Combined effect of temperature and zinc on Caenorhabditis elegans wild type and daf-21 mutant strains

Heavy metal pollution in aquatic ecosystems is a far reaching environmental problem. The possible influences of heavy metal exposure and the potential harm to organisms when combined with other environmental stressors such as temperature have been largely unexplored. An aquatic toxicity test of Caenorhabditis elegans was performed to estimate the 24 h median lethal concentration (LC₅₀) of different zinc concentrations at different temperatures (15 °C, 20 °C, 25 °C, and 30 °C). We also examined the time course thermotolerance on wild type (N2) and daf-21 null (JT6130) adults exposed to 6.1 mM zinc at 37 °C. Hsp90 protein expression level in response to the combined effect of temperature and zinc toxicity was also investigated by both Western blots and ELISA. Our results show that C. elegans wild type nematodes exhibit severe lethal toxicity after a 24 h exposure to zinc at higher temperatures. In addition, the expression level of Hsp90 was highly inhibited in adult worms subjected to zinc stress. This toxicity assay at different temperatures provides insight into organism response to combined effects of temperature and zinc toxicity.     

Preparation,characterization of silver phyto nanoparticles and their impact on growth potential of Lupinus termis L. seedlings

The current study reports rapid and easy method for synthesis of eco-friendly silver nanoparticles (AgNPs) using Coriandrum sativum leaves extract as a reducing and covering agent. The bio-reductive synthesis of AgNPs was monitored using a scanning double beam UV-vis spectrophotometer. Transmission electron microscopy (TEM) was used to characterize the morphology of AgNPs obtained from plant extracts. X-ray diffraction (XRD) patterns of AgNPs indicate that the structure of AgNPs is the face centered cubic structure of metallic silver. The surface morphology and topography of the AgNPs were examined by scanning electron microscopy and the energy dispersive spectrum revealed the presence of elemental silver in the sample. The silver phyto nanoparticles were collected from plant extract and tested growth potential and metabolic pattern in (Lupinus termis L.) seedlings upon exposure to different concentrations of AgNPs. The seedlings were exposed to various concentrations of (0, 0.1, 0.3 and 0.5 mg L^?1) AgNPs for ten days. Significant reduction in shoot and root elongation, shoot and root fresh weights, total chlorophyll and total protein contents were observed under the higher concentrations of AgNPs. Exposure to 0.5 mg L^?1 of AgNPs decreased sugar contents and caused significant foliar proline accumulation which considered as an indicator of the stressful effect of AgNPs on seedlings. AgNPs exposure resulted in a dose dependent decrease in different growth parameters and also caused metabolic disorders as evidenced by decreased carbohydrates and protein contents. Further studies needed to find out the efficacy, longevity and toxicity of AgNPs toward photosynthetic system and antioxidant parameters to improve the current investigation.     

Extracellular biosynthesis of silver nanoparticles using Rhizopus stolonifer

Synthesis of silver nanoparticles (AgNPs) has become a necessary field of applied science. Biological method for synthesis of AgNPs by Rhizopus stolonifer aqueous mycelial extract was used. The AgNPs were identified by UV–visible spectrometry, X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared spectrometry (FT-IR). The presence of surface plasmon band around 420 nm indicates AgNPs formation. The characteristic of the AgNPs within the face-centered cubic (fcc) structure are indicated by the peaks of the X-ray diffraction (XRD) pattern corresponding to (1 1 1), (2 0 0) and (2 2 0) planes. Spherical, mono-dispersed and stable AgNPs with diameter around 9.47 nm were prepared and affirmed by high-resolution transmission electron microscopy (HR-TEM). Fourier Transform Infrared (FTIR) shows peaks at 1426 and 1684 cm⁻¹ that affirm the presence of coat covering protein the AgNPs which is known as capping proteins. Parameter optimization showed the smallest size of AgNPs (2.86 ± 0.3 nm) was obtained with 10⁻² M AgNO₃ at 40 °C. The present study provides the proof that the molecules within aqueous mycelial extract of R. stolonifer facilitate synthesis of AgNPs and highlight on value-added from R. stolonifer for cost effectiveness. Also, eco-friendly medical and nanotechnology-based industries could also be provided. Size of prepared AgNPs could be controlled by temperature and AgNO₃ concentration. Further studies are required to study effect of more parameters on size and morphology of AgNPs as this will help in the control of large scale production of biogenic AgNPs.     

