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This study presents the fabrication and characterization of mixed matrix membranes (MMMs) incorporating green-synthesized silver nanoparticles (AgNPs) using Hibiscus Rosa sinensis extract within a polyethersulfone (PES) matrix for nanofiltration (NF) application. The membranes were evaluated for their pure water permeability, salt rejection, dye removal, and antifouling performance. Results showed that the membrane with 0.75 wt% AgNPs exhibited the highest pure water permeability of 36 L/m 2 h -1 bar -1 attributed to increased porosity and enhanced hydrophilicity. Addition of 0.75wt% AgNPs resulted in significant improvements, with NaCl rejection increased from 32 to 57%, MgSO 4 from 26 to 67%, and CaCl 2 from 27 to 41%. Antifouling tests revealed that the 0.75 wt% AgNPs membrane had the lowest irreversible fouling and highest flux recovery due to the antimicrobial action and improved surface properties of AgNPs. Importantly, the performance of the fabricated membranes align with loose nanofiltation characteristcs, as evidence by high dye rejection rates coupled with moderate rejection of salts. This study highlights the potential of green-synthesized AgNPs as effective nanofillers for developing high-performance and environmentally sustainable membranes into wastewater treatment., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Glutathione Peroxidase 1 (GPx1) gene has been reported for its role in cellular redox homeostasis, and the dysregulation of its expression is linked with the progression of diverse cancers. Non-synonymous single nucleotide polymorphism (nsSNPs) have been emerged as the crucial factors, playing their role in GPx1 overexpression. To understand the deleterious mutational effects on the structure and function of GPx1 enzyme, we delved deeper into the exploration of possibly damaging nsSNPs using in-silico based approaches. Eight widely utilized computational tools were employed to roughly shortlist the deleterious nsSNPs. Their damaging effects on structure and function of the genes were evaluated by using different bioinformatics tools. Subsequently, the three final proposed deleterious mutants including mutations rs373838463, rs2107818892, and rs763687242, were docked with their reported binder, TNF receptor-associated factor 2 (TRAF2). The lowest binding affinity and stability of the docked mutant complexes as compared to the wild type GPx1 were validated by molecular dynamic simulation. Finally, the comparison of RMSD, RMSF, RoG and hydrogen bond analyses between wild-type and mutant's complexes validated the deleterious effects of proposed nsSNPs. This study successfully identified and verified the possibly damaging nsSNPs in GPx1 enzyme, which may be linked the progression of various types of cancer. Our findings underscore the value of in-silico approaches in mutational analysis and encourage further preclinical and clinical trials., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests. Ethical consideration Not applicable., (© 2024. The Author(s).)

Introduction: This research aims to explore the molecular composition, antioxidant capabilities, and antibacterial effects of the essential oils from Tetraclinis articulateata and Juniperus phoenicea ., Methods: Essential oils were extracted using hydrodistillation. Gas chromatography combined with mass spectrometry was used to determine the chemical makeup of essential oils. Two methods are used to assess the antioxidant activity of essential oils: the reduction of iron (ferric reducing antioxidant power or frap) and the trapping of the free radical 2,2-Diphenyl-1-Picrylhydrazyl (DPPH). The antimicrobial potential of essential oils was assessed using the diffusion method on a solid-state disk in comparison to nine bacterial and seven fungal souches., Results and Discussion: The essential oil yields from Tetraclinis articulata and Juniperus phoenicea are 0.46% ± 0.02% and 0.83% ± 0.05%, respectively. According to CG/SM's chromatographic analyses, the predominant constituent in the essential oil of J. Phoenicea is α-pinène (59.51%), while the main constituents in the essential oil of T. Articulata ? are Bornyle acetate (18.91%) and camphor (28.48%). The assessment of antioxidant activities reveals intriguing antioxidant qualities in the essential oils of the species under investigation. T. Articulata essential oils yield the greatest results in the DPPH and FRAP tests, with CI50 values of around 266.9 ± 5.4 μg/mL and EC50 values of 433.16 ± 4.13 μg/mL, respectively. Except for Staphylococcus epidermidis, Staphylococcus aureus BLACT, and Pseudomonas aeroginosa, the two essential oils have demonstrated significant bactericidal activity against all bacterial and fungal souches (MIC <2 mg/mL et MBC <3.5 mg/mL). The inhibiting effect of these oils on bacterial and fungal development raises potential application areas in the food, cosmetic, and pharmaceutical industries. In addition, the current study investigated the potential antifungal, antibacterial, and antioxidant activities of the essential oils from Juniperus phoenicea and Tetraclinis articulate plants via the Glide molecular docking methodology, and most of these constituents were observed to be potent therapeutic agents., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Asbabou, Hanane, Gourich, Siddique, Drioiche, Remok, Saidi, Adadi, Khamar, Almaary, Mekonnen, Bourhia, Bouzoubaa and Zair.)

Sugarcane smut is the most damaging disease that is present almost across the globe, causing mild to severe yield losses depending upon the cultivar types, pathogen races and climatic conditions. Cultivation of smut-resistant cultivars is the most feasible and economical option to mitigate its damages. Previous investigations revealed that there is a scarcity of information on early detection and effective strategies to suppress etiological agents of smut disease due to the characteristics overlapping within species complexes. In this study, 104 sugarcane cultivars were screened by artificial inoculation with homogenate of all possible pathogen races of Sporisorium scitamineum during two consecutive growing seasons. The logistic smut growth pattern and the disease intrinsic rate were recorded by disease growth curve. Variable levels of disease incidence i.e., ranging from 0 to 54.10% were observed among these sugarcane cultivars. Besides, pathogen DNA in plant shoots of all the cultivars was successfully amplified by PCR method using smut-specific primers except 26 cultivars which showed an immune reaction in the field trial. Furthermore, the plant germination and tillering of susceptible sugarcane cultivars were greatly influenced by pathogen inoculation. In susceptible cultivars, S. scitamineum caused a significant reduction in setts germination, coupled with profuse tillering, resulting in fewer millable canes. Correlation analysis demonstrated that there was a positive relationship between reduction in setts germination and increase in the number of tillers. The present study would be helpful for the evaluation of smut resistance in a wide range of sugarcane germplasm, especially from the aspects of setts germination and tillers formation, and it also screened out several excellent germplasm for potential application in sugarcane breeding., (© 2024. The Author(s).)

