Commercial Extraction Research Articles

Real-time monitoring of cattle (Bos taurus) tissue using a novel point-of-care (POC) polymerase spiral reaction (PSR) colorimetric assay

Adulteration of cattle meat with other species raises critical public health concerns, as existing detection methods lack sensitivity, specificity and practicality. We address this gap by introducing a novel Point-of-Care (POC) polymerase spiral reaction (PSR) assay. This rapid, on-site tool enables highly specific and sensitive identification of cattle tissue, even in resource-limited settings. This innovation paves the way for improved food safety and consumer protection. The cattle-specific PSR assay employs COX3 gene-targeting primers and a chromogenic indicator triad (HNB, Malachite Green and Phenol Red) for rapid visual detection. Cross-amplification absence with other meats (buffalo, sheep, goat and pig) and concordance with SYBR Green I and agarose gel electrophoresis confirm specificity. PSR at 62 °C for 60minutes exhibits a 100 fg detection limit and identifies 0.1% meat admixing. Efficacy is demonstrated across a diverse range of cattle meat samples, blind samples and processed cattle muscle samples. Compatibility with both conventional (snap chill) and commercial DNA extraction kits enables on-site analysis within 90minutes, facilitating rapid field sample analysis.

Inside the Atacama Desert: uncovering the living microbiome of an extreme environment.

The Atacama Desert in Chile is one of the driest and most inhospitable places on Earth. To analyze the diversity and distribution of microbial communities in such an environment, one of the most important and challenging steps is DNA extraction. Using commercial environmental DNA extraction protocols, a mixture of living, dormant, and dead cells of microorganisms is extracted, but separation of the different DNA pools is almost impossible. To overcome this problem, we applied a novel method on soils across a west-east moisture transect in the Atacama Desert to distinguish between extracellular DNA (eDNA) and intracellular DNA (iDNA) at the cell extraction level. Here, we show that a large number of living and potentially active microorganisms, such as Acidimicrobiia, Geodermatophilaceae, Frankiales, and Burkholderiaceae, occur in the hyperarid areas. We observed viable microorganisms involved as pioneers in initial soil formation processes, such as carbon and nitrogen fixation, as well as mineral-weathering processes. In response to various environmental stressors, microbes coexist as generalists or specialists in the desert soil environment. Our results show that specialists compete in a limited range of niches, while generalists tolerate a wider range of environmental conditions. Use of the DNA separation approach can provide new insights into different roles within viable microbial communities, especially in low-biomass environments where RNA-based analyses often fail.IMPORTANCEThe novel e- and iDNA separation technique offers insights into the living community at the cell extraction level in the hyperarid Atacama Desert. This approach provides a new framework for analyzing the composition and structure of the potentially active part of the microbial communities as well as their specialization, ecological network and community assembly process. Our findings underscore the significance of utilizing alternative genomic techniques in low-biomass environments where traditional DNA- and RNA-based analyses may not be feasible. The results demonstrate the viability of the proposed study framework and show that specialized microorganisms are important in initial soil formation processes, including microbial-driven mineral weathering, as well as the fixation of carbon and nitrogen.

Comparative evaluation of commercial DNA isolation approaches for nanopore-only bacterial genome assembly and plasmid recovery.

The advent of Oxford Nanopore Technologies has undergone significant improvements in terms of sequencing costs, accuracy, and sequencing read lengths, making it a cost-effective, and readily accessible approach for analyzing microbial genomes. A major challenge for bacterial whole genome sequencing by Nanopore technology is the requirement for a higher quality and quantity of high molecular weight DNA compared to short-read sequencing platforms. In this study, using eight pathogenic bacteria, we evaluated the quality, quantity, and fragmented size distribution of extracted DNA obtained from three different commercial DNA extraction kits, and one automated robotic platform. Our results demonstrated significant variation in DNA yield and purity among the extraction kits. The ZymoBIOMICS DNA Miniprep Kit (ZM) provided a higher purity of DNA compared to other kit-based extractions. All kit-based DNA extractions were successfully performed on all twenty-four samples using a single MinION flow cell, with the Nanobind CBB Big DNA kit (NB) yielding the longest raw reads. The Fire Monkey HMW-DNA Extraction Kit (FM) and the automated Roche MagNaPure 96 platform (RO) outperformed in genome assembly, particularly in gram-negative bacteria. Based on our finding, we recommend a minimum read coverage and raw read N50, obtained from the appropriate DNA extraction kit for each bacterial species, to optimize genome assembly and plasmid recovery. This approach will assist end-users in selecting the most effective kit-based extraction method for bacterial whole-genome assembly using only long-read nanopore sequences.

