Degradability of atrazine, cyanazine, and dicamba in methanogenic enrichment culture microcosms using sediment from the Pearl River of Southern China

Degradation of three herbicides, atrazine, cyanazine and dicamba, was assessed in laboratory microcosms incubated under simulated methanogenic conditions using sediment from Pearl River of Southern China as an inoculum. Atrazine was much more resistant to degradation than cyanazine and dicamba over 300 days of incubation. Biodegradation of cyanazine and dicamba was further substantiated by establishment of enrichment transfer cultures in which the degradation of the respective herbicide was accelerated by the active microorganisms. Degradation of cyanazine initially involved the removal of chlorine and the two side chains, while that of dicamba was O-demethylation reaction forming 3,6-dichlorosalicyclic acid. Results suggest that biodegradation of xenobiotics can be established through enrichment culture transfer technique, and further mechanism of degradation and microorganisms involved can be elucidated.     

Sequential Microaerophilic-Oxic Phase Mineralization of Azo Dyes by a Monoculture of Pseudomonas Aeruginosa Strain AWF Isolated from Textile Wastewater

A novel dye degrading bacterium capable of decolorizing and mineralizing four different dyes (Methyl red, Orange II, Direct red 80, and Direct blue 71) was isolated from textile industrial wastewater using the selective enrichment technique. The bacterium was identified as Pseudomonas aeruginosa. More than 80 % decolorization of Direct red 80 was obtained under microaerophilic conditions in 48 h, whereas only 10 % color removal was obtained under oxic conditions at the same time. Subsequent aeration of the decolorized medium resulted in the mineralization of the metabolic intermediates generated after azo bond cleavage by P. aeruginosa as confirmed by total organic carbon content and high-performance liquid chromatography analyses. The degradation products were characterized by Fourier transform infrared spectrometer and nuclear magnetic resonance techniques whereas the biotoxicity profile of the samples were evaluated using the brine shrimp lethality test assay. Data from this study provide evidence of dye mineralization and detoxification by a monoculture of P. aeruginosa in successive microaerophilic/oxic stages.     

Study Potential of Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Bioremediation of Diesel-Contaminated Water

Petroleum products which are used in a wide variety of industries as energy sources and raw materials have become a major concern in pollution of terrestrial and marine environments. The purpose of this study was to assess the potential of indigenous microbial isolates for degradation of diesel fuel. Two most proficient bacterial strains among five isolated strains from polluted soil of an industrial refinery were studied. The isolates then were identified as Pseudomonas aeruginosa and Bacillus subtilis using biochemical tests and 16S rRNA gene sequence analyses. P. aeruginosa showed higher biodegradation efficiency than B. subtilis in shaking flask containing diesel-contaminated water. P. aeruginosa and B. subtilis degraded about 87 and 75% of total hydrocarbons, respectively, in flasks containing 2% diesel and 98% water. The biodegradation efficiency of the isolates decreased as diesel contamination increased from 2 to 5%. The isolates showed significantly higher efficiency on degradation of short-chain hydrocarbons in 20 days, i.e., by using P. aeruginosa, removal efficiency of C₁₀ hydrocarbons was near 90%, while about 69% of C₂₀₊ hydrocarbons and 47% of aromatic hydrocarbons were removed. Therefore, the isolates showed high capability in biodegradation of diesel contamination of the refinery.     

Biodegradation of the Anionic Surfactant Linear Alkylbenzene Sulfonate (LAS) by Autochthonous Pseudomonas sp.

