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1.
AIMS: The objective of this study was to determine whether Cu-amendment of field plots affects the frequency of Cu resistance, and antibiotic resistance patterns in indigenous soil bacteria. METHODS AND RESULTS: Soil bacteria were isolated from untreated and Cu-amended field plots. Cu-amendment significantly increased the frequency of Cu-resistant isolates. A panel of isolates were characterized by Gram-reaction, amplified ribosomal DNA restriction analysis and resistance profiling against seven antibiotics. More than 95% of the Cu-resistant isolates were Gram-negative. Cu-resistant Gram-negative isolates had significantly higher incidence of resistance to ampicillin, sulphanilamide and multiple (> or =3) antibiotics than Cu-sensitive Gram-negative isolates. Furthermore, Cu-resistant Gram-negative isolates from Cu-contaminated plots had significantly higher incidence of resistance to chloramphenicol and multiple (> or =2) antibiotics than corresponding isolates from control plots. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this field experiment show that introduction of Cu to agricultural soil selects for Cu resistance, but also indirectly selects for antibiotic resistance in the Cu-resistant bacteria. Hence, the widespread accumulation of Cu in agricultural soils worldwide could have a significant effect on the environmental selection of antibiotic resistance.  相似文献   

2.
ABSTRACT: BACKGROUND: Copper mining has led to Cu pollution in agricultural soils. In this report, the effects of Cu pollution on bacterial communities of agricultural soils from Valparaiso region, central Chile, were studied. Denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA genes was used for the characterization of bacterial communities from Cu-polluted and non-polluted soils. Cu-resistant bacterial strains were isolated from Cu-polluted soils and characterized. RESULTS: DGGE showed a similar high number of bands and banding pattern of the bacterial communities from Cu-polluted and non-polluted soils. The presence of copA genes encoding the multi-copper oxidase that confers Cu-resistance in bacteria was detected by PCR in metagenomic DNA from the three Cu-polluted soils, but not in the non-polluted soil. The number of Cu-tolerant heterotrophic cultivable bacteria was significantly higher in Cu-polluted soils than in the non-polluted soil. Ninety two Cu-resistant bacterial strains were isolated from three Cu-polluted agricultural soils. Five isolated strains showed high resistance to copper (MIC ranged from 3.1 to 4.7 mM) and also resistance to other heavy metals. 16S rRNA gene sequence analyses indicate that these isolates belong to the genera Sphingomonas, Stenotrophomonas and Arthrobacter. The Sphingomonas sp. strains O12, A32 and A55 and Stenotrophomonas sp. C21 possess plasmids containing the Cu-resistance copA genes. Arthrobacter sp. O4 possesses the copA gene, but plasmids were not detected in this strain. The amino acid sequences of CopA from Sphingomonas isolates (O12, A32 and A55), Stenotrophomonas strain (C21) and Arthrobacter strain (O4) are closely related to CopA from Sphingomonas, Stenotrophomonas and Arthrobacter strains, respectively. CONCLUSIONS: This study suggests that bacterial communities of agricultural soils from central Chile exposed to long-term Cu-pollution have been adapted by acquiring Cu genetic determinants. Five bacterial isolates showed high copper resistance and additional resistance to other heavy metals. Detection of copA gene in plasmids of four Cu-resistant isolates indicates that mobile genetic elements are involved in the spreading of Cu genetic determinants in polluted environments.  相似文献   

3.
Abstract: A total of 132 different fluorescent Pseudomonas strains were isolated from several agricultural and industrial soils. The bacteria from the two different soil environments were compared for species and biotype variation, antibiotic and heavy metal resistance profiles, ability to degrade polyaromatic hydrocarbons, and plasmid incidence. Irrespective of the soil type, the isolates belonged to Pseudomonas fluorescens biotypes I–VI and Pseudomonas putida biotype B. Except for a streptomycin resistant isolate from one of the industrial soils, all the strains had the same antibiotic resistance profile. However, there was a higher incidence of heavy metal resistance and polyaromatic hydrocarbon degradation phenotypes in the isolates from industrial soils than from the agricultural soils. Only 2 out of 68 strains from agricultural soil were found to carry plasmids, while 28 out of 64 strains from industrial soil had plasmids. A majority of the plasmids (56%) were estimated to be larger than 50 kb, indicating that they could encode transfer functions. However, transferability as indicated by the ability to mobilize an IncQ plasmid (tra, mob+), was observed with only one plasmid. None of the plasmid(s) containing isolates hybridized to a 32P-labelled repP probe suggesting that none of the indigenous plasmids in the soil fluorescent Pseudomonas strains was related to the IncP group of conjugative plasmids commonly associated with resistance and catabolic genes.  相似文献   