Origanum grosii (L.) and Thymus pallidus (L.) are medicinal plants recognized for their uses in traditional medicine. In this context, the aim of this article is to highlight the results of a phytochemical analysis (HPLC), with particular emphasis on the antioxidant (DPPH, TAC, and FRAP), analgesic, anti-inflammatory, haemagglutinin-test-related, and hemolytic activities of the total extracts of these plants. Phytochemical analysis via HPLC revealed that licoflavone C (30%) is the main compound in Origanum grosii , while hesperidin (43%) is found in T. pallidus . Evaluation of the antioxidant capacity of Origanum grosii and Thymus pallidus using the DPPH, TAC, and FRAP methods revealed an IC50 of the order of 0.085 mg/mL and 0.146 mg/mL, an EC50 of the order of 0.167 mg/mL and 0.185 mg/mL, and a total antioxidant capacity of between 750 mg EQ/g and 900 mg EQ/g, respectively. Analgesic evaluations revealed writhes inhibition of the order of 97.83% for O. grosii and 90% for T. pallidus . In addition, both plant extracts showed limited hemolytic activity, not exceeding 30% at a concentration of 100 mg/mL. Evaluation of the anti-inflammatory potential showed edema inhibition of the order of 94% (800 mg/kg) for O. grosii and 86% (800 mg/kg) for T. pallidus . These results highlight the potential applications of these extracts in pharmacological research.

Saprolegnia parasitica is responsible for devastating infections in fish and poses a tremendous threat to the global aquaculture industry. Presently, no safe and effective control measures are available, on the contrary, use of banned toxic compounds against the pathogen is affecting humans via biomagnification routes. This pioneering study aims to design an effective multi-epitope multi-target vaccine candidate against S. parasitica by targeting key proteins involved in the infection process. The proteins were analyzed and linear B-cell epitopes, MHC class I, and class II epitopes were predicted. Subsequently, highly antigenic epitopes were selected and fused to a highly immunogenic adjuvant, 50S ribosomal protein L7/L12, to design a multi-epitope chimeric vaccine construct. The structure of the vaccine was generated and validated for its stereochemical quality, physicochemical properties, antigenicity, allergenicity, and virulence traits. Molecular docking analyses demonstrated strong binding interactions between the vaccine and piscine immune receptors (TLR5, MHC I, MHC II). Molecular dynamics simulations and binding energy calculations of the complexes, further, reflected the stability and favorable interactions of the vaccine and predicted its cytosolic stability. Immune simulations predicted robust and consistent kinetics of the immune response elicited by the vaccine. The study posits the vaccine as a promising solution to combat saprolegniasis in the aquaculture industry., (© 2024. The Author(s).)

Apple vinegar is highly recommended for nutrition due to its health benefits and bioactive components. However, the apple cultivar greatly influences the quality of the vinegar. In this research, our focus was on examining the impact of four different apple cultivars on the physicochemical attributes, chemical composition, as well as biological properties-including antidepressant and anti-inflammatory activities-of vinegar. Interestingly, the physicochemical properties of vinegar and the contents of acetic acid and polyphenols depend on the apple cultivars. HPLC chromatographic analysis showed that citric acid (820.62-193.63 mg/100 g) and gallic acid (285.70-54.40 µg/g) were mostly abundant in the vinegar samples. The in vivo results showed that administration of Golden Delicious apple vinegar (10 mL/kg) to adult Wistar rats reduced carrageenan-induced inflammation by 37.50%. The same vinegar sample exhibited a significant antidepressant effect by reducing the rats' immobility time by 31.07% in the forced swimming test. Due to its high acidity, Golden Delicious vinegar was found to be more effective against bacteria, particularly Bacillus subtilis and Candida albicans , resulting in a MIC value of 31.81 mg/mL. Furthermore, the antioxidant activity of various vinegar samples was found to be powerful, displaying optimal values of IC 50 = 65.20 mg/mL, 85.83%, and 26.45 AAE/g in the DPPH, β-carotene decolorization and TAC assays, respectively. In conclusion, the apple cultivars used in this study impact the chemical composition and biological activities of vinegar, which may help demonstrate the importance of raw material selection for the production of vinegar.

The present study aimed to explore the phytochemical profile, and evaluate the antioxidant, antimicrobial, and insecticidal properties, of Moroccan Mentha longifolia L. essential oil (ML-EO) using in vitro and in silico assays. Noteworthily, as chromatography (GC-MS/MS) revealed that ML-EO is majorly composed of piperitenone oxide (53.43%), caryophyllene (20.02%), and (-) germacrene D (16.53%). It possesses excellent antioxidant activity with an IC 50 of 1.49 ± 0.00 for DPPH and 0.051 ± 0.06 μg/mL for ABTS. Moreover, the RP and TAC activities were 0.80 ± 0.01 μg/mL and 315.532 ± 0.00 mg EAA/g, respectively. ML-EO exhibited a potent antimicrobial effect, specifically against Pseudomonas aeruginosa . It also exhibited strong antifungal ability, especially against Candida albicans . Regarding insecticidal activity, for ML-EO, a dose of 20 µL/mL produced a complete reduction in fecundity, fertility, and emergence of adult C. maculatus with mortality rates reaching 100%. In silico results showed that the antioxidant activity is mostly attributed to α-Cadinol, the antibacterial efficiency is attributed to piperitenone oxide, and antifungal capacity is related to cis-Muurola-4(15),5-diene and piperitenone oxide. Accordingly, ML-EO has high potential to be used as an alternative for preserving food and stored grain and protecting them against microbes and insect pests in the food and pharmaceutical sectors.