Anionic surfactants, the earliest and the most common surfactants in detergent and cosmetic product formulations contribute significantly to the pollution profile of the ecosystem. Linear alkylbenzene sulfonates (LAS), a major chemical constituent of detergents, forms an imperative group of anionic surfactants. Bioremediation of LAS by conventional processes such as activated sludge is ineffective due to the low kinetics of degradation by unsuitable organisms and foam production. Hence this study was focused on isolating and characterizing indigenous LAS-degrading bacteria from soil. Twenty different LAS-degrading bacteria were isolated from detergent-contaminated soil by enrichment culture technique and degradation efficiency was assessed by Methylene Blue Active Substances (MBAS) assay and by reverse-phase high-performance liquid chromatography (HPLC) analysis. The most efficient LAS-degrading isolates, L9 (81.33?±?0.7) and L12 (81.81?±?0.8), were selected and identified as Pseudomonas nitroreducens (MTCC 10463) and Pseudomonas aeruginosa (MTCC 10462). The 16S rDNA sequences of the isolates were deposited in NCBI GenBank under the accession numbers HQ 271083 (L9) and HQ 271084 (L12). The isolates were capable of degrading 0.05?g/l LAS at 25?°C and pH 7.0?C7.5. Presence of a solid support caused biofilm formation which in turn enhanced LAS degradation. The isolates tend to display diauxic growth with alternate carbon source such as dextrose. These isolates also have the capability to degrade other xenobiotics like hydrocarbons and pesticides. Since xenobiotic pollutants in nature occur as a mixture of compounds rather than single pollutants, the potential of these two indigenous LAS-degrading isolates to degrade multiple xenobiotics gains relevance.     

Plant Growth Promoting Traits of Indigenous Phosphate Solubilizing Pseudomonas aeruginosa Isolates from Chilli (Capsicumannuum L.) Rhizosphere

Phosphorus (P) is the second key nutrient for plants and it affects several attributes of plant growth. Identification of a potent phosphate solubilizing microorganism capable of transforming the insoluble P into soluble and plant-accessible forms is considered as the best eco-friendly option for providing inexpensive P to plants. Hence, this study was focused to assess the growth enhancement traits of the phosphate solubilizing bacteria (PSB) isolated from chili rhizosphere. Twelve PSB were isolated by enrichment culture technique and its P solubilization efficiency was checked using Vanadomolybdate phosphoric yellow color method. Among them, two potent strains PS2 and PS3, identified as Pseudomonas aeruginosa KR270346 and KR270347 based on biochemical and molecular characterization, were selected for further study. The Pseudomonas aeruginosa isolates interestingly showed the presence of various potential plant growth-promoting properties including indole acetic acid and siderophore production. The growth enhancement effect of Pseudomonas aeruginosa isolates on chilli showed promising results, and the growth parameters were found to be statistically signi?cant when compared to control. The results demonstrated an eloquent impact on various aspects, namely microbial count and PSB population, phosphatase and dehydrogenase activity, available phosphorous in the soil, plant nutrient uptake, and yield parameters. Inoculation of these two isolates together with the addition of rock phosphate increased comparable amount of available P and these treatments were statistically at par throughout the growth period. The results con?rmed the growth-promoting potential of the isolates to develop as biofertilizers either alone or as components of integrated nutrient management systems.     

Arbuscular mycorrhizae and phosphate solubilising bacteria of the rhizosphere of the mangrove ecosystem of Great Nicobar island, India

Mangroves form an important ecosystem of Great Nicobar, a continental island in the Bay of Bengal with luxuriant tropical rainforests. The rhizosphere of the mangrove plants of Great Nicobar was investigated for the presence of arbuscular mycorrhizal fungus (AMF) and phosphate solubilising bacteria (PSB). The soils of the Great Nicobar mangroves were silt–clays and were poor in phosphate content. Five species of AMF belonging to the genus Glomus were isolated. The %AMF colonization in the mangrove plants was between 0 and 17%, and the presence of AMF in the aerenchymatous cortex suggests that the mangrove plants may be aiding in AMF survival by providing oxygen. Two strains of phosphate solubilising Pseudomonas aeruginosa were found in the mangrove soils of Great Nicobar. Phosphate solubilisation by the two isolated strains was almost 70% under in vitro conditions. PSB may play a role in the mangrove ecosystems of Great Nicobar by mobilising insoluble phosphate. The plant roots could pick up the released phosphate directly or with the aid of AMF hyphae.     

