Bioremediation of Heavy Metals in Liquid Media Through Fungi Isolated from Contaminated Sources

Wastewater particularly from electroplating, paint, leather, metal and tanning industries contain enormous amount of heavy metals. Microorganisms including fungi have been reported to exclude heavy metals from wastewater through bioaccumulation and biosorption at low cost and in eco-friendly way. An attempt was, therefore, made to isolate fungi from sites contaminated with heavy metals for higher tolerance and removal of heavy metals from wastewater. Seventy-six fungal isolates tolerant to heavy metals like Pb, Cd, Cr and Ni were isolated from sewage, sludge and industrial effluents containing heavy metals. Four fungi (Phanerochaete chrysosporium, Aspegillus awamori, Aspergillus flavus, Trichoderma viride) also were included in this study. The majority of the fungal isolates were able to tolerate up to 400 ppm concentration of Pb, Cd, Cr and Ni. The most heavy metal tolerant fungi were studied for removal of heavy metals from liquid media at 50 ppm concentration. Results indicated removal of substantial amount of heavy metals by some of the fungi. With respect to Pb, Cd, Cr and Ni, maximum uptake of 59.67, 16.25, 0.55, and 0.55 mg/g was observed by fungi Pb3 (Aspergillus terreus), Trichoderma viride, Cr8 (Trichoderma longibrachiatum), and isolate Ni27 (A. niger) respectively. This indicated the potential of these fungi as biosorbent for removal of heavy metals from wastewater and industrial effluents containing higher concentration of heavy metals.     

Performance of rose scented geranium (Pelargonium graveolens) in heavy metal polluted soil vis-à-vis phytoaccumulation of metals

An investigation was carried out to evaluate the effect of heavy metal toxicity on growth, herb, oil yield and quality and metal accumulation in rose scented geranium (Pelargonium graveolens) grown in heavy metal enriched soils. Four heavy metals (Cd, Ni, Cr, and Pb) each at two levels (10 and 20 mg kg–1 soil) were tested on geranium. Results indicated that Cr concentration in soil at 20 mg kg–1 reduced leaves, stem and root yield by 70, 83, and 45%, respectively, over control. Root growth was significantly affected in Cr stressed soil. Nickel, Cr, and Cd concentration and accumulation in plant increased with higher application of these metals. Chromium, nickel and cadmium uptake was observed to be higher in leaves than in stem and roots. Essential oil constituents were generally not significantly affected by heavy metals except Pb at 10 and 20 ppm, which significantly increased the content of citronellol and Ni at 20 ppm increased the content of geraniol. Looking in to the higher accumulation of toxic metals by geranium and the minimal impact of heavy metals on quality of essential oil, geranium can be commercially cultivated in heavy metal polluted soil for production of high value essential oil.     

Dual augmentation for aerobic bioremediation of MTBE and TCE pollution in heavy metal-contaminated soil

In this work we isolated from soil and characterized several bacterial strains capable of either resisting high concentrations of heavy metals (Cd²⁺ or Hg²⁺ or Pb²⁺) or degrading the common soil and groundwater pollutants MTBE (methyl-tert-butyl ether) or TCE (trichloroethylene). We then used soil microcosms exposed to MTBE (50 mg/l) or TCE (50 mg/l) in the presence of one heavy metal (Cd 10 ppm or Hg 5 ppm or Pb 50 or 100 ppm) and two bacterial isolates at a time, a degrader plus a metal-resistant strain. Some of these two-membered consortia showed degradation efficiencies well higher (49–182% higher) than those expected under the conditions employed, demonstrating the occurrence of a synergetic relationship between the strains used. Our results show the efficacy of the dual augmentation strategy for MTBE and TCE bioremediation in the presence of heavy metals.     

Assessment of heavy metal induced stress responses in pea (Pisum sativum L.)