The drug resistance of bacterial pathogens causes considerable morbidity and death globally, hence there is a crucial necessity for the development of effective antibacterial medicines to address the antibacterial resistance issue. The bioprepared zinc oxide nanoparticles (ZnO-NPs) were prepared utilizing the flower extract of Hibiscus sabdariffa and then characterized using different physicochemical techniques. The antibacterial effectiveness of the bioprepared ZnO-NPs and their synergism with fosfomycin were evaluated using disk diffusion assay against the concerned pathogens. Transmission electron microscopy (TEM) investigation of the bioprepared ZnO-NPs showed that their average particle size was 18.93 ± 2.65 nm. Escherichia coli expressed the highest sensitivity to the bioinspired ZnO-NPs with a suppressive zone of 22.54 ± 1.26 nm at a concentration of 50 µg/disk, whereas the maximum synergistic effect of the bioinspired ZnO-NPs with fosfomycin was noticed against Klebsiella pneumoniae strain with synergism ratio of 100.29%. In conclusion, the bioinspired ZnO-NPs demonstrated significant antibacterial and synergistic efficacy with fosfomycin against the concerned nosocomial bacterial pathogens, highlighting the potential of using the ZnO NPs-fosfomycin combination for effective control of nosocomial infections in intensive care units (ICUs) and health care settings. Furthermore, the biogenic ZnO-NPs' potential antibacterial action against food pathogens such as Salmonella typhimurium and E. coli indicates their potential usage in food packaging applications.

Soil salinity is the major abiotic stress that disrupts nutrient uptake, hinders plant growth, and threatens agricultural production. Plant growth-promoting rhizobacteria (PGPR) are the most promising eco-friendly beneficial microorganisms that can be used to improve plant responses against biotic and abiotic stresses. In this study, a previously identified B. thuringiensis PM25 showed tolerance to salinity stress up to 3 M NaCl. The Halo-tolerant Bacillus thuringiensis PM25 demonstrated distinct salinity tolerance and enhance plant growth-promoting activities under salinity stress. Antibiotic-resistant Iturin C ( ItuC ) and bio-surfactant-producing ( sfp and srfAA ) genes that confer biotic and abiotic stresses were also amplified in B. thuringiensis PM25. Under salinity stress, the physiological and molecular processes were followed by the over-expression of stress-related genes (APX and SOD) in B. thuringiensis PM25. The results detected that B. thuringiensis PM25 inoculation substantially improved phenotypic traits, chlorophyll content, radical scavenging capability, and relative water content under salinity stress. Under salinity stress, the inoculation of B. thuringiensis PM25 significantly increased antioxidant enzyme levels in inoculated maize as compared to uninoculated plants. In addition, B. thuringiensis PM25-inoculation dramatically increased soluble sugars, proteins, total phenols, and flavonoids in maize as compared to uninoculated plants. The inoculation of B. thuringiensis PM25 significantly reduced oxidative burst in inoculated maize under salinity stress, compared to uninoculated plants. Furthermore, B. thuringiensis PM25-inoculated plants had higher levels of compatible solutes than uninoculated controls. The current results demonstrated that B. thuringiensis PM25 plays an important role in reducing salinity stress by influencing antioxidant defense systems and abiotic stress-related genes. These findings also suggest that multi-stress tolerant B. thuringiensis PM25 could enhance plant growth by mitigating salt stress, which might be used as an innovative tool for enhancing plant yield and productivity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ali, Hafeez, Ahmad, Javed, Sumaira, Afridi, Dawoud, Almaary, Muresan, Marc, Alkhalifah and Selim.)

In South Asia, the high costs of synthetic fertilizers have imposed research on alternative nutrient inputs. We aimed to identify potential trace elements (PTE) present in some organic manure that might be a source of environmental pollutions and risk to public health following consumption. The study aims to evaluate how different organic waste (poultry waste, PW; press mud, PM; and farmyard manure, FYM) influences the heavy metal migration in soil, the accumulation in garlic, and their potential health risks. Organic waste caused a higher accumulation of certain metals (Zn, Cu, Fe, and Co), whereas Mn, Cd, Cr, and Pb were in lower concentrations in soil. Amendments of soil with PM revealed a higher accumulation of Cd, Cr, Fe, and Pb, whereas PW resulted in Cu and Zn accumulation in garlic. Treatment of soil with FYM exhibited higher metal concentration of Co and Mn. An environmental hazard indices study revealed that pollution load index (PLI) was highest for Cu following treatment with PM. The health risk index (HRI) was greater for Cd following amendment with PM. Maximum daily intake of metals (DIM) was observed for Zn after treatment with PW. The Pb exhibited maximum bioconcentration factor (BCF) in PM-amended plants. Based on these findings, we concluded that garlic grown on contaminated soil with organic waste may pose serious health hazards following consumption.

