Seven strains of Gram-positive microorganisms capable of forming endospores have been isolated from soils of the region of potassium salt mining (Berezniki, Perm krai) by the method of enrichment cultures. Their typing by means of REP- and BOX-PCR has shown that four strains—I23 (= VKM V-2504), I26 (= VKM V-2505), I27, and I28—are of the same genogroup (type I), while strains SN501 (= VKM V-2508), I10b (= VKM V-2507), and I12b (= VKM V-2506) represent three different genogroups (types II, III, and IV, respectively). A phylogenetic analysis of 16S rRNA gene sequences has shown that strains I10b and SN501 belong to the genus Paenibacillus, and the remaining strains, to the genus Bacillus. These bacteria are tolerant of high salinity and alkalinity: they can grow at pH 7.0–9.0 and NaCl content of 90–110 g/l in the culture medium. They are also capable of utilizing diesel fuel and individual aromatic and aliphatic hydrocarbons.     

Long-term field application of sewage sludge increases the abundance of antibiotic resistance genes in soil

Sewage sludge and manure are common soil amendments in crop production; however, their impact on the abundance and diversity of the antibiotic resistome in soil remains elusive. In this study, by using high-throughput sequencing and high-throughput quantitative PCR, the patterns of bacterial community and antibiotic resistance genes (ARGs) in a long-term field experiment were investigated to gain insights into these impacts. A total of 130 unique ARGs and 5 mobile genetic elements (MGEs) were detected and the long-term application of sewage sludge and chicken manure significantly increased the abundance and diversity of ARGs in the soil. Genes conferring resistance to beta-lactams, tetracyclines, and multiple drugs were dominant in the samples. Sewage sludge or chicken manure applications caused significant enrichment of 108 unique ARGs and MGEs with a maximum enrichment of up to 3845 folds for mexF. The enrichment of MGEs suggested that the application of sewage sludge or manure may accelerate the dissemination of ARGs in soil through horizontal gene transfer (HGT). Based on the co-occurrence pattern of ARGs subtypes revealed by network analysis, aacC, oprD and mphA-02, were proposed to be potential indicators for quantitative estimation of the co-occurring ARGs subtypes abundance by power functions. The application of sewage sludge and manure resulted in significant increase of bacterial diversity in soil, Proteobacteria, Acidobacteria, Actinobacteria and Chloroflexi were the dominant phyla (> 10% in each sample). Five bacterial phyla (Chloroflexi, Planctomycetes, Firmicutes, Gemmatimonadetes and Bacteroidetes) were found to be significantly correlated with the ARGs in soil. Mantel test and variation partitioning analysis (VPA) suggested that bacterial community shifts, rather than MGEs, is the major driver shaping the antibiotic resistome. Additionally, the co-occurrence pattern between ARGs and microbial taxa revealed by network analysis indicated that four bacterial families might be potential hosts of ARGs. These results may shed light on the mechanism underlining the effects of amendments of sewage sludge or manure on the occurrence and dissemination of ARGs in soil.     

Effect of copper-tolerant rhizosphere bacteria on mobility of copper in soil and copper accumulation by Elsholtzia splendens

The role of rhizosphere bacteria in facilitating the solubility of copper (Cu) in contaminated soil and Cu accumulation in plant were studied. The bacteria strains were isolated from the rhizosphere of Elsholtzia splendens, a Cu accumulator growing on Tonglu Mountain copper mines. After the sandy soils containing 237 mg kg(-1) were incubated with the bacteria strains, it was indicated that rhizosphere microbes played an important role in influencing the availability of water-soluble Cu in soils. Soils had greater concentrations of water-extractable Cu compared with axenic soils inoculated with different bacterial strains. Further evidence for bacterial facilitation of increased solubility of Cu in the soil was obtained using the antibiotic ampicillin (0.1 mg g(-1)). There were 36% decreases in Cu concentration in the presence of bacterial strain MS12 and ampicillin together compared with bacterial inoculation alone. Different bacterial strains had different abilities on soil water-soluble Cu. To achieve the highest rates of plant Cu accumulation, it was necessary for bacteria to be present in the rhizosphere of E. splendens. Inoculated plants supplied with 20 micromol L(-1) CuSO4 had significantly greater concentrations of Cu in shoots and roots than uninoculated plants and bacterial strain MS2 was the most effective strain in promoting plant Cu uptake. There were 2.2-fold and 2.5-fold increases in Cu accumulation in the shoots and roots of plants inoculated with strain MS2 compared to axenic controls. Furthermore, when ampicillin and the bacterial strains were added together to the nutrient solution, the Cu concentrations in roots and shoots of ampicillin-treated plants were lower than those in inoculated plants. When ampicillin was added to the nutrient solution, Cu accumulation was inhibited by about 24-44% in shoots and 20-44% in roots. The above results provided a new insight into the phytoremediation of Cu-contaminated soil.     

