Assessment of growth, reproduction, and vermi-remediation potentials of Eisenia fetida on heavy metal exposure.

Heavy metal pollution is a significant environmental concern with detrimental effects on ecosystems and human health, and traditional remediation methods may be costly, energy-intensive, or have limited effectiveness. The current study aims were to investigate the impact of heavy metal toxicity in Eisenia fetida, the growth, reproductive outcomes, and their role in soil remediation. Various concentrations (ranging from 0 to 640 mg per kg of soil) of each heavy metal were incorporated into artificially prepared soil, and vermi-remediation was conducted over a period of 60 days. The study examined the effects of heavy metals on the growth and reproductive capabilities of E. fetida, as well as their impact on the organism through techniques such as FTIR, histology, and comet assay. Atomic absorption spectrometry demonstrated a significant (P < 0.000) reduction in heavy metal concentrations in the soil as a result of E. fetida activity. The order of heavy metal accumulation by E. fetida was found to be Cr > Cd > Pb. Histological analysis revealed a consistent decline in the organism's body condition with increasing concentrations of heavy metals. However, comet assay results indicated that the tested levels of heavy metals did not induce DNA damage in E. fetida. FTIR analysis revealed various functional group peaks, including N-H and O-H groups, CH2 asymmetric stretching, amide I and amide II, C-H bend, carboxylate group, C-H stretch, C-O stretching of sulfoxides, carbohydrates/polysaccharides, disulfide groups, and nitro compounds, with minor shifts indicating the binding or accumulation of heavy metals within E. fetida. Despite heavy metal exposure, no significant detrimental effects were observed, highlighting the potential of E. fetida for sustainable soil remediation. Vermi-remediation with E. fetida represents a novel, sustainable, and cutting-edge technology in environmental cleanup. This study found that E. fetida can serve as a natural and sustainable method for remediating heavy metal-contaminated soils, promising a healthier future for soil.

Characterization of heavy metal-associated bacteria from petroleum-contaminated soil and their resistogram and antibiogram analysis.

The aim of the current study was to screen and identify heavy metal (chromium, cadmium, and lead) associated bacteria from petroleum-contaminated soil of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan to develop ecofriendly technology for contaminated soil remediation. The petroleum-contaminated soil was collected from 99 different localities of district Muzaffarabad and the detection of heavy metals via an atomic absorption spectrometer. The isolation and identification of heavy metals-associated bacteria were done via traditional and molecular methods. Resistogram and antibiogram analysis were also performed using agar well diffusion and agar disc diffusion methods. The isolated bacteria were classified into species, i.e., B. paramycoides, B. albus, B. thuringiensis, B. velezensis, B. anthracis, B. pacificus Burkholderia arboris, Burkholderia reimsis, Burkholderia aenigmatica, and Streptococcus agalactiae. All heavy metals-associated bacteria showed resistance against both high and low concentrations of chromium while sensitive towards high and low concentrations of lead in the range of 3.0 ± 0.0 mm to 13.0 ± 0.0 mm and maximum inhibition was recorded when cadmium was used. Results revealed that some bacteria showed sensitivity towards Sulphonamides, Norfloxacin, Erythromycin, and Tobramycin. It was concluded that chromium-resistant bacteria could be used as a favorable source for chromium remediation from contaminated areas and could be used as a potential microbial filter.

Efficacy of cow and buffalo dung on vermiremediation and phytoremediation of heavy metals via Fourier-transform infrared spectroscopy and comet assay.