Diclofenac is an anti-inflammatory drug and has been frequently detected from the wastewater. In the present study, factors affecting diclofenac adsorption on sewage sludge was evaluated. At 1 mg/L initial diclofenac concentration, more than 80% diclofenac removal was achieved. Adsorption increased at higher concentration (100 mg/L concentration) and more than 99% diclofenac was adsorbed from the wastewater. Significant removal of diclofenac was observed after 5 min contact time. The adsorption efficacy was more than 98% after 50 and 60 min. Pseudo-first and second order kinetics revealed reasonable regression value (0.9) indicated that the model is best fitted. Diclofenac adsorption was extremely high at acidic pHs than alkaline range. The sludge samples showed the presence of multi drug resistant bacteria. Vancomycin-resistant enterococcus stains were 27%, Methicillin-resistant Staphylococcus aureus positive strains were 16.5% and Extended-spectrum betal-lactamase-harbouring Enterobacteriacea were 65.4% in the sludge. The drug resistance Enterobacteriaceae revealed 14 Klebsiella pneumonia strains, 11 strains from E. coli and two from the genus Enterobacter. To conclude, the activated sludge could be effectively utilized for the removal of diclofenac from wastewater., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Citrus is a valuable crop in Pakistan. It is rich in vitamin C, other nutrients and antioxidants. Huanglongbing (HLB) caused by a bacterium "Candidatus liberibacter asiaticus" (CLas), africanus and americanus has an influence on citrus production around the world. Beside HLB there exist several other bacterial species in citrus groves in Pakistan. The structure and diversity of bacterial species in various ecosystems can be quickly examined using NGS. This approach is considerably quicker and more precise than outdated methods. Healthy or citrus greening infected leaf samples of Grapefruit (Citrus paradisi), C. aurantifolia, and C. reticulata Blanco were used for diversity analysis. In this study high throughput, NGS technique was used to access the population of both cultivable and non-cultivable bacterial endophytes from citrus leaves, by using PCR amplicons of 16S rDNA sequences (V5-V7 regions) with Illumina Hi seq. As a result, a total number of 68,722 sequences were produced from the test samples. According to the NGS-based diversity classification, the most common genera of exploited bacterial endophytes were Proteobacteria, Firmicutes, Bacteroides, Cyanobacteria, and Actinobacteria. C. aurantifolia and C. paradisi showed almost equal diversity, whereas C. reticulata Blanco had a higher proportion of Proteobacteria and Cyanobacteria in their leaves. To determine alpha diversity (AD), additional data was analyzed using statistical indices such as Shannon, Chao1, and Simpson. According to the inverse Simpson diversity index, the abundance of the microbial population in six different citrus samples was 0.48, 0.567, and 0.163, respectively. The metagenomics of microbiota in plant tissues was successfully recorded by NGS technology, which can help us learn more about the interactions between plants and microbes. This research is the first step toward a better understanding of 16SrRNA-based metagenomics from citrus in Pakistan using Illumina (Hi seq) Technology., Competing Interests: The authors have declared that no competing interests exist.

In this paper, Magnesium Zinc Ferrite (MZF) nanoparticles (Mg 0.8-x Zn x Fe 2 O 4 , where x = 0.2, 0.4 and 0.6) are successfully fabricated by combustion process. The prepared nanoparticles are characterized through XRD, FTIR, UV, SEM, EDS and TEM. It has been confirmed that the samples produced cubic spinel structure with crystal size in the range of 13-15 nm. From the ultraviolet spectrum, the optical band gap is calculated which ranges from 5.6 to 4.6 eV. TEM micrographs confirm the nanocrystalline nature of combustion derived ferrite nanoparticles with average particle diameter of 7-28 nm. Antibacterial studies confirmed that the nanoparticles are toxic to Pseudomonas aeruginosa consists of greatest zone of inhibition of 25 mm. The antibacterial and photocatalytic studies exhibited improved activity which is strongly influenced by the zinc doping. Photocatalytic degradation study reveal that the prepared nanoparticles function as perfect catalyst for degradation of Methylene Blue (MB) dye and Textile Dyeing Waste Water (TDWW) under UV light, thus revealing their potential usage on organic pollutants., (Copyright © 2021. Published by Elsevier Inc.)