Alginate Formulation for Wound Healing Applications.

Significance: Alginate, sourced from seaweed, holds significant importance in industrial and biomedical domains due to its versatile properties. Its chemical composition, primarily comprising β-D-mannuronic acid and α-L-guluronic acid, governs its physical and biological attributes. This polysaccharide, extracted from brown algae and bacteria, offers diverse compositions impacting key factors such as molecular weight, flexibility, solubility, and stability. Recent Advances: Commercial extraction methods yield soluble sodium alginate essential for various biomedical applications. Extraction processes involve chemical treatments converting insoluble alginic acid salts into soluble forms. While biosynthesis pathways in bacteria and algae share similarities, differences in enzyme utilization and product characteristics are noted. Critical Issues: Despite its widespread applicability, challenges persist regarding alginate's stability, biodegradability, and bioactivity. Further understanding of its interactions in complex biological environments and the optimization of extraction and synthesis processes are imperative. Additionally, concerns regarding immune responses to alginate-based implants necessitate thorough investigation. Future Directions: Future research endeavors aim to enhance alginate's stability and bioactivity, facilitating its broader utilization in regenerative medicine and therapeutic interventions. Novel approaches focusing on tailored hydrogel formations, advanced drug delivery systems, and optimized cellular encapsulation techniques hold promise. Continued exploration of alginate's potential in tissue engineering and wound healing, alongside efforts to address critical issues, will drive advancements in biomedical applications.

A novel ionic liquid-based approach for DNA and RNA extraction simplifies sample preparation for bacterial diagnostics.

DNA- and RNA-based diagnostics play a pivotal role in accurately detecting and characterizing health-relevant bacteria, offering insights into bacterial presence, viability and treatment efficacy. Herein, we present the development of a novel extraction protocol for both DNA and RNA, designed to enable simple and rapid molecular diagnostics. The extraction method is based on the hydrophilic ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate and silica-coated magnetic beads. First, we developed an IL-based cell lysis protocol for bacteria that operates at room temperature. Subsequently, we established a magnetic bead purification procedure to efficiently and reproducibly extract DNA and RNA from the IL-lysates. The IL not only lyses the cells, but also facilitates the adsorption of nucleic acids (NAs) onto the surface of the magnetic beads, eliminating the need for a chaotropic binding buffer and allowing for purification of NAs without significant effort and materials required. Lastly, we combined the cell lysis step and the purification step and evaluated the novel IL-based extraction method on periopathogenic bacterial cultures, comparing it to commercial DNA and RNA extraction kits via (RT)-qPCR. In comparison to the reference methods, the IL-based extraction protocol yielded similar or superior results. Furthermore, costs are lower, required materials and equipment are minimal and the process is fast (30min), simple and automatable. These characteristics favour the developed method for use in routine and high-throughput testing as well as in point-of-care, on-site and low-resource settings, thereby advancing the field of molecular diagnostics.

Do commercial dog extracts cross-react with Felis domesticus allergen 1.

Background: Half of U.S. households own a dog despite 10% of individuals being sensitized to dog. Assessment and treatment options for dog allergy include the use of commercial dog extracts which have inconsistent performance, making diagnosing and managing dog allergy challenging. Contamination of dog extracts with other allergens has previously been reported. Objective: We sought to determine whether commercial dog extracts contain other aeroallergens. Methods: An extract purity and quantification study on acetone precipitated dog hair and dander extract (AP dog) was performed, 6 aeroallergens; Alternaria (Alt a 1), Ragweed (Amb a 1), German Cockroach (Bla g 2), Dust Mite (Der p t), Cat (Fel d 1), and Rye Grass (Lol p 1). Following, conventional dog hair and dander extract (CV dog) and the new ultrafiltered dog hair and dander extract (UF dog) were also assessed based on the initial results of AP dog. SDS-PAGE was performed on all three dog extracts to compare allergen content. Lastly, serology results and aeroallergen immunotherapy prescriptions were compared. Results: The ELISA trays with Alt a 1, Amb a 1, Bla g 2, Der p 1, and Lol p 1 antibodies did not capture AP dog, while the ELISA tray with Fel d 1 antibody captured AP dog, CV dog, and UF dog. SDS-PAGE results of the 3 dog extracts did not reveal a band at the molecular weight for Fel d 1. Conculsion: Contamination of commercial dog extracts was not found. However, our findings are supportive of commercial dog extracts containing a Fel d 1-like dog allergen that is cross-reactive to Fel d 1. Cross-reactivity between commercial dog extracts and Fel d 1 could be responsible for double positivity to cat and dog in serology. Additional studies are needed to better illustrate this Fel d 1-like dog allergen.

