Comparative study on removal of enteric pathogens from domestic wastewater using Typha latifolia and Cyperus rotundus along with different substrates

A comparative study was carried out to evaluate the efficiency of different substrate materials along with macrophytes Typha latifolia and Cyperus rotundus in treating domestic wastewater intended for reuse in agriculture. The study was conducted over a period of 6 months with different retention times, and observations were taken twice per month. One-way analysis of variance and Tukey's Honest Significant Difference (HSD) tests were used to determine statistical significant differences between experimental columns. Treatment with T. latifolia planted in sand and mix substrate with 4-day retention time remarkably reduced the concentration of all bacterial pathogens. Log reductions observed were approximately 5.01 and 4.82 for total coliform (TC), 4.46 and 3.93 for Escherichia coli, and 5.52 and 5.48 for Shigella, respectively. Moreover, these treatments were also efficient in completely removing fecal coliform (FC) and Salmonella.Maximum parasites were removed by the treatment having sand alone as a substrate containing C. rotundus, but the difference was not significant from those planted with T. latifolia in the same substrate. The results suggest that T. latifolia aids in bacterial pathogens removal, while C. rotundus aids in parasites removal. Thus, wastewater treatment through constructed wetland having mix plantation of these species along with sand can eliminate some of the major enteric pathogens.     

Antifungal efficacy of some ethyl acetate extract fractions of Cyperus rotundus rhizomes against spore germination of some fungi

Plant products play an important role as safe and ecofriendly method in controlling various plant diseases. Ethyl acetate fractions of Cyperus rotundus rhizomes have been found highly effective against some species of Alternaria (A. alternata, A. brassicola, A. solani, Alternaria chearanthi), Colletotrichum (C. musae, Colletotrichum sp.), Curvularia (C. lunata, C. maculans, C. pallescens, C. pennisetti), Helminthosporium (H. pennissetti, H. spiciferum, H. echinoclova and Heterosporium colocasiae) in vitro. A. brassisicola was highly sensitive to all the fractions at all the concentrations. Fractions 4–7 inhibited complete spore germination at 2000 and 3000 μg/ml as compared to control. However, 100% spore germination inhibition was found in Colletotrichum species in all the fractions at 2000 and 3000 μg/ml but at lower concentrations 50–60% spore germination and 90% reduction of germ tube elongation were observed. Curvularia species was highly sensitive to all concentrations of all the fractions as 50–100% spore germination inhibition was recorded at 500 μg/ml dose. Some species of Helminthosporium were sensitive at 3000 μg/ml in some fractions but other test fractions showed least efficacy. Fractions 4–7 were 100% inhibitory for H. colocasiae. Germ tube elongation was also affected by 60–90%. The germ tube branching and their elongation were affected in almost all species at 30 to 95%. The high efficacy of ethyl acetate fractions of rhizomes of C. rotundus against some fungi indicates that they can be very well tried under field conditions against some important plant diseases as an ecofriendly method of plant disease control.     

Contaminated Soil Phytoremediation by Cyperus Laxus Lam. Cytochrome P450 Erod-Activity Induced by Hydrocarbonsin Roots

Laboratory and greenhouse experiments with Cyperus laxus Lam were conducted to determine the rate and extent of phytoremediation and the effect of hydrocarbons on the cytochrome P450 EROD (7-ethoxyresorufin-O-deethylase) enzymatic activity in roots. Plants were cultivated on hydrocarbon-contaminated soil (HCS) and spiked perlite. Phytoremediation was evaluated using 6.5 kg HCS (173 ± 15 mg total petroleum hydrocarbons [TPH] g^?1 of dry soil) pots at different moisture contents; the average removal rate was 3.46 ± 0.25 mg TPH g^?1 dry soil month^?1 and 48% was removed when moisture was kept at 60%. The aromatic hydrocarbon fraction was the mostly removed, 60%; aliphatic, 51%; and polar 24% after 24-month experiments. In unplanted pots, TPH concentration did not exhibit significant differences with respect to the initial concentration. We confirmed that the presence of hydrocarbons induced ERODactivity up to 6.5-fold. Moreover, short-term experiments (up to 13 d) with spiked perlite demonstrated that two EROD activities in roots contributed to the total detected; 60% was found in the cytosolic and 40% in the microsomal fraction. To our knowledge, this is the first work that tries to build links between the hydrocarbon-inducible character of ERODactivity in roots and the phytoremediation ability of C. laxus in highly contaminated soils.     