4.
Animal manures and municipal biosolids recycled onto crop production land carry antibiotic-resistant bacteria that can influence the antibiotic resistome of agricultural soils, but little is known about the contribution of bacteriophage to the dissemination of antibiotic resistance genes (ARGs) in this context. In this work, we quantified a set of ARGs in the bacterial and bacteriophage fractions of agricultural soil by quantitative PCR. All tested ARGs were present in both the bacterial and phage fractions. We demonstrate that fertilization of soil with dairy manure or human biosolids increases ARG abundance in the bacterial fraction but not the bacteriophage fraction and further show that pretreatment of dairy manure can impact ARG abundance in the bacterial fraction. Finally, we show that purified bacteriophage can confer increased antibiotic resistance to soil bacteria when combined with selective pressure. The results indicate that soilborne bacteriophage represents a substantial reservoir of antibiotic resistance and that bacteriophage could play a significant role in the horizontal transfer of resistance genes in the context of an agricultural soil microbiome. Overall, our work reinforces the advisability of composting or digesting fecal material prior to field application and suggests that application of some antibiotics at subclinical concentrations can promote bacteriophage-mediated horizontal transfer of ARGs in agricultural soil microbiomes.  相似文献   

5.
Heavy metal contents of agricultural and industrial soils were determined by atomic absorption spectrophotometry. The analysis of the samples collected from two different locations revealed significantly high levels of Fe, Zn, Cu, Cr and Ni. Certain microbiological parameters (total aerobic heterotrophs, asymbiotic N2-fixers, total Actinomycetes and fungi) were also monitored from these soils. A total of 70 bacterial isolates from agricultural and industrial soils were examined for plasmid DNA content and resistance to the antibiotics amoxycillin, cloxacillin, chloramphenicol, doxycycline methicillin, nalidixic acid, and tetracycline. Minimum inhibitory concentrations (MICs) of Cu, Cr, Pb, Cd, Hg, Zn, and Ni for each isolate were also determined. Resistance was most frequent to methicillin (48.5%), cloxacillin (45.7%), and nalidixic acid (40%) for all isolates of bacteria. The highest MICs observed were 100 g/ml for mercury, 800 g/ml for Ni and 1600 g/ml for other metals. The incidences of metal resistance and MICs of metals for bacteria from industrial soil were significantly different to those of agricultural soil. On a percentage basis, 91.4% of the total bacterial isolates from industrial soil were found to harbour plasmids whereas 40% of the isolates from agricultural soil contained plasmids.  相似文献   

6.
The usage of antibiotics in animal husbandry has promoted the development and abundance of antibiotic resistance in farm environments. Manure has become a reservoir of resistant bacteria and antibiotic compounds, and its application to agricultural soils is assumed to significantly increase antibiotic resistance genes and selection of resistant bacterial populations in soil. The genome location of resistance genes is likely to shift towards mobile genetic elements such as broad-host-range plasmids, integrons, and transposable elements. Horizontal transfer of these elements to bacteria adapted to soil or other habitats supports their environmental transmission independent of the original host. The human exposure to soil-borne resistance has yet to be determined, but is likely to be severely underestimated.  相似文献   