Heavy metal contamination raised significant concerns throughout the world. The current research aimed to evaluate the impact of organic manure (cow dung and buffalo dung) on vermiremediation and phytoremediation and to remediate heavy metals, i.e., cadmium, lead, and chromium, from artificial contaminated soil via both remediation techniques. The impact of livestock manure was evaluated for the first time which could be effective in in situ as well as ex situ studies. Eisenia fetida, Pheretima lignicola, and Spinacia oleracea were used for the remediation process. Results revealed that E. fetida tolerated lead at 280 mg, cadmium at 150 mg, and chromium at 860 mg compared to P. lignicola. The growth and reproduction of E. fetida were efficient in the cow dung manure compared to buffalo dung. Similarly, seed germination and growth of Spinacia oleracea were better in cow dung media compared to buffalo dung. Bioaccumulation factor showed that E. fetida showed a higher accumulation of heavy metals in their tissues when vermi + phytoremediation was jointly applied (9.50 mg/l of Pb, 24.166 of Cd, and 6.695 of Cr). Fourier-transform infrared spectroscopy indicated that heavy metals had no drastic effects on E. fetida and S. oleracea. Similarly, comet assay revealed that heavy metals had no genotoxic effect on the E. fetida and S. oleracea. It was concluded that both E. fetida and S. oleracea are appropriate for heavy metals remediation in cow dung manure.

Phylogenetic Illustration of Eisenia fetida Associated Vermi-bacteria Involved in Heavy Metals Remediation and Retaining Plant Growth Promoting Traits.

Heavy metals contamination in the soil is a major threat to wildlife, the environment, and human health. Microbial remediation is an emerging and promising technology to reduce heavy metals toxicity. Therefore, the present research aimed to isolate and to identify the heavy metals tolerated bacteria from the Eisenia fetida for the first time, and to screen the bacto-remediation capabilities and plant growth promoting traits of vermi-bacterial isolates. Vermi-bacteria was isolated from the gut of E. fetida, identified through staining, culturing, biochemical tests, and ribotyping. Plant growth-promoting traits were also evaluated. Phylogenetic results revealed that isolated Vermi-bacterial strains showed resemblance with Bacillus thuringiensis, Bacillus aryabhattai, Staphylococcus hominis, Bacillus toyonensis, Bacillus cabrialesii, Bacillus tequilensis, Bacillus mojavensis, Bacillus amyloliquefaciens, Bacillus toyonensis, Bacillus anthracis, and Bacillus paranthracis. All identified Vermi-bacterial species are Gram-positive (rod and cocci) in nature, not only indicated the efficient biosorption of lead, cadmium, and chromium but also produce all plant growth stimulating traits such as indole acetic acid (IAA), amylase, protease, lipase, hydrogen cyanide, ammonia, and siderophore production, and also act as a phosphate solubilizers. Bacillus anthracis showed significant production of siderophore (33.0±0.0 mm), phosphate solubilizing (33.0±0.0 mm), proteolytic (15.0±0.0 mm), and lipolytic activities (20.0±0.0 mm) compared to other vermi-bacterial isolates. Bioaccumulation factor results revealed that Bacillus anthracis showed more accumulation of Cd (12.00±0.01 ppm), Cr (5.38±0.01 ppm), and Pb (4.38±0.01 ppm). Therefore, the current findings showed that all identified vermi-bacteria could be used as potential bactoremediation agents in heavy metals polluted environments and could be used as microbial biofertilizers to enhance crop production in a polluted area.

Molecular characterization of plant growth-promoting vermi-bacteria associated with Eisenia fetida gastrointestinal tract.

Earthworms are highly productive invertebrates and play a vital role in organic farming and improving soil structure and function. The gastrointestinal tract of earthworms possessed agricultural important bacteria. So, the current research aimed was to examine, screen, and identify the plant growth promoting bacteria existing in the digestive tract of Eisenia fetida called plant growth promoting vermi-bacteria. The plant growth promoting traits such as siderophore, phytohormone, and hydrolytic enzymes production, and phosphate solubiliation were assessed. Eleven vermi-bacteria i.e. Bacillus mycoides, B. aryabhattai, B. megaterium, Staphylococcus hominis, B. subtilis, B. spizizenii, B. licheniformis, B. mojavensis, B. toyonensis, B. anthracis, B. cereus, B. thuringiensis, and B. paranthracis were isolated and identified based on microscopic studies, biochemical tests, ribotyping, and agricultural traits. All vermi-bacteria are Gram-positive rods except Staphylococcus hominis and produce different compounds such as siderophore, indole acetic acid, catalase, oxidase, proteases, amylases, and lipases. All vermi-bacteria also act as phosphate solubilizers. Therefore, all isolated vermi-bacteria could be used as potential microbial biofertilizers to enhance crops production in Pakistan.