Responses of Cyperus brevifolius (Rottb.) Hassk. and Cyperus kyllingia Endl. to varying soil water availability

Different responses of Cyperus brevifolius and Cyperus kyllingia to varying soil water regimes were examined to explain their successful existence in a diverse range of habitats throughout the year in Indonesia. Thirty 43-day sprouts of each species were grown in three soil conditions, namely drought, field capacity, and flooding under greenhouse conditions. Plant height, leaf length, tiller number, and flower number were measured twice a week, from 45 to 98 days after sowing (DAS), while the other 12 traits were recorded at the end of the observation time. Ten out of the twelve traits were substantially influenced by the soil water content. Both species exhibited their best growth, production, and reproduction under field capacity conditions, and these traits were greatly subdued under drought conditions. Under drought conditions, both species manifested reduced growth and leaf expansion; however, stomatal aperture and frequency did not exhibit strong response to the soil water content. C. brevifolius showed a significantly greater biomass production and reproductive traits in field capacity and flooded conditions, but under drought conditions, growth was greatly hampered and only vegetative propagation occurred. On the other hand, C. kyllingia showed a higher tolerance to drought conditions, indicated by both a higher biomass and a higher number of flowers. The results obtained suggest that survival ability when faced with drought conditions was more apparent in C. kyllingia than in C. brevifolius, but where there was sufficient soil water, C. brevifolius was more prolific. This could be the explanation for the dominance of C. brevifolius in flooded areas and during the rainy season, and the occurrence of C. kyllingia in a wider range of habitats throughout the year.     

Phytodegradation of Ethanolamines by Cyperus alternifolius: Effect of Molecular Size

Our screening of plants showed that Cyperus alternifolius (Umbrella papyrus) had the highest efficiency removal in real wastewater containing monoethanolamine—higher than Echinodorus cordifolius (Creeping Burrhead), Thalia geniculata (Alligator Flag), Acorus calamus (Sweet Flag), and Dracaena sanderiana (Lucky Bamboo). Therefore, this research studied the degradation of monoethanolamine (MEA), diethanolamine (DEA), and triethanolamine (TEA) by C. alternifolius. Plants could degrade TEA into DEA, then into MEA, and then further into acetic acid. The accumulation of ethanolamines was found mainly in plant stems, which had the highest biomass. This demonstrated that the molecular size is closely related to a diffusion coefficient that affects the removal rate through plant bodies. A smaller molecular weight—MEA (MW = 61.08 g mol^?1)—was taken up the fastest, followed by DEA (MW = 105.14 g mol^?1) and TEA (MW = 149.19 g mol^?1), the highest molecular weight. The plants’ toxicity when exposed to ethanolamines elucidated that MEA had the highest toxicity, followed by DEA and TEA. In addition, the application of C. alternifolius in monoethanolamine-contaminated wastewater revealed that plant could completely uptake MEA at day 5 from an initial MEA concentration of 18 mM. The result indicated that C. alternifolius has the potential to remove ethanolamines and can be applied to ethanolamine-contaminated wastewater.     