Pyrethroids in house dust from the homes of farm worker families in the MICASA study

Indoor pesticide exposure is a growing concern, particularly for pyrethroids, a commonly used class of pesticides. Pyrethroid concentrations may be especially high in homes of immigrant farm worker families, who often live in close proximity to agricultural fields and are faced with poor housing conditions, potentially causing high pest infestation and pesticide use. We investigate levels of pyrethroids in the house dust of farm worker family homes in a study of mothers and children living in Mendota, CA, within the population-based Mexican Immigration to California: Agricultural Safety and Acculturation (MICASA) Study. We present pesticide use data and levels of pyrethroid pesticides in indoor dust collected in 2009 as measured by questionnaires and a GC/MS analysis of the pyrethroids cis- and trans-permethrin, cypermethrin, deltamethrin, esfenvalerate and resmethrin in single dust samples collected from 55 households. Cis- and trans-permethrin had the highest detection frequencies at 67%, with median concentrations of 244 and 172 ng/g dust, respectively. Cypermethrin was detected in 52% of the homes and had a median concentration of 186 ng/g dust. Esfenvalerate, resmethrin and deltamethrin were detected in less than half the samples. We compared the pyrethroid concentrations found in our study to other studies looking at both rural and urban homes and daycares. Lower detection frequencies and/or lower median concentrations of cis- and trans-permethrin and cypermethrin were observed in our study as compared to those studies. However, deltamethrin, esfenvalerate and resmethrin were detected more frequently in the house dust from our study than in the other studies. Because households whose children had higher urinary pyrethroid metabolite levels were more likely to be analyzed in this study, a positive bias in our estimates of household pyrethroid levels may be expected. A positive association was observed with reported outdoor pesticide use and cypermethrin levels found in the indoor dust samples (r_s = 0.28, p = 0.0450). There was also a positive association seen with summed pyrethroid levels in house dust and the results of a pesticide inventory conducted by field staff (r_s = 0.32, p = 0.018), a potentially useful predictor of pesticide exposure in farm worker family homes. Further research is warranted to fully investigate the utility of such a measure.     

Plant uptake of radiocaesium from artificially contaminated soil monoliths covering major European soil types

Uptake of ¹³⁷Cs was measured in different agricultural plant species (beans, lettuce, barley and ryegrass) grown in 5 undisturbed soil monoliths covering major European soil types. The first cultivation was made three years after soil contamination and plants were grown during 3 successive years. The plant–soil ¹³⁷Cs transfer factors varied maximally 12-fold among soils and 35-fold among species when grown on the same soil. Single correlations between transfer factors and soil properties were found, but they varied widely with plant type and can hardly be used as a predictive tool because of the few soils used. The variation of ¹³⁷Cs concentrations in plants among soils was related to differences in soil solution ¹³⁷Cs and K concentrations, consistent with previous observations in hydroponics and pot trials. Absolute values of transfer factors could not be predicted based on a model validated for pot trials. The ¹³⁷Cs activity concentration in soil solution decreased significantly (11- to 250-fold) for most soils in the 1997–1999 period and is partly explained by decreasing K in soil solution. Transfer factors of lettuce showed both increasing and decreasing trends between 2 consecutive years depending on soil type. The trends could be explained by the variation in ¹³⁷Cs and K concentrations in soil solution. It is concluded that differences in ¹³⁷Cs transfer factors among soils and trends in transfer factors as a function of time can be explained from soil solution composition, as shown previously for pot trials, although absolute values of transfer factors could not be predicted.     