Effects of biofertilizers and cycocel on some physiological and biochemical traits of wheat (Triticum aestivum L.) under salinity stress

In order to study the effects of biofertilizers and cycocel on some physiological and biochemical characteristics of wheat (Triticum aestivum L.) under salinity condition, a factorial experiment was conducted based on randomized complete block design with three replications under greenhouse condition in 2015. Treatments were included salinity in four levels [no salt (control or S₀), salinity 30 (S₁), 60 (S₂) and 90 (S₃) mM NaCl equivalent of 2.76, 5.53 and 8.3 dS m^?1, respectively], four biofertilizers levels [no biofertilizer (F₀), seed inoculation by Azotobacter chrocoocum strain 5 (F₁), Pseudomonas putida strain 186 (F₂), both inoculation Azotobacter + Pseudomonas (F₃)] and three cycocel levels [without cycocel as control (C₀), application of 600 (C₁) and 1000 (C₂) mg L^?1]. Results showed that salinity severe stress (90 mM) decreased chlorophyll content, relative water content (RWC), total chlorophyll, photochemical efficiency of PSII and yield of wheat. Whereas, soluble sugars and proline content, electrical conductivity (EC), the activity of catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO) enzymes were increased. Similar results were observed in CAT, POD and PPO activities due to inoculation by biofertilizers and cycocel. Salinity at 30 mM increased the photochemical efficiency of PSII and chlorophyll content in plants grown under biofertilizer and cycocel treatment but with increasing salinity up to 90 mM mentioned parameters were decreased. The highest proline and soluble carbohydrate at all salinity levels were observed in plants treated in the highest cycocel level and Azotobacter+ Pseudomonas application. Generally, it was concluded that biofertilizers and cycocel can be used as a proper tool for increasing wheat yield under salinity condition.     

Pseudomonas spp. isolates with high phosphate-mobilizing potential and root colonization properties from agricultural bulk soils under no-till management

Seven phosphate-mobilizing pseudomonads were isolated, identified, and characterized in terms of their biofertilizer potential and root-colonizing properties. Pseudomonas protegens (ex-fluorescens) CHA0 was used for comparative purposes. Four isolates (LF-MB1, LF-P1, LF-P2, and LF-P3) clustered with members of the “Pseudomonas fluorescens complex,” whereas the other three (LF-MB2, LF-V1, and LF-V2) clustered with members of the “Pseudomonas putida/Pseudomonas aeruginosa complex.” Assays in buffered liquid growth medium supplemented with tricalcium phosphate enabled the separation of the isolates into two groups: group A (LF-P1, LF-P2, LF-P3, and LF-V1) solubilized P from 151 up to 182?μg?mL^?1, and group B (LF-MB1, LF-MB2, and LF-V2) solubilized less than 150?μg?P?mL^?1. All isolates displayed acid and alkaline phosphatase activities. With the exception of LF-MB2, all isolates were able to degrade phospholipids from lecithin. Additionally, all isolates exhibited extracellular protease activity, and four isolates produced hydrogen cyanide, two traits that are related to biocontrol of phytopathogens. To study root colonization in non-sterile soil, isolates were doubly tagged with gfp and a tetracycline resistance cassette. After 15?days of competition with the indigenous bacterial flora, all tagged isolates colonized soybean roots at counts ranging from 7.6?×?10⁵ to 1.7?×?10⁷?CFU?g^?1. The results indicate that there are already efficient phosphate-mobilizing pseudomonads adapted to agricultural bulk soils under no-till management in Argentina and thus having excellent potential for use as biofertilizers.     

Isolation of a selected microbial consortium from a pesticide-contaminated mix-load site soil capable of degrading the herbicides atrazine and alachlor

Contaminated soil from a 100-year-old mix-load site located in Reading, PA was evaluated for its potential to provide indigenous microorganisms capable of degrading two widely utilized herbicides, atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) and alachlor (2-chloro-2′,6′-diethyl-N-[methoxymethyl]-acetanilide; AL). Three different locations from the site were chosen for experimentation based on herbicide handling activities. Standard enrichment techniques were used to isolate a selective microbial consortium (SCM) with the desirable degrading capabilities. Three enrichment treatment schemes were evaluated; AT and AL, AL alone, and only AT. Degradative organisms were isolated from only one of the sample locations. Considerable differences in the soil parameters of the three sample locations were found that might have had an effect on the ability of the indigenous microbial populations within the soil to degrade AT and AL. In the initial cultures from this location, degradation occurred in the AT and AL treatment only. Because the AT and AL were the only sources of carbon and nitrogen (N) for the microbes, these results suggest that AL alone was not a sufficient N source. In general, the ability to degrade AL by the SMC was dependent on AT degradation. Alachlor degradation did not begin until approximately 15% of the AT was transformed. Once all of the AT was removed very little further AL degradation occurred. The average half-life (t_1/2) of AT was 7.5 d, while average t_1/2 for AL degradation was 11 d. Individual colonies from the SMC were identified by fatty acids methyl ester (FAME) analysis. Five strains were identified with similarity indexes above 70%. These isolates included the following: Alcaligenes xylosoxydans subsp. denitrificans, Alcaligenes xylosoxydans subsp. xylosoxydans, Pseudomonas putida, Pseudomonas marginalis, and Providencia rustigianii.     