In recent years, many research on the quantity of lignocellulosic waste have been developed. The production, partial purification, and characterisation of ligninolytic enzymes from various fungi are described in this work. On the 21st day of incubation in Potato Dextrose (PD) broth, Hypsizygus ulmarius developed the most laccase (14.83 × 10 -6 IU/ml) and manganese peroxidase (24.11 × 10 -6 IU/ml), while Pleurotus florida produced the most lignin peroxidase (19.56 ×  -6 IU/ml). Laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP), all generated by selected basidiomycetes mushroom fungi, were largely isolated using ammonium sulphate precipitation followed by dialysis. Laccase, lignin peroxidase, and manganese peroxidase purification findings indicated 1.83, 2.13, and 1.77 fold purity enhancements, respectively. Specific activity of purified laccase enzyme preparations ranged from 305.80 to 376.85 IU/mg, purified lignin peroxidase from 258.51 to 336.95 IU/mg, and purified manganese peroxidase from 253.45 to 529.34 IU/mg. H. ulmarius laccase (376.85 IU/mg) with 1.83 fold purification had the highest specific activity of all the ligninolytic enzymes studied, followed by 2.13 fold purification in lignin peroxidase (350.57 IU/mg) and manganese peroxidase (529.34 IU/mg) with 1.77-fold purification. Three notable bands with molecular weights ranging from 43 to 68 kDa and a single prominent band with a molecular weight of 97.4 kDa were identified on a Native PAGE gel from mycelial proteins of selected mushroom fungus. The SDS PAGE profiles of the mycelial proteins from the selected mushroom fungus were similar to the native PAGE. All three partially purified ligninolytic isozymes display three bands in native gel electrophoresis, with only one prominent band in enzyme activity staining. The 43 kDa, 55 kDa, and 68 kDa protein bands correspond to laccase, lignin peroxidase, and manganese peroxidase, respectively., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Background: In recent decades focus of research has been toward an alternative antibacterial agent because of growing bacterial resistance and side effects of antibacterial agents. In the current study, the antibacterial activity of gold nanoparticles has been evaluated on selected human pathogens., Methods: In this study, we used panchagavya (PG) to synthesize gold nanoparticles, and the resulting nanoparticles (PG-AuNPs) were characterized by several spectroscopic techniques. In addition, antibacterial activity of PG-AuNPs against Escherichia coli, Bacillus subtilis, and Klebsiella pneumoniae were studied by well diffusion method., Results: The synthesis of AuNPs was affirmed by a colour change, which was further validated by UV-vis spectra with a maximum absorption peak at 527 nm. Bandgap energy was calculated as 2.13 eV by Tauc method from the UV result. The presence of amino acids and proteins in PG was responsible for the conversion of gold ions to AuNPs, according to FTIR analysis. (111), (200), (220), and (311) crystallographic planes were observed by XRD; further crystalline nature was validated by SAED analysis. The size and zeta value were found to be 53.29 nm and -9.8 mV respectively. Spherical shaped nanoparticles and elemental structure of PG-AuNPs were confirmed by HRTEM and EDS analysis. The antibacterial activity of PG-AuNPs showed the maximum and minimum zone of inhibition against K. pneumoniae (17.12 ± 0.14 mm) and B. subtilis (11.42 ± 0.58 mm)., Conclusion: Antibacterial activity of PG-AuNPs was found to be strong against gram negative bacteria and moderate against gram positive bacteria. Based on the result, it was concluded that PG-AuNPs could be used to combat antibiotic drug resistance. Besides, in vitro and in vivo toxicity studies of PG-AuNPs should be conducted., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Progressive decline of immune response in HIV patients makes them susceptible to frequent bacterial infections. High usage of antibiotics influences the emergence of multidrug-resistant bacteria and worsens the clinical outcomes. In this study, the occurrence of drug-resistant genes in Gram-negative bacterial isolates from HIV patients in South India was analyzed., Methods: A total of 173 Gram-negative bacterial (GNB) isolates from HIV patients were screened for antibiotic susceptibility profile using the Kirby-Bauer diskdiffusion method. Positivity of drug-resistant genes was analyzed using polymerase chain reaction method., Results: In this study, 72.8% of bacterial isolates were obtained from urine specimens, and Escherichia coli (47.4%) was the predominantly isolated bacterium. Overall, 87.3% and 83.2% of GNB were resistant to 3rd generation cephalosporin antibiotics such as cefotaxime and ceftazidime, respectively, 56.6% were resistant to cephamycin (cefoxitin) and 43% to carbapenem (imipenem) antibiotics. Extended-spectrum β-lactamases (ESBL) production was noted among 79.5% of GNB isolates, followed by AmpC (57.1%) and Metallo β-lactamases (37.3%). Molecular analysis revealed that ESBL genes such as blaTEM (94.1%), blaCTX-M (89.2%), and blaSHV (24.2%) were detected at higher levels among GNB isolates. Carbapenemase-producing genes such as blaOXA-48 (20%), blaOXA-23 (2.6%), and both blaOXA-23 and blaOXA-51 like genes (2.6%) and AmpC producing genes such as blaCIT (26.7%), blaDHA (3.6%), and blaACC (1.8%) were detected at low-level., Conclusions: This study concludes that ESBL producing genes are detected at high level among gram-negative bacterial isolates from HIV patients in South India., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Herbal and traditional medicines can play a pivotal role in combating cancer and neglected tropical diseases. Ajuga bracteosa , family Lamiaceae, is an important medicinal plant. The genetic transformation of A. bracteosa with rol genes of Agrobacterium rhizogenes further enhances its metabolic content. This study aimed at undertaking the molecular, phytochemical, and in vitro biological analysis of A. bracteosa extracts. We transformed the A. bracteosa plant with rol genes and raised the regenerants from the hairy roots. Transgenic integration and expression of rolB were confirmed by conventional polymerase chain reaction (PCR) and qPCR analysis. The methanol: chloroform crude extracts of wild-type plants and transgenic regenerants were screened for in vitro antibacterial, antihemolytic, cytotoxic, anticancer, and leishmanial activity. Among all plants, transgenic line 3 (ABRL3) showed the highest expression of the rolB gene. Fourier transform infra-red (FTIR) analysis confirmed the enhanced number of functional groups of active compounds in all transgenic lines. Moreover, ABRL3 exhibited the highest antibacterial activity, minimum hemolytic activity (CC 50 = 7293.05 ± 7 μg/mL) and maximum antileishmanial activity (IC 50 of 56.16 ± 2 μg/mL). ABRL1 demonstrated the most prominent brine shrimp cytotoxicity (LD 50 39.6 ± 4 μg/mL). ABRL3 was most effective against various human cancer cell lines with an IC 50 of 57.1 ± 2.2 μg/mL, 46.2 ± 1.1 μg/mL, 72.4 ± 1.3 μg/mL, 73.3 ± 2.1 μg/mL, 98.7 ± 1.6 μg/mL, and 97.1 ± 2.5 μg/mL against HepG2, LM3, A549, HT29, MCF-7, and MDA-MB-231, respectively. Overall, these transgenic extracts may offer a cheaper therapeutic source than the more expensive synthetic drugs.