Effects of soil amendments on lead uptake by two vegetable crops from a lead-contaminated soil from Anhui, China

Previous studies have documented that phosphate compounds of lead (Pb) [e.g., pyromorphite Pb5(PO4)3-(X), where X=OH, F or Cl] are comparatively insoluble, and their formation in Pb-contaminated soil may be a means of reducing the bioavailability and chemical lability of Pb in soil. In this study, the effect of phosphate compound amendments on the bioavailability of Pb in a polluted alkaline soil was examined. A Pb-contaminated soil was treated with hydroxyapatite (HA), phosphate rock (PR), water-soluble P fertilizer (single superphosphate, SSP) and the combination of HA with SSP. The bioavailability of Pb was determined in plant uptake studies with vegetables (Brassica campetris L. var. communis, BC) and Brassica oleracea L. var. acephala, BO) and sequential extraction. The results indicated that the Pb concentrations in both shoots and roots of two vegetable plants decreased with increasing quantities of added P compound, and the HA treatment had the best effect at the level of 5000 mg of P kg(-1)as compared with other treatments in which the Pb concentrations in shoots of BO and BC decreased 51.9% and 65.5%, respectively, and the Pb concentrations in roots of BO and BC decreased 67.3% and 57.2%, respectively, as compared with the control treatment. The SSP treatment had little effect on the Pb concentrations in plant tissues. Sequential extraction results indicated that the addition of soil amendments transform soil Pb from nonresidual fractions to residual fraction substantially. The effect of treatments followed this order at the equivalent P addition: HA>PR>HA+SSP>SSP. The results suggested that HA amendments can lower the bioavailability and increase the geochemical stability of soil Pb, so it has the potential for in situ remediation in Pb-contaminated soils.     

Characteristics of microorganisms isolated from technogenic soils of the Kama region

Microorganisms capable of degrading monocyclic and polycyclic aromatic hydrocarbons and several chlorinated aromatic compounds were isolated from soils polluted with industrial waste from chemical plants. They were identified as representatives of the genera Pseudomonas, Flavobacterium, Alcaligenes, Rhodococcus, Microbacterium, Cellulomonas, Arthrobacter, and Brevibacterium. Among them, bacteria capable of utilizing xenobiotics in a wide range of ambient temperatures and pH and in the presence of high sodium chloride concentrations were revealed.     

Multi-elemental EDXRF mapping of polluted soil from former horticultural land

The distribution of major and trace elements was systematically investigated by use of energy dispersive X-ray fluorescence spectrometry (EDXRF) on a former horticultural soil. The purpose of the study was to combine mapping of soil element concentration levels with multivariate statistics for characterisation of soil metal pollution in relation to previous and present land use. A 1-ha study site was chosen from a former horticulture where a previous preliminary survey indicated increased concentration levels of toxic elements. The soil was sampled from the top 20 cm of the soil surface in a 10 x 10-m grid-like pattern covering the 1-ha study area. In addition, three soil profiles were studied. The elemental composition of the soil samples was investigated by EDXRF while the composition of aqueous soil extracts was determined by total reflection X-ray fluorescence spectrometry (TXRF). Based on mapping and multivariate statistically analysis of the data obtained by EDXRF, most elements were found in almost constant concentration levels in the top soil throughout the investigated site. However, the contents of the toxic elements Zn, Cu, As, and Pb were found to vary significantly within the area. Hence, the samples with high accumulations of As also contained relatively high amounts of Zn, Cu, and Pb, which indicates that toxic-element-containing pesticides have been applied to the soil surface in the area of the former green houses at the study site. The Pb/As mass ratio in the soil indicates that PbHAsO3 was the preferential lead arsenate used for pest management at the investigated site, while Cu as Bordeaux liquid (CuSO4) and Zn were applied to minimize the leaf damaging effect from the former compounds. Calculations indicated that As annually was applied to the soil in the former greenhouses in doses up to 4 kg As/ha while Pb had been annually applied in doses up to 12 kg Pb/ha. The enrichment of Zn, Cu, As and Pb was greatest in the top 20 cm of the soil and no anthropogenic enrichment of these elements occurred below a depth of 50 cm, indicating that the toxic elements are rather immobile in this soil. The results of this investigation suggest that EDXRF used in combination with multivariate statistics is a strong tool for multi-element mapping of elemental contents, sources and mobility in the terrestrial environment.     

