Unraveling the impact of human perturbation on nitrogen cycling in terrestrial ecosystems of lower Himalaya, Pakistan.

Terrestrial ecosystems are under the enormous pressure of land use management regimes through human disturbances, resulting in the disruption of biogeochemical cycles and associated ecosystem services. Nitrogen (N) in soil ecosystems is of vital importance for primary productivity, hence estimating the extent of these human interventions on N-cycling processes becomes imperative from economic and environmental perspectives. This work investigated the impacts of variable anthropogenic activities on N cycling in three different terrestrial ecosystems (arable, grassland, and forest) in three regions of lower Himalaya, Pakistan. Potential nitrification (PNA) and denitrification (DEA) enzyme activities, relative distribution of inorganic N species (NH4, NO3), and the role of inherent edaphic factors were assessed. Results revealed high nitrification potentials and increased nitrous oxide (N2O) emissions in the incubated soil microcosms, in the order as arable > grassland > forest ecosystems. Notably, higher rates of both studied processes (~ 30-50%) and elevated soil mineral nitrogen pool were observed in arable ecosystems. Forest soils, assumed as pristine ecosystems relying mainly on natural N fixation, produced (de)nitrification rates relatively lower than grasslands, followed by arable soils which were moderately disturbed through long-term fertilization and intensive land-use regimes. Linear regression modeling revealed that the inorganic N species (particularly NO3), and inherent edaphic factors were the key determinants of high (de)nitrification rates, hence warn of accelerated N losses in these ecosystems. The study highlights that elevated PNA and DEA being proxies for the altered N cycling in the studied terrestrial ecosystems are of great ecological relevance in view of predicted N2O budget in the lower Himalaya.

Gene expression and biochemical response of giant reed under Ni and Cu stress.

Giant reed (Arundo donax) has proved to be effective in detoxification, accumulation and tolerance of toxic metals. The present study explored the stress response of giant reed against Cu and Ni stress. The effect of metal stress was studied on dry weight, chlorophyll pigments antioxidant enzymes production and selected genes expression. The accumulation of heavy metals increased in a concentration-dependent manner and depicted toxicity symptoms in leaves beyond 75 mg/L of Cu or Ni. Oxidative stress was evident in giant reed under highest exposure of Ni and Cu which increased antioxidants activities (SOD, POD and CAT). It was observed that metal transport and detoxification were possible due to the expression of glutathione reductase, Natural Resistance-Associated Macrophage Protein (NRAMP) and Yellow Stripe-Like (YSL) genes. These insights into the genetic basis of a successful remediating plant species will be useful in understanding heavy metals tolerance in giant reed.

Assessment of flood-induced changes in soil heavy metal and nutrient status in Rajanpur, Pakistan.

Flood events around the globe have severely impaired the soil functioning resulting in compromised food security in several parts of the world. The current study was aimed to explore the impacts of floods on soil heavy metals and nutrients status at three locations; Tibbi Solgi (TS), Vinri Khosa (VK), and Noshehra West (NW-control) in the district Rajanpur of Punjab, Pakistan. TS and VK sites were under regular influence of flooding over the last many years, but no flood event was reported on NW site during the same tenure; hence, it served as control. Sampling was carried out before and after flooding on the experimental sites. Vegetation cover was monitored through remote sensing techniques. Results revealed varying effects of floods on soil heavy metals; Cd, Cr, Pb, and soil phosphorous and nitrates. Flood events increased the Cd while lowered Pb concentration at VK site; however, flooding did not influence the status of Cr in soil. Similar to the trend observed in case of Cd, soil phosphorous and nitrates were reduced after flood events. Correlation analyses of soil physicochemical properties with soil heavy metals and nutrients indicated that after flood events, soil texture and organic carbon content seem to be the major factors driving the shift in soil heavy metals and nutrient concentrations. Although pollution indices indicated a marginally low contamination levels, but as projected in empirical studies, regular flood events in the studied sites may contaminate the whole ecosystem rendering it unfit for agricultural productivity.