Carbon mineralization of Chinese fir (Cunninghamia lanceolata) soils under different temperature and humidity conditions

Burned and unburned mineral soils (0–10 cm) from a 40-year-old Chinese fir (Cunninghamia lanceolata) forest in Nanping, Fujian, China were incubated for 90 days at different temperatures (25 °C and 35 °C) and humidity [25%, 50%, and 75% of water holding capacity (WHC)] conditions. Carbon (C) mineralization of all soils was determined using CO₂ respiration method. The results showed that CO₂ evolution rates of the burned and control soils exhibited similar temporal patterns, and similar responses to temperature and moisture. CO₂ evolution rates for all soil samples decreased with incubation time. At different humidity conditions, average rate of C mineralization and cumulative mineralized C from burned and control soils were significantly higher at 35 °C than at 25 °C. This implied that C mineralization was less sensitive to soil moisture than to temperature. In both soils at 25 °C or 35 °C, the amount of soil evolved CO₂ over the 90 days incubation increased with increasing moisture content from 25% to 75% WHC. A temperature coefficient (Q₁₀) varied with soil moisture contents. The maximum values recorded for Q₁₀ were 1.7 in control soil and 1.6 in burned soil both at 25% WHC. However, there were no significant differences in Q₁₀ values between the control and burned soils over all moisture ranges (P > 0.05). The data of cumulative C–CO₂ released from control and burned soils were fitted to two different kinetic models. The two simultaneous reactions model described mineralization better than the first-order exponential model, which reflected the heterogeneity of substrate quality. Based on these results, it is possible to conclude that temperature and moisture are important in the controls of C mineralization, and the combined effects of these variables need to be considered to understand and predict the response of CO₂ release in subtropical ecosystems to climate change.     

Effect of tertiary sewage effluent additions on Prymnesium parvum cell toxicity and stable isotope ratios

We investigated the ability of the ichthyotoxic haptophyte Prymnesium parvum to use sewage-originated nutrients applying stable carbon (C) and nitrogen (N) isotope techniques. P. parvum was cultured under N and phosphorus (P) sufficient and deficient conditions in either sewage effluent-based medium or in a nitrate- and phosphate-based control. Cell densities and toxicities were monitored and stable carbon N isotopes signatures (δ¹³C and δ¹⁵N) of P. parvum and the sewage effluent analysed. Nitrogen and P sufficient cultures achieved the highest biomass followed by P and N deficient cultures, regardless of sewage effluent additions. The P deficient cultures with sewage effluent had higher toxicity, estimated as haemolytic activity (9.4 ± 0 × 10^?5 mg Saponin equiv. cell^?1) compared to the P deficient control and to all N deficient and NP sufficient cultures. Nutrient deficient conditions had no effect on the cell δ¹⁵N, but a decreasing effect on δ¹³C in the inorganic N deficient treatment. Growth in sewage-based media was followed by a substantial increase in the cell δ¹⁵N (10.4–16.1‰) compared to the control treatments (2.4–4.9‰), showing that P. parvum is capable of direct use of sewage-originated N, inorganic as well as organic. Uptake of terrestrial derived C in the sewage treatments was confirmed by a decrease in cell δ¹³C, implying that P. parvum is able to utilize organic nutrients in sewage effluent.     

Earthworms as bioindicator of metals (Zn,Fe, Mn,Cu, Pb and Cd) in soils: Is metal bioaccumulation affected by their ecological category?