Exposure of plants to heavy metals severely affects their growth and physiological processes. Nevertheless, different plants show variable responses to different heavy metals, generally in a concentration-dependent manner. In this study, phytotoxic effects of cadmium (Cd), cobalt (Co) and lead (Pb) applied as chlorides at concentration 500, 750, 1000 and 1250 ppm were evaluated on seed germination, early seedling growth and dry biomass of pea (Pisum sativum L.). A lower concentration (500 ppm) of Pb promoted seed germination but declined other growth parameters. Higher concentration had a phytotoxic influence on the pea. Cd and Co severely affected germination and seedling growth of pea resulting in complete failure of germination and seedling growth at higher metal concentration. Tolerance index (TI) calculated for seed germination and dry biomass indicated that tested plant had zero tolerance to 1250 ppm of Cd as well as 750 ppm and higher concentrations of Co. The order of heavy metals for their phytotoxic effects was Co > Cd > Pb. The study suggests that P. sativum is relatively tolerant to Pb but highly sensitive to Co and Cd.     

Bioaccumulation and translocation of heavy metals by Plantago major L. grown in contaminated soils under the effect of traffic pollution

The present study was performed at a heavy-traffic affected soil to examine the efficacy of bioaccumulation and translocation potentials of heavy metals by the naturally growing weed Plantago major. Heavy metals were analyzed in soil as well as in plant below- and above-ground parts along different distances from a heavy-traffic highway. All the investigated soil heavy metals, except Cd, varied significantly, while pH and E.C had no significant difference, with increasing distance from the highway. Likewise, there was a significant decrease of heavy metals in plant below- and aboveground parts. In addition, no significant difference between most soil and root heavy metals at 20 and 100 m as well as those at 500 and 750 m distance from the highway. The bioaccumulation factor (BF) of all heavy metals, except Cd and Sr, were less than unity at most distances. However, Cd showed relative BF decline with the distance in contrast to Sr, which increases as distance from the highway increases. On the other hand, the translocation factors (TF) of Cd, Co, Cu, Pb and Zn were higher at the distances far from the highway, while that of Fe, Cr and Sr were higher near the highway. Furthermore, the enrichment factor (EF) showed small variations, among the investigated heavy metals, with varying distances from the pollution source. It was found that soil Fe, Al, Cr, Ni, Sr, V and Zn had significant positive correlation with all investigated heavy metals in P. major roots. The higher TFs of Cd, Fe and Pb in P. major shoots makes it suitable for phytoextraction from soil, while the lower ratios of Al, Mn, V, Co, Ni, Cr, Zn, Cu and Sr make it suitable for their phytostabilization. Therefore, this plant can be used as a bioindicator and biomonitor for traffic related heavy metals.     

Screening of plant growth promoting traits in heavy metals resistant bacteria: Prospects in phytoremediation

Phytoremediation is considered as a novel environmental friendly technology, which uses plants to remove or immobilize heavy metals. The use of metal-resistant plant growth-promoting bacteria (PGPB) constitutes an important technology for enhancing biomass production as well as tolerance of the plants to heavy metals. In this study, we isolated twenty seven (NF1-NF27) chromium resistant bacteria. The bacteria were tested for heavy metals (Cr, Zn, Cu, Ni, Pb and Co) resistance, Cr(VI) reduction and PGPB characters (phosphate solubilization, production of IAA and siderophores). The results showed that the bacterial isolates resist to heavy metals and reduce Cr(VI), with varying capabilities. 37.14% of the isolates have the capacity of solubilizing phosphate, 28.57% are able to produce siderophores and all isolates have the ability to produce IAA. Isolate NF2 that showed high heavy metal resistance and plant growth promotion characteristics was identified by 16S rDNA sequence analysis as a strain of Cellulosimicrobium sp.. Pot culture experiments conducted under greenhouse conditions showed that this strain was able to promote plant growth of alfalfa in control and in heavy metals (Cr, Zn and Cu) spiked soils and increased metal uptake by the plants. Thus, the potential of Cellulosimicrobium sp. for both bioremediation and plant growth promotion has significance in the management of environmental pollution.     