Comparative Analysis of Pharmaceutical Persistence in Canadian Wastewater Treatment Plants Utilizing Molecularly Imprinted Polymers and Commercial Solid Phase Extraction Sorbents

In this study, the performances of a synthesized Molecularly Imprinted Polymer (MIP) and a commercial Solid Phase Extraction (SPE) cartridge were compared in terms of the removal efficiency and quantification of pharmaceutical compounds in wastewater samples. Concentration analysis of Emtricitabine, Tenofovir, Naproxen, Diclofenac, Ibuprofen, and Efavirenz was conducted in influent, primary effluent, secondary effluent, and post-chlorination water samples using liquid chromatography-UV detection. Regressions analysis revealed high linearity (R2 values from 0.9980 to 0.9999) for the compounds, alongside recoveries ranging from 90.9% to 100%. The method detection limits (MDLs) were also determined. Removal efficiencies were computed, with Naproxen and Ibuprofen demonstrating complete removal (100%), while other compounds displayed different of removal efficiency levels. The MIP was found to perform slightly better than traditional SPE cartridges in terms of selectivity, efficiency, and overall performance. This study provides valuable insights into the concentrations of target compounds at various treatment stages, emphasizing the importance of choosing extraction cartridges in wastewater treatment processes.

Comparative evaluation of PCR and loop-mediated isothermal amplification (LAMP) assays for detecting Pasteurella multocida in poultry

ABSTRACT Aims To develop a colourimetric loop-mediated isothermal amplification (LAMP) assay for the detection of Pasteurella multocida in clinical poultry samples and compare the performance of this assay with PCR. A secondary aim was to evaluate a simple DNA extraction method that could enable LAMP-based testing in the field without the need for specialised laboratory equipment. Methods Primer sets for both LAMP and PCR were designed to amplify the KMT1 gene of P. multocida. DNA was extracted from 12 P. multocida isolates using a commercial extraction kit, and subjected to analysis using both LAMP and PCR. The analytical specificity of the LAMP assay was evaluated by testing it against a panel of 12 unrelated bacterial species, and the analytical sensitivity (limit of detection) was determined through testing of serial dilutions of the target DNA and compared to that of PCR. Subsequently, cloacal swabs (n = 40) from a commercial turkey flock were subjected to analysis using both LAMP and PCR assays, using a rapid DNA extraction method and a commercial extraction kit. Clinical sensitivity and specificity of the LAMP assay were calculated in comparison to PCR. Results A single DNA fragment of the expected size (∼ 200 base pairs), was amplified by PCR from 12 P. multocida isolates, which were also all positive by the LAMP assay. The identity of all PCR amplicons was confirmed by sequencing. Both PCR and LAMP showed similar analytical sensitivity, with a LOD of 20 pg of target DNA. As neither PCR nor LAMP assays produced positive results with 12 non-related bacterial species, the analytical specificity was assessed as 100%. However, LAMP demonstrated lower clinical specificity (94.74%) compared to PCR (100%) when 40 clinical samples were tested. None of the DNA samples extracted using the simplified DNA extraction method were amplified by either LAMP or PCR. Conclusion The LAMP assay developed in this study exhibits comparable performance to PCR in detecting P. multocida. Clinical relevance The use of a rapid and portable DNA extraction method, in conjunction with LAMP assays, could create opportunities for point-of-care testing for fowl cholera in field settings.