ESIDE: A computationally intelligent method to identify earthworm species (E. fetida) from digital images: Application in taxonomy.

Earthworms (Crassiclitellata) being ecosystem engineers significantly affect the physical, chemical, and biological properties of the soil by recycling organic material, increasing nutrient availability, and improving soil structure. The efficiency of earthworms in ecology varies along with species. Therefore, the role of taxonomy in earthworm study is significant. The taxonomy of earthworms cannot reliably be established through morphological characteristics because the small and simple body plan of the earthworm does not have anatomical complex and highly specialized structures. Recently, molecular techniques have been adopted to accurately classify the earthworm species but these techniques are time-consuming and costly. To combat this issue, in this study, we propose a machine learning-based earthworm species identification model that uses digital images of earthworms. We performed a stringent performance evaluation not only through 10-fold cross-validation and on an external validation dataset but also in real settings by involving an experienced taxonomist. In all the evaluation settings, our proposed model has given state-of-the-art performance and justified its use to aid earthworm taxonomy studies. We made this model openly accessible through a cloud-based webserver and python code available at https://sites.google.com/view/wajidarshad/software and https://github.com/wajidarshad/ESIDE.

Biological Activities and Secondary Metabolite Screening of Rumex hastatus Extract through Fourier Transform Infrared and Raman Spectroscopy.

BACKGROUND: Human infectious diseases are caused by various pathogens including bacteria, fungi, viruses, parasites, and protozoans. These infectious agents are controlled by using synthetic drugs as well as natural sources. OBJECTIVE: The aim of current study was to evaluate the antibacterial effect of Rumex hastatus against clinical bacterial pathogens. METHODS: In current research antibacterial effect of Rumex hastatus was analyzed against seven clinical pathogenic bacteria such as Escherichia coli, Serratia marcescens, Streptococcus pyogenes, Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa through agar well diffusion method. The boiled extract was used for the phytochemical screening, antioxidant potential, thin layer chromatography, bio-autography, and spot screening. Genomic DNA was extracted to find the DNA protection effect of R. hastatus. RESULTS: Antibacterial results showed that diethyl ether extract has the maximum inhibition of S. pyogenes (9.66 ± 0.57 mm). Acetone and diethyl ether extracts showed moderate inhibition of K. pneumoniae (6.33 ± 1.52 mm and 5.66 ±1.15 mm) and S. aureus (6.33 ± 1.52 mm and 5.66 ± 0.57 mm). Similarly, chloroform extract indicated moderate inhibition of S. pyogenes (5.66 ± 1.15 mm). Ethanol extract had low or even no effect on the growth of bacteria. Genomic DNA extraction also encouraged the antibacterial effect of R. hastatus. Various phytochemical constituents such as ketoses, oligosaccharides, amino acids, amines, sugars, flavonoids, and antioxidant constituents were detected. TLC-Bioautography and spot screening results revealed the potential use of R. hustatus as an antibacterial agent. CONCLUSION: It was concluded that most of the tested fractions appeared as an important source for the discovery of new antimicrobial drugs.

Phytochemical Screening, Alpha-Glucosidase Inhibition, Antibacterial and Antioxidant Potential of Ajuga bracteosa Extracts.