Efficiency of various sewage sludges and their biochars in improving selected soil properties and growth of wheat (Triticum aestivum).

Due to increasing demand of P fertilizers and gradual decrease in P resources, recyclable P is the focus of researchers in recent years. Sewage sludge (SS) is a municipal waste that contains appreciable amounts of P and probably other nutrients. In present study, the effects of various SS and their biochars (450 °C for 2 h) were investigated on soil properties and P uptake in wheat (Triticum aestivum) with and without P fertilizer. The biomass of plants and grain yield were significantly increased with application of SS and their biochars as compared to the control treatment either without or with P application. Moreover, there was significant interaction between treatments and P application for the concentration of K, and P in shoots and roots of wheat. Shoot P concentration was not significantly affected with SS than biochars whereas root P concentration was higher in SS treatments than respective biochars. Higher increase in Olsen's P concentration was observed in populated area sludge applied-soil as compared to disposal sludge and their biochars. Overall, it is observed that SS application increased the wheat yield and P concentrations in plants than control depending upon SS types whereas biochar application decreased the P concentration in roots. Grain yield and P concentration in shoots were not significantly affected for the treatment with P fertilizers than without P. Sewage sludge and their biochars might be a potential source of P but further research is needed to recommend the use of modified SS-biochars as source of available P for crops.

Fractionation and Characterization of Metallic Elements in Soils in Land Use Systems.

Land use has a great impact on soil dynamics. The soils of various land use systems in Central Karakoram have been under immense pressure in the recent past due to certain anthropogenic activities such as land use practices and land use cover changes. These influences have an impact on the spatial distribution of metallic elements (MEs) in the soils of various land uses. Herein, we investigated the occurrence of the MEs, copper (Cu), zinc (Zn), and nickel (Ni), in soils of various land uses such as the permafrost, pasture, forest, and agricultural lands of the Central Karakorum region. The MEs were extracted in exchangeable, adsorbed, organically bound, carbonated, precipitated, and residual forms. The concentrations of MEs showed a significant dependence on the extraction method used, and the extraction trend followed the order of EDTA > HNO3 > KNO3 > NaOH > H2O. Zn showed the highest concentration compared to Ni and Cu in all extractions, whereas the land uses' ME concentration followed the order of agricultural land > permafrost > forest > pasturelands. The highest values of total Zn, Ni, and Cu were 712 ± 01 mg/kg, 656 ± 02 mg/kg, and 163 ± 02 mg/kg, respectively, in agricultural soil. The ME concentration showed significant variations between different land uses, and the highest concentration was noted in agricultural soil. Zn was found to be a dominant ME compared to Ni and Cu. We believe this effort will provide opportunities for scholars to investigate MEs around the globe.

Improved photocatalytic decolorization of reactive black 5 dye through synthesis of graphene quantum dots-nitrogen-doped TiO2.

Graphene quantum dots (GQDs), a new solid-state electron transfer material was anchored to nitrogen-doped TiO2 via sol gel method. The introduction of GQDs effectively extended light absorption of TiO2 from UV to visible region. GQD-N-TiO2 demonstrated lower PL intensity at excitation wavelengths of 320 to 450 nm confirming enhanced exciton lifespan. GQD-N-TiO2-300 revealed higher surface area (191.91m2 g-1), pore diameter (1.94 nm), TEM particle size distribution (4.88 ± 1.26 nm) with lattice spacing of 0.45 nm and bandgap (2.91 eV). In addition, GQDs incorporation shifted XPS spectrum of Ti 2p to lower binding energy level (458.36 eV), while substitution of oxygen sites in TiO2 lattice by carbon were confirmed through deconvolution of C 1 s spectrum. Photocatalytic reaction followed the pseudo first order reaction and continuous reductions in apparent rate constant (Kapp) with incremental increase in RB5 concentration. Langmuir-Hinshelwood model showed surface reaction rate constants KC = 1.95 mg L-1 min-1 and KLH = 0.76 L mg-1. The active species trapping, and mechanism studies indicated the photocatalytic decolorization of RB5 through GQD-N-TiO2 was governed by type II heterojunction. Overall, the photodecolorization reactions were triggered by the formation of holes and reactive oxygen species. The presence of •OH, 1O2, and O2• during the photocatalytic process were confirmed through EPR analysis. The excellent photocatalytic decolorization of the synthesized nanocomposite against RB5 can be ascribed to the presence of GQDs in the TiO2 lattice that acted as excellent electron transporter and photosensitizer. This study provides a basis for using nonmetal, abundant, and benign materials like graphene quantum dots to enhance the TiO2 photocatalytic efficiency, opening new possibilities for environmental applications.

