RESUMO
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.
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Recuperação e Remediação Ambiental , Metais Pesados , Oligoquetos , Reprodução , Poluentes do Solo , Oligoquetos/efeitos dos fármacos , Metais Pesados/toxicidade , Animais , Poluentes do Solo/toxicidade , Reprodução/efeitos dos fármacos , Recuperação e Remediação Ambiental/métodos , Ensaio Cometa , Espectroscopia de Infravermelho com Transformada de Fourier , Dano ao DNA , Solo/químicaRESUMO
BACKGROUND: With increased urbanization and industrialization, modern life has led to an anthropogenic impact on the biosphere. Heavy metals pollution and pollutants from black liquor (BL) have caused severe effects on environment and living organisms. Bacterial biofilm has potential to remediate heavy metals and remove BL from the environment. Hence, this study was planned to investigate the potential of microbial biofilms for the bioremediation of heavy metals and BL polluted environments. METHODS AND RESULTS: Eleven biofilm forming bacterial strains (SB1, SB2, SC1, AF1, 5A, BC-1, BC-2, BC-3, BC-4, BC-5 and BC-6) were isolated and identified upto species level via 16S rRNA gene sequencing. Biofilm strains belonging to Bacillus and Lysinibacillus sphaericus were used to remediate heavy metals (Pb, Ni, Mn, Zn, Cu, and Co). Atomic absorption spectroscopy showed significantly high (P ≤ 0.05) bioremediation potential by L. sphaericus biofilm (1462.0 ± 0.67 µgmL-1) against zinc (Zn). Similarly, Pseudomonas putida biofilm significantly (P ≤ 0.05) decolourized (65.1%) BL. Fourier transform infrared (FTIR) analysis of treated heavy metals showed the shifting of major peaks (1637 & 1629-1647, 1633 & 1635-1643, and 1638-1633 cm-1) corresponding to specific amide groups due to C = O stretching. CONCLUSION: The study suggested that biofilm of the microbial flora from tanneries and pulp paper effluents possesses a strong potential for heavy metals bioremediation and BL decolourization. To our knowledge, this is the first report showing promising biofilm remediation potential of bacterial flora of tanneries and pulp-paper effluent from Kasur and Sheikhupura, Punjab, Pakistan, against heavy metals and BL.
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Bacillus , Metais Pesados , Pseudomonas putida , Biodegradação Ambiental , RNA Ribossômico 16S/genética , Metais Pesados/análise , Zinco/análise , Pseudomonas putida/genética , Bacillus/genética , BiofilmesRESUMO
Water salinity causes less production of agricultural productivity, low economic returns, soil destructions, less sustainability, and reduction in the germination rate. The current study was aimed to understand the combined potential of halophilic bacteria and rice husk in treating water salinity. In total, 10 halophilic bacterial isolates were isolated from Khewra Mines, Pakistan. Bacterial isolates were characterized by biochemical tests. 16S rRNA gene sequencing identified the isolate SO 1 as Bacillus safensis (accession number ON203008) being the promising halophilic bacteria tolerating upto 3 M NaCl concentration. Next, rice husk was used as carbon source for bacterial biofilm formation, growth and propagation. For saline water treatment, the experimental setting comprising glass wool, rice husk and artificial sea water (3 M) was set. B. safensis biofilm was developed in test samples to desaline the saline water containing 3 M NaCl concentration. Following NaCl decline, flame photometric analysis was used to check the desalination extent of treated saline water. Results showed decreased sodium level in sea water in the presence of rice husk and glass wool. The eluted water used for the germination of Zea mays seeds showed improved growth performance. Also, decreased photosynthetic pigments (chlorophyll "a" = 18.99, and chlorophyll "b" = 10.65), sugar contents (0.7593), and increased carotenoid (1526.91), protein contents (0.4521) were noted compared to control. This eco-friendly approach for bioremediation of salt-affected soils to optimize crop yields under stress through halophilic bacteria and rice husk may overcome the problem of the reduced yield of cash crops/agriculture and water shortage by salinity.