The importance of earthworms in metal pollution monitoring is widely recognized in terrestrial ecosystems. Metal bioaccumulation by soil-dwelling earthworms can be used as an ecological indicator of metal availability in soils. In this study, we quantify the level of DTPA extractable metals in casts and tissues of earthworms (endogeic: Metaphire posthuma (Vaillant) and anecic: Lampito mauritii Kinberg) and ingesting soils, collected form cultivated land, urban garden and sewage soils. Soil and worm casts collected from sewage and cultivated land showed the greater metal concentrations. The concentration of Zn, Fe, Pb and Mn in earthworm casts was in the order: sewage soil > cultivated land > urban garden, while for Cu and Cd the order was cultivated land > sewage soil > urban garden. Data suggested that the level of DTPA extractable metals was higher than that of surrounding soils. We got close relationships between metal concentration in worm tissues and surrounding soils: Zn (r² = 0.94 and 0.89, P < 0.01 for both), Fe (r² = 0.95 and 0.97, P < 0.01 for both), Cu (r² = 0.93 and 0.96, P < 0.01), Pb (0.63, P < 0.01 and 0.57, P > 0.05), and Cd (r² = 0.15, P > 0.01 and 0.75, P < 0.01), respectively, for M. posthuma and L. mauritii. The study clearly indicates that earthworms have efficient potentials for bioaccumulation of metals in their tissues which can be used as an ecological indicator of soil contaminations. A species-specific metal accumulation pattern was observed in studied earthworms. Comparatively, endogeic showed the higher metal contents in their tissues than anecic (t-test: P < 0.05); collected form different habitats studied. Data suggested that species-specific feeding behaviour, earthworm niche structure, ecological category of inhabiting earthworm and even horizontal distribution of contaminants in soil layers are some major determinant for metal accumulation patterns in soil dwelling earthworms. The difference in burrowing patterns can influence the patterns of metal bioaccumulations between endogeic and anecic, although other factors are also contributory. Further more detailed study is still required to elaborate the proposed hypothesis.     

Silver nanopaticles synthesized using the seed extract of Trigonella foenum-graecum L. and their antimicrobial mechanism and anticancer properties

Background

Synthesis of silver nanoparticles (AgNPs) through biological route plays an important role in their applications in the medical field, especially in the prevention of disease causing microbial pathogens and arresting the propagation of cancer cells. The stable, green synthesis of AgNPs is very much welcomed in the medical field because of their low toxicity. Therefore, the demands of AgNPs synthesised biologically is on the rise. The present study aimed to investigate the antimicrobial mechanisms and anticancer properties of the AgNPs synthesized using the seed extract of Trigonella foenum-graecum L. The AgNPs were characterized by UV–vis, SEM, XRD, FTIR and EDAX analysis. The minimum inhibitory concentrations (MIC) of the AgNPs were determined by the broth micro dilution method.

Antifungal potentiality of mycogenic silver nanoparticles capped with chitosan produced by endophytic Amesia atrobrunnea

This research reports the fabrication of silver nanoparticles (AgNPs) from endophytic fungus, Amesia atrobrunnea isolated from Ziziphus spina-christi (L.). Influencing factors for instance, thermal degree of incubation, media, pH, and silver nitrate (AgNO₃) molarity were optimized. Then, the AgNPs were encapsulated with chitosan (Ch-AgNPs) under microwave heating at 650 W for 90 s. Characterization of nanoparticles was performed via UV–visible (UV–vis) spectrophotometer, Fourier-transform infrared spectrophotometer (FTIR), zeta potential using dynamic-light scattering (DLS), and field-emission-scanning electron microscope (FE-SEM). Anti-fungal activity of Ch-AgNPs at (50, 25, 12.5, 6.25 mg/L) was tested against Fusarium oxysporum, Curvularia lunata, and Aspergillus niger using the mycelial growth inhibition method (MGI). Results indicated that Czapek-dox broth (CDB) with 1 mM AgNO₃, an acidic pH, and a temperature of 25–30 °C were the optimum for AgNPs synthesis. (UV–vis) showed the highest peak at 435 nm, whereas Ch-AgNPs showed one peak for AgNPs at 405 nm and another peak for chitosan at 230 nm. FTIR analysis confirmed that the capping agent chitosan was successfully incorporated and interacted with the AgNPs through amide functionalities. Z-potential was −19.7 mV for AgNPs and 38.9 mV for Ch-AgNPs, which confirmed the significant stability enhancement after capping. FES-SEM showed spherical AgNPs and a reduction in the nanoparticle size to 44.65 nm after capping with chitosan. The highest mycelial growth reduction using fabricated Ch-AgNPs was 93% for C. lunata followed by 77% for A. niger and 66% F. oxysporum at (50 mg/L). Biosynthesis of AgNPs using A. atrobrunnea cell-free extract was successful. Capping with chitosan exhibited antifungal activity against fungal pathogens.     