BACKGROUND: Ajuga bracteosa, a medicinal herb, is used by local community to cure a number of diseases such as inflammation, jaundice bronchial asthma, cancer and diabetes. OBJECTIVES: The aim of present work was to evaluate the antioxidant potential, in vitro antidiabetic and antimicrobial effects of A. bracteosa. METHODS: n-hexane, ethyl acetate, chloroform, acetone, methanol and aqueous extracts of Ajuga bracteosa roots, were prepared via maceration. Antibacterial activity was carried out by agar well diffusion method. Quantitative and qualitative phytochemical screening was done. The antioxidant activity was determined by iron (II) chelating activity, iron reducing power, DPPH, and ABTS free radical scavenging methods, Antidiabetic activity was evaluated through inhibition of α-glucosidase assay. RESULTS: Phytochemical analysis showed the presence of phenols, flavonoids, tannins, saponins, quinines, terpenoids, xanthoproteins, glycosides, carbohydrates, steroids, phytosterols and amino acids. DPPH and ABTS potential values were recorded as 61.92% to 88.84% and 0.11% to 38.82%, respectively. Total phenolic and total flavonoid contents were expressed as gallic acid and rutin equivalents. Total iron content was expressed as FeSO4 equivalents. Chloroform and n-hexane extracts showed significant enzyme inhibition potential with IC50 values of 29.92 µg/ml and 131.7 µg/ml respectively. Aqueous extract showed maximum inhibition of E. coli, S. typhimurium, E. amnigenus, S. pyogenes, and S. aureus, (18.0±1.0 mm, 12.5±0.7 mm, 17.0±0.0 mm, 11.0±0.0 mm and 15.3±2.0 mm mm), respectively. Similarly, n-hexane extract showed maximum inhibition of E. coli, E. amnigenus, S. aureus (11.6±1.5 mm; 11.3±1.5 mm; 13.3±0.5 mm). This study also shows that n-hexane, chloroform, ethyl acetate and aqueous extracts of A. bracteosa root possess α-glucosidase inhibitory activities and therefore it may be used as hypoglycemic agents in the management of postprandial hyperglycemia. CONCLUSION: Ajuga bracteosa root extracts may provide a basis for development of antioxidant, antimicrobial and antidiabetic drugs.

Introducing a structured prescription form improves the quality of handwritten prescriptions in limited resource setting of developing countries.

INTRODUCTION: Incomplete or illegible prescriptions can lead to serious errors in administration of the prescribed medication, which can become hazardous. OBJECTIVE: Our aim is to determine if a structured prescription form can improve the quality of handwritten prescription in terms of completeness and legibility. METHODS: We conducted a prospective, non-randomized, time series study of quality of written prescriptions of general practitioners at a tertiary teaching hospital in Peshawar, Pakistan. The study involved an intervention, composed of the introduction of a pre-printed structured prescription form. The data were collected within 4 weeks including a 2-week pre-intervention phase and 2-week post-intervention phase. Completeness, quality of prescriptions and legibility were compared before and after the intervention of the pre-printed structured prescription form. RESULTS: A total of 463 prescriptions were obtained (260 in the pre-intervention phase and 203 in the post-intervention phase). Between pre-intervention phase and post-intervention phase, the Pakistan Medical and Dental Council registration number presence in prescriptions improved from 73.1% to 100% (P < 0.0005). The presence of prescriber's signature improved from 92.7% to 99% (P = 0.001). Drug duration was not missing in 99.5% in post-intervention phase as compared with 90.4% in pre-intervention phase (P < 0.0005). Prescriptions with no legibility problems improved from 76.2% to 94.1% (P < 0.0005). Although not statistically significant, prescriptions in which drug dosage was not missing improved from 85% to 90.6% (P = 0.07). LIMITATIONS: We have a limited single-center study. A larger study in multiple settings is needed to develop adequate evidence for such interventions. Subjective nature of prescription legibility can also be considered as a limitation. CONCLUSION: Structuring a prescription form alone may improve certain aspects of quality of written prescription in terms of completeness and legibility.