Anaerobic microbial degradation of the herbicide propanil

The fate of propanil (3,4-dichloropropionanilide) in an anaerobic soil environment was studied. Two mineral salts media, one amended with 0.05% yeast extract and 0.05% tryptone, and both with 33 μg propanil ml^?1, were inoculated with 10% (v/v) soil to establish enrichment cultures. Cultures were incubated at 30°C under an atmosphere of 95% N₂ and 5% CO₂. There was a complete loss of propanil in 15 days in soil enrichment cultures. One degradation product was detected and identified by HPLC and TLC co-chromatography as 3,4-dichloroaniline. Propanil was also degraded in a soil-free medium containing 0.05% yeast extract and 0.05% tryptone and inoculated with supernatant from a soil enrichment culture. After 100 days, propanil concentration decreased by 81%; 3,4-dichloroaniline and meta-chloroaniline were produced. No decrease in propanil concentration was detected in soil-free cultures without yeast extract and tryptone. No labeled CO₂ or volatile products were detected after 80 days in soil-free enrichment cultures containing ¹⁴C-ring-labeled propanil.     

Biodegradation of hexachlorocyclohexane-isomers in contaminated soils

Several sites that are contaminated with isomers of the chlorinated insecticide hexachlorocyclohexane (HCH) are present across the globe and cause toxicity. For their bioremediation, we studied the degradation of HCH-isomers in contaminated soils by an isolate Pseudomonas aeruginosa ITRC-5. The degradation is optimal at 2 mg technical-HCH (t-HCH)/g soil, 15% water content, pH 8.0, temperature 28 °C and inoculum density 10⁶ colony forming unit/g soil. Under these conditions, from 5 kg soil, >98% α- and γ-HCH, 17% β-HCH and 76% δ-HCH are degraded after 15 days of incubation, which is accompanied with the release of 600 μg chloride/mg t-HCH. Concomitant to the degradation, a four-fold reduction in the toxicity of HCH-isomers to earthworm, Eisenia foetida, is also observed. Addition of ITRC-5 enhanced the degradation of soil-applied HCH-isomers in ‘open field’ conditions as well, and 97%, 43%, 94% and 77% of α-, β-, γ- and δ-HCH, respectively, are degraded after 12 weeks of incubation. Thus, the bacterium causes microbial degradation and detoxification of HCH-isomers, and can be used for the bioremediation of contaminated soils.     

Cinnamaldehyde Induces PCD-Like Death of Microcystis aeruginosa via Reactive Oxygen Species

In recent years, Microcystis bloom occurs frequently and causes a wide range of social, environmental, and economic problems. In this study, dose-dependent inhibitory effect of cinnamaldehyde on the growth of Microcystis aeruginosa was investigated. It was found that cinnamaldehyde with the concentration more than 0.6 mM showed algicide activity against M. aeruginosa. When M. aeruginosa was exposed to 0.6 mM cinnamaldehyde, considerable reactive oxygen species (ROS) were generated followed by lipid peroxidation and decrease in the content of both chlorophyll a and soluble protein. Although superoxide dismutase had made response to the stress caused by cinnamaldehyde, activity increasing after a time of lag could not prevent the lysis of M. aeruginosa cells. Interestingly, the addition of antioxidants glutathione and l-ascorbic acid (Vc) could prevent the lysis of M. aeruginosa cells. All the results suggested that cinnamaldehyde induced the death of M. aeruginosa cells via inducing ROS burst. Further understanding of the mechanism of cinnamaldehyde-induced M. aeruginosa cell death would contribute to the control of cyanobacteria pollution.     

