Synthesis and characterization of marine seagrass (Cymodocea serrulata) mediated titanium dioxide nanoparticles for antibacterial, antibiofilm and antioxidant properties.

Cymodocea serrulata mediated titanium dioxide nanoparticles (TiO2 NPs) were successfully synthesized. The XRD pattern and FTIR spectra demonstrated the crystalline structure of TiO2 NPs and the presence of phenols, flavonoids and alkaloids in the extract. Further SEM revealed that TiO2 NPs has uniform structure and spherical in shape with their size ranged from 58 to 117 nm. Antibacterial activity of TiO2 NPs against methicillin-resistant Staphylococcus aureus (MRSA) and Vibrio cholerae (V. cholerae), provided the zone of inhibition of 33.9 ± 1.7 and 36.3 ± 1.9 mm, respectively at 100 µg/mL concentration. MIC of TiO2 NPs against MRSA and V. cholerae showed 84% and 87% inhibition at 180 µg/mL and 160 µg/mL respectively. Subsequently, the sub-MIC of V. cholerae demonstrated minimal or no impact on bacterial growth at concentration of 42.5 µg/mL concentration. In addition, TiO2 NPs exhibited their ability to inhibit the biofilm forming V. cholerae which caused distinct morphological and intercellular damages analysed using CLSM and TEM. The antioxidant properties of TiO2 NPs were demonstrated through TAA and DPPH assays and exposed its scavenging activity with IC50 value of 36.42 and 68.85 µg/mL which denotes its valuable antioxidant properties with potential health benefits. Importantly, the brine shrimp based lethality experiment yielded a low cytotoxic effect with 13% mortality at 100 µg/mL. In conclusion, the multifaceted attributes of C. serrulata mediated TiO2 NPs encompassed the antibacterial, antioxidant and anti-biofilm inhibition effects with low cytotoxicity in nature were highlighted in this study and proved the bioderived TiO2 NPs could be used as a promising agent for biomedical applications.

Morphological modification of silver nanoparticles against multi-drug resistant gram-negative bacteria and cytotoxicity effect in A549 lung cancer cells through in vitro approaches.

In the present study, the individual cultures of Proteus mirabilis (P. mirabilis) and Klebsiella pneumoniae (K. pneumoniae) were treated with morphologically modified silver nanoparticles (Ag NPs) and were found to display zones of inhibition of ~ 8 mm, 16 mm, 20 mm, and 22 mm (P. mirabilis) and 6 mm, 14 mm, 20 mm, and 24 mm (K. pneumoniae) at concentrations of 25 µg/ml, 50 µg/mL, 75 µg/mL, and 100 µg/mL, respectively. In addition, turbidity tests were performed based on O. D. values, which exhibited 92% and 90% growth inhibitions at 100 µg/mL concentration for P. mirabilis and K. pneumoniae, respectively. Furthermore, the IC50 concentration of Ag NPs was established for A549 lung cancer cells and found to be at 500 µg/mL. Evidently, the morphological variation of Ag NPs treated A549 lung cancer cells was exhibited with differential morphology studied by phase-contrast microscopy. The results demonstrated that the synthesized Ag NPs was not only efficient against gram-positive bacteria but also against gram-negative bacteria and A549 cancer cells, suggesting that the potential of these biosynthesized Ag NPs is a future drug discovery source for inhibiting bacteria and cancer cells.

Metal tolerance and biosorption of Pb ions by Bacillus cereus RMN 1 (MK521259) isolated from metal contaminated sites.