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Oryza , Cloreto de Sódio/metabolismo , RNA Ribossômico 16S/genética , Archaea/genética , Clorofila/metabolismo , Solo/química , Águas Salinas , Biofilmes , SalinidadeRESUMO
Diabetes has become a critical challenge to the global health concerns. Cytotoxicity and development of resistance against available drugs for management of diabetes have shifted the focus of global scientific researchers from synthetic to herbal medications. Therefore, the current study was conducted to investigate the possible anti-hyperglycemic potential of Dryopteris stewartii using Swiss albino mice. To evaluate any possible toxic effect of the plant, acute oral toxicity test was performed while the anti-diabetic effects of aqueous and ethanol extracts at 500 mg/kg, positive, negative and normal control were assessed simultaneously. The anti-diabetic study revealed that aqueous extract has higher anti-diabetic potential than ethanol extract while lowered blood glucose level at second week reaching 150 mg/dL, exerting stronger anti-diabetic effects, compared to ethanol extract (190 mg/dL). Oral glucose tolerance findings revealed that aqueous extract decreased blood glucose level by -0.41-fold, compared to ethanol extract showing a decrease by only -0.29-folds. The histopathological evaluation of liver and pancreas of all groups revealed normal cell architecture with no morphological abnormalities. These results suggested the possible use of D. stewartii as anti-diabetic herbal drug in near future. However, these recommendations are conditioned by deep mechanistic studies.
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Diabetes Mellitus Experimental , Dryopteris , Gleiquênias , Camundongos , Animais , Aloxano/efeitos adversos , Diabetes Mellitus Experimental/patologia , Glicemia , Hipoglicemiantes/efeitos adversos , Extratos Vegetais/efeitos adversos , Etanol/efeitos adversosRESUMO
This study was aimed at investigating the effect of cultured gut microbiota (GM) from obese humans coupled HFD in inducing metabolic endotoxemia in humanized mice. In total, 30 strains were isolated from 10 stool samples of obese patients. Following morphological and biochemical characterization, 16S rRNA gene sequencing of six abundant isolates identified these Klebsiella aerogenes, Levilactobacillus brevis, Escherichia coli, Staphylococcus aureus, Bacillus cereus and Bacillus subtilis (MZ052089-MZ052094). In vivo trial using above isolates, known as human gut microbiota (HGM), was performed for six months. Sixteen mice were distributed into four groups, i.e., G1 (control) mice fed with chow diet, group 2 (G2) with HFD, group 3 (G3) with HFD + HGM and group 4 (G4) with chow diet + HGM. Body mass index (BMI) and plasma endotoxins were measured pre- and post-experiment. In vivo study revealed that HFD + HGM caused significant increase (3.9 g/cm at 20 weeks) in the body weight and BMI (0.4 g/cm post-experiment) of G3 mice compared to the other groups. One-way ANOVA showed significantly higher level of endotoxins (2.41, 4.08 and 3.7 mmol/L) in mice groups G2, G3 and G4, respectively, indicating onset of metabolic endotoxemia. Cecal contents of experimental mice groups showed a shift in microbial diversity as observed by all isolates belonging to either Firmicutes or Bacteroidetes phyla, respectively. In conclusion, current study reported that minor alteration in GM composition through HFD feeding and cultured GM transfer has significant impact in development of metabolic endotoxemia, possibly via modified intestinal permeability.
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Endotoxemia , Microbioma Gastrointestinal , Animais , Dieta Hiperlipídica , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Obesidade , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: The educational efficacy in neonatal resuscitation relies on the subject and teaching strategies. Therefore, it is imperative to test diverse educational methods if they are more instructive to engage students in active learning and practicing knowledge. Hence, the present study aims to investigate the efficacy of a pedagogical framework in neonatal resuscitation skill learning among nursing students in a resource-limited setting. METHODS: A single-blind randomized controlled trial was conducted between October 2020 to March 2021. Sixty nursing students in the 3rd and 4th year of professional training were randomly allocated to the pedagogy and the traditional group. The pedagogy group learned via 6-step LSPPDM (Learn, See, Practice, Prove, Do, Maintain) pedagogy including lectures, video, clinical observation, skill sessions under supervision, and self-directed practice. The traditional group learned through 2-step (Learn, Practice) method that included lectures and skill sessions under supervision. The outcomes measured included technical and non-technical skills in neonatal resuscitation. The technical skill deals with steps such as stimulation, ventilation, oxygenation, intubation, chest compression, medications, and reporting. Non-technical skills refer to teamwork skills that focus on the interaction between leader and helper. Both skills were measured through previously published validated tools two times before and after the intervention by blinded assessors in a simulated delivery room. RESULTS: Overall, the skill was significantly improved in both groups after intervention. Yet, the results showed that the mean difference of technical skill score in the pedagogy group (24.3 ± 3.5) was significantly higher (p < 0.001) compared to the traditional group (16.2 ± 2.4). Likewise, the mean difference of non-technical skill score in the pedagogy (36.9 ± 1.9) was highly significant (p < 0.001) compared to the traditional group (31.2 ± 1.7). CONCLUSIONS: The LSPPDM pedagogy was found more effective in enhancing technical and non-technical skills in neonatal resuscitation compared to the traditional method. The results of this study support the efficacy of the 6-step LSPPDM pedagogy in the education of nursing students regarding neonatal resuscitation in a resource-limited setting. TRIAL REGISTRATION: Prospectively registered at ClinicalTrials.gov ( NCT04748341 ).