EFFECTS OF SALINITY AND CALCIUM ON THE GROWTH AND CHEMICAL COMPOSITION OF PISTACHIO SEEDLINGS

Poor quality of irrigation water (high salinity) has reduced the yields of pistachio over recent years, especially in Kerman. The effects of four salinity levels [0, 30, 60, and 90 mM sodium chloride (NaCl)] and three calcium (Ca) levels [0, 0.5, and 1 mM Ca as calcium nitrate (Ca(NO₃)₂.4H₂O)] on growth and chemical composition of pistachio seedlings cv. ‘Badami’ were studied in sand culture under greenhouse conditions in completely randomized design (CRD) with four replications. After 170 days, leaf area, leaf number, shoot and root dry weights were determined. Also shoot and root sodium (Na), potassium (K), Ca, and magnesium (Mg) concentrations were measured. Results showed salinity decreased all growth parameters. Ca application increased shoot and root Ca concentrations and root K concentration, while Ca application decreased shoot K concentration and shoot and root Mg concentrations. Salinity decreased shoot Ca, root K, and root Mg concentrations, while salinity increased shoot and root total sodium uptake, and shoot and root Cl concentrations.     

Natural occurrence of Pseudomonas aeruginosa,a dominant cultivable diazotrophic endophytic bacterium colonizing Pennisetum glaucum (L.) R. Br.

Endophytic microbial communities can have strong influence on the growth of their host plants. The present study reports a diazotrophic endophytic bacterial species that colonizes predominantly Pennisetum glaucum (pearl millet) and remains stabilized throughout the latter's various growth stages under field conditions. Qualitative and quantitative changes in the endophytic bacterial population structure were examined during the plant growth period at regular intervals of 7 days up to harvesting. DNA fingerprinting (ERIC-PCR) was used as a biomarker to track the identity of various isolates obtained at different time intervals. Identification of representative bacterial species corresponding to different ERIC types was made on the basis of 16S rRNA gene sequence analysis. Based on the colony forming unit (cfu) count of bacterial isolates observed at various growth stages, Pseudomonas aeruginosa PM389 was found to be the dominant diazotrophic species among the cultivable endophytes colonizing pearl millet. The population of P. aeruginosa was detected in the host 21 days after sowing (DAS), indicating its entry in plant roots from soil and suggesting its non-vertical transfer in pearl millet. Moreover, an upward migration of this dominant diazotrophic bacterial species to shoots was observed with the plant growth. Further evaluation of P. aeruginosa PM389 revealed its various plant growth promoting properties viz. nitrogen fixation, mineral phosphate solubilization, siderophore production and antagonistic properties.     

海洋细菌生物膜用于生物修复陆地生态系统中土壤多环芳烃污染

Polycyclic aromatic hydrocarbons(PAHs) in soil retain for a quite long period due to their hydrophobicity and aggregation properties. Biofilm-forming marine bacterial consortium(named as NCPR), composed of Stenotrophomonas acidaminiphila NCW702,Alcaligenes faecalis NCW402, Pseudomonas mendocina NR802, Pseudomonas aeruginosa N6P6, and Pseudomonas pseudoalcaligenes NP103, was used for the bioremediation of PAHs in a soil microcosm. Phenanthrene and pyrene were used as reference PAHs. Parameters that can affect PAH degradation, such as chemotaxis, solubility of PAHs in extracellular polymeric substances(EPS), and catechol2,3-dioxygenase(C23O) activity, were evaluated. P. aeruginosa N6P6 and P. pseudoalcaligenes NP103 showed chemotactic movement towards both the reference PAHs. The solubility of both the PAHs was increased with an increase in EPS concentration(extracted from all the 5 selected isolates). Significantly(P 0.001) high phenanthrene(70.29%) and pyrene(55.54%) degradation was observed in the bioaugmented soil microcosm. The C23O enzyme activity was significantly(P 0.05) higher in the bioaugmented soil microcosm with phenanthrene added at 173.26 ± 2.06 nmol min~(-1) mg~(-1) protein than with pyrene added at 61.80 ± 2.20 nmol min~(-1) mg~(-1) protein. The C23O activity and gas chromatography-mass spectrometer analyses indicated catechol pathway of phenanthrene metabolism. However, the metabolites obtained from the soil microcosm added with pyrene revealed both the catechol and phthalate pathways for pyrene degradation.     