7.
Heavy metal contamination of agricultural soils resulting from rapid industrialization and urbanization is of great concern because of potential health risk due to dietary intake of contaminated vegetables. The present study aims to evaluate the status of heavy metals contamination of agricultural soils and food crops around an urban-industrial region in India. Transfer factor values of Cu, Cr, Pb, Cd, Zn, and Ni from soil to vegetable was estimated. The mean heavy metal concentrations (mg/kg) in agricultural soils (Cu: 17.8, Cr: 27.3, Pb: 29.8, Cd: 0.43, Zn: 87, Mn: 306.6, Fe: 16984, and Ni: 53.8) were within allowable concentrations for Indian agricultural soil. The concentrations of Pb, Cd, Zn, and Ni in crops/vegetables exceeded the World Health Organization/Food and Agriculture Organization safe limits. Relative orders of transfer of metals from soil to edible parts of the crops/vegetables were Cd > Pb > Ni > Zn > Cu > Cr. The enrichment factors of heavy metals in soil indicated minor to moderately severe enrichment for Pb, Cd, and Ni; minor to moderate enrichment for Zn; no enrichment to minor enrichment for Mn; and no enrichment to moderate enrichment for Cu at different sites. Ecological risk index of soil showed considerable contamination in one of the wastewater irrigated sites.  相似文献   

8.
Pseudomonas putida CR30RNS (pADPTel) is an antibiotic-resistant strain with a recombinant plasmid that confers resistance to tellurite and the ability to catabolize atrazine. The survival of this strain as well as its ability to transfer genes for atrazine degradation and tellurite resistance to indigenous soil bacteria were tested in both fallow soil and canola (Brassica napus) rhizosphere by the use of parallel field and laboratory releases. Culturable CR30RNS (pADPTel) were enumerated in field and microcosm soils at 7- to 14-day intervals over 49 d. Strain CR30RNS (pADPTel) survived for up to 7 weeks in microcosm soils at a density of 10(4) CFU/g soil, whereas in field soils the population declined to 10(3) CFU/g soil by the fourth week. In contrast, when CR30RNS (pADPTel) was introduced into the soil as a seed coating of canola (B. napus 'Karoo'), the bacterium established at higher cell densities in the rhizosphere (10(6)-10(5) CFU/g fresh root mass), with no subsequent decrease in numbers. The presence of selective pressure (i.e., atrazine) had no significant effect on the survival of CR30RNS (pADPTel) in either field or microcosm soils. One year postinoculation field sites were examined for the presence of CR30RNS (pADPTel) and no evidence of culturable parental cells was observed when samples were plated onto selective media. However, the atzC and telAB gene segments were amplified from the field soils at that time. Under laboratory conditions, indigenous soil bacteria were capable of receiving and expressing the engineered plasmid construct at frequencies ranging from 1 to 10(-3) transconjugants per donor. However, no plasmid transfer to indigenous soil bacteria was detected in the field or microcosm soils regardless of the presence of canola rhizosphere and (or) the application of atrazine. Our results show that the survival and population size of P. putida CR30RNS (pADPTel) might be sufficient for degradation of environmental pollutants but that the transfer frequency was too low to be detected under the conditions of this study.  相似文献   

9.
The biodegradable chelating agent (S,S)-Ehylenediaminedisuccinic acid (EDDS), autochthonous acidophilic bacteria, and a combination of the two means were investigated for the removal of pseudo-total and ethylenediaminetetraacetic acid (EDTA)-available content of Cu from surface layers of three soil categories in the Bor copper mining area. Their efficiencies were compared at mine overburden, flotation tailings, and agricultural land sites in order to determine the potential role of these approaches in the soil remediation process. The most effective removal of Cu was achieved on flotation tailings, where combined treatment showed significant reduction of pseudo-total and EDTA-available concentrations of Cu (40.5?±?27.3% and 99.6?±?0.2%, respectively). Acidophilic bacteria treatment showed high efficiency on flotation tailings, removing 94.1?±?1.2% of EDTA-available Cu. EDDS treatment showed discernible results in the removal of EDTA-available Cu from agricultural land soil (44.4?±?13.9%). In the case of overburden soil material, selected agents did not have statistically significant results in the removal of pseudo-total or EDTA-available fraction of Cu. Chosen remediation approaches showed diverse efficiency for soil categories on investigated sites. Combined approach showed synergistic results in the case of EDTA-available Cu removal from flotation tailings soils, suggesting that this combination deserves further attention as a potentially promising environmentally friendly remediation option.  相似文献   