Phytoremediation of fuel oil and lead co-contaminated soil by Chromolaena odorata in association with Micrococcus luteus

Phytoremediation is widely promoted as a cost-effective technology for treating heavy metal and total petroleum hydrocarbon (TPH) co-contaminated soil. This study investigated the concurrent removal of TPHs and Pb in co-contaminated soil (27,000 mg kg^?1 TPHs, 780 mg kg^?1 Pb) by growing Siam weed (Chromolaena odorata) in a pot experiment for 90 days. There were four treatments: co-contaminated soil; co-contaminated soil with C. odorata only; co-contaminated soil with C. odorata and Micrococcus luteus inoculum; and co-contaminated soil with M. luteus only. C. odorata survived and grew well in the co-contaminated soil. C. odorata with M. luteus showed the highest Pb accumulation (513.7 mg kg^?1) and uptake (7.7 mg plant^?1), and the highest reduction percentage of TPHs (52.2%). The higher TPH degradation in vegetated soils indicated the interaction between the rhizosphere microorganisms and plants. The results suggested that C. odorata together with M. luteus and other rhizosphere microorganisms is a promising candidate for the removal of Pb and TPHs in co-contaminated soils.     

Earthworm-assisted bioremediation of petroleum hydrocarbon–contaminated soils from motorcar mechanic workshops in Ibadan,Oyo State,southwestern Nigeria

Enhanced microbial bioremediation of petroleum hydrocarbon–contaminated (PHC) soils with the earthworm Alma millisoni and the bacterium Bacillus spp. was conducted. The petroleum-contaminated topsoils (PCTS) (0–15 cm) collected from motorcar mechanic workshops were thoroughly mixed, sieved, and air dried for 7 days. The pH, water holding capacity (WHC), total nitrogen (N), organic carbon (OC), heavy metal (HM), and bacteriological analysis of the soil samples were evaluated. The indigenous bacterial isolates were subjected to 1%, 5%, and 50% of spent engine oil (SEO), incubated for 7 days at 37°C, and the isolate with the highest tolerance pattern was used for the remediation. Out of four indigenous bacteria isolated, Bacillus spp. had the highest tolerance to SEO. Preliminary exposure assessments of A. millisoni to PHC soils (100%, 60%, 50%, and 40% PHC) were carried out using 48-h avoidance response, coiling exhibition, swollen clitelium, 14-day survival tests, and antioxidant enzyme activities such as catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), and glutathione peroxidase (GPx). Subsequently, four treatments of 1 kg soil mixed with 100%, 75%, 50%, and 0% PCTS were designed and spiked with 20 g of dried cow dung. Each of the treatments consisted of four setups, viz., A. millisoni alone, A. millisoni and Bacillus spp., Bacillus spp. alone, and control. The bacterial counts, total petroleum hydrocarbon (TPH), total and bioavailable HM, and total OC and N of the soils were evaluated every 7 days for 35 days. Significant increases in the activities of CAT, SOD, GPx, and GST compared with control were recorded in A. millisoni exposed to the various treatments. Treatment with combined A. millisoni and Bacillus spp. resulted in significant (p < .05) reduction in TPH, reduction in total and bioavailable heavy metals, and increased total OC and N of the soil compared with other treatments. The percentage reduction in TPH and heavy metals with concomitant increase in total OC and total N recorded in the 50% PHC soils followed the order A. millisoni and Bacillus spp. > A. millisoni alone > Bacillus spp. alone. Hence, enhanced bioremediation using A. millisoni and Bacillus spp. may be a good biocatalyst in the remediation of petroleum hydrocarbon–contaminated soils.     

Bioremediation of Soils Contaminated with Nigerian Petroleum Products Using Composted Municipal Wastes