Phytoremediation of Pb in the sediment of a mangrove ecosystem

Purpose

Coal-fuelled power plants can discharge hazardous materials, particularly heavy metals such as lead (Pb). An alternative way of reducing Pb concentration from contaminated sediments is through phytoremediation. Presently, there are few research findings on the phytoremediation potential of mangroves on metals like Pb. The study was conducted to survey and identify mangroves that thrive near the coal-fired power plant and to assess the phytoremediation potential of mangroves on Pb in sediment.

Materials and methods

The study sites were located in the mangrove ecosystems of Sitio Oyon and Sitio Asinan in Masinloc, Zambales, Philippines. The first stage of our study was to survey and identify the mangrove species. The second stage was to assess the levels of Pb in the sediments, water, and tissues of mangrove trees. The diversity assessment of the mangrove species was done through the use of 10?×?12 m quadrat technique. Water and sediment samples from each mangrove ecosystem were collected using composite sampling methods.

Results and discussion

Three mangrove species were identified in the study sites: Avicennia marina, Rhizophora stylosa, and Sonneratia alba. The order of importance of the mangrove trees in the two sampling locations, based on an importance value index (IVI), were as follows: SA (IVI?=?171.20)?>?AM (77.79)?>?RS (51.01). The total uptake of Pb from sediments near the power plants varied significantly (p?≤?0.001) among the three mangrove species. S. alba had the highest Pb uptake of 48.4 kg ha^?1 followed by A. marina (23.1 kg ha^?1), and R. stylosa (2.4 kg ha^?1). These three mangrove species have the potential to phytoremediate Pb in the sediment.

Conclusions

The three mangrove species present in the coastal ecosystem near the electric power plant—A. marina, R. stylosa, and S. alba—were potential phytoremediators of sediment Pb. The present study indicated that the mangroves possess beneficial characteristics that remove Pb from contaminated sediments in areas directly affected by coal-fired power plants, and thus have potential phytoremediation properties.     

Promotion of plant growth and in situ degradation of phenol by an engineered Pseudomonas fluorescens strain in different contaminated environments

We show that Pseudomonas fluorescens strain P13, a plant growth-promoting bacterium, enhanced the growth of corn in uncontaminated soil but not in contaminated soil, perhaps because of its inability to reduce phytotoxicity. Another bacterial strain, Pseudomonas aeruginosa strain SZH16, showed in situ phenol-degrading activity and contained a plasmid loaded with a gene encoding for catechol 2, 3-dioxygenase, an important enzyme in the degradation pathway of aromatic compounds. We implanted this biodegradation ability into strain P13, using horizontal gene transfer techniques using strain SZH16 as the donor and P13 as the recipient, to generate a phenol-degrading transconjugant which obtained the effective plasmid from strain SZH16. Introduction of the transconjugant P13 strain into an artificially phenol-spiked soil promoted the growth of corn and in situ phenol degradation, and the increase in plant biomass correlated with the decrease in soil phenol content. Furthermore, the transconjugant P13 strain was also found to stimulate corn growth and reduce phenol concentration in water containing phenol and in historically contaminated field soils, indicating that the transconjugant strain could promote plant growth in both contaminated and uncontaminated environments. The transconjugant P13 strain was more efficient than either strain P13 or SZH16, and shows how plant growth-promoting bacteria which show no, or only limited, ability to degrade organic pollutants may be modified. This technique is attractive for many environmental remediation and agronomic applications.     

Europium chelate and fluorescent brightener staining of soil propagules and their photomicrographic counting—II. Efficiency

The efficiency of staining Lipomyces starkeyi and Pseudomonas aeruginosa cells added to agar and soil agar films and soil smears was examined using 2 mM europium(111) thenoyltrifluoroacetonate together with 25 μM disodium salt of 4,4'-bis (4-anilino-6-bis(2-hydroxyethyl) amino-S-triazin-2-ylamino) 2,2'-stilbene disulphonic acid. abbreviated to Eu(TTA)₃ and FB. respectively. The combined stains, or DFS (differential fluorescent stain), gave recoveries of added cells close to the expected values, but prestained cells lost their Eu(TTA)₃ fluorescence when added to agar at 55–60°C. Aqueous extracts of agar powder quenched the Eu(TTA)₃ fluorescence of cells, an effect which was attributed to water-soluble organic or inorganic ions in agar. It is suggested that the method of preparing soil-agar films and the long staining period required, effectively overcomes the quenching effect. Pre-stained cells were not affected by 10^?2 dilutions in soil.Photomicrographically recorded counts were not significantly different from visual counts made on the same fields.Comparative counts with phenol-aniline blue. fluorescein iso-thiocyanate and DFS showed the latter stain to be more efficient in differentiating soil propagules on and in soil organic matter.     