10.
Aims:  To analyse the arsenic-resistant bacterial communities of two agricultural soils of Bangladesh, to isolate arsenic-resistant bacteria, to study their potential role in arsenic transformation and to investigate the genetic determinants for arsenic resistance among the isolates.
Methods and Results:  Enrichment cultures were performed in a minimal medium in the presence of As(III) and As(V) to isolate resistant bacteria. Twenty-one arsenic-resistant bacteria belonging to different genera of Gram-positive and Gram-negative bacteria were isolated. The isolates, with the exception of Oceanimonas doudoroffii Dhal Rw, reduced 2 mmol l−1 As(V) completely to As(III) in aerobic conditions. Putative gene fragments for arsenite efflux pumps were amplified in isolates from Dhal soil and a putative arsenate reductase gene fragment was amplified from a Bacillus sp. from Rice soil.
Conclusions:  Phylogenetically diverse arsenic-resistant bacteria present in agricultural soils of Bangladesh are capable of reducing arsenate to arsenite under aerobic conditions apparently for detoxification purpose.
Significance and Impact of the Study:  This study provides results on identification, levels of arsenic resistance and reduction of arsenate by the bacterial isolates which could play an important role in arsenic cycling in the two arsenic-contaminated soils in Bangladesh.  相似文献   

11.
Transfer of plasmid RP4 to indigenous bacteria in bulk soil could only be detected in soil with nutrient amendment. Lack of physiological active donor and recipient cells was apparently one of the limiting factors in un-amended bulk soil. Plasmid transfer was detected both in the spermosphere and rhizosphere of barley seedlings. Transfer occured from seed coated donor bacteria (i) to introduced recipient bacteria and (ii) to indigenous bacteria present in soil. Plasmid transfer was also detected from donor bacteria introduced to the soil to seed coated recipient bacteria. Transfer efficiencies in the rhizosphere were significantly below the transfer efficiencies obtained in the spermosphere. The transfer efficiencies detected in the barley spermosphere were among the highest reported from any natural environment.  相似文献   

12.
Understanding the soil bacterial resistome is essential to understanding the evolution and development of antibiotic resistance, and its spread between species and biomes. We have identified and characterized multi-drug resistance (MDR) mechanisms in the culturable soil antibiotic resistome and linked the resistance profiles to bacterial species. We isolated 412 antibiotic resistant bacteria from agricultural, urban and pristine soils. All isolates were multi-drug resistant, of which greater than 80% were resistant to 16–23 antibiotics, comprising almost all classes of antibiotic. The mobile resistance genes investigated, (ESBL, bla NDM-1, and plasmid mediated quinolone resistance (PMQR) resistance genes) were not responsible for the respective resistance phenotypes nor were they present in the extracted soil DNA. Efflux was demonstrated to play an important role in MDR and many resistance phenotypes. Clinically relevant Burkholderia species are intrinsically resistant to ciprofloxacin but the soil Burkholderia species were not intrinsically resistant to ciprofloxacin. Using a phenotypic enzyme assay we identified the antibiotic specific inactivation of trimethoprim in 21 bacteria from different soils. The results of this study identified the importance of the efflux mechanism in the soil resistome and variations between the intrinsic resistance profiles of clinical and soil bacteria of the same family.  相似文献   

13.
Adsorption of Cu, Cd, Ni, and Zn in single and multi-metal solutions by agricultural and forest soils was investigated in batch sorption experiments. The results showed significant differences in sorption capacities of the studied soils. The selectivity order was as follows: agricultural soil? top forest soil > bottom forest soil. The adsorption sequence Cu > Zn > Ni > Cd was established for the agricultural and bottom forest soil, while the order for the top forest soil was Cu > Ni > Zn > Cd. The experimental isotherms for the metal sorption were described satisfactorily by the Freundlich and Langmuir models. The competitive adsorption experiment indicated a reduction in the amount of metals adsorbed by the soils from the multi-metal solution compared to the single metal solution. Properties of the soils, such as pH, content of clay and organic matter, exchangeable bases and hydrolytic acidity, showed a significant influence on adsorption capacities of the studied soils.  相似文献   