The influence of heavy metals upon the growth of sitka-spruce in South Wales forests

Summary Experiments on sitka-spruce seedlings grown in acidic peaty gley soils under green-house conditions, where the soils where doped with increasing amounts of Cd, Cu and Pb up to maximum levels of metal added of 16 ppm, 32 ppm and 400 ppm respectively, showed that the levels of Cd and Pb in shoots and roots increased with increasing levels in the soil, whereas levels of copper appeared to be independent. The addition of these three metals to the soils did not influence the uptake of other heavy metals, or of the nutrients potassium or calcium. Increases in the shoot cadmium levels significantly reduced the yields of the plant shoots. However, the plant yields were only affected by the highest level of lead that was added to the soil (400 ppm Pb) and unaffected by all the copper treatments (0–32 ppm Cu in the soil). The lengths of the sitka-spruce roots were reduced when cadmium and lead levels in the soil exceeded certain threshold concentrations (2.5 ppm total Cd, where 0.3 ppm was extractable with 0.5 M acetic acid; and 48 ppm total Pb, where 1.7 ppm was extractable). However, root lengths were not reduced by copper. This was probably related to the fact that copper appears to be relatively unavailable in the type of soil used, as only 1.1. ppm Cu was extractable from a total of 32 ppm Cu added. Root branching was apparently reduced by increases in the soil levels of cadmium, copper and lead. The roots of some control plants had symbiotic mycorrhizal associations (4 out of 19 plants), whereas the roots of all the plants grown in the soils with added heavy metals did not develop these.     

Biochemical responses of the aquatic moss Fontinalis antipyretica to Cd,Cu, Pb and Zn determined by chlorophyll fluorescence and protein levels

Biochemical reactions to Cu, Cd, Zn and Pb in the aquatic moss Fontinalis antipyretica were studied in order to characterize the physiological background of the metal response. Chlorophyll fluorescence and intracellular metal localization and stress protein levels were measured. Exposure to 25 or 100 μM Cu over a 7-day period resulted in a decline of chlorophyll fluorescence to about 70% and 52%, respectively. Up to 100 μM Cd caused a decrease in chlorophyll fluorescence to 75%. With all metals used at 25–100 μM concentrations, the intracellular uptake increased. For all metals investigated at 25–100 μM, the intracellular uptake increased. Maximum values were reached at 100 μM Cu, Pb, Zn or Cd exposure. As shown by analytical electron microscopy (EDX, EELS) moss material treated with 50 μM Cu exhibited reduced sulphur levels in the cytoplasm and an increase in phosphate in vacuolar dense particles. EEL-spectra indicated that Cu is chelated in the cytoplasm by SH-groups and coprecipitated with orthophosphate in vacuoles. To monitor the stress response at the protein level, heavy metal induced heat shock protein 70 (hsp70) was measured. An antibody was raised against conserved plant metallothionein p2 motifs derived from Brassica juncea. In all metal-treated samples the antibody bound to proteins of about 8 kDa. However, sequencing failed to reveal significant homologies to known proteins. These experiments provide for the first time results on protein level after heavy metal stress in the aquatic bioindicator moss.     

Water quality index,Labeo rohita,and Eichhornia crassipes: Suitable bio-indicators of river water pollution

The present study investigated the water quality index (WQI) of the Kshipra river at Dewas, Madhya Pradesh, India, using native fish Labeo rohita, and plant Eichhornia crassipes. The temperature, pH, dissolved oxygen, alkalinity, turbidity, and dissolved solids were found to be within the prescribed limits. However, heavy metals concentration exceeded the limit except for Cu and Zn. Their occurrence in river water was as follows: Ni > Fe > Cd > Cr > Mn > Zn > Cu. Among these heavy metals, Cd was found to be highly bioavailable, whereas Zn was the least bioavailable metal. Based on WQI, the water was found to be unfit for drinking, and the high WQI value was due to the presence of Cr and Cd. In fish tissues (muscle, liver, gut, gills, and kidney), the highest and lowest metal pollution index was found in gills (45.03) and kidneys (12.21), respectively. Bioaccumulation of these metals resulted in significant depletion of energy reserves (protein, glucose, and glycogen) and also altered hematological parameters. Moreover, liver function tests showed hepatic damage in the exposed fish. In-plant, both the bioaccumulation and mobility factor exceeded 1 for all these metals. On the other hand, the translocation factor was found to be beyond 1 for Fe, Ni, and Zn. These high values make this plant fit for phytoextraction of Mn, Fe, Cu, Zn, and Cd and phytostabilization of Cr in water. Moreover, consumption of L. rohita from the Kshipra River does not pose a non-cancer risk as the target hazard quotient was below 1, but it may pose cancer risk because of the presence of Cr in the range of 1.402 × 10^?3 to 1.599 × 10^?3.     