The efficiency of ready-to-use, source-separated, composted municipal organic wastes of Nigerian origin on degradation of soil total petroleum hydrocarbons (TPHs) in soils polluted with petroleum products (crude oil, diesel, and spent engine oil) was assessed in screen house experiments. The effect of compost:soil ratios and combined effect of compost-phytoremediation technique were also studied. TPH was determined spectrophotometrically, after extraction with 1:1 acetone-dichloromethane mixture at 425 nm. Soil pH, electrical conductivity, and phytotoxicity to seed germination and growth of maize (Zea mays L.) served as risk assessments on soil quality and evidence of recovery for the oil-impacted soil. Results showed that the treatments increased soil pH and electrical conductivity but reduced TPH. Reductions in TPH by compost technology ranged from 40% to 75.87%. Toxicity to seed germination reduced from 100% to 16.12%. Positive correlations were obtained for plant agronomical parameters and growth period, for all treatments, with coefficients in the range of .905 to .996, p < .05. This study revealed that ready-to-use composted waste has the potential for bioremediation of soils polluted with petroleum and petroleum products. This study is a contribution to the data bank of relatively simple bioremediation methods, suitable for workers in the developing countries, where there is no easy access to high-technology facilities. However, further development of this technique to achieve zero residual TPH is recommended.     

Plants‐associated microflora and the remediation of oil‐contaminated soil

The use of plants and their rhizospheric microorganisms is a promising emerging technology for remediating contaminated soils. The degradation of total petroleum hydrocarbon (TPH) in the rhizospheric and nonrhizospheric soil of three domestic plants, namely, alfalfa (Medicaga sativa) broad beans (Vicia faba) and ryegrass (Lolium perenne) was investigated. The experimental data from the studies of plantmicrobe‐soil interactions implicated the enhancement of TPH degradation by the rhizospheric microbial community. Although the three domestic plants exhibited normal growth in the presence of ～1.0% TPH, the degradation was more profound in the case of leguminous plants. The TPH degradation in the soil cultivated with broad beans and alfalfa was 36.6 and 35.8%, respectively, compared with 24% degradation in case of ryegrass. Such a high correlation between plant type and TPH degradation rates indicate that selection for enhanced rhizosphere degradation may be accomplished by selecting leguminous plants.     

Phytoremediation of petroleum hydrocarbon-contaminated saline-alkali soil by wild ornamental Iridaceae species

As a green remediation technology, phytoremediation is becoming one of the most promising methods for treating petroleum hydrocarbons (PHCs)-contaminated soil. Pot culture experiments were conducted in this study to investigate phytoremediation potential of two representative Iridaceae species (Iris dichotoma Pall. and Iris lactea Pall.) in remediation of petroleum hydrocarbon-contaminated saline-alkali soil from the Dagang Oilfield in Tianjin, China. The results showed that I. lactea was more endurable to extremely high concentration of PHCs (about 40,000 mg/kg), with a relatively high degradation rate of 20.68%.The degradation rate of total petroleum hydrocarbons (TPHs) in soils contaminated with 10,000 and 20,000 mg/kg of PHCs was 30.79% and 19.36% by I. dichotoma, and 25.02% and 19.35% by I. lactea, respectively, which improved by 10–60% than the unplanted controls. The presence of I. dichotoma and I. lactea promoted degradation of PHCs fractions, among which saturates were more biodegradable than aromatics. Adaptive specialization was observed within the bacterial community. In conclusion, phytoremediation by I. dichotoma should be limited to soils contaminated with ≤20,000 mg/kg of PHCs, while I. lactea could be effectively applied to phytoremediation of contaminated soils by PHCs with at least 40,000 mg/kg.     

Phytoremediation Effect and Growth Responses of Cynodon spp. and Agropyron desertorum in a Petroleum-Contaminated Soil