Recent years, metal pollution is an alarming factor to know about protects the environmental ecosystem due to the toxic, persistent and abundant in nature. Metals are present everywhere in the biotic and abiotic samples including soil, water, and microbes. The rate of bioaccumulation and biotransformation are very high. The excess concentration of the metals causes heavy metal pollution or contamination. Due to these defects, the removal of metals using biological sources is heightened in the current research. In this current investigation, the biosorption potential ability of the metal tolerable Bacillus cereus on Pb and Cu rich environment was chosen and thoroughly monitored. The 16s rRNA of the Bacillus cereus was sequenced, and named as Bacillus cereus RMN 1 (MK521259). The various test concentration (10-60 mg/mL) of Pb and Cu was exhibited the maximum removal percentages of 85.2% and 60.2%. The result of bisorption factors exhibited, 300 mg/mL of the biosorbent potency, 60 min contact time and pH 7, and they found to be optimal to remove the maximum of Pb ion from the solution. In the regression coefficients, the Freundlich and Langmuir isotherm models were used to study the adsorption kinetics of metal ions. In addition, the isotherm model confirmed that the of B. cereus biomass medicated metal adsorption was more favourable reaction for metal degradation. With the above evidences, the results of the present investigation proved that B. cereus derived biomass was actively adsorbing the metals ions. Thus we are recommending for the implementation of effective waste water treatment.

Preparation of antibacterial Zn and Ni substituted cobalt ferrite nanoparticles for efficient biofilm eradication.

Zinc (Zn) and, alternatively, nickel (Ni) substituted cobalt ferrite (CF) nanoparticles (NPs) were prepared by sol-gel method. X-ray diffraction analysis revealed the formation of cubic structure of cobalt ferrite. FTIR analysis confirmed the vibrational band located at 550-580 cm-1 that belongs to the M - O bond (M = Ni, and Zn). The alteration of the surface morphology of CF after the addition of Zn and Ni ions was observed from scanning electron microscopic images. The additional peaks in the energy dispersive X-ray diffraction (EDX) analysis spectra were found to correspond to Zn and Ni. The presence of Zn and, alternatively, Ni ions enhanced the biocidal properties of CF NPs against gram negative organisms, in a concentration and time-dependent manner. Furthermore, exposure to CF, CF-Zn and CF-Ni NPs decreased metabolic activity due to the damage of extra polymorphic substances, live/dead cell variation, architecture and surface integrity of the cells. Altogether, the present investigation provides the basis of metal ion substituted metal oxide NPs as anti-biofilm agents against gram-positive and gram-negative bacteria.

Biosorption and adsorption isotherm of chromium (VI) ions in aqueous solution using soil bacteria Bacillus amyloliquefaciens.

This study looked at the development of effective biosorbents to recover the most toxic elements from industrial water. B. amyloliquefaciens was isolated from marine soils showing extreme resistance to Chromium (Cr(VI)) ions. During the 60 min of contact time, 79.90% Cr(VI) was adsorbed from the aqueous solution. The impact of important factors such as biomass concentration, pH of the medium, and initial metal ions concentration on biosorption rate was also examined. The desorption study indicated that 1 M HCl (91.24%) was superior to 0.5 M HCl (74.81%), 1 M NaOH (64.96%), and distilled water (3.66%). Based on the Langmuir model, the maximum adsorption capacity of the bio-absorbent was determined to be 48.44 mg/g. The absorption mechanism was identified as monolayer, and 1/n from the Freundlich model falls within 1, thus indicating favorable adsorption. Based on the findings of the present study, the soil bacterium B. amyloliquefaciens was found to be the best alternative and could be used to develop strategies for managing existing environmental pollution through biosorption.

Effective removal of heavy metals in industrial wastewater with novel bioactive catalyst enabling hybrid approach.

Recent years, heavy metal reduction of contaminated atmosphere using microbes is heightened worldwide. In this context, the current study was focused on heavy metal resistant actinomycete strains were screened from effluent mixed contaminated soil samples. Based on the phenotypic and molecular identification, the high metal resistant actinomycete strain was named as Nocardiopsis dassonvillei (MH900216). The highest bioflocculent and exopolysaccharide productions of Nocardiopsis dassonvillei (MH900216) was confirmed by various invitro experiments result. The heavy metal degrading substances was characterized and effectively confirmed by Fourier transform infrared spectroscopy (FT-IR), X-Ray Diffraction (XRD), Scanning electron microscope (SEM). Further, the heavy metal sorption ability of actinomycete substances bioflocculent was exhibited 85.20%, 89.40%, 75.60%, and 51.40% against Cd, Cr, Pb and Hg respectively. Altogether, the bioflocculent produced actinomycete Nocardiopsis dassonvillei (MH900216) as an excellent biological source for heavy metal reduction in waste water, and it is an alternative method for effective removal of heavy metals towards sustainable environmental management.