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Ressuscitação , Estudantes de Enfermagem , Competência Clínica , Humanos , Recém-Nascido , Aprendizagem , Método Simples-CegoRESUMO
Colebrookea oppositifolia is a highly used medicinal plant and an enriched source of essential oils. Therefore, the present study was designed with the aim to extract the chemical constituents and to evaluate its antioxidant potential. Fresh plant parts were subjected to the extraction of volatile chemical constituents by maceration using n-hexane as the menstruum. The resulting n-hexane fractions were purified and then subjected to GC-MS and FTIR analysis. In-vitro antioxidant abilities were evaluated by, DPPH, total phenolic content (TPC), total flavonoid content (TFC) method against the standard solutions of (Gallic acid, Quercetin) as a positive control. The GC-MS analysis of leaves, stem and inflorescence showed a total of 100, 98 and 48 components out of which 47, 16 and 17 peaks were identified representing the 67.64 %, 73.16 % and 61.93 % of the total oily fractions, respectively. The FTIR spectrum indicated the presence of various functional groups. In-vitro antioxidant results exhibited that leaves showed the highest antioxidant potential by DPPH (3.365 ± 0.002), and the highest total phenolic content by FC method (203.00 ± 0.091). Foliar micromorphological features were found significant in the authentication of C. oppositifolia. Further pharmacognostic studies of this plant are recommended to evaluate its therapeutic potential.
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Plantas Medicinais , Antioxidantes , Flavonoides , Fenóis , Extratos VegetaisRESUMO
Nanoparticles (NPs) are being recognized as antibacterial agents due to their rapidly increasing multidrug resistance in bacterial pathogens. Hence, there is an unmet need to identify the natural antibacterial agent. The present study aimed to evaluate the antibacterial activity of sericin-conjugated silver NPs synthesized by using sericin as a reducing and capping agent. Synthesized NPs were characterized by scanning electron microscope, nanolaser particle size analyzer (BT-90), Fourier-transform infrared analysis, and energy-dispersive X-ray. The biogenic NPs significantly inhibited the growth of Escherichia coli (12-15 mm zone of inhibition), Staphylococcus aureus (14.6-15.4 mm zone of inhibition), and Klebsiella pneumoniae (12.5-18 mm zone of inhibition). The stability of naturally synthesized NPs was examined at various temperatures (i.e., 4°C, 37°C, and 55°C) and pH (i.e., 3, 7, and 11). Temperature variability did not significantly affect the efficacy of NPs. However, NPs performed better at higher pH levels. This study suggested that the sericin-based silver NPs are not only effective against bacteria, but they also maintain the stability at different ranges of temperature and pH. We concluded that the sericin-conjugated silver NPs possess the remarkable antibacterial potential, which suggests their large-scale use as a cheap and stable antimicrobial agent in the future.