14.
The community structure in two different agricultural soils has been investigated. Phenotypic diversity was assessed by applying BIOLOG-profiles on a total of 208 bacterial isolates. Diversity indices were calculated from cluster analysis of the BIOLOG data. The bacterial isolates were also evaluated for resistance towards six different antibiotics, mercury resistance and the presence of plasmids. The presence of tetracycline-resistant determinants class A to E among Gram-negative bacteria was analysed with DNA probes. The distribution of tetracycline resistance markers among colonies growing on non-selective and tetracycline-selective plates were compared. The phenotypic approach demonstrated some difference in the diversity within the two soils. The frequency of antibiotic resistance isolates was high in both soils, whereas the frequency of mercury resistance differed significantly. We found no correlation between plasmid profiles and antibiotic resistance patterns. We found all the tetracycline resistance determinants except class B, indicating that the diversity of the tetracycline resistance determinants was complex in populations of resident soil bacteria under no apparent selective pressure for the genes in question.  相似文献   

15.
High Cu contamination in agricultural soil can cause toxicity, leading to ecological damages. Thus, we need to understand the concentration level, contaminated area, and spatial distribution of Cu in agricultural soils on a regional or national scale. This paper reviewed the studies on Cu concentrations throughout Chinese agricultural soils, based on relevant 482 published papers from 2004 to 2017. The results showed that the average Cu concentration was 29.85 mg/kg, higher than its background of 22.60 mg/kg, indicating that Cu has been introduced into soil from exterior sources. A large high spatial cluster was observed in southern China, including Hunan, Fujian, Guangdong, and Guangxi provinces, while low spatial clusters were mainly found in Chongqing and Shandong provinces. About 3.08% of the agricultural areas faced a high Cu risk, higher than the pollution rates of the whole land uses, based on the spatial distribution of Cu concentrations throughout China. The remaining areas were within the ranges of moderate and low Cu risks.  相似文献   

16.
 Soils from valley oak (Quercus lobata Nee) riparian areas of the Cosumnes River Nature Conservancy Preserve near Sacramento, California were added to growth medium of valley oak seedlings grown in a greenhouse or in agricultural fields at Cosumnes which probably once supported valley oak trees and are now replanted with native riparian vegetation or allowed to revegetate naturally. Agricultural field soil from the Cosumnes River Preserve was presumed to be low or lacking in ectomycorrhizal inoculum. The study was designed to (1) determine whether valley oak stand soil transfer could cause mycorrhizal infection on valley oak seedlings in an agricultural field and in a greenhouse, (2) describe ectomycorrhizal morphological types formed on valley oak seedlings, and (3) determine whether seedling growth is enhanced more by transfer of natural valley oak stand soil than agricultural field soil. In the field study, transfer of forest soil increased average ectomycorrhizal diversity (2.4 types) more than transfer of agricultural field soil (1.2 types). Valley oak seedlings were responsive to ectomycorrhizal infection in the field study. With increase in mycorrhizal infection there was an increase in shoot growth at the expense of root growth. In the greenhouse study, both percent mycorrhizal infection and mycorrhizal diversity were increased more by transfer of oak forest and woodland soils than agricultural field soil. Eight morphotypes occurred on seedlings in forest and woodland soils but only three morphotypes in agricultural soil. This result strongly suggests that the agricultural field also harbors ectomycorrhizal propagules but forest and woodland soils support a more abundant and diverse ectomycorrhizal flora. Accepted: 17 August 1997  相似文献   

17.
Few studies have been done to evaluate the transfer of catabolic plasmids from an introduced donor strain to indigenous microbial populations as a means to remediate contaminated soils. In this work we determined the effect of the conjugative transfer of two 2,4-D degradative plasmids to indigenous soil bacterial populations on the rate of 2,4-D degradation in soil. We also assessed the influence of the presence of 2,4-D on the number of transconjugants formed. The two plasmids used, pEMT1k and pEMT3k, encode 2,4-D degradative genes (tfd) that differ in DNA sequence as well as gene organisation, and confer different growth rates to Ralstonia eutropha JMP228 when grown with 2,4-D as a sole carbon source. In an agricultural soil (Ardoyen) treated with 2,4-D (100 ppm) there were ca. 107CFU of transconjugants per gram bearing pEMT1k as well as a high number of pEMT3k bearing transconjugants (ca. 106 CFU/g). In this soil the formation of a high number of 2,4-D degrading transconjugants resulted in faster degradation of 2,4-D as compared to the uninoculated control soil. In contrast, only transconjugants with pEMT1k were detected (at a level of ca. 103 CFU/g soil) in the untreated Ardoyen soil. High numbers of transconjugants that carried pEMT1k were also found in a second experiment done using forest soil (Lembeke) treated with 100 ppm 2,4-D. However, unlike in the Ardoyen soil, no transconjugants with pEMT3k were detected and the transfer of plasmid pEMT1k to indigenous bacteria did not result in a higher rate of decrease of 2,4-D. This may be because 2,4-D was readily metabolised by indigenous bacteria in this soil. The results indicate that bioaugmentation with catabolic plasmids may be a viable means to enhance the bioremediation of soils which lack an adequate intrinsic ability to degrade a given xenobiotic.  相似文献   