Re-emission of heavy water vapour from soil to the atmosphere

The re-emission process of tritiated water (HTO) deposited on a soil surface is an important process to assess tritium doses to the general public around nuclear fusion facilities in future. A field experiment using heavy water (HDO) as a substitute for HTO was carried out in the summertime to investigate the re-emission process of HTO from soil to the atmosphere. In the experiment, the time variations of depth profiles of HDO concentrations in soil exposed to HDO vapour and soil mixed with HDO were measured during the re-emission process on the field. The HDO concentrations in soil water in top soil layers of both the exposed and mixed soil rapidly decreased with time during the re-emission. However, the decrease of exposed soil was much greater than that of mixed soil. The re-emission process was analysed using a model including the evaporation of HDO from soil, the exchange between soil HDO and air H2O, and the diffusion of HDO in soil. It was found that the model is applicable to calculating the time variations of detailed depth profiles of HDO concentration in soil water in surface soil layers, using an estimated exchange velocity.     

Transfer factors of Polonium from soil to parsley and mint

Transfer factors of ²¹⁰Po from soil to parsley and mint have been determined. Artificial polonium isotope (²⁰⁸Po) was used as a tracer to determine transfer factor of Po from soil to plant in pot experiments. Two plant growing systems were used for this study namely, an outdoor system and a sheltered system by a polyethylene tent. ²⁰⁸Po and ²¹⁰Po were determined in soil and different parts of the studied plants (stem and leaf), using alpha spectroscopy. The results have shown that there was a clear uptake of ²⁰⁸Po by roots to leaves and stems of both plants. Higher values of transfer factors using the ²¹⁰Po activity concentrations than the ²⁰⁸Po activity concentration were observed. Transfer factors of ²¹⁰Po from soil to parsley varied between 20 × 10⁻² and 50 × 10⁻² and 22 × 10⁻³ and 67 × 10⁻³ in mint, while ²⁰⁸Po transfer factors varied between 4 × 10⁻² and 12 × 10⁻² for parsley and 10 × 10⁻² and 22 × 10⁻² in mint. Transfer factors of Po were higher in those plants grown in the sheltered system than in the open system; about 75% of Po was transferred from atmosphere to parsley parts using the two systems. Ratios of transferred Po from soil to mint stem and leaf in the sheltered system were higher by 2 times from those in the open system.     

Kinetics of radiocesium released from contaminated soil by fertilizer solutions

(137)Cs is one of the major artificial radionuclides found in environments; but the mechanisms behind fertilizer-induced (137)Cs desorption from soil remain unknown. This study aimed to investigate the kinetics and mechanisms underlying the various cations and anions that cause Cs release from soil under acidic conditions. NH(4)H(2)PO(4) (1M), 0.5M (NH(4))(2)SO(4), 1M NH(4)Cl, 1M KCl or 1M NaCl solutions were added to (137)Cs-contaminated soil. The power function model well described the short term (137)Cs desorption with the solutions. The rate coefficients for (137)Cs release from soil in NH(4)H(2)PO(4), (NH(4))(2)SO(4), NH(4)Cl, and KCl solutions were 7.7, 7.3, 6.8, and 6.1 times higher than the rate observed in a NaCl solution, respectively. The NH(4)H(2)PO(4) and (NH(4))(2)SO(4) solutions induced significantly greater (137)Cs release from the contaminated soil than the NH(4)Cl, KCl and NaCl solutions. After four times repeated extractions with the fertilizer solutions, the total amount of (137)Cs extracted by (NH(4))(2)SO(4) and NH(4)Cl solutions reached equilibrium, while that extracted using an NH(4)H(2)PO(4) solution continued to increase. The combined effect of phosphate and protons was the major mechanism behind (137)Cs release from contaminated soils, when an NH(4)H(2)PO(4) solution was used.     