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Antibacterianos/farmacologia , Nanopartículas Metálicas/química , Sericinas/química , Prata/química , Animais , Antibacterianos/química , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bombyx , Estabilidade de Medicamentos , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Nanoconjugados/química , Prata/farmacologia , TemperaturaRESUMO
Chronic infections caused by gram negative bacteria are the mains reasons to have morbidity and death in patients, despite using high doses of antibiotics applied to cure diseases producing by them. This study was designed to identify the role of flagella in biofilm formation Ten pure strains were collected from our lab. Morphological variation and motility assays led us to study two strains in detail. They were characterized biochemically, physiologically and genetically. Biofilm formation analysis was performed using test tube assay, congo red assay and liquid-interface coverslip assay. In order to disrupt flagella of studied strains, blending was induced for 5, 10 and 15 minutes followed by centrifugation and observing motility using motility test. Biofilm quantification of wild type (parental) and blended strains was done using test tube and liquid interface coverslip assays. 16S rRNA sequencing identified strains as Pseudomonas aeroginosa and Enterobacter cloacae. Significant biofilm formation (p>0.05) by was observed after 72 and 18 hours using test tube and liquid-interface coverslip assays respectively. Flagellar disruption showed that 15 minutes blending caused significant reduction in both strains, hence demonstrated that flagellar mediated motility could be a potent strategy to stabilize aggregate and invest resources for biofilm formation in P. aeruginosa and E. cloacae.
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Biofilmes/crescimento & desenvolvimento , Enterobacter cloacae/fisiologia , Flagelos/fisiologia , Pseudomonas aeruginosa/fisiologia , Enterobacter cloacae/citologia , Enterobacter cloacae/genética , Concentração de Íons de Hidrogênio , Pseudomonas aeruginosa/citologia , Pseudomonas aeruginosa/genética , RNA Ribossômico 16S , Temperatura , Fatores de TempoRESUMO
Bacteria live either independently as planktonic cells or in organized surface associated colonies called as biofilms. Biofilms play an important role in increased pathogenesis of bacteria and it is assumed that motility is one of the contributing factors towards biofilm initiation. This study was planned to identify the role of flagella in biofilm formation by constructing flagellated (wild type) and physically disrupted variants (non-motile). Total 10 clinical bacterial strains were isolated and characterized. Morphological and biochemical study identified these strains as Enterobacter spp., Pseudomonas spp., Yersinia spp., Escherichia spp., Salmonella spp., Proteus spp., Staphylococcus spp., Streptococcus spp., Lactobacillus spp. and Bacillus spp. Among all strains, two strains including Yersinia spp and Bacillus spp. showed higher antibiotic resistance, hence studied at molecular and physiological level. Biofilm formation capacity of strains was analyzed using three methods including Congo red assay, Test tube assay and Liquid-interface coverslip assay. Afterwards, flagellar disintegration was induced by blending and centrifugation for 5, 10 and 15 minutes. 16S rRNA sequencing showed two strains as Bacillus cereus and Yersinia enterocolitica. Both strains produced significant biofilm by all three above mentioned methods. A motility test of these blended variants showed partial/diminished motility with increased blending time. The significant loss in biofilm formation after 15 minutes blending confirmed the important flagellar contribution to the initiation of biofilm formation. This biofilm defect observed in flagella paralysed/minus variants presumably may be due to defects in attachments to surface at early stages. This study indicated that flagellar motility is crucial initially for surface attachment and subsequently for biofilm formation.
Assuntos
Bacillus cereus/fisiologia , Biofilmes/crescimento & desenvolvimento , Movimento Celular/fisiologia , Flagelos/microbiologia , Flagelos/fisiologia , Yersinia enterocolitica/fisiologia , Bacillus cereus/isolamento & purificação , Humanos , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , Yersinia enterocolitica/isolamento & purificaçãoRESUMO
BACKGROUND: Recently, we reported high in vitro antibacterial efficacy of Althaea officinalis, Ziziphus jujuba, Cordia latifolia and Thymus vulgaris out of a total 21 plants against wide range of bacteria including MRSA. This study was therefore, designed to confirm efficacy of these four herbs against MRSA in an animal model. METHODS: A pilot study was conducted to establish the dose of S. aureus (KY698020) required to induce clinical infection. Afterword, in main trial, efficacy of aforementioned plant extracts on the course of sore throat was checked by evaluating general health, gross lesion score, bacterial load and hematology in mice. RESULTS: Pilot study revealed that 40 µl dose of 107 CFU/ml could induce infection which persist upto 08 days post infection. Mice treated with T. vulgaris and Z. jujuba showed reduction in gross lesion score of both heart and lungs. Treatment with only some plants could significantly decrease bacterial load of throat (T. vulgaris) heart, blood and joint (C. latifolia, and T. vulagris). Hematological indicators confirmed in vivo control of MRSA infection in all treatment groups except A. officinalis. CONCLUSION: This is first report confirming in vivo anti-MRSA potential of C. latifolia and T. vulgaris and highlight the need to explore bioactive constituents of these plants. Moreover, previously reported in vitro antibacterial efficiency of A. officinalis could not be validated in current study.