Strategies to reduce the culture medium costs for a high‐yield and high‐selectivity bio‐based 2,3‐butanediol production

AbstractBio‐based 2,3‐butanediol (2,3‐BDO) production on a large scale depends on critical factors, such as culture medium, oxygen supply, pH and biosafety. In this study, three strategies for reducing culture medium costs were investigated: carbon/nitrogen (C/N) ratio; low‐cost nitrogen sources (crude yeast extract, brewer's yeast extract, corn steep liquor, urea, sodium nitrate, ammonium chloride, ammonium sulfate and dibasic ammonium phosphate); and microbial pH autoregulation. Batch fermentations were performed in a microaerobic environment using wild‐type and safe Paenibacillus peoriae NRRL BD 62. The yield and selectivity of 2,3‐BDO were used as control variables. A ratio between 2,3‐BDO production and glucose consumption (YP/S) of almost 80% and an optical purity of 87% levo‐2,3‐BDO, with no acetoin accumulation, were achieved in an NH4Cl‐based medium at C/N = 8.5 g/g and without external pH control, considering an initial glucose of 10 g/L. Based on Free on Board prices, a 63% savings in culture medium costs was achieved by replacing commercial yeast extract at pH 5. Validation assays with higher initial glucose concentrations showed a YP/S of 0.40 g/g and an optical levo‐/meso‐2,3‐BDO ratio of 1:0.8, with negligible acetoin accumulation. Therefore, the NH4Cl‐based medium at C/N = 8.5 g/g and without pH control was considered economically promising for high‐yield and high‐selectivity bio‐based 2,3‐BDO production.

Association of HPA Antigens with Immune Thrombocytopenia: A Case-Control Study by PCR-SSP Method

Background: Human platelet antigens (HPAs) play a clinically significant role in alloimmunization and the development of immune-mediated disorders such as immune thrombocytopenia (ITP), fetal and neonatal alloimmune thrombocytopenia (FNAIT), and post-transfusion purpura (PTP). Understanding the genetic profiles of HPAs is critical for preventing and treating these conditions. Given the limitations of serological methods in determining HPA genotypes, this study aims to investigate the association between the genotypes of HPA1, HPA2, HPA3, HPA4, and HPA15 antigens and autoimmune thrombocytopenia in Lorestan Province, utilizing the PCR-SSP method. Materials and Methods: This case-control study involved 80 individuals diagnosed with ITP and 120 healthy controls. DNA samples were extracted using a commercial DNA extraction kit, with concentrations quantified via a Nanodrop spectrophotometer. Genotyping was performed using the PCR-SSP method with specific primers for each HPA gene. The genotype data were verified using previously established sample sets. The frequencies of each HPA genotype were recorded, and a comparative analysis was conducted between the patient and control groups to evaluate the study hypothesis. Results: The results revealed that individuals carrying the HPA2b allele had a 5.31-fold increased risk of developing ITP, a statistically significant finding (P < 0.05, OR = 5.31). Similarly, the presence of the HPA15b allele was associated with a 6.54-fold increased risk (P < 0.05, OR = 6.54). Conclusion: These findings, in conjunction with previous studies, suggest the need for larger-scale investigations across different populations. Such research could aid in the early diagnosis and prediction of thrombocytopenia severity, inform treatment strategies, and facilitate the removal of pathogenic antibodies from circulation.

Multispecies Trichoderma in Combination with Hydrolyzed Lignin Improve Tomato Growth, Yield, and Nutritional Quality of Fruits

The application of biological pesticides as alternatives to chemical phytosanitary products is a natural and innovative method to improve environmental protection and sustainable agricultural production. In this work, the compatibility between Trichoderma spp. and a commercial lignin extract was assessed in vitro and in vivo. The beneficial effects of lignin in combination with different Trichoderma consortia were evaluated in terms of improved growth and quantitative and qualitative tomato productivity. T. virens GV41 + T. asperellum + T. atroviride + lignin formulation was the most effective in growth promotion and increased root and stem dry weight compared to control (45.4 and 43.9%, respectively). This combination determined a 63% increase in tomato yield compared to the control, resulting in the best-performing treatment compared to each individual constituent. Consistent differences in terms of lycopene, GABA, ornithine, total, essential, and branched-chain amino acids were revealed in fruits from tomato plants treated with Trichoderma–lignin formulations (T. asperellum + T. virens GV41 + lignin) or with the microbial consortia (T. asperellum + T. virens GV41, T. atroviride + T. virens GV41). The developed bioformulations represent a sustainable biological strategy to increase yield and produce nutritional compound-enriched vegetables.

Streamlined boiling lysis DNA extraction for Gram-positive aquaculture pathogen Streptococcus agalactiae.