Soundness of in situ lipid biomarker analysis: Early effect of heavy metals on leaf fatty acid composition of Lactuca serriola

In previous studies we proposed the plant leaf fatty acid composition, and more precisely the ratio C18:3/(C18:0 + C18:1 + C18:2) as a tool to diagnose soil contamination ex situ. These studies led us to normalize this parameter as a biomarker of soil quality using Lactuca sativa growing under controlled conditions as the plant model species. The evaluation of the toxic effects of pollutants is more difficult to grasp in a real and more complex environment. Hence we developed a study to test the hypothesis that plant leaf fatty acid composition could be used also as a reliable tool to diagnose soil contamination in situ. We tested whether Lactuca serriola (prickly lettuce) could be used as a plant model species for such studies, primarily because it is widely distributed in various European countries and recorded at different habitats, and regarded as an ancestor of the cultivated lettuce L. sativa that was used for ex situ experiments. The results show that the lipid biomarker values were significantly lower in leaves of L. serriola grown on a metallurgic landfill soil than in leaves of plants harvested on surrounding countryside areas. To determine whether the value of the lipid biomarker in L. serriola leaves could be correlated to the soil metal content, we also measured the metal content of the soil corresponding to the rhyzospheric areas from where the prickly lettuce leaves were harvested. The results of the Principal Component Analysis (PCA) show that the lipid biomarker is negatively correlated with both Ni and Cr content. Moreover, these results show that the lipid biomarker values measured in situ were repeatable from year to year, confirm that this biomarker measured in situ would provide an early indication of a plant's exposure to metal, and could be therefore used to facilitate or strengthen the diagnosis of soil contamination.     

Instant green synthesis of silver nanoparticles using Terminalia chebula fruit extract and evaluation of their catalytic activity on reduction of methylene blue

A novel green approach for the synthesis and stabilization of silver nanoparticles (AgNPs) using water extract of Terminalia chebula (T. chebula) fruit under ambient conditions is reported in this article. The instant formation of AgNPs was analyzed by visual observation and UV–visible spectrophotometer. Further the effect of pH on the formation of AgNPs was also studied. The synthesized AgNPs were characterized by FT-IR, XRD, HR-TEM with EDS and DLS with zeta potential. Appearance of brownish yellow color confirmed the formation of AgNPs. In the neutral pH, the stability of AgNPs was found to be high. The stability of AgNPs is due to the high negative values of zeta potential and capping of phytoconstituents present in the T. chebula fruit extract which is evident from zeta potential and FT-IR studies. The XRD and EDS pattern of synthesized AgNPs showed their crystalline structure, with face centered cubic geometry oriented in (1 1 1) plane. HR-TEM and DLS studies revealed that the diameter of stable AgNPs was approximately 25 nm. Moreover the catalytic activity of synthesized AgNPs in the reduction of methylene blue was studied by UV–visible spectrophotometer. The synthesized AgNPs are observed to have a good catalytic activity on the reduction of methylene blue by T. chebula which is confirmed by the decrease in absorbance maximum values of methylene blue with respect to time using UV–visible spectrophotometer and is attributed to the electron relay effect.     