Effect of tetracycline residues in pig manure slurry on tetracycline-resistant bacteria and resistance gene tet(M) in soil microcosms

Effects of tetracycline residues from pig manure slurry on the prevalence of tetracycline-resistant bacteria and the tetracycline resistance gene, tet(M), were studied in soil microcosms. Four types of soil microcosms were established for a period of 152 days, supplemented with combinations of pig manure slurry and a tetracycline-resistant Enterococcus faecalis, CG110, containing the tetracycline resistance gene tet(M) (on the conjugative transposon, Tn916). The prevalence of both tetracycline-resistant aerobic bacteria and tetracycline-resistant enterococci declined rapidly until day 45 where no significant differences in the levels of tetracycline-resistant bacteria in any of the four types of microcosms could be detected. tet(M) could be detected in microcosms supplemented with either pig manure slurry and/or E. faecalis CG110 (tet(M)) for the whole period (152 days). tet(M) could be detected longer than tetracycline-resistant enterococci could be isolated (limit of detection 100 CFU/g soil) probably due to viable but not culturable (VBNC) bacteria with tet(M), horizontal gene transfer of tet(M) to indigenous soil bacteria or presence of "free" DNA. The concentration of chlortetracycline and oxytetracycline were almost stable through out the experimental period, but the tetracycline concentrations had no effect on prevalence of tetracycline-resistant bacteria. The presented microcosm approach simulated natural farmland conditions well and supported results from previous field studies.     

Characterisation, dissemination and persistence of gentamicin resistant Escherichia coli from a Danish university hospital to the waste water environment

The aim of the study was to investigate the potential spread of gentamicin resistant (GEN(R)) Escherichia coli isolates or GEN(R) determinants from a Danish university hospital to the waste water environment. Waste water samples were collected monthly from the outlets of the hospital bed wards and the inlet of the related waste water treatment plant (WWTP) from October 2002 to August 2003. Waste water samples were also collected monthly from a residential area in the same period to be able to compare the prevalence of GEN(R)E. coli isolates from hospital related and residential waste water. The waste water isolates were compared to GEN(R)E. coli isolates obtained consecutively from September 2002 to September 2003 from patients mainly with urinary tract infections at the hospital with respect to Pulsed Field Gel Electrophoresis (PFGE) profiles. All isolates were investigated for GEN(R) mechanisms (aac(3)-II, aac(3)-IV, ant(2')-I, armA), phenotypic resistance pattern, and virulence genes (hlyA, chuA, sfaS, fogG, malX, traT, iutA, fyuA, iroN, cnf1) to investigate if the hospital and waste water could be reservoirs of antimicrobial resistance and virulence. The ability for GEN(R) determinants to transfer horizontally was investigated by mating experiments. A total of 38, 15, 21, and two GEN(R)E. coli were isolated from patients, the hospital outlets, the inlet of the WWTP, and the residential area, respectively. GEN(R)E. coli were more prevalent in waste water from the hospital and the WWTP than in waste water from the residential area. PFGE profiling revealed no spread of specific patient isolates to the waste water. The aac(3)-II gene was detected both in patient and waste water isolates. Furthermore horizontal transfer of the aac(3)-II gene of patient origin to a recipient was shown in vitro, indicating a potential spread of the gene from patient isolates to waste water isolates. Regardless of origin, most isolates exhibited multi-resistance and contained several virulence genes. In conclusion, our study showed a possible spread of aac(3)-II from the hospital to the waste water. Most of the GEN(R)E. coli isolates from both patients and waste water had a multi-resistant phenotype and contained virulence genes and should therefore be considered reservoirs of antimicrobial resistance and virulence genes.     

Leaching of nickel and copper from soil contaminated by metallurgical dust

The paper presents the results of the laboratory percolation experiment simulated soil contamination by emissions from a Ni-Cu smelter. Humus (Ao horizon) columns were transferred to lysimeters from an illuvial, humic, ferriferous forest podzol site. Fine metallurgical dust containing Ni and Cu was layered on the columns and irrigated with sulphuric acid solutions at pH 3, 4, 5, and 6. Irrigation for 19 months indicated that the leaching of metals down the humus column was greatest at pH 6. Calculations indicated that it would take 160-270 years for complete leaching of Ni from the Ao layer, and 100-200 years for Cu, depending on the dust composition. Natural decontamination of affected soils will take centuries.