Effects of Wood Amendments on the Degradation of Terbuthylazine and on Soil Microbial Community Activity in a Clay Loam Soil

The herbicide terbuthylazine is widely used within the EU; however, its frequent detection in surface and groundwater, together with its intrinsic toxicological properties, may pose a risk both for human and environmental health. Organic amendments have recently been proposed as a possible herbicide sorbent in soil, in order to limit herbicide movement from soil to water. The environmental fate of terbuthylazine depends not only in its mobility but also in its persistence. The latter is directly dependent on microbial degradation. For this reason, the effects of pine and oak residues on terbuthylazine soil microbial community functioning and on the potential of this community for terbuthylazine degradation were studied. For this purpose, degradation kinetics, soil dehydrogenase activity and the number of live bacteria were assessed in a clay loam soil treated with terbuthylazine and either amended with pine or oak wood or unamended (sterilised and non-sterilised). At day 65, 85?% of the herbicide applied still persisted in the sterile soil, 73?% in the pine-amended one and 63?% in the oak-amended and unamended ones. Pine residues increased the sorption of terbuthylazine to soil and hampered microbial degradation owing to its high terbuthylazine sorption capacity and a decrease in the bioavailability of the herbicide. On the contrary, in the presence of oak residues, the herbicide sorption did not increase significantly. The overall results confirm the active role of the soil microbial community in terbuthylazine degradation in amended and unamended soils and in a liquid enrichment culture performed using an aliquot of the same soil as the inoculum. In this clay loam soil, in the absence of amendments, the herbicide was found to be quite persistent (t _1/2?>?95?days), while in the enrichment culture, the same natural soil bacterial community was able to halve terbuthylazine in 24?days. The high terbuthylazine persistence in this soil was presumably ascribable to its texture and in particular to the mineralogy of the clay fraction.     

Growth stimulation and fruit yield improvement of greenhouse tomato plants by inoculation with Pseudomonas putida or Trichoderma atroviride: Possible role of indole acetic acid (IAA)

Five bacteria (Pseudomonas fluorescens, P. fluorescens subgroup G strain 2, P. marginalis, P. putida subgroup B strain 1 and P. syringae strain 1) and three fungi (Penicillium brevicompactum, P. solitum strain 1 and Trichoderma atroviride) were evaluated to determine their promoting effect on the growth of mature healthy tomato plants grown under hydroponic conditions. P. putida and T. atroviride were shown to improve fruit yields in rockwool and in organic medium. The production or degradation of indole acetic acid (IAA) by the two microorganisms was investigated as possible mechanisms for plant growth stimulation. Both P. putida and T. atroviride were shown to produce IAA. The production of IAA by the two microorganisms was stimulated in vitro by the addition of l-tryptophan, tryptamine and tryptophol (200 μg ml⁻¹) in the culture medium. P. putida and T. atroviride also increased the fresh weight of both the shoot and the roots of tomato seedlings grown in the presence of increasing concentrations of l-tryptophan (up to 0.75 mM). Both microorganisms showed partial degradation of IAA in vitro when grown in a minimal medium with or without sucrose. In addition, the capacity of these microorganisms to reduce the deleterious effect of exogenous IAA was investigated using tomato seedlings. The results showed that the roots of tomato seedlings grown in the presence of increasing concentrations of IAA (0-10 μg ml⁻¹) were significantly longer when seeds were previously treated with P. putida or T. atroviride. The reduction in the detrimental effect of IAA on root elongation could be associated with a reduced ethylene production resulting from a decrease of its precursor 1-aminocyclopropane-1-carboxylic acid (ACC) by microbial degradation of IAA in the rhizosphere and/or by ACC deaminase activity present in both microorganisms.