Green biosynthesis of silver nanoparticles from Annona squamosa leaf extract and its in vitro cytotoxic effect on MCF-7 cells

The biological method for the synthesis of silver nanoparticles (AgNPs) using Annona squamosa leaf extract and its cytotoxicity against MCF-7 cells are reported. The synthesized AgNPs using A. squamosa leaf extract was determined by UV–visible spectroscopy and it was further characterized by FT-IR, X-ray diffraction (XRD), Transmission electron microscopy (TEM), Zeta potential and energy dispersive spectrometric (EDS) analysis. The UV–visible spectrum showed an absorption peak at 444 nm which reflects surface plasmon resonance (SPR) of AgNPs. TEM photography showed biosynthesized AgNPs were predominantly spherical in shape with an average size ranging from 20 to 100 nm. The Zeta potential value of ?37 mV revealed the stability of biosynthesized AgNPs. Furthermore, the green synthesized AgNPs exhibited a dose-dependent cytotoxicity against human breast cancer cell (MCF-7) and normal breast epithelial cells (HBL-100) and the inhibitory concentration (IC₅₀) were found to be 50 μg/mL, 30 μg/mL, and 80 μg/mL, 60 μg/ml for AgNPs against MCF-7 and normal HBL-100 cells at 24 h and 48 h incubation respectively. An induction of apoptosis was evidenced by (AO/EtBr) and DAPI staining. Application of such eco-friendly nanoparticles makes this method potentially exciting for the large scale synthesis of nanoparticles.     

Short-term response of soil enzyme activities in a chlorpyrifos-treated mesocosm: Use of enzyme-based indexes

Soil enzyme activities have been long used as indicators of soil contamination, and their integration into numerical indexes of microbial functional diversity is a practical approach in the environmental risk assessment of soil pollutants. However, suitable numerical indexes need to be developed and standardized for monitoring deterioration of soil quality by agrochemicals. Herein, a mesocosm study was performed to examine short-term responses of selected soil enzyme activities to chlorpyrifos (Lorsban^® 4E). Hydrolases (carboxylesterase, acid phosphatase, β-glucosidase, urease and protease) and oxidoreductases (dehydrogenase and catalase) were measured in Andisols 14 d after an application with two doses (4.8 and 24 kg a.i. ha⁻¹) of chlorpyrifos. Both application rates caused a strong inhibition of carboxylesterase (62–78% of controls), acid phosphatase (56–60%) and β-glucosidase (43–58%) activities. Soil microbial activity was also reduced in pesticide-sprayed soils as indicated by the decreased dehydrogenase (47%) and catalase (38%) activities compared with control soils. However, only carboxylesterase activity showed a dose-dependent response with the chlorpyrifos application rate. An in vitro trial was further performed to provide evidence of a direct interaction between the enzyme (carboxylesterase, acid phosphatase and β-glucosidase) and the pesticide (chlorpyrifos and its main metabolites chlorpyrifos-oxon and 3,5,6-trichloro-2-pyridinol). Results of these in vitro assays showed that the activity of carboxylesterase was directly affected by chlorpyrifos-oxon and, at less extend, by chlorpyrifos, whereas variations of both acid phosphatase and β-glucosidase activities were likely dependent on changes in microbial activity. Urease and protease activities did not change in pesticide-treated soils compared with pesticide-free soils. Despite the absence of response in these two N-cycling enzyme activities, four enzymatic indexes (geometric mean, weighted mean, “treated-soil quality index” [T-SQI] and “integrated biological response” [IBRv2] index) were significantly lower in the chlorpyrifos-sprayed soils compared with controls. Moreover, there was a significant (r² = 0.87, P < 0.0001) correlation between T-SQI and IBRv2 scores, which suggested that the IBRv2 index (an index used for assessing animal’s health inhabiting contaminated sites) may be a complementary index in soil quality assessment.     