Comparison of the toxicity of heavy metals to cabbage growth

Summary Cabbage plants were grown for 55 days with a nutrient solution containing 1 and 10 ppm of V, Cr(III), Cr(VI), Mn, Fe, Co, Ni, Cu, Zn, Cd, Hg(I), orHg(II). A comparison of the plant growth and chemical analysis revealed that Cr(VI), Cu, Cd, and Hg(II) in the solution are most toxic to the plant growth (hence detrimental to the cabbage-head formation) and Mn, Fe, and Zn are less toxic than other heavy metals, and that Mn, Zn, Co, Ni, and Cd and translocated into all the plant organs while V, Cr(III), Cr(VI), Fe, Cu, Hg(I), and Hg(II) are accumulated in the roots.     

Detecting the heavy metal tolerance level in ectomycorrhizal fungi <Emphasis Type="Italic">in vitro</Emphasis>

Summary Eight isolates of ectomycorrhizal fungi namely, Laccaria fraterna (EM-1083),Laccaria laccata (EM-1191), Pisolithus tinctorius (EM-1081),Pisolithus tinctorius (EM-1293), Scleroderma cepa (EM-1233),Scleroderma flavidum (EM-1235),Scleroderma verucosum, (EM-1283) and Hysterangium incarceratum (EM-1185) were grown on specially designed cocktail media prepared by adding various concentrations of different heavy metals namely Al, As, Cd, Cr, Ni and Pb. The heavy metals were selected keeping in view their relative abundance in coal ash and potential toxicity. The fungal isolates were grown on such designed cocktail media. The colony diameter was used for the measurement of the fungal growth. Total heavy metal accumulated in the mycelia was assayed by atomic absorption spectrophotometry.In relation to metal tolerance ability in general,Hysterangium incarceratum (EM-1185) showed maximum tolerance with respect to growth, Laccaria fraterna (EM-1083) and Pisolithus tinctorius (EM-1293) also showed considerable tolerance to the heavy metals tested. In relation to metal uptake in particular, Pisolithus tinctorius (EM-1293), has reported maximum uptake of Al (34642.58 ppm), Cd (302.12 ppm) and Pb (3501.96 ppm). In Laccaria fraterna (EM-1083), As (130.57 ppm) and Cr (402.38 ppm) uptake was recorded maximum; and Hysterangium incarceratum (EM-1185) has recorded maximum Ni (2648.59 ppm) uptake among the three suitable isolates documented here.     

Protective effect of jasmonic acid and potassium against cadmium stress in peas (Pisum sativum L.)

A combination of mineral nutrients and plant growth regulators should be assessed to improve crop performance under various abiotic stresses. There is a need to include plant growth regulators in fertilization regime of various crops along with essential mineral nutrients, especially when they are irrigated with polluted water with higher levels of heavy metals. The performance of pea was evaluated under cadmium (Cd) stress coupled with potassium (K) and jasmonic acid (JA) supplementation. The Cd stress (50 μM) was applied to soil (sandy loam) grown pea plants as basal dose after a month of sowing. The control and stressed plants were then supplemented with K (5 M), JA (0.5 mM) and their collective application along with control as distilled water. Cd stress showed a marked reduction in growth pattern, however, the collective supplementation sufficiently improved the growth pattern of stressed peas plants as evidenced by improvement in shoot length (cm), root length (cm), number of leaves per plant, leaf area (cm²), plant fresh and dry weight (gm). Potassium application under Cd stress significantly enhanced internodal distance (cm) while the number of seeds per pod and relative water contents remained nonsignificant. The applied treatment (JA + K) under Cd stress prominently improved enzymatic activities, which were measured as nitrate reductase activity (NRA), nitrite reductase activity (NiRA), superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Cd stress impacted the biochemical profile by enhancing antioxidant capacity (AC), antioxidant activity (AA), total phenols (TP), while reducing total soluble protein (TSP), chlorophyll ‘a’, chlorophyll ‘b’ and carotenoids. The combined application of JA and K under Cd stress enhanced AC, AA, TP, Chl a and b, TSP and carotenoids. The results indicate that foliar application of JA and K efficiently negated the harmful effects of Cd stress on peas.     