In order to compare the petroleum tolerance and phytoremediation ability of a native grass, Agropyron desertorum (desert Wheatgrass) with Cynodon spp. (Bermuda grass) in a petroleum hydrocarbon-contaminated soil, a 7-month greenhouse experiment was performed. There were 4 soil treatments with 0% (uncontaminated soil), 2%, 4%, and 12% (w_oil/w_soil) petroleum concentration. Parameters including shoot and root fresh weight and dry weight, root penetration depth and root density depth, soil respiration, and total petroleum hydrocarbons (TPH) degradation were measured during and after experiments. The results showed an increase in shoot fresh weight of A. desertorum in soil polluted with 2% petroleum sludge compared to the uncontaminated soil, whereas the growth of Bermuda grass significantly decreased in corresponding treatment. Root growth of A. desertorum was decreased in 2% and 4% petroleum sludge, whereas it was increased in Bermuda grass species. Overall, root fresh weight of Bermuda grass was higher than that of A. desertorum in all treatments. Significant increase in microorganisms' activity was observed in the presence of petroleum sludge and plants in soil compared with uncontaminated soil without plants, and the highest soil respiration (37.6 mg C-CO₂/kg soil day) has been observed in the rhizosphere of Bermuda grass in treatment with 12% petroleum sludge. Plants had a significant role in the degradation of soil contaminants as TPH degradation in planted soils was significantly higher than that in unplanted soil (TPH degradation (%) was 30.4 and 38.9 in A. desertorum and Bermuda grass, respectively, whereas it was just 13.3 in unplanted soil). The rhizosphere of Bermuda grass had significantly less residual TPHs compared to A. desertorum. The results indicated that both Cynodon spp. and A. desertorum had a peculiar tolerance to petroleum pollution. Therefore, as Bermuda grass has already been suggested to be a typical and efficient species for phytoremediating petroleum-contaminated sites, A. desertorum may also prove to be a suitable native alternative.     

Dactylaria higginsii,a Fungal Bioherbicide Agent for Purple Nutsedge (Cyperus rotundus)

Ten different fungi recovered from diseased purple nutsedge and yellow nutsedge plants collected in several locations in Florida and southeastern United States were screened for pathogenicity to purple nutsedge (Cyperus rotundus) plants. Only Dactylaria higginsii, recovered from diseased purple nutsedge plants collected in Gainesville, caused disease in greenhouse trials. Based on the results of pathogenicity and host-range tests conducted in a greenhouse, D. higginsii was determined to have potential as a bioherbicide agent for purple nutsedge. It was highly pathogenic to purple nutsedge, yellow nutsedge (C. esculentus), annual sedge (C. compressus), globe sedge (C. globulosus), rice flatsedge (C. iria), and green kyllinga (Kyllinga brevifolia [=Cyperus brevifolius]). Initial symptoms on inoculated plants consisted of moist, dark-brown leaf spots that appeared 4 days after inoculation. The spots later coalesced into larger lesions and blotches, killing the leaves and sometimes the entire aerial parts. The fungus sporulated on the infected leaves and caused secondary infections on the emerging leaves and shoots within 20 to 28 days after inoculation. Inoculation with conidial suspensions of D. higginsii resulted in significant reductions in shoot numbers (72%), shoot dry weight (73%), and tuber dry weight (67%) of greenhouse-grown purple nutsedge plants 45 days after inoculation. The fungus did not infect any of the crop plants or weedy grasses (Poaceae) tested.     

Kinetic Modelling and Half-Life Study on Bioremediation of Soil Co-Contaminated with Lubricating Motor Oil and Lead Using Different Bioremediation Strategies

The focus of this study was to investigate the effect of nutrient supplement (urea fertilizer) and microbial species augmentation (mixed culture of Aeromonas, Micrococcus, and Serratia sp.) on biodegradation of lubricating motor oil (LMO) and lead uptake by the autochthonous microorganism in LMO and lead-impacted soil were investigated. The potential inhibitory effects of lead on hydrocarbon utilization were investigated over a wide range of lead concentrations (25–200 mg/kg) owing to the complex co-contamination problem frequently encountered in most sites. Under aerobic conditions, total petroleum hydrocarbons (TPH) removal was 45.3% in the natural attenuation microcosm while a maximum of 72% and 68.2% TPH removal was obtained in biostimulation and bioaugmentation microcosms, respectively. Lead addition, as lead nitrate, to soil samples reduced the number of hydrocarbon degraders in all samples by a wide range (11–52%) depending on concentration and similarly, the metabolic activities were affected as observed in mineralization of LMO (3–60%) in soils amended with various lead concentrations. Moreover, the uptake of lead by the autochthonous microorganisms in the soil reduced with increase in the initial lead concentration. First-order kinetics described the biodegradation of LMO very well. The biodegradation rate constants were 0.015, 0.033, and 0.030 day^?1 for LMO degradation in natural attenuation, biostimulation and bioaugmentation treatment microcosms, respectively. The presence of varying initial lead concentration reduced the biodegradation rate constant of LMO degradation in the biostimulation treatment microcosm. Half-life times were 46.2, 21, and 23 days for LMO degradation in natural attenuation, biostimulation and bioaugmentation treatment microcosms, respectively. The half-life time in the biostimulation treatment microcosm was increased with a range between 10.7 and 39.2 days by the presence of different initial lead concentration. The results have promising potential for effective remediation of soils co-contaminated with hydrocarbons and heavy metals.     