Diversity,complexity and transmission of double-stranded RNA elements in Chalara elegans (synanam. Thielaviopsis basicola)

Double-stranded (ds) RNA banding patterns were determined in 21 wild-type strains of the soilborne plant pathogen Chalara elegans originating from different geographic regions worldwide. Five strains, each with a unique dsRNA pattern, were selected for cDNA cloning, northern blot analysis and dsRNA transmission experiments. Four strains contained multiple (up to 6) dsRNA elements (2.0 kbp to 12 kbp in size) and one strain contained a single 2.8 kbp fragment. These five strains were distinguished from one another by their unique RAPD-PCR patterns. Seven partial cDNA clones were derived from the predominant 2.8, 5.3, and 12 kbp dsRNA elements. Nucleotide sequence analysis and northern blot hybridizations revealed a high degree of genetic dissimilarity among the different molecular-size dsRNA elements, even those found within a single strain. Four clones from the 5.3 kbp dsRNA fragment showed a 23-43 % amino acid identity to either the coat protein or RNA-dependent RNA polymerase regions of viruses in the Totiviridae. One clone from the 2.8 kbp dsRNA fragment had a 55-57 % amino acid identity to the RdRp region of viruses in the Narnaviridae. Two clones from the 12 kbp dsRNA fragment showed no significant homology to any known virus group. Colonies derived from 100 single-conidia isolates of C. elegans strains with the 2.8, 5.3 and 12 kbp elements all contained the corresponding dsRNA element, indicating that dsRNA transmission through conidia was highly efficient, regardless of molecular size. However, transmission of dsRNA between the mycelium of strains of C. elegans could not be achieved in this study. Genetically unique strains carrying diverse dsRNA elements appear to have evolved within populations of C. elegans. Based on our findings, there are at least 3 groups of viruses present in C. elegans.     

Assessment of soil-quality index based on microarthropods in corn cultivation in Northern Italy

The aim of this paper is to assess of the stability and repeatability of the biological soil-quality (BSQ) index over time and at short/long distances in agricultural soils of the Po Valley, Northern Italy, to support its correct use in soil-quality monitoring and is based on soil microarthropods representative of the structure, function and composition of ecological systems, but avoiding classification at the species level. Microarthropods are separated from the soil and weighted using the biological form approach to evaluate their level of adaptation to the soil environment. The field test was organized to assess time and scale effects in a landscape within the coordinates 45°00′ to 45°16′N and 9°50′ to 10°15′E for an estimated area of 200 km². The BSQ had, in different soils cropped with corn and fertilized with sewage sludge, an average value of 76.4 ± 23.5; it appears to be reproducible both over short distances as well as over short periods of time, but no significant differences (p > 0.05) between years nor between farms were observed. In the landscape conditions tested, it appears reproducible and stable at a field and basin scale, though very sensitive to seasonal climatic variations. From a practical point of view, the information reported in this paper contributes to the standardization of the application of BSQ and provides reference values for this index in soils of the Northern Italy cropped with corn.     

Stress responses and specific metal exclusion on mine soils based on germination and growth studies by Australian golden wattle

We reported the Australian golden wattle as a copper stabilizer in abandoned copper mine soils earlier. Here we investigate to confirm this plant’s suitability to grow on metal contaminated mine soils based on stress indication. The seeds of Acacia pycnantha collected from mining area were germinated after heat and no heat treatment on two types of irrigation. The daily irrigated and heat treated seeds gave up to 85% germination on sandy soil. The A. pycnantha was grown under greenhouse condition in six different soils collected from abandoned copper mine at Kapunda in South Australia. Among the six soil samples, soil-1 with the highest copper concentration produced 2.05 mmol g⁻¹ tissue of proline. Proline expression was prominent in more saline soils (1, 5 and 6) having electrical conductivity (EC) 1184, 1364 and 1256 μS, respectively. Chlorophyll a, b and carotenoid levels in plants showed a gradually decreasing trend in all the soils as experiment progressed. The plants grown on soil sample-1, containing 4083 ± 103 mg kg⁻¹ of copper resulted in 18 ± 2 mg kg⁻¹ accumulation in its leaf. The calcium accumulation was significant up to 11648 ± 1209 mg kg⁻¹ in leaf. Although pore water samples showed higher Cu concentration in soils, an increased mobility of arsenic and lead was observed in all the soil samples. Our experiment points out the need for proper monitoring of revegetation processes to avoid revegetation and reclamation failure.     