植物抗旱和耐重金属基因工程研究进展

干旱和重金属污染严重影响植物的生长发育.植物耐逆相关基因的克隆和功能鉴定研究,为通过基因工程途径提高植物的抗逆性奠定了理论基础.水分亏缺、高盐、低温和重金属胁迫都能诱导LEA(late embryogenesis abundant protein)基因的表达.转基因研究表明,LEA蛋白具有抗旱保护作用、离子结合特性以及抗氧化活性;水孔蛋白存在于细胞膜和液泡膜上,在细胞乃至整个植物体水分吸收和运输过程中发挥重要作用.干旱和盐胁迫促进水孔蛋白基因转录物的积累.过量表达水孔蛋白可增强水分吸收和运输,提高植物的抗旱能力.金属转运蛋白参与重金属离子的吸收、运输和累积等过程.这些蛋白基因在改良草坪草植物的抗旱节水和耐重金属能力等方面具有潜在的应用价值.     

Biosorption of Chromium(VI) and Lead(II): Role of Spirulina platensis in the Treatment of Industrial Effluent

Biosorption is the process of removal of any chemical molecules by the treatment of biological material. Industrialization resulted in the discharge of various toxic heavy metals into water bodies, which poses serious health hazards to humans and animals. In the present study, live Spirulina platensis was used as a biosorbent for the removal of the heavy metals chromium (Cr(VI)) and lead (Pb(II)) from the aqueous samples. S. platensis were cultured in the presence of different concentrations of heavy metals. The growth of the algal cells was found to be decreased by 59% and 36% in media containing 50 ppm Cr(VI) and Pb(II), respectively. To assess the biosorption of heavy metals, at different time intervals, the spent culture media were used to detect Cr(VI) by atomic absorption spectroscopy method and Pb(II) by 4-(2-pyridylazo)resorcinol indicator method. Results suggested that there was a significant uptake of Cr(VI) and Pb(II) from the medium by S. platensis, with corresponding decrease of metals in the medium. When metal salt solutions or industrial effluent samples were passed through the column containing immobilized live S. platensis in calcium alginate beads, the concentration of Cr(VI) was found to be reduced drastically. The present study indicates the application of S. platensis for the bioremediation of heavy metals from the samples obtained from industrial effluents.     

Effect of two biodegradable chelates on metals uptake,translocation and biochemical changes of Lantana Camara growing in fly ash amended soil

The present work had two purposes firstly to evaluate the potential of Lantana Camara for phytoextraction of heavy metals from fly ash amended soil and to assess the suitability of a proper biodegradable chelating agent for chelate assisted phytoextraction. Plants were grown in manure mixed soil amended with various concentration of fly ash. Two biodegradable chelating agents were added (EDDS and MGDA) in the same dose separately before maturation stage. Sampling was done at different growing stages. The plant took up metal in different plant parts in the following order: for Cu, and Zn leaf >root >stem, for Cr and Mn leaf>stem >root, for Ni root >leaf>stem and for Pb root≈leaf>stem respectively. For Cu, Zn, Cr and Mn Lantana camara acted as phytoextractor. Translocation factor and bioaccumulation coefficient was>1 signifying enrichment and translocation of metals in the plant. Morphological studies showed no toxicity symptom in the plant. Among biochemical parameters protein and nitrate reductase activity decreased, whereas, chlorophyll and peroxidise activity increased with the growth stages. Finally, it was evident from the results that Lantana Camara can be used as efficient phytoextractor of metals, with proper harvesting cycle and both chelate were proved as effective chelators for phytoextraction of metals.     

