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 bla_CTX, 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, bla_CTX, bla_TEM 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.

Cedrus deodara is economically and ethnobotanically an important forest tree and is shown to be at decline in Northern areas of Pakistan in recent years mainly due to high concentration of Nitrogen in forests. Ectomycorrhizal (ECM) association forming fungi enables the forest trees to develop optimally by absorbing water from the rhizosphere through their absorptive hyphae and by making available the nutrients by mobilization of N and P from the organic substrates. This study was conducted to identify the ECM strains from C. deodara rhizosphere and to analyse the impact of high N load on the C. deodara seedlings to establish N critical load value for coniferous forests of Pakistan. Six new fungal strains were identified from the rhizosphere of C. deodara and were registered at GenBank (NCBI) as Emmia latemarginata strain ACE1, Aspergillus terreus strain ACE2, Purpureocillium lilacinum strain ACE3, Talaromyces pinophilus strain ACE4, A. fumigatus strain ACE5 and T. pinophilus strain ACE6 with accession numbers MH145426, MH145427, MH145428, MH145429, MH145430 and MH547115. Four out of six isolated strains were inoculated with seedlings of C. deodara singly and in consortium (CN) in combination with nitrogen load of 0 (C), 25 (T1), 50 (T2), 100 kg N ha^-1 yr^-1 (T3). Agronomic, physiological and gene expression studies for ExpansinA4 (EXPA4) and Cystatins (Cys) were made to analyse the impact of fungal strains in relation to high N stress. This study suggests a positive impact of T1 (25 kg N ha^-1 yr^-1) Nitrogen load and a negative impact of T3 (100 kg N ha^-1 yr^-1) on growth parameters and expression patterns of EXPA4 and Cys genes. Peroxidase (POX) activity decreased in the order ACE5 > ACE2 > C > ACE3 > ACE1 > CN. However, the results of Superoxide dismutase (SOD) showed decreasing trend in the order ACE5 > C > CN > ACE1 > ACE2 > ACE3. Strain ACE3 was shown to have a positive impact on the seedlings in terms of growth, physiology and expression of genes. Present study suggests that newly identified fungal strains showing positive impact on the growth and physiology of C. deodara could be used for the propagation of this economically important plant in Pakistan after pathogenicity test.

Plants evolve complex interactions with diverse soil mutualist organisms to enhance P mobilization from the soil. These strategies are particularly important when P is poorly available. It is still unclear how the soil P source (e.g., mineral P versus recalcitrant organic P) and its mobility in the soil (high or low) affect soil mutualist biological (ectomycorrhizal fungi, bacteria and bacterial-feeding nematodes) richness—plant P acquisition relationships. Using a set of six microcosm experiments conducted in growth chamber across contrasting P situations, we tested the hypothesis that the relationship between the increasing addition of soil mutualist organisms in the rhizosphere of the plant and plant P acquisition depends on P source and mobility. The highest correlation (R² = 0.70) between plant P acquisition with soil rhizosphere biological richness was found in a high P-sorbing soil amended with an organic P source. In the five other situations, the relationships became significant either in soil conditions, with or without mineral P addition, or when the P source was supplied as organic P in the absence of soil, although with a low correlation coefficient (0.09 < R² < 0.15). We thus encourage the systematic and careful consideration of the form and mobility of P in the experimental trials that aim to assess the role of biological complexity on plant P nutrition.

Lignocellulosic biomass needs attention as an alternative energy source to overcome the adverse impacts of fossil fuels. Diversified Galiyat forests of Lower Himalaya may represent the potential source of lignocellulose degrading microbiota, particularly the lytic bacteria. Therefore, soil and wood samples were collected from different sites of Nathiagali and Thandiani of Galiyat forests. The soil samples collected were clayey, with a pH between 6.7 and 7.0, and with an organic matter of 2.8%–2.9% in Nathiagali and 2.1%–2.2% in Thandiani. The soils were enriched with more diversified cultivable bacteria (9 Log CFU/g) than the respective wood samples (7.4–8.6 Log CFU/g). Out of 90 bacteria, 22 isolates were efficient for cellulose degradation, 14 for xylanase activity, and 10 for laccase production. Cluster analysis showed that lignocellulolytic bacteria were grouped based on the sample medium (soil–wood) rather than the sampling site (Thandiani–Nathiagali). Efficient bacteria were also sequenced, and we found that cellulase production was prevalent in Pseudomonas spp. while laccase activity was diverse among taxonomically varied bacteria. Moreover, Stenotrophomonas sp. TS2B1 performed the best for corncob xylose degradation. Overall, the results suggest that Galiyat forests represent diverse lignocellulolytic microbial populations which should be further evaluated for applications in lignocellulosic waste management and for potential consequent fuel production.

Functioning of ecosystems depends on the nutrient dynamics across trophic levels, largely mediated by microbial interactions in the soil food web. The present study investigated the use of phosphate solubilizing bacteria (PSB) and poultry manure (PM) for maintaining labile P in the soil for an extensive fertility enhancement and as a substitution of chemical fertilizers. Based on the different P solubilizing capabilities of Bacillus and Pseudomonas, a quadruple consortium of Bacillus subtilis, Bacillus cereus, Bacillus thuringiensis and Pseudomonas fluorescens, and their grazer nematodes (soil free living) supplemented with PM were studied. This study was carried out on the trophic levels of soil communities to assess the growth and availability of P to the wheat plants. Experiment was performed for 90 days. Comparing the unamended and amended predator results showed that nematode addition beyond bacterial treatment substantially increased the net available P by ≈2 times, and alkaline phosphatase (ALP) activity by 3.3 times. These results demonstrated the nematodes association with increasing nutrient availability or P mineralization. The interactive effect of PM as substrate and biological drivers was more noticeable on plant dry biomass (1.6 times) and plant P concentration (3.5times) compared to the similar unamended treatment. It is concluded that the biological drivers significantly enhanced the soil ALP and available P while the substrate and biological drivers enhanced dry biomass and plant P concentration. Bacterivore nematodes enhanced the effect of PSB for P mineralization via microbial loop and could be used for the enhancement of wheat production.