The effects of applying sewage sludge into Jiangxi red soil on the growth of vegetables and the migration and enrichment of Cu and Zn

Jiangxi red soil was used as the tested soil and water spinach (Ipomoea aquatic) and Chinese chive (Allium tuberosum) were used as the tested vegetables in this study to investigate the effects of different amounts of sewage-sludge application on the growth of vegetables and the migration and enrichment patterns of Cu and Zn in vegetables using the potted method. The results indicated that the application of sewage sludge could improve the properties of red soil and promote vegetable growth. The dry weight of water spinach and Chinese chive reached the maximal levels when treated with the amount of sewage sludge at 4% and 10%, which was 4.38 ± 0.82 g and 1.56 ± 0.31 g, respectively. The dry weights after the application of sewage sludge were all larger than control treatment (CK) without sludge application. With increases in the applied amount of sewage sludge, the concentrations of Cu and Zu in red soil continued to increase, and the peak value was not reached. After the two vegetables were planted, the concentrations of Cu and Zn in red soil decreased by different degrees. The degrees of decrease of Zn were generally higher than those of Cu. The enrichment coefficient of water spinach on Cu showed a trend of increase followed by a decrease and reached the peak value of 1.04 ± 0.38 when the applied amount was 4%. The enrichment coefficient of Chinese chive on Cu overall showed a decreasing trend and did not reach the peak value under the treatment levels used in this experiment. The enrichment pattern of Chinese chive on Zn was not obvious, and the differences among all treatment levels were not significant (p < 0.05). However, the enrichment coefficient after the application of sewage sludge was significantly lower than that without the application of sludge.     

Comparative assessment of in situ bioremediation potential of cadmium resistant acidophilic Pseudomonas putida 62BN and alkalophilic Pseudomonas monteilli 97AN strains on soybean

A study was undertaken to examine the effects of the acidophilic strain 62BN (pH 5.5) and alkalophilic strain 97AN (pH 9.0) on remediation of cadmium and their subsequent effects on soybean (Glycine max var. PS-1347) in acidic and alkaline soils, respectively. The effect of cadmium on soybean plants was studied in acidic (pH 6.3 ± 0.2) and alkaline (8.5 ± 0.2) soil amended with 124 μM CdCl₂ concentration, respectively, and the cadmium toxicity was evident from stunted growth, poor rooting, and cadmium accumulation in each case. Furthermore, 16S ribosomal DNA (rDNA) sequencing identified 62BN as a Pseudomonas putida strain and 97AN as a Pseudomonas monteilli strain. In situ studies showed that on seed bacterization, both the P. putida 62BN strain and P. monteilli 97AN strain were able to enhance plant growth in terms of agronomical parameters, in the presence of cadmium in acidic and alkaline soils, respectively. Apart from this, strain 62BN and 97AN reduced cadmium concentration in plant and soil significantly (p < 0.05) in their respective soil types. Further comparative analysis revealed that P. putida 62BN was more effective than P. monteilli 97AN strain in remediation of cadmium. The bacterial strains offer promise as inoculants to improve the growth of plants in the presence of toxic Cd concentrations in the environment with their optimum pH.     