Assuntos
Carga Bacteriana/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Althaea/química , Animais , Cordia/química , Modelos Animais de Doenças , Testes Hematológicos , Camundongos , Projetos Piloto , Extratos Vegetais/química , Infecções Estafilocócicas/mortalidade , Thymus (Planta) , Distribuição Tecidual , Ziziphus/químicaRESUMO
Present study has been undertaken to isolate and identify the bacterial flora in raw, boiled and pasteurized milk. Agar disc diffusion method was used to determine their sensitivity using medicinal plants, antibiotics and heavy metals. Methylene blue reduction test was used to test the quality of milk samples. Total 10 pathogenic strains were isolated, five strains were isolated from raw milk, three from boiled milk and 2 two from pasteurized milk. To determine optimum conditions for growth, these pathogenic microorganisms were incubated at various temperatures and pH. Gram's staining and biochemical tests revealed that these pathogenic bacteria include Lactobacillus sp., E. coli, Salmonella sp., Pseudomonas sp., Streptococcus sp. and Staphylococcus. Ribotyping revealed S2 as Pseudomonas fluorescens, S5 as Lactococcus lactis and S9 as Lactobacillus acidophilus. Prevalence of pathogenic organisms provided the evidence that contamination of milk arises during milking, transportation and storage of milk. Raw milk is more contaminated than other two types of milk because it contains highest percentage of pathogenic organisms and pasteurized milk was found to be of best quality among three types. So it is recommended to drink milk after proper boiling or pasteurization. Proper pasteurization and hygienic packing of milk is essential to minimize contamination in milk which can save human beings from many milk borne diseases. Our study suggests that antimicrobial use in animal husbandry should be minimized to reduce the hazard of antibiotic resistance. Plant extracts are better alternative against pathogenic bacteria in milk.
Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Microbiologia de Alimentos , Temperatura Alta , Metais Pesados/farmacologia , Leite/microbiologia , Pasteurização/métodos , Extratos Vegetais/farmacologia , Plantas Medicinais , Temperatura de Transição , Animais , Antibacterianos/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Extratos Vegetais/isolamento & purificação , Plantas Medicinais/química , RibotipagemRESUMO
OBJECTIVE: To investigate the role of glutathione peroxidase 1 (GPX1) C/T polymorphism (rs1800668) in modulating the chances of Rheumatoid arthritis (RA) in Pakistani population. METHODS: A total of 400 individuals including 200 controls and 200 patients of RA, were genotyped. Detection of rs1800668 polymorphism was carried out using PCR based amplification strategy (allele specific). RESULTS: The results for Hardy Weinberg Equilibrium (HWE) indicated that the allele frequencies for GPX1 polymorphism were not deviant from HWE in whole population under observation. The statistical analysis indicated that significant association existed between rs1800668 polymorphism and RA (p<0.01). CT genotype increased the risk of RA development by 1.8582 times (OR: 1.8582; 95% CI 1.2154 to 2.8409). CC genotype was found to have protective effect against the disease development (OR: 0.5133; 95% CI 0.3403 to 0.7742) while TT genotype was found to have association with RA development but the risk level was marginal (OR: 1.5319; 95% CI 0.6124 to 3.8322). CONCLUSION: The present finding suggests the importance of GPX1 C/T polymorphism (rs1800668) in development of RA in Pakistani population. The protective role of CC genotype against the development of RA in local population was also observed.