The disintegration of natural water sources signals out the scarcity of adam's ale and will be hurdle for the human physical state. So it is necessary to decrease waste loads and hence pressure on the ecology for the sustainability of fishery and dye industry. Herein, TiO 2 nanoparticles doped with Sn and F are synthesized and the influence of simultaneous doping on the optical, surface morphological, structural, photocatalytic and antibacterial activities are investigated. Doping of TiO 2 with Sn and F suppress the growth of both anatase and rutile phase because of the dissimilar boundaries. All the prepared doped and undoped samples are found to possess tetragonal structure. The influence of F and Sn in TiO 2 lattice is recognized with the XRD and FT-IR spectra of the prepared particles The size of the obtained nanoparticles decreases as increasing concentration of F and Sn. TiO 2 is showing the presence of spherical and ellipsoidal nanoparticles whereas doped samples showing nanobulk, pentagons and rods. The absorption edge of the doped samples are blue shifted with increasing concentration of dopants indicates the control of optical absorption property of TiO 2. The visible light assisted photocatalytic degradation of fish processing waste water by doped and undoped samples are found to be established as 0.0076/min and 0.0071/min respectively. Visible light assisted degradation of commercially available dyes and fish processing waste water is assessed. Methyl blue showed enhanced photocatalytic activity under visible light irradiation compared to Methyl orange. It is observed that all the prepared particles show good antimicrobial activity against Staphylococcus aureus., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Klebsiella pneumoniae is predominantly exists in the pus of the human wounds and it creates massive infections in the skin and causes serious health associated infections. Modern antibiotics are highly active in the treatment of wound infections. In this study was aimed to determine resistance of K. pneumoniae screened from wound specimens of patients. Sample was collected from the pus of the patients associated with secondary infection., Methods: Samples were serially diluted and the isolated bacterial strains were characterized by biochemical tests, colony morphology and Gram's staining methods. Resistance of K. pneumoniae was tested using antibiotics such as, Gentamycin, Ampicillin, Tetracycline, Cefurooxime, Oxacillin, Ofloxacin, Erythromycin, Nalidic acid, Cefepine, Piperacillin, Norfloxacin, Imipenem, Nitrofurantoin, Amikacin, Ciprofloxacin, Vancomycin, Meropeneum and Cefotaxime with Kirby-Bauer disc diffusion method., Results: Among the 73 K. pneumoniae strains, four strains produced AmpC and ESBLs, 42 strains produced ESBLs and 7 bacterial strains synthesized only AmpC enzyme. Four stains produced ESBLs and showed multidrug resistance against various antibiotics. Most of the strains synthesized extracellular polysaccharides and mediated biofilm formation. Among the K. pneumoniae strains, K. pneumoniae PS02 showed multidrug resistant against most of the tested antibiotics. It produced ESBLs and AmpC enzyme. To produce secondary metabolites, actinomycetes were isolated and characterized as Streptomyces sp. AC14. The secondary metabolite was effective against Klebsiella strains., Conclusions: To conclude, secondary metabolites extracted from Streptomyces sp. AC14 was found to be effective against multidrug resistant bacterium. Further studies are warranted to analyze the drug hydrolyzing pathways of bacteria and to identify the mechanism of action of secondary metabolites from Streptomyces sp. AC14., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In the present study, Allium cepa leaf extract was utilized to reduce the silver nitrate into the nanoscale range of silver ions (Ag NPs). The biosynthesized Ag NPs were extensively characterized by X-ray diffraction analysis (XRD), Dynamic light scattering analysis (DLS), UV-Visible spectroscopy (UV-vis), Transmission electron microscopy (TEM), Energy dispersive X-ray analysis (EDX) and Fourier transform infrared spectroscopy (FTIR). The antioxidant activity of synthesized Ag NPs was verified by DPPH assay. From the results obtained from XRD and DLS studies, the size of Ag NPs was determined to be around 54.3 nm. The measured zeta potential value of -19.1 mV confirms the excellent stability of biosynthesized Ag NPs. TEM analyses reveal that the biosynthesized Ag NPs have a spherical structure of 13 nm in size. The presence of various functional groups was confirmed through FTIR studies and EDAX verifies the weight percentage of silver content in biosynthesized nanoparticles to be 30.33%. In the present study, anti-cancer activity was carried out by using breast cancer cell line MCF-7. Further, silver nanoparticles exhibited antimicrobial effectiveness against gram-positive Bacillus cereus and gram-negative Escherichia coli. The MTT assay also showed better cytotoxic activity against the MCF- 7 cell line., (Copyright © 2021 Elsevier Inc. All rights reserved.)

We find out the antimicrobial potential of partially purified 2,4-diacetylphloroglucinol (DAPG) against Ralstonia solanacearum and fungal plant pathogens isolated from tomato rhizobacterium Pseudomonas fluorescens VSMKU3054. The present study is mainly focused on the control of wilt disease of tomato by our isolate VSMKU3054 and DAPG. The cell free culture filtrate of P. fluorescens VSMKU3054 was significantly arrested the growth of R. solanacearum and fungal pathogens such as Rhizoctonia solani , Sclerotium rolfsii , Macrophomina phaseolina and Fusarium oxysporum compared to control. The existence of DAPG from the crude metabolites of P. fluorescens VSMKU3054 was confirmed on TLC with Rf value 0.34, which is coincide with that of authentic phloroglucinol. The partially purified DAPG exhibited much higher activity against R. solanacearum at 30 µg/ml than the fungal plant pathogens compared to control. The antimicrobial partially purified compound was identified as DAPG by UV, FT-IR and GC-MS analysis. The percentage of live cells of R. solanacearum when supplemented with DAPG at 30 µg/ml, significantly controlled the living nature of R. solanacearum up to 68% compared to tetracycline and universal control observed under high content screening analysis. The selected isolate P. fluorescens VSMKU3054 and DAPG significantly controlled wilt disease of tomato up to 59.5% and 42.12% on 3rd and 7th days compared to positive and negative control by detached leaf assay. Further, in silico analysis revealed that high interaction of DAPG encoding protease with lectin which is associated with R. solanacearum . Based on our findings, we confirmed that P. fluorescens VSMKU3054 and DAPG could be used a potential bio inoculants for the management of bacterial wilt disease of tomato., Competing Interests: The authors declared that there is no conflict of interest., (© 2021 The Author(s).)

Plant viral diseases are the foremost threat to sustainable agriculture, leading to several billion dollars in losses every year. Many viruses infecting several crops have been described in the literature; however, new infectious viruses are emerging frequently through outbreaks. For the effective treatment and prevention of viral diseases, there is great demand for new techniques that can provide accurate identification on the causative agents. With the advancements in biochemical and molecular biology techniques, several diagnostic methods with improved sensitivity and specificity for the detection of prevalent and/or unknown plant viruses are being continuously developed. Currently, serological and nucleic acid methods are the most widely used for plant viral diagnosis. Nucleic acid-based techniques that amplify target DNA/RNA have been evolved with many variants. However, there is growing interest in developing techniques that can be based in real-time and thus facilitate in-field diagnosis. Next-generation sequencing (NGS)-based innovative methods have shown great potential to detect multiple viruses simultaneously; however, such techniques are in the preliminary stages in plant viral disease diagnostics. This review discusses the recent progress in the use of NGS-based techniques for the detection, diagnosis, and identification of plant viral diseases. New portable devices and technologies that could provide real-time analyses in a relatively short period of time are prime important for in-field diagnostics. Current development and application of such tools and techniques along with their potential limitations in plant virology are likewise discussed in detail.