Novel microalgae strains from selected lower Himalayan aquatic habitats as potential sources of green products.

Microalgal biomass provides a renewable source of biofuels and other green products. However, in order to realize economically viable microalgal biorefinery, strategic identification and utilization of suitable microalgal feedstock is fundamental. Here, a multi-step suboptimal screening strategy was used to target promising microalgae strains from selected freshwaters of the study area. The resulting strains were found to be affiliated to seven closely-related genera of the family Scenedesmaceae, as revealed by both morphologic and molecular characterization. Following initial screening under upper psychrophilic to optimum mesophilic (irregular temperature of 14.1 to 35.9°C) cultivation conditions, superior strains were chosen for further studies. Further cultivation of the selected strains under moderate to extreme mesophilic cultivation conditions (irregular temperature of 25.7 to 42.2°C), yielded up to 74.12 mgL-1day-1, 19.96 mgL-1day-1, 48.56%, 3.34 µg/mL and 1.20 µg/mL, for biomass productivity, lipid productivity, carbohydrate content, pigments content and carotenoids content respectively. These performances were deemed promising compared with some previous, optimum conditions-based reports. Interestingly, the fatty acids profile and the high carotenoids content of the studied strains revealed possible tolerance to the stress caused by the changing suboptimal cultivation conditions. Overall, strains AY1, CM6, LY2 and KL10 were exceptional and may present sustainable, promising feedstock for utilization in large-scale generation of green products, including biodiesel, bioethanol, pigments and dietary supplements. The findings of this study, which exposed promising, eurythermal strains, would expand the current knowledge on the search for promising microalgae strains capable of performing under the largely uncontrolled large-scale cultivation settings.

Potential of nanocomposites of zero valent copper and magnetite with Eleocharis dulcis biochar for packed column and batch scale removal of Congo red dye.

The current study is the first attempt to prepare nanocomposites of Eleocharis dulcis biochar (EDB) with nano zero-valent Copper (nZVCu/EDB) and magnetite nanoparticles (MNPs/EDB) for batch and column scale sequestration of Congo Red dye (CR) from synthetic and natural water. The adsorbents were characterized with advanced analytical techniques. The impact of EDB, MNPs/EDB and nZVCu/EDB dosage (1-4 g/L), pH (4-10), initial concentration of CR (20-500 mg/L), interaction time (180 min) and material type to remove CR from water was examined at ambient temperature. The CR removal followed sequence of nZVCu/EDB > MNPs/EDB > EDB (84.9-98% > 77-95% > 69.5-93%) at dosage 2 g/L when CR concentration was increased from 20 to 500 mg/L. The MNPs/EDB and nZVCu/EDB showed 10.9% and 20.1% higher CR removal than EDB. The adsorption capacity of nZVCu/EDB, MNPs/EDB and EDB was 212, 193 and 174 mg/g, respectively. Freundlich model proved more suitable for sorption experiments while pseudo 2nd order kinetic model well explained the adsorption kinetics. Fixed bed column scale results revealed excellent retention of CR (99%) even at 500 mg/L till 2 h when packed column was filled with 3.0 g nZVCu/EDB, MNPs/EDB and EDB. These results revealed that nanocomposites with biochar can be applied efficiently for the decontamination of CR contaminated water.