Chemical Constituents and Anti-Gastric Ulcer Activity of Essential Oils of Alpinia officinarum (Zingiberaceae), Cyperus rotundus (Cyperaceae), and Their Herbal Pair

The essential oil (EO) of the herbal pair (HP), Alpinia officinarum-Cyperus rotundus (HP G-X) has been conventionally used in traditional Chinese medicine (TCM) for ‘warming the stomach’ and relieving pain. However, its pharmacologically active compounds, as well as the mechanism of its anti-gastric ulcer properties remain unclear. In this study, the EOs obtained from HP G-X and its corresponding single herbs were analyzed using GC/MS. A total of 74, 56, and 85 compounds were detected in A. officinarum (GLJ), C. rotundus (XF), and HP G-X, accounting for 93.2 %, 89.5 %, and 92.0 % of the total content, respectively. GLJ mainly contains 1,8-cineol (22.0 %) and α-terpineol (11.8 %), whereas cyperenone (22.4 %) and cyperene (12.3 %) were the major constituents in XF. These four compounds were also detected in the HP G-X with relatively high composition as 11.8 %, 5.5 %, 11.8 %, and 10.6 %, respectively. Although no new compounds were detected in HP G-X, the relative concentration of some compounds increased, while others decreased or even disappeared. HP G-X showed the lowest toxicity (TC₅₀ >800 μg/mL) against human gastric mucosal epithelial cells (GES-1) and had the best protective effect against ethanol-induced GES-1 cell damage compared to the individual herbs. In vitro studies demonstrated that HP G-X and the corresponding single herbs significantly reduced IL-6, TNF-α, and COX-2. In addition, in vivo investigations indicated that HP G-X can protect the gastric mucosa of mice from ethanol-induced damage by inhibiting the inflammatory reaction and providing analgesia. It can also inhibit the expression of NF-κBp65, COX-2, and TRPV1 protein, reduce the concentrations of IL-6 and TNF-α, and relieve heat-induced pain. This study further substantiated the traditional application of HP G-X against gastric ulcers through both in vivo and in vitro investigations.     

Use of Rice Husk as Bulking Agent in Bioremediation of Automobile Gas Oil Impinged Agricultural Soil

Evaluation of rice husk (RH) as bulking agent in bioremediation of automobile gas oil (AGO) hydrocarbon polluted agricultural soil using renewal by enhanced natural attenuation (RENA) as control was the subject of the present investigation. The effect of different parameters such as total petroleum hydrocarbon (TPH), dehydrogenase activity (DHA), optical density and pH on bioremediation performance were evaluated. The studied parameters such as microbial dynamics, percentage degradation and DHA were found to be higher in RH-amended system and differed significantly with control at P < 0.05. RH resulted in high removal efficiency of 97.85 ± 0.93% under a two-month incubation period, while RENA had lesser removal efficiency of 53.15 ± 3.81%. Overall hydrocarbon biodegradation proceeded very slowly in the RENA particularly from week 0 to 4. Experimental data perfectly fitted into the first-order kinetic and generated high r² values (0.945), first-order degradation constant (0.47 day^?1), and shorter degradation half-life (1.50 d)—t_1/2 = Ln2/K and Ln2 numerically equals to 0.693 and hence written as 0.693/K. Micrococcus luteus and Rhizopus arrhizus were isolated in the present study, which displayed extreme AGO hydrocarbon biodegradative abilities. The use of RH in hydrocarbon-polluted soil significantly increased biodegradation rate and resulted in effective AGO cleanup within 2 months period. Therefore, RH provides an alternative source of bioremediation material in field application for abundant petroleum hydrocarbon soil pollution.     