18.
生物耐铜的分子机理及铜污染环境的生物联合修复   总被引:2,自引:0,他引:2  
李杰  贺纪正  马延和  朱永官  张蕾 《生态学报》2007,27(6):2615-2626
铜是动植物和人类必需的微量元素,缺乏或过多都将产生不良影响。随着社会经济的发展,人类活动对环境的干扰日益加剧,工业和农业生产活动常可导致土壤铜污染,铜已成为土壤重金属污染的主要元素之一。总结了铜在植物体内的自发内稳态调节机制,在细菌和真菌体内的吸收、分布、解毒和调节因子,同时以蚯蚓为例简要阐述了土壤动物对铜的解毒机理;从分子生物学角度对重金属铜在生物体内的代谢机理及生物对环境中过量铜的联合修复研究进展进行了综述,以期为铜污染环境的植物、微生物和动物联合修复的分子机理研究提供借鉴。  相似文献   

19.
Urbanization and agriculture cause alterations in soil biodiversity. As arthropods quickly respond to soil changes, they have been recognized as good indicators of soil quality. This study aimed to evaluate if (1) soil physico-chemical properties act as environmental filters of the functional trait distribution of soil arthropod community; (2) the abundance of certain functional traits is linked to different land uses; (3) the taxonomical and functional approaches provide complementary information about the evaluation of biological soil quality. The study was performed at forest, agricultural and urban sites in downtown and surroundings of Naples. The soils were characterized for texture, pH, organic matter content, water content, total Cd, Cr, Cu, Ni, Pb and Zn concentrations. The forest soils showed the highest organic matter and water contents, and the lowest metal concentrations; the agricultural soils showed the lowest organic matter and water contents and high Cd and Cu concentrations, whereas the urban soils showed high Pb and Zn concentrations. The forest soils showed the highest arthropod density, taxa richness and diversity. The distribution of the functional traits in the arthropod communities of the different land uses highlighted similarity only for the preference to occupy soil and for the percentage contribution of detritivores. The abundance of arthropods living above litter and omnivores were positively correlated to Pb and Zn concentrations, whereas predators were positively correlated to Cu concentrations. The findings suggest that the arthropod identification at order level could be enough to detect differences in functional traits among forest, urban, and agricultural soils.  相似文献   

20.
The role of a hemiparasitic life-style in plant resistance to toxic trace elements in polluted soils is unclear. Restriction of metal uptake by the host, restriction of metal transfer from host to parasite, or transformation of metals into a less toxic form may play a role. This study analysed the transfer of selected mineral elements from soil to host ( Cistus spp . ) and from host to hemiparasite ( Odontites lutea) at locations with different metal burdens: a Cu-rich serpentine site, Pb–Ba mine spoil and an unpolluted soil. Highest soil-to-host transfer factors for K, Mg, Ca, Zn, Cu and Pb were observed on the unpolluted soil. Statistically significant differences among locations of host-to-parasite transfer factors were only found for Ca and Pb. Restriction of transfer of unfavourable Ca/Mg ratios, characteristic at the serpentine site, and of high Pb and Zn concentrations at the Pb–Ba mine occurred mainly at the soil–host, and not at the host–parasite, level. Odontites lutea was able to withstand enhanced Zn and Pb concentrations and low Fe/Cu ratios in shoot tissue without developing toxicity symptoms. This could be caused by specific metal resistance mechanisms in this hemiparasite and/or the transformation and transfer of these metals into a less toxic form by the metal-tolerant host.  相似文献   

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