Green synthesis and characterization of silver nanoparticles from Acacia nilotica and their anticancer, antidiabetic and antioxidant efficacy.

Both cancer and diabetes mellitus are serious health issues, accounting more than 11 million deaths worldwide annually. Targeted use of plant-mediated nanoparticles (NPs) in treatment of ailments has outstanding results due to their salient properties. The current study was designed to investigate the safe production of silver nanoparticles (AgNPs) from Acacia nilotica. Different concentrations of AgNO3 were tested to optimize the protocol for the synthesis of AgNPs from the bark extract. It was demonstrated that 0.1 M and 3 mM were found to be the optimum concentrations for the synthesis of AgNPs. Standard characterization techniques such as UV-vis spectrophotometry, SEM, SEM-EDX micrograph, spot analysis, elemental mapping and XRD were used for the conformation of biosynthesis of AgNPs. Absorption spectrum of plant-mediated AgNPs under UV-vis spectrophotometer showed a strong peak at 380 nm and 420 nm for AgNPs synthesized at 0.1 M and 3 mM concentration of salt. The SEM results showed that AgNPs were present in variable shapes within average particle size ranging from (20-50 nm). Anticancer, antidiabetic and antioxidant potential of green AgNPs was investigated and they showed promising results as compared to the positive and negative controls. Hence, AgNPs were found potent therapeutic agent against the human liver cancer cell lines (HepG2), strong inhibitor for α-glucosidase enzyme activity and scavenging agent against free radicals that cause oxidative stress. Further studies are however needed to confirm the molecular mechanism and biochemical reactions responsible for the anticancer and antidiabetic activities of the particles.

Removal of Antibiotic Resistance Genes, Class 1 Integrase Gene and Escherichia coli Indicator Gene in a Microalgae-Based Wastewater Treatment System.

Microalgae-based wastewater treatment systems (AWWTS) have recently shown promise in the mitigation of antibiotic resistance genes (ARGs) from municipal wastewater (MWW). However, due to the large number of ARGs that exist in MWW, the use of indirect conventional water quality parameters to monitor ARGs reduction in wastewater would make the process less burdensome and economically affordable. In order to establish a robust relationship between the ARGs and water quality parameters, the current study employed different microalgae strains in monoculture (CM2, KL10) and multi-species combinations (CK and WW) for the MWW treatment under outdoor environmental conditions. The studied genes were quantified in the MWW influents and effluents using real-time PCR. All the cultures substantially improved the physicochemical qualities of the MWW. Out of the 14 genes analyzed in this study, tetO, tetW, tetX and ermB were decreased beyond detection within the first 4 days of treatment in all the cultures. Other genes, including blaCTX, sul1, cmlA, aadA, int1 and uidA were also decreased beyond a 2 log reduction value (LRV). The mobile genetic element, int1, correlated positively with most of the ARGs, especially sul1 (r ≤ 0.99, p < 0.01) and aadA (r ≤ 0.97, p < 0.01). Similarly, the Escherichia coli indicator gene, uidA, correlated positively with the studied genes, especially with aadA, blaCTX, blaTEM and cmlA (r ≤ 0.99 for each, p < 0.01). Some of the studied genes also correlated positively with total dissolved solids (TDS) (r ≤ 0.98, p < 0.01), and/or negatively with total suspended solids (TSS) (r ≤ −0.98, p < 0.01) and pH (r ≤ −0.98, p < 0.01). Among the tested cultures, both monocultures, i.e., KL10 and CM2 were found to be more consistent in gene suppression than their multi-species counterparts. The findings revealed water quality parameters such as TDS, TSS and E. coli as reliable proxies for ARGs mitigation in AWWTS and further highlight the superiority of monocultures over multi-species cultures in terms of gene suppression from the MWW stream.

Role of Wheat Phosphorus Starvation Tolerance 1 Genes in Phosphorus Acquisition and Root Architecture.