Accurate genetic analysis is essential for the detection of pathogens as it necessitates suitable DNA extraction methods tailored to specific microorganisms such as Gram-positive bacteria. This study examined several commercial and simplified DNA extraction methods for their suitability in isothermal downstream applications. Extracted DNA was assessed using spectrophotometry, electrophoresis, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) while its stability was inspected after five months of storage. The findings revealed variations in DNA yield, purity and integrity among the extraction methods. While extraction kits demonstrated high yield and purity, the in-house extraction techniques showed incoherent correlation between yield and purity, yet showed promise for a streamlined extraction process. The DNA acquired from all methods yielded positive amplification in PCR and LAMP. DNA extracted by kits exhibits prolonged stability than those obtained via boiling lysis. Both methods offer distinct advantages, with commercial kits providing longer stability and high-quality DNA while boiling lysis stands out for its simplicity, with shorter handling and processing periods. This study emphasizes selecting ideal extraction methods for Streptococcus agalactiae, in the prospect of aquaculture settings. In particular, successful LAMP reaction suggests that boiled extracts are feasible enough for detection, and suited for point-of-care (POC) testing where prompt detection of aquatic pathogens is often critical. Ultimately, the choice of method should be contemplated on a case-by-case basis such as the study goals, intended settings, and type of samples.

Efficient recovery of organic phase from crud in solvent extraction unit of Sarcheshmeh copper complex in Iran by powerful water-absorbent reagents

The formation of stable multi-phase emulsions (crud) is a serious problem in almost all commercial solvent extraction (SX) plants. Crud is a mixture of organic and aqueous phases, air, and organic or inorganic solid particles, often found to distribute itself randomly throughout the separating phases of SX plants. Crud formation is encountered in almost all SX units in metal extraction plants. It causes the loss of solvent and thus increases the process costs. Therefore, the removal of crud and recovery of the expensive organic phase including diluent and extracting reagents from it is of great importance. In this research, the applicability of a range of powerful water absorbent reagents was studied for the recovery of organic phase from the crud formed in copper extraction/electrowining unit of Sarchashmeh copper complex (IRAN). The reagents included gypsum, carbomer, sodium polyacrylate, sodium sulfate, magnesium sulfate and calcium chloride. The results showed that gypsum, carbomer and sodium polyacrylate are effective additives which let the organic phase to be recovered with up to 95 % yield. To ensure that gypsum does not cause a negative effect on the copper loading and stripping of the recovered solvent, the extraction/stripping properties of the organic phase separated with gypsum was compared with the organic phase separated by centrifugation without additives. Surface tension analysis of the gypsum-recovered organic phase also confirmed its suitability for recovery of organic phase from crud. Finally, the calcium content of gypsum-recovered organic phase was measured and compared with that of the organic phase separated by centrifugation without any additive.

Biosynthesis and Characterization of Iron Oxide Nanoparticles Using Chenopodium quinoa Extract.

In this study, we achieved the biosynthesis of novel 7-8 nm iron-oxide nanoparticles in the presence of different concentrations (5 to 50% w/v) of commercial white quinoa extract. Initially, quinoa extract was prepared at various concentrations by a purification route. The biosynthesis optimization was systematically monitored by X-ray diffraction, and the Rietveld quantitative analysis showed the presence of goethite (5 to 10 wt.%) and maghemite phases. The first phase disappeared upon increasing the organic loading (40 and 50% w/v). The organic loading was corroborated by thermogravimetric measurements, and it increased with quinoa extract concentration. Its use reduces the amount of precipitation agent at high quinoa extract concentrations with the formation of magnetic nanoparticles with hard ferrimagnetic character (42 and 11 emu g-1). The enrichment of hydroxyl groups and the negative zeta potential above pH = 7 were corroborated by a reduction in the point of zero charge in all the samples. For alkaline values, the zeta potential values were above the stability range, indicating highly stable chemical species. The evidence of hydroxyl and amide functionalization was qualitatively observed using infrared analysis, which showed that the carboxyl (quercetin/kaempferol), amide I, and amide III chemical groups are retained after biosynthesis. The resultant biosynthesized samples can find applications in environmental remediation due to the affinity of the chemical agents present on the particle surfaces and easy-to-handle them magnetically.