Soil microbial biomass influence on growth and biocontrol efficacy of Trichoderma harzianum

Hyphal growth and biocontrol efficacy of Trichoderma harzianum may depend on its interactions with biotic components of the soil environment. Effects of soil microbial biomass on growth and biocontrol efficacy of the green fluorescent protein transformant T. harzianum ThzID1-M3 were investigated using different levels of soil microbial biomass (153, 328, or 517 μg biomass carbon/g of dry soil). Hyphal growth of T. harzianum was significantly inhibited in soil containing 328 or 517 μg biomass carbon/g of dry soil compared with soil containing 153 μg biomass carbon/g. However, when ThzID1-M3 was added to soil as an alginate pellet formulation, recoverable populations of ThzID1-M3 varied, with the highest populations in soil containing 517 μg biomass carbon/g. When sclerotia of Sclerotinia sclerotiorum were added to soils (10 sclerotia per 150 g soil) with ThzID1-M3 (20 pellets per 150 g soil), colonization of sclerotia by ThzID1-M3 was significantly lower in the soil containing the highest level of biomass. Addition of alginate pellets of ThzID1-M3 to soils (10 pellets per 50 g) resulted in increased indigenous microbial populations (total fungi, bacteria, fluorescent Pseudomonas spp., and actinomycetes). Our results suggest that higher levels of microbial soil biomass result in increased interactions between introduced T. harzianum and soil microorganisms, and further that microbial competition in soil favors a shift from hyphal growth to sporulation in T. harzianum, potentially reducing its biocontrol efficacy.     

Cytotoxic effect of Green synthesized silver nanoparticles using Melia azedarach against in vitro HeLa cell lines and lymphoma mice model

This communication explains the biosynthesis of stable silver nanoparticles (AgNPs) from Melia azedarach and its cytotoxicity against in vitro HeLa cells and in vivo Dalton's ascites lymphoma (DAL) mice model. The AgNPs synthesis was determined by UV–visible spectrum and it was further characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS) and X-ray diffraction (XRD) analysis. Zeta potential analysis revealed stable AgNPs at ?24.9 mV. UV visible spectrum indicated an absorption peak at 436 nm which reflects its specific Surface Plasmon Resonance (SPR). Biosynthesized AgNPs were predominantly cubical and spherical with an average particle size of 78 nm approximately as observed through SEM and DLS analysis, respectively. Cytotoxicity of biosynthesized AgNPs against in vitro Human epithelial carcinoma cell line (HeLa) showed a dose–response activity. Lethal dose (LD₅₀) value was found to be 300 μg/mL of AgNPs against HeLa cell line. Cytotoxicity against normal continuous cell line human breast lactating, donor 100 (HBL 100) was found only in increased concentration of both AgNPs and 5-FU. In addition, in vivo DAL mice model showed significant increase in life span, induction of apoptosis was evidenced by acridine orange and ethidium bromide (AO and EB) staining.     

A self-evaluation method to attribute sound bio-values based on in situ ecotoxicological monitoring

Microbial, plant and animal species were examined within the framework of the ADEME Bioindicator Program in France that involves the monitoring, characterization and risk assessment of soils from various sites. Whenever any of the selected species under study were not found in all of the sampling areas of a given site, the data were difficult to analyze. It is explained how discrepancies in the data analysis may arise depending on the way the values of the biological parameters are taken into account: as a biomarker to assess values for each species in different areas, or as bioindicator to evaluate the number of species impacted in each area. We present a method to prevent discrepancies in interpretation in which a bio-value (“soil quality”) is attributed to various areas, allowing them to be ranked. The method also provides an evaluation of the soundness of the results and the resulting ranking, and no areas need to be assigned a priori as “control” or “contaminated” areas.In the framework of the ADEME Bioindicator Program, the leaf C18:3/(C18:0 + C18:1 + C18:2) fatty acid ratio was used as biological parameter. The experimental data obtained from two sites, in which not all plant species under study were found in all sampling areas, were analyzed by the above-mentioned method. The method proved usable to calculate a sound bio-value for each area in both sites and to rank them. Comparison of data demonstrated lower bio-values for the soil in the four areas within the industrial wasteland than for that in the two areas located just outside it. The soundness of the scoring obtained in the second site demonstrates that coherent results could also be obtained with plant species harvested in areas several kilometers apart and displaying varying soil properties.