The wheat plant requires elevated phosphorus levels for its normal growth and yield, but continuously depleting non-renewable phosphorus reserves in the soil is one of the biggest challenges in agricultural production worldwide. The Phosphorus Starvation Tolerance 1 (PSTOL1) gene has been reported to play a key role in efficient P uptake, deeper rooting, and high yield in rice. However, the function of the PSTOL1 gene in wheat is still unclear. In this study, a total of 22 PSTOL1 orthologs were identified in the wheat genome, and found that wheat PSTOL1 orthologs are unevenly distributed on chromosomes, and these genes were under strong purifying selection. Under different phosphorus regimes, wheat PSTOL1 genes showed differential expression patterns in different tissues. These results strengthen the classification of Pakistan-13 as a P-efficient cultivar and Shafaq-06 as a P-inefficient cultivar. Phenotypic characterization demonstrated that Pakistan-13 wheat cultivar has significantly increased P uptake, root length, root volume, and root surface area compared to Shafaq-06. Some wheat PSTOL1 orthologs are co-localized with phosphorus starvation's related quantitative trait loci (QTLs), suggesting their potential role in phosphorus use efficiency. Altogether, these results highlight the role of the wheat PSTOL1 genes in wheat P uptake, root architecture, and efficient plant growth. This comprehensive study will be helpful for devising sustainable strategies for wheat crop production and adaptation to phosphorus insufficiency.

Alleviation of cadmium (Cd) toxicity and minimizing its uptake in wheat (Triticum aestivum) by using organic carbon sources in Cd-spiked soil.

Cadmium (Cd)-contamination of agricultural soils has been receiving attention worldwide due to its entry into food crops such as wheat (Triticum aestivum L.). Little is known regarding the use of organic carbon (OC) sources in alleviating Cd toxicity in cereals. The current experiment was aimed to study the effects of different OC sources on the Cd accumulation by wheat. A pot study was conducted to determine the effects of rice husk biochar (RHB), farmyard manure (FYM), and lignite (LT) either alone or in combination on crop growth, Cd bioavailability and health risk assessment. The results proved that the application of OC sources like RHB, FYM, and LT either alone or in combination were highly effective in enhancing the wheat growth and yield as well as in minimizing the phyto-available fraction of Cd and its transfer to edible tissue of wheat. The RHB was the most efficient source in enhancing the plant growth and reducing the Cd concentration in wheat tissues. RHB increased grain yield by 91% and decreased Cd concentration in shoot, roots, grains, and bioavailable fraction of Cd by 67, 69, 62.5, and 74% than control, respectively. The RHB reduced the daily Cd uptake and health risk index in adults in comparison to control. Overall, where un-amended soil resulted in diminished plant productivity, the application of other OC sources also significantly proved their potential to enhance the dry weight and grain yield, suggesting that these OC sources may be used aiming to minimize the Cd concentration in crops. However, there is still a need to explore the potential of different OC sources in combination with other frequently available amendments for their large scale implementation in metal-contaminated soils.

Biochar soil amendment on alleviation of drought and salt stress in plants: a critical review.

Drought and salt stress negatively affect soil fertility and plant growth. Application of biochar, carbon-rich material developed from combustion of biomass under no or limited oxygen supply, ameliorates the negative effects of drought and salt stress on plants. The biochar application increased the plant growth, biomass, and yield under either drought and/or salt stress and also increased photosynthesis, nutrient uptake, and modified gas exchange characteristics in drought and salt-stressed plants. Under drought stress, biochar increased the water holding capacity of soil and improved the physical and biological properties of soils. Under salt stress, biochar decreased Na+ uptake, while increased K+ uptake by plants. Biochar-mediated increase in salt tolerance of plants is primarily associated with improvement in soil properties, thus increasing plant water status, reduction of Na+ uptake, increasing uptake of minerals, and regulation of stomatal conductance and phytohormones. This review highlights both the potential of biochar in alleviating drought and salt stress in plants and future prospect of the role of biochar under drought and salt stress in plants.