A-302 Evaluation of real-time PCR assays for detection of sexually-transmitted infections

Abstract Background Sexually transmitted infections (STIs) have serious negative consequences for reproductive health worldwide. They are associated with infertility, premature birth and neonatal infections. Five STI pathogens, namely Trichomonas vaginalis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum were selected for the evaluation of three potential real-time PCR assays that can be adopted for diagnostic testing. Methods A total of 36 samples were included, consisting of genital swabs, urine, abscesses and samples provided by quality assurance programmes. DNA extraction was performed using an automated nucleic acid extraction system, abGenixTM. Commercial genomic controls and DNA extracts donated by external laboratories were used as well. The extracted nucleic acids were tested using three different assays, namely Seegene Anyplex II STI-5 Detection, Thermo Fisher Scientific customized STI Panel assay, and TIB MOLBIOL STI LightMix Modular kits, according to the manufacturers’ instructions. Results A total of 67% and 11% of the samples showed equivalent positive and negative results, respectively. However, 22% of the samples had non-concordant results. TIB MOLBIOL STI LightMix Modular kit was unable to differentiate between Ureaplasma parvum and Ureaplasma urealyticum in 9 samples. Genomic controls were tested, and Seegene Anyplex II STI-5 Detection was unable to detect lower concentrations of DNA for Trichomonas vaginalis, Mycoplasma genitalium and Mycoplasma hominis. All assays were able to detect lower concentrations of Ureaplasma parvum DNA, but detectable concentrations were higher for Ureaplasma urealyticum. Conclusions In conclusion, the performance of each assay differed according to pathogen. None of the assays offered equal performance for all pathogens. We have adopted Thermo Fisher Scientific customized STI Panel assay in our diagnostic testing due to its advantage of being user friendly and cost effective.

Preservative Effect of a Gelatin-Based Film Including a Gelidium sp. Flour Extract on Refrigerated Atlantic Mackerel

This research evaluated the preservative properties of flour from the alga Gelidium sp., which is a waste substrate resulting from commercial phycocolloid extraction. Gelatin-based biofilms, which included two different concentrations of red alga flour, were developed and used as packaging systems during refrigerated storage (up to 9 days at 4 °C) of Atlantic mackerel (Scomber scombrus) muscle. In all batches tested, a progressive decrease in quality could be observed in the muscle of the fish as the storage time increased. Compared with the control fish, the Gelidium alga flour extract had an inhibitory effect (p < 0.05) on microbial activity (total aerobes, psychrotrophs, and proteolytic bacteria), lipid oxidation (peroxide, thiobarbituric acid, fluorescence and polyene indices), lipid hydrolysis (formation of free fatty acids) and pH increase in refrigerated mackerel muscle. In contrast, no significant effect (p > 0.05) was observed on trimethylamine formation, Enterobacteriaceae, or lipolytic bacteria counts. A preservative effect resulting from the incorporation of Gelidium alga flour into the gelatin-based biofilm was observed, indicating both quality and safety enhancement. In accordance with current global interest in the search for natural and waste sources, a novel and beneficial use of Gelidium flour for enhancing the quality of refrigerated fish has been proposed.

Sustainable Exploitation of Wine Lees as Yeast Extract Supplement for Application in Food Industry and Its Effect on the Growth and Fermentative Ability of Lactiplantibacillus plantarum and Saccharomyces cerevisiae

Wine lees, the residue left behind after racking or bottling of wine, are predominantly composed of dead yeast cells, ethanol, phenolic compounds, and tartrates. Yeast extract (i.e., commercial yeast extract), a highly nutritious powder derived from commercially cultivated yeast biomass, is commonly used in nutrient media as a nitrogen source. In the context of by-product valorization, wine lees could potentially be used to produce a substitute for commercial yeast extract (CYE). In our study we investigated the growth and fermentative ability of two major winemaking microorganisms, Lactiplantibacillus plantarum and Saccharomyces cerevisiae, in culture media containing a wine lees yeast extract (WLYE) and a CYE. The effects of yeast extract type, concentration, and initial cell concentration (y0) on key kinetic parameters—maximum specific growth rate (μmax), lag phase duration (λ), and maximum cell concentration (ymax)—were evaluated. For L. plantarum, the results showed that using a WLYE led to similar kinetic parameters to those obtained with a CYE, with λ being unaffected by y0 in samples containing a WLYE. For S. cerevisiae, simultaneous addition of both yeast extracts led to increased μmax values (up to 0.136 h−1) compared to individually added yeast extracts, although this negatively affected λ and ymax. Current research on wine lees is mainly focused on using them as a substrate to produce valuable metabolites through fermentation, overlooking the potential industrial applications of the nutrient-rich autolysate. The findings of this study appear promising for the holistic valorization of wine lees, contributing towards the concepts of sustainability and circular economy.