Acanthamoeba is an opportunistic protozoan pathogen which is found in diverse environment worldwide. Being ubiquitous nature of this amoeba we come across it in our daily life. Acanthamoeba species are recognized as human pathogens; that may cause blinding keratitis and rare but fatal granulomatous encephalitis involving central nervous system. To date, there is not a single report in literature demonstrating anti-Acanthamoeba antibodies among the Saudi population, and thus aim of the present study. Using ELISA, we identified the antibody level in the local population. Our results represent the secretory IgA antiAcanthamoeba in mucosal secretions from 133 individuals aged 15-60 years. The antiAcanthamoeba antibody prevalence rate was > 80%, and no considerable differences were observed between prevalence in males (80.28%) and that in females (80.64%). In addition, environmental sources (soil and water) from the environment of the participants in our study were evaluated for amoeba incidence. The amoeba was identified by morphological characteristics of cysts or trophozoites on non-nutrient agar plates grown with E. coli. Overall, 58.75% of samples from water and 32.85% of those from soil were culture positive for outgrowth of amoeba on non-nutrient agar plates. Furthermore, PCR was carried out with genus-specific primers to confirm the presence of Acanthamoeba DNA. Our results revealed that about 68% of cultures from water and 43% of those from soil were successfully amplified and proved to be amoeba DNA. Interestingly, a few samples yielded more than one product, which suggests that some other amoebic species may be present in the same sample (MAC-W1 and MADW1). To the best of our knowledge, we described for the first time the amoeba isolation from the participant's close environment and antibodies level among Saudi population. Our future studies will be focused on additional molecular characterization of isolated amoeba and their pathogenic potential which could be a possible threat for the community.

Plants polysaccharides are an infinite stock of drug composites with varying pharmacological and biological activities. The present investigation aimed to examine the antibacterial, anti-scavenging and cytotoxic potential of garden cress (GC) polysaccharides. The antibacterial effects vs Escherichia coli and as well as Staphylococcus aureus of GC polysaccharides were examined by means of agar diffusion assay, minimum inhibitory concentration (MIC), outer and inner cell membrane permeability. Antioxidant potential of the GC polysaccharides were performed by free radical DPPH scavenging, superoxide anion scavenging, hydroxyl radical scavenging, reducing power potential assay, and hydrogen peroxide method. Cytotoxicity potential of GC polysaccharides were evaluated by MTT assay in human cervical (HeLa) and liver carcinoma (HepG2) cell lines. The findings showed that GC polysaccharides MIC were 1.06 and 0.56 mg mL -1 against E. coli and S. aureus, respectively. Compared to the standard inhibitor, the GC polysaccharides showed essential inhibitor assays in a very dose dependent approach, and notable actions to scavenge reactive oxygen species (ROS) are also due to the large quantities of hydrophilic polyphenols. The IC 50 values of all tested parameters were measured against standard ascorbic acid antioxidant agent. The GC polysaccharides diminish the cell viability percentage of HeLa and HepG2 in a concentration dependent manner. GC polysaccharides at a dose of 500 µg ml -1 exhibited higher anti-tumor activity in both HeLa (65.33 ± 3.75%) and HepG2 (60.33 ± 3.48%). The findings obtained in this study indicate that GC polysaccharides has antibacterial and has a possible source of natural antioxidant and also has cytotoxic effect on different carcinoma cell lines., (© 2020 The Author(s).)

Seed-borne fungus Penicillium duclauxii was examined in this study to investigate its capability of synthesizing silver nanoparticles (Ag-NPs). In vitro experiments were conducted using corn-grain contaminating fungal isolate. Ag-NPs detection and characterization were assayed by the aid of spectroscopic techniques. Spectroscopy (energy dispersive), X-ray diffraction, transmission electron-microscope and optical absorption dimensions were employed. Ag-NPs with biosynthesized were used to test invitro against Bipolaris sorghicola ; the cause of target leaf spot disease on sorghum plants. The myco-synthesis of Ag NPs using Penicillium duclauxii was proved in this study. Moreover, Bipolaris sorghicola was successfully inhibited by such Ag NPs in vitro ., Competing Interests: The authors declared that there is no Conflict of Interest., (© 2019 Published by Elsevier B.V. on behalf of King Saud University.)

Prevention of food spoilage and food poisoning pathogens is usually achieved by use of chemical preservatives which have negative impacts including: human health hazards of the chemical applications, chemical residues in food & feed chains and acquisition of microbial resistance to the used chemicals. Because of such concerns, the necessity to find a potentially effective, healthy safer and natural alternative preservatives is increased. Within these texts, Plant extracts have been used to control food poisoning diseases and preserve foodstuff. Antimicrobial activity of five plant extracts were investigated against Bacillus cereus , Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa and Salmonella typhi using agar disc diffusion technique. Ethanolic extracts of Punica granatum, Syzygium aromaticum , Zingiber officinales and Thymus vulgaris were potentially effective with variable efficiency against the tested bacterial strains at concentration of 10 mg/ml while extract of Cuminum cyminum was only effective against S. aureus respectively. P. granatum and S. aromaticum ethanolic extracts were the most effective plant extracts and showed bacteriostatic and bactericidal activities against the highly susceptible strains of food borne pathogenic bacteria ( S. aureus and P. aeruginosa ) with MIC's ranged from 2.5 to 5.0 mg/ml and MBC of 5.0 and 10 mg/ml except P . aeruginosa which was less sensitive and its MBC reached to 12.5 mg/ml of S. aromaticum respectively. These plant extracts which proved to be potentially effective can be used as natural alternative preventives to control food poisoning diseases and preserve food stuff avoiding healthy hazards of chemically antimicrobial agent applications.