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The quorum sensing (QS) dependent behaviour of micro-organisms, in particular expression of virulence genes, biofilm formation and dispersal, have provided impetus for investigating practical approaches to interfere with microbial QS. This study tests Halomonas pacifica and Marinobacter hydrocarbonoclasticus, two halophilic marine micro-organism, for their AI-2 dependent QS signalling and the effect of two well-known quorum-sensing inhibitors (QSIs), patulin and penicillic acid, on biofilm formation. We report, for the first time, the successful amplification of a putative luxS gene in H. pacifica using degenerated primers and AI-2 dependent QS as well as inhibition using QSIs. Penicillic acid had a strong inhibitory effect on AI-2 induction of H. pacifica at non-growth inhibitory concentrations, while patulin has an adverse effect only at the highest concentration (25 µM). QSIs effect on biofilm forming capability was isolate specific, with maximum inhibition at 25 µM of patulin in H. pacifica. In M. hydrocarbonoclasticus, no adverse effects were noted at any tested concentration of either QSIs. Detection of bioluminescence and the presence of a putative luxS gene provide biochemical and genetic evidence for the production of a signalling molecule(s) which is the essential first step in characterizing H. pacifica QS. This study highlights the importance of AI-2 dependent QS in a marine setting, not previously reported. It further suggests that QSI compounds must be selected in the specific system in which they are to function, and they cannot easily be transferred from one QS system to another.
Assuntos
Organismos Aquáticos/fisiologia , Biofilmes/crescimento & desenvolvimento , Halomonas/fisiologia , Marinobacter/fisiologia , Percepção de Quorum , Organismos Aquáticos/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Halomonas/efeitos dos fármacos , Homosserina/análogos & derivados , Homosserina/metabolismo , Lactonas/metabolismo , Marinobacter/efeitos dos fármacos , Patulina/metabolismo , Ácido Penicílico/metabolismoRESUMO
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.
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The order Hymenoptera holds great significance for humans, particularly in tropical and subtropical regions, due to its role as a pollinator of wild and cultivated flowering plants, parasites of destructive insects and honey producers. Despite this importance, limited attention has been given to the genetic diversity and molecular identification of Hymenopteran insects in most protected areas. This study provides insights into the first DNA barcode of Hymenopteran insects collected from Hazarganji Chiltan National Park (HCNP) and contributes to the global reference library of DNA barcodes. A total of 784 insect specimens were collected using Malaise traps, out of which 538 (68.62%) specimens were morphologically identified as Hymenopteran insects. The highest abundance of species of Hymenoptera (133/538, 24.72%) was observed during August and least in November (16/538, 2.97%). Genomic DNA extraction was performed individually from 90/538 (16.73%) morphologically identified specimens using the standard phenol-chloroform method, which were subjected separately to the PCR for their molecular confirmation via the amplification of cytochrome c oxidase subunit 1 (cox1) gene. The BLAST analyses of obtained sequences showed 91.64% to 100% identities with related sequences and clustered phylogenetically with their corresponding sequences that were reported from Australia, Bulgaria, Canada, Finland, Germany, India, Israel, and Pakistan. Additionally, total of 13 barcode index numbers (BINs) were assigned by Barcode of Life Data Systems (BOLD), out of which 12 were un-unique and one was unique (BOLD: AEU1239) which was assigned for Anthidium punctatum. This indicates the potential geographical variation of Hymenopteran population in HCNP. Further comprehensive studies are needed to molecularly confirm the existing insect species in HCNP and evaluate their impacts on the environment, both as beneficial (for example, pollination, honey producers and natural enemies) and detrimental (for example, venomous stings, crop damage, and pathogens transmission).
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Himenópteros , Parques Recreativos , Humanos , Animais , Abelhas/genética , Paquistão , Código de Barras de DNA Taxonômico/métodos , Insetos/genética , Himenópteros/genética , Plantas/genéticaRESUMO
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.