Oil industry waste: a potential feedstock for biodiesel production.

The worldwide rising energy demands and the concerns about the sustainability of fossil fuels have led to the search for some low-cost renewable fuels. In this scenario, the production of biodiesel from various vegetable and animal sources has attracted worldwide attention. The present study was conducted to evaluate the production of biodiesel from the oil industry waste following base-catalysed transesterification. The transesterification reaction gave a yield of 83.7% by 6:1 methanol/oil molar ratio, at 60°C over 80 min of reaction time in the presence of NaOH. The gas chromatographic analysis of the product showed the presence of 16 fatty acid methyl esters with linoleic and oleic acid as principal components representing about 31% and 20.7% of the total methyl esters, respectively. The fourier transform infrared spectroscopy spectrum of oil industry waste and transesterified product further confirmed the formation of methyl esters. Furthermore, the fuel properties of oil industry waste methyl esters, such as kinematic viscosity, cetane number, cloud point, pour point, flash point, acid value, sulphur content, cold filter plugging point, copper strip corrosion, density, oxidative stability, higher heating values, ash content, water content, methanol content and total glycerol content, were determined and discussed in the light of ASTM D6751 and EN 14214 biodiesel standards. Overall, this study presents the production of biodiesel from the oil industry waste as an approach of recycling this waste into value-added products.

Cadmium stress in rice: toxic effects, tolerance mechanisms, and management: a critical review.

Cadmium (Cd) is one of the main pollutants in paddy fields, and its accumulation in rice (Oryza sativa L.) and subsequent transfer to food chain is a global environmental issue. This paper reviews the toxic effects, tolerance mechanisms, and management of Cd in a rice paddy. Cadmium toxicity decreases seed germination, growth, mineral nutrients, photosynthesis, and grain yield. It also causes oxidative stress and genotoxicity in rice. Plant response to Cd toxicity varies with cultivars, growth condition, and duration of Cd exposure. Under Cd stress, stimulation of antioxidant defense system, osmoregulation, ion homeostasis, and over production of signaling molecules are important tolerance mechanisms in rice. Several strategies have been proposed for the management of Cd-contaminated paddy soils. One such approach is the exogenous application of hormones, osmolytes, and signaling molecules. Moreover, Cd uptake and toxicity in rice can be decreased by proper application of essential nutrients such as nitrogen, zinc, iron, and selenium in Cd-contaminated soils. In addition, several inorganic (liming and silicon) and organic (compost and biochar) amendments have been applied in the soils to reduce Cd stress in rice. Selection of low Cd-accumulating rice cultivars, crop rotation, water management, and exogenous application of microbes could be a reasonable approach to alleviate Cd toxicity in rice. To draw a sound conclusion, long-term field trials are still required, including risks and benefit analysis for various management strategies.

Distribution of bacteria and nitrogen-cycling microbial communities along constructed Technosol depth-profiles.

Technosol construction through assemblage of treated soil and recycled wastes is an innovative option for the restoration of degraded lands and re-use of industrial wastes. Recent studies have evidenced that Technosols could support soil functions such as primary production but the knowledge about other ecosystemic services, such as nutrient cycling, is limited. We investigated how the total bacterial community and key functional microbial communities involved in nitrogen cycling were influenced by the depth and type of Technosol. We found that despite being artificially constructed, Technosols exhibited a gradual change in microbial activity and abundance along the soil profile. Both nitrification and denitrification rates decreased with increasing depth as previously observed in other soil systems. Potential denitrification and nitrification were correlated with Technosol physico-chemical properties but also with abundances of nirS denitrifiers and bacterial amoA gene, respectively. The correlation between nitrification rates and bacterial ammonia-oxidizers suggests that bacteria are driving nitrification in Technosols. A-RISA fingerprints showed a distinct community structure along different Technosol layers. Technosol properties affected denitrification strongly than nitrification underlining the importance of better understanding the microbial communities in Technosols to maximize their potential for nutrient cycling, an essential ecosystem function.