Seed priming with methyl jasmonate mitigates copper and cadmium toxicity by modifying biochemical attributes and antioxidants in Cajanus cajan

Contamination of agricultural soils with heavy metals (HMs) has posed major threat to the environment as well as human health. The aim of this study was to appraise the efficiency of key-antioxidant enzymes in enhancing plants’ tolerance to HMs (heavy metals) like copper (Cu) and Cadmium (Cd), under the action of methyl jasmonate (Me-JA) in Cajanus cajan L. Seeds of C. cajan treated with Me-JA (0, 1 nM) were discretely subjected to noxious concentrations of Cu and Cd (0, 1, 5 mM) and raised for 12 days under controlled conditions in plant growth chamber for biochemical analysis. In contrast to Cd, Cu triggered oxidative stress more significantly (44.54% in 5 mM Cu increase in MDA as compared to control) and prominently thereby affecting plants’ physiological and biochemical attributes. By activating the antioxidant machinery, Me-JA pre-treatment reduced HMs-induced oxidative stress, increased proline production, glutathione (41.95% under 5 mM Cu when treated with 1 nM Me-JA treatment) and ascorbic acid content by 160.4 % under aforemtioned treatments thus improving the redox status. Thus, in light of this our results put forward a firm basis of the positive role that Me-JA might play in the mitigation of oxidative stress caused due to HMs stress by stimulating antioxidant defense system leading to overall improvement of growth of C. cajan seedlings.     

Microtubule organizing centers in plant cells: localization of a 49 kDa protein that is immunologically cross-reactive to a 51 kDa protein from sea urchin centrosomes in synchronized tobacco BY-2 cells

Summary A 49 kDa protein in tobacco BY-2 cells has been found to be cross-reactive with antibodies raised against a 51 kDa protein that was isolated from sea urchin centrosomes and identified as a microtubule-organizing center (MTOC) in animal cells. Tracing the fate of the 49 kDa protein during progression of the cell cycle in highly synchronized tobacco BY-2 cells revealed that this protein was colocalized with plant microtubules (MTs): the location of the 49 kDa protein coincided with preprophase bands (PPBs), mitotic spindles and phragmoplasts. Furthermore, between the M and G₁ phases, the 49 kDa protein was observed in the perinuclear regions, in which the initials of MTs are organizing to form cortical MTs. At the G₁ phase the location of the 49 kDa protein in the cell cortex coincided with that of the cortical MTs. It appeared that the 49 kDa protein in the cell cortex was transported as granules from the perinuclear regions. Thus, it is highly probable that the 49 kDa protein, which reacts with antibodies against the 51 kDa protein in sea urchin centrosomes, plays the role of an MTOC in plant cells. Thus, the mechanisms for organizing MTs in higher organisms appear to share a common protein, even though the organization of MTs is superficially very different in plant and animal cells.Abbreviations DAPI 4,6-diamidino-2-phenyl indole - MT microtubule - MTOC microtubule-organizing center - PAGE polyacrylamide gel electrophoresis - PBS phosphate-buffered saline - PPB preprophase band - SDS sodium dodecylsulfate     

Evaluation of some heavy metals toxicity in Channa punctatus and riverine water of Kosi in Rampur,Uttar Pradesh,India

This study was aimed to assess the bioaccumulation and concentration level of three heavy metals such as Chromium (Cr), Cadmium (Cd) and Lead (Pb) in the riverine water and edible fish, Channa punctatus obtained from River Kosi, Rampur, Uttar Pradesh, India. These toxic heavy metals are released into the environment because of E-waste, industrial activities, municipal urban runoff, coal burning, fertilizers etc., then paved the way into the aquatic system due to direct input, atmospheric deposition, and erosions caused by rain. There is every apprehension of aquatic animals getting exposed to elevated levels of heavy metals, thus possessing harmful effects both to flora and fauna. The concentrations of Cd, Cr and Pb in water were found to be 0.051 ± 0.026, 1.091 ± 0.408, and 0.019 ± 0.002 whereas in the kidneys of Channa punctatus, as 0.076 ± 0.208, 0.482 ± 0.059, and 0.127 ± 0.705 respectively.