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Metais Pesados , Oligoquetos , Poluentes do Solo , Animais , Feminino , Bovinos , Esterco , Cádmio , Búfalos , Biodegradação Ambiental , Ensaio Cometa , Solo/química , Metais Pesados/análise , Cromo , Análise Espectral , Poluentes do Solo/análiseRESUMO
A low temperature alkali (LTA) pretreatment method was used to treat wheat straw. In order to obtain good results, different factors like temperature, incubation time, NaOH concentration and solid to liquid ratio for the pretreatment process were optimized. Wheat straw is a potential biomass for the production of monomeric sugars. The objective of the current study was to observe the saccharification (%) of wheat straw with immobilized magnetic nanoparticles (MNPs). For this purpose, immobilized MNPs of purified ß-xylanase enzyme was used for hydrolysis of pretreated wheat straw. Wheat straw was pretreated using the LTA method and analyzed by SEM analysis. After completion of the saccharification process, saccharification% was calculated by using a DNS method. Scanning electron micrographs revealed that the hemicellulose, cellulose and lignin were partially removed and changes in the cell wall structure of the wheat straw had caused it to become deformed, increasing the specific surface area, so more fibers of the wheat straw were exposed to the immobilized ß-xylanase enzyme after alkali pretreatment. The maximum saccharification potential of wheat straw was about 20.61% obtained after pretreatment with optimized conditions of 6% NaOH, 1/10 S/L, 30 °C and 72 hours. Our results indicate the reusability of the ß-xylanase enzyme immobilized magnetic nanoparticles and showed a 15% residual activity after the 11th cycle. HPLC analysis of the enzyme-hydrolyzed filtrate also revealed the presence of sugars like xylose, arabinose, xylobiose, xylotriose and xylotetrose. The time duration of the pretreatment has an important effect on thermal energy consumption for the low-temperature alkali method.
RESUMO
Antibiotic drug resistance is a global public health issue that demands new and novel therapeutic molecules. To develop new agents, animal secretions or products are used as an alternative agent to overcome this problem. In this study, earthworm (Pheretima posthuma) coelomic fluid (PCF), and body paste (PBP) were used to analyze their effects as antibiofilm agents against four bacterial isolates MH1 (Pseudomonas aeruginosa MT448672), MH2 (Escherichia coli MT448673), MH3 (Staphylococcus aureus MT448675), and MH4 (Klebsiella pneumoniae MT448676). Coelomic fluid extraction and body paste formation were followed by minimum inhibitory concentrations (MICs), biofilm formation time kinetics, and an antibiofilm assay, using heat and cold shock, sunlight exposure auto-digestion, and test tube methods. The results showed that the MIC values of PCF and PBP against S. aureus, P. aeruginosa, K. pneumoniae, and E. coli bacterial isolates ranged from 50 to 100 µg/mL, while, the results related to biofilm formation for P. aeruginosa, S. aureus, and K. pneumoniae strains were observed to be highly significantly increased (p < 0.005) after 72 h. E. coli produced a significant (p < 0.004) amount of biofilm after 48 h. Following time kinetics, the antibiofilm activity of PCF and PBP was tested at different concentrations (i.e., 25-200 µg/mL) against the aforementioned four strains (MH1-MH4). The findings of this study revealed that both PBP (5.61 ± 1.0%) and PCF (5.23 ± 1.5%) at the lowest concentration (25 µg/mL) showed non-significant (p > 0.05) antibiofilm activity against all the selected strains (MH1-MH4). At 50 µg/mL concentration, both PCF and PBP showed significant (p < 0.05) biofilm inhibition (<40%) for all isolates. Further, the biofilm inhibitory potential was also found to be more significant (p < 0.01) at 100 µg/mL of PCF and PBP, while it showed highly significant (p < 0.001) biofilm inhibition at 150 and 200 µg/mL concentrations. Moreover, more than 90% biofilm inhibition was observed at 200 µg/mL of PCF, while in the case of the PBP, <96% biofilm reduction (i.e., 100%) was also observed by all selected strains at 200 µg/mL. In conclusion, earthworm body fluid and paste have biologically active components that inhibit biofilm formation by various pathogenic bacterial strains. For future investigations, there is a need for further study to explore the potential bioactive components and investigate in depth their molecular mechanisms from a pharmaceutical perspective for effective clinical utilization.
RESUMO
Circular single-stranded DNA viruses of the family Geminiviridae encode replication-associated protein (Rep), which is a multifunctional protein involved in virus DNA replication, transcription of virus genes, and suppression of host defense responses. Geminivirus genomes are replicated through the interaction between virus Rep and several host proteins. The Rep also interacts with itself and the virus replication enhancer protein (REn), which is another essential component of the geminivirus replicase complex that interacts with host DNA polymerases α and δ. Recent studies revealed the structural and functional complexities of geminivirus Rep, which is believed to have evolved from plasmids containing a signature domain (HUH) for single-stranded DNA binding with nuclease activity. The Rep coding sequence encompasses the entire coding sequence for AC4, which is intricately embedded within it, and performs several overlapping functions like Rep, supporting virus infection. This review investigated the structural and functional diversity of the geminivirus Rep.