Bioremediation and phytoremediation of total petroleum hydrocarbons (TPH) under various conditions

Remediation of contaminated soils is often studied using fine-textured soils rather than low-fertility sandy soils, and few studies focus on recontamination events. This study compared aerobic and anaerobic treatments for remediation of freshly introduced used motor oil on a sandy soil previously phytoremediated and bioacclimated (microorganisms already adapted in the soil environment) with some residual total petroleum hydrocarbon (TPH) contamination. Vegetated and unvegetated conditions to remediate anthropogenic fill containing residual TPH that was spiked with nonaqueous phase liquids (NAPLs) were evaluated in a 90-day greenhouse pot study. Vegetated treatments used switchgrass (Panicum virgatum). The concentration of aerobic bacteria were orders of magnitude higher in vegetated treatments compared to unvegetated. Nevertheless, final TPH concentrations were low in all saturated soil treatments, and high in the presence of switchgrass. Concentrations were also low in unvegetated pots with fertilizer. Acclimated indigenous microbial communities were shown to be more effective in breaking down hydrocarbons than introducing microbes from the addition of plant treatments in sandy soils. Remediation of fresh introduced NAPLs on pre-phytoremediated and bioacclimated soil was most efficient in saturated, anaerobic environments, probably due to the already pre-established microbial associations, easily bioavailable contaminants, and optimized soil conditions for microbial establishment and survival.     

Phytoremediation of Total Petroleum Hydrocarbons From Highly Saline and Clay Soil Using Sorghum halepense (L.) Pers. and Aeluropus littoralis (Guna) Parl

This study evaluated the effects of native plants (Sorghum halepense and Aeluropus littoralis), total petroleum hydrocarbons (TPH) concentrations, and nutrients on the removal of TPHs from a highly saline clay soil. For a period of 180 days, rhizosphere microbial number, plant biomass, and residual TPHs were determined monthly. Results showed that TPH removal from soil in the rhizosphere was 13% higher than that in the control (unplanted soil). In addition, the number of heterotrophic bacteria in the rhizosphere and non-rhizosphere soil was 7.407 and 6.629 log₁₀CFU/g, respectively. The maximum TPH removal, microbial numbers, and plant biomass were measured in the treated soil, polluted with 0.86% (w/w) of TPH. The high clay and salinity of the experimental soil had a negative effect on the phytoremediation efficiency. Hence, it was necessary to improve the physicochemical properties of the soil to provide a good condition for plants and microbes, thereby increasing the phytoremediation efficiency.     

Microbial Activity and Community Composition during Bioremediation of Diesel-Oil-Contaminated Soil: Effects of Hydrocarbon Concentration,Fertilizers, and Incubation Time