Separation of Co(II), Mn(II), and Ni(II) by solvent extraction with Cyanex 272 and D2EHPA from the sulfuric acid leaching solution of spent lithium-ion batteries

Spent lithium-ion batteries contain valuable metals such as cobalt, lithium, manganese and nickel. Smelting reduction of spent LIBs at high temperature results in metallic alloys. Therefore, it is necessary to recover the valuable metal ions present in the sulfuric acid leaching solutions of the metallic alloys. In this work, solvent extraction experiments were performed to separate the three metal ions, Co(II), Mn(II) and Ni(II) present in the leaching solutions of the metallic alloys. Synthetic sulfate solutions were prepared and commercial organophosphorous extractants were employed. First, both Co(II) and Mn(II) were completely extracted in one stage by using 40% saponified 0.3 M Cyanex 272, leaving Ni(II) in the raffinate. The small amount of Ni(II) co-extracted into Cyanex 272 was removed by two stages of cross-current scrubbing using pure CoSO4 solution. The effect of pH and CoSO4 concentration of the scrubbing solution on the scrubbing of Ni(II) was investigated. The Co(II) and Mn(II) present in the scrubbed Cyanex 272 were completely stripped by 0.05 M H2SO4 solution. Batch simulation experiments for the three stage of counter-current extraction with 1 M D2EHPA verified that Mn(II) and Co(II) were completely separated by selective extraction of Mn(II) into D2EHPA, leaving Co(II) in the raffinate. By consecutive extraction with Cyanex 272 and D2EHPA, pure solutions of Co(II), Mn(II) and Ni(II) with purity higher than 99.9% can be recovered from the starting solutions. A process was proposed for the treatment of sulfuric acid leaching solutions of spent LIBs containing Co(II), Mn(II) and Ni(II) by solvent extraction.

Evaluation of commercial RNA extraction kits for long-read metatranscriptomics in soil

Metatranscriptomic analysis of the soil microbiome has the potential to reveal molecular mechanisms that drive soil processes regulated by the microbial community. Therefore, RNA samples must be of sufficient yield and quality to robustly quantify differential gene expression. While short-read sequencing technology is often favoured for metatranscriptomics, long-read sequencing has the potential to provide several benefits over short-read technologies. The ability to resolve complete transcripts on a portable sequencing platform for a relatively low capital expenditure makes Oxford Nanopore Technology an attractive prospect for addressing many of the challenges of soil metatranscriptomics. To fully enable long-read metatranscriptomic analysis of the functional molecular pathways expressed in these diverse habitats, RNA purification methods from soil must be optimised for long-read sequencing. Here we compare RNA samples purified using five commercially available extraction kits designed for use with soil. We found that the Qiagen RNeasy PowerSoil Total RNA Kit performed the best across RNA yield, quality and purity and was robust across different soil types. We found that sufficient sequencing depth can be achieved to characterise the active community for total RNA samples using Oxford Nanopore Technology, and discuss its current limitations for differential gene expression analysis in soil studies.

Synergistic effect of compost and moringa leaf extract biostimulants on the remediation of gold mine tailings using chrysopogon zizanioides

Phytoremediation is a cost-effective and environmentally friendly method for restoring degraded land. However, optimizing phytoremediation conditions and using locally sourced materials can further improve the sustainability of this approach. A 3×2×2 factorial design was used in determining the optimal parameters in the remediation of gold mine tailings using vetiver grass. The treatments consisted of three compost concentrations (0 %, 30 % and 60 %), two biostimulant types (laboratory-extracted and commercial biostimulants) and two application regimens (once and twice a week) in a randomized complete block design. The biomass and metal concentrations in the vetiver grass were measured after 16 weeks. All the vetiver that was planted in 0 % compost amendment died within four weeks regardless the MLE treatment. There was no significant difference in vetiver biomass between vetiver grown in 30 % and that grown in 60 % compost amendment. Biostimulant application frequency led to no significant differences. A difference on vetiver biomass due to the type of moringa leaf extract was eminent, with laboratory moringa leaf extract resulting in significantly higher biomass in vetiver grown on 60 % compost yet the commercial moringa leaf extract producing significantly higher vetiver biomass on vetiver grown on 30 % compost amendment. These findings suggest that the optimal combination for field study would be 30 % compost amendment with commercial biostimulant for a more cost-effective option.