Salmonella is recognized as a common foodborne pathogen, causing major health problems in Saudi Arabia. Herein, we report epidemiology, antimicrobial susceptibility and the genetic basis of resistance among S. enterica strains isolated in Saudi Arabia. Isolation of Salmonella spp. from clinical and environmental samples resulted in isolation of 33 strains identified as S. enterica based on their biochemical characteristics and 16S-rDNA sequences. S. enterica serovar Enteritidis showed highest prevalence (39.4%), followed by S. Paratyphi (21.2%), S. Typhimurium (15.2%), S. Typhi and S. Arizona (12.1%), respectively. Most isolates were resistant to 1st and 2nd generation cephalosporin; and aminoglycosides. Moreover, several S. enterica isolates exhibited resistance to the first-line antibiotics used for Salmonellosis treatment including ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol. In addition, the results revealed the emergence of two S. enterica isolates showing resistance to third-generation cephalosporin. Analysis of resistance determinants in S. enterica strains (n=33) revealed that the resistance to β-lactam antibiotics, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline, was attributed to the presence of carb-like, dfrA1, floR, tetA gene, respectively. On the other hand, fluoroquinolone resistance was related to the presence of mutations in gyrA and parC genes. These findings improve the information about foodborne Salmonella in Saudi Arabia, alarming the emergence of multi-drug resistant S. enterica strains, and provide useful data about the resistance mechanisms., (Copyright © 2017. Published by Elsevier Editora Ltda.)

Synthesis of silver nanoparticles (AgNPs) has become a necessary field of applied science. Biological method for synthesis of AgNPs by Rhizopus stolonifer aqueous mycelial extract was used. The AgNPs were identified by UV-visible spectrometry, X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared spectrometry (FT-IR). The presence of surface plasmon band around 420 nm indicates AgNPs formation. The characteristic of the AgNPs within the face-centered cubic (fcc) structure are indicated by the peaks of the X-ray diffraction (XRD) pattern corresponding to (1 1 1), (2 0 0) and (2 2 0) planes. Spherical, mono-dispersed and stable AgNPs with diameter around 9.47 nm were prepared and affirmed by high-resolution transmission electron microscopy (HR-TEM). Fourier Transform Infrared (FTIR) shows peaks at 1426 and 1684 cm -1 that affirm the presence of coat covering protein the AgNPs which is known as capping proteins. Parameter optimization showed the smallest size of AgNPs (2.86 ± 0.3 nm) was obtained with 10 -2  M AgNO 3 at 40 °C. The present study provides the proof that the molecules within aqueous mycelial extract of R. stolonifer facilitate synthesis of AgNPs and highlight on value-added from R. stolonifer for cost effectiveness. Also, eco-friendly medical and nanotechnology-based industries could also be provided. Size of prepared AgNPs could be controlled by temperature and AgNO 3 concentration. Further studies are required to study effect of more parameters on size and morphology of AgNPs as this will help in the control of large scale production of biogenic AgNPs.

Background: Tuberculosis (TB) is an infectious disease that is caused by Mycobacterium tuberculosis (M.tb). TB has high morbidity and mortality around the world., Objective: To evaluate molecular-based methods performed directly on stool samples for the diagnosis of pulmonary tuberculosis (PTB) and to determine the susceptibility to Rifampicin (RMP) and Isoniazid (INH)., Study Design: This is a descriptive study evaluating the performance of the PCR-based method for direct PTB diagnosis and to determine the susceptibility of RMP and INH using stool samples from PTB patients. The study was conducted between March 2011 and March 2014., Methods: Three stool samples and three sputum samples (n=300 stool and 300 sputum) were collected from 100 PTB patients (75 pretreatment and 25 follow up). Stool samples (n=60) were also collected from 20 healthy individuals to serve as controls. DNA was extracted from stool samples using Chelex ® -100. PTB was diagnosed using a Genekam ® kit. RMP and INH susceptibility testing was performed using the Genotype MTBDRplus ® assay. The sputum Lowenstein Jensen (LJ) culture and agar proportion method (PM) of drug susceptibility testing for INH and RMP were considered to be the gold standard methods for comparing the results of the molecular methods., Results: The Genekam kit showed 100% sensitivity and 95.24% specificity for diagnosing new patients and showed 100% sensitivity and 80% specificity for follow-up patients. The Genotype MTBDR-plus assay showed 86.4% and 100% sensitivity and 98.1% and 97.8% specificity for determining INH and RMP sensitivity, respectively, in newly diagnosed patients and 85.7% and 94.4% sensitivity and specificity, respectively, for both INH and RMP in follow-up patients., Conclusion: Molecular-based methods are promising techniques for the diagnosis and susceptibility testing of PTB when the ease of sample collection and the speed of diagnosis are taken into consideration. However, they are not as useful for assessing follow-up patients., (© 2016 by the Association of Clinical Scientists, Inc.)

The stability and reusability of soluble enzymes are of major concerns, which limit their industrial applications. Herein, alkaline protease from Bacillus sp. NPST-AK15 was immobilized onto hollow core-mesoporous shell silica (HCMSS) nanospheres. Subsequently, the properties of immobilized proteases were evaluated. Non-, ethane- and amino-functionalized HCMSS nanospheres were synthesized and characterized. NPST-AK15 was immobilized onto the synthesized nano-supports by physical and covalent immobilization approaches. However, protease immobilization by covalent attachment onto the activated HCMSS-NH₂ nanospheres showed highest immobilization yield (75.6%) and loading capacity (88.1 μg protein/mg carrier) and was applied in the further studies. In comparison to free enzyme, the covalently immobilized protease exhibited a slight shift in the optimal pH from 10.5 to 11.0, respectively. The optimum temperature for catalytic activity of both free and immobilized enzyme was seen at 60 °C. However, while the free enzyme was completely inactivated when treated at 60 °C for 1 h the immobilized enzyme still retained 63.6% of its initial activity. The immobilized protease showed higher V(max), k(cat) and k(cat)/K(m), than soluble enzyme by 1.6-, 1.6- and 2.4-fold, respectively. In addition, the immobilized protease affinity to the substrate increased by about 1.5-fold. Furthermore, the enzyme stability in various organic solvents was significantly enhanced upon immobilization. Interestingly, the immobilized enzyme exhibited much higher stability in several commercial detergents including OMO, Tide, Ariel, Bonux and Xra by up to 5.2-fold. Finally, the immobilized protease maintained significant catalytic efficiency for twelve consecutive reaction cycles. These results suggest the effectiveness of the developed nanobiocatalyst as a candidate for detergent formulation and peptide synthesis in non-aqueous media.