We investigated the influence of three factors—diesel oil concentration [2500, 5000, 10,000, 20,000 mg total petroleum hydrocarbons (TPH) kg⁻¹ soil], biostimulation (unfertilized, inorganic fertilization with NPK nutrients, or oleophilic fertilization with Inipol EAP22), and incubation time—on hydrocarbon removal, enzyme activity (lipase), and microbial community structure [phospholipid fatty acids (PLFA)] in a laboratory soil bioremediation treatment. Fertilization enhanced TPH removal and lipase activity significantly (P ≤ 0.001). The higher the initial contamination, the more marked was the effect of fertilization. Differences between the two fertilizers were not significant (P > 0.05). Microbial communities, as assessed by PLFA patterns, were primarily influenced by the TPH content, followed by fertilization, and the interaction of these two factors, whereas incubation time was of minor importance. This was demonstrated by three-factorial analysis of variance and multidimensional scaling analysis. Low TPH content had no significant effect on soil microbial community, independent of the treatment. High TPH content generally resulted in increased PLFA concentrations, whereby a significant increase in microbial biomass with time was only observed with inorganic fertilization, whereas oleophilic fertilization (Inipol EAP22) tended to inhibit microbial activity and to reduce PLFA contents with time. Among bacteria, PLFA indicative of the Gram-negative population were significantly (P ≤ 0.05) increased in soil samples containing high amounts of diesel oil and fertilized with NPK after 21–38 days of incubation at 20°C. The Gram-positive population was not significantly influenced by TPH content or biostimulation treatment.     

Using Maize (Zea mays L.) and Sewage Sludge to Remediate a Petroleum-Contaminated Calcareous Soil

Phyto-stimulation, the use of plants to stimulate activity of microorganisms in a root zone, has been proposed as an approach to promote the degradation of petroleum hydrocarbons and thus the remediation of petroleum-polluted soils. In this study, we investigated the potential use of sewage sludge to enhance phyto-stimulating effects of maize (Zea mays L.) on the elimination of an aged petroleum contamination in a calcareous soil. In a pot experiment, maize was grown on the experimental soil for two months at three levels of sewage sludge application (0, 20, and 50 g dry matter of sludge per kg soil). The amendments increased root and shoot growth of the experimental plants approximately by a factor of two at the lower sludge treatment level and by a factor of five at the higher sludge treatment level. In a separate incubation experiment, sludge application also led to an immediate stimulation of soil respiration, which then further increased over time. The initial stimulation was three times larger at the higher than at the lower treatment level, but the rate of subsequent increase was similar in both treatments. The two sludge treatments also accelerated TPH elimination in the contaminated soil, and again the effect was approximately three times stronger at the higher than at the lower treatment level. The sludge effect on TPH elimination was much stronger than the effect of the plants. More than half of the initial contamination was reduced in combined treatment with maize and sludge application at the highest rate. The results show that sewage sludge can substantially enhance the remediation of petroleum-contaminated soil, especially when applied in conjunction with a suitable plant such as maize.     

Assessment of hydrocarbon degradation potentials in a plant–microbe interaction system with oil sludge contamination: A sustainable solution

A pot culture experiment was conducted for 90 days for the evaluation of oil and total petroleum hydrocarbon (TPH) degradation in vegetated and non-vegetated treatments of real-field oil-sludge-contaminated soil. Five different treatments include (T1) control, 2% oil-sludge-contaminated soil; (T2), augmentation of microbial consortium; (T3), Vertiveria zizanioides; (T4), bio-augmentation along with V. zizanioides; and (T5), bio-augmentation with V. zizanioides and bulking agent. During the study, oil reduction, TPH, and degradation of its fractions were determined. Physico-chemical and microbiological parameters of soil were also monitored simultaneously. At the end of the experimental period, oil content (85%) was reduced maximally in bio-augmented rhizospheric treatments (T4 and T5) as compared to control (27%). TPH reduction was observed to be 88 and 89% in bio-augmented rhizospheric soil (T4 and T5 treatments), whereas in non-rhizospheric and control (T2 and T1), TPH reduction was 78 and 37%, respectively. Degradation of aromatic fraction after 90 days in bio-augmented rhizosphere of treatments T4 and T5 was found to 91 and 92%, respectively. In microbial (T2) and Vertiveria treatments (T3), degradation of aromatic fraction was 83 and 68%, respectively. A threefold increase in soil dehydrogenase activity and noticeable changes in organic carbon content and water-holding capacity were also observed which indicated maximum degradation of oil and its fractions in combined treatment of plants and microbes. It is concluded that the plant–microbe soil system helps to restore soil quality and can be used as an effective tool for the remediation of oil-sludge-contaminated sites.