Partially purified actinomycetes compounds enhance the intracellular damages in multi-drug resistant P. aeruginosa and K. pneumoniae.

Based on the excellent nutrient level, the current study was focused on isolation and anti-bacterial activity of the actinomycetes from marine mangrove soil samples. As result, 10 different strains of actinomycetes strains were identified on actinomycetes isolation agar plates. The identified strains were shown with white, clear, uncontaminated well matured spore producing ability. Based on the initial observation, the isolated colonies were actinomycetes. The partially extracted crude compound shown excellent anti-bacterial activity against P. aeruginosa and K. pneumoniae with 15 mm and 13 mm zone of inhibitions were observed at 500 µL concentrations. The minimum inhibition concentration result was also confirmed the 500 µL concentration against both the tested concentration with high inhibition rate. Then, the intracellular damages, decreased cell growth of the crude actinomycetes extract treated bacterial strains were clearly observed by confocal laser scanning electron microscope. The extracellular damages of bacterial cell wall and shape of the both the pathogens were clearly shown by scanning electron microscope. Therefore, all the results were clearly supported to the partially extracted crude compound and it has excellent anti-bacterial activity against tested multi drug resistant bacteria.

Isolation and molecular identification of extended spectrum beta-lactamase producing bacteria from urinary tract infection.

BACKGROUND: Recent years, the treatment of multi-drug resistant bacteria and their effect are very difficult due to the virulence factors modification. Based on the world wide thread, we have tried to identify the ESBLs producing bacteria from urinary tract infection patients. In result, the highly antibiotic resistant effect of Pseudomonas aeruginosa and Klebsiella pneumoniae were identified. METHODS: Initially, Hexa disc diffusion method was performed to detect the multi-drug resistant bacteria using respective antibiotics of HX066, HX033 and HX077, HX012 discs. Consecutively, the ESBL producing ability of confirmed multi drug resistant bacteria was performed to detect their ESBL producing ability using specific extended spectrum beta lactamase (ESBLs) detection discs of Hexa G-minus 24. Furthermore, the ESBL producing ability of the bacteria was confirmed by ESBLs detection Ezy MIC™ E-test stripe method. RESULTS AND CONCLUSIONS: In result, 10, 5 and 4 mm and 10, 14 and 8 mm zone of inhibition against imipenem (IPM), Ticarcillin/Clavulanic acid (TCC), Cefoperazone (CPZ) and Ampicillin (AMP), Norfloxacin (NX), Nalidixic acid (NA) antibiotics for P. aeruginosa and 16, 22 and 10, 18 mm zone of inhibition against ceftazidime (CAZ), methicillin (MET), ampicillin amoxyclav (AMC), co-trimoxazole (COT) of the HX077 HX012 for K. pneumoniae were observed. Based on the Clinical & Laboratory Standards Institute (CLSI) guidelines, both the bacteria were more resistant to tested antibiotics and it could be developed more resistant against all the tested antibiotics. In addition, the phenotypic detection of ESBL production effect was also performed against both the selected uropathogens, and the results were shown ≥22 mm, ≥27 zone of inhibition against all the tested antibiotics. Further, the genetic identification of multi plux PCR result was shown TEM, SHV and CTX-m genes were present in both the selected uropathogens. Finally, our results were correlated each other and concluded that the selected uropathogens were multi drug resistant effect and also ESBLs producer.

Isolation and molecular identification of biofilm producing P. aeruginosa and K. pneumoniae from urinary tract infections patient urine sample.

BACKGROUND: Recent years, multi drug resistant pathogens and their pathogenicity were increased worldwide due to unauthorized consumption of antibiotics. In addition, correlation between multi drug resistant bacteria and biofilm formation is heightened due to the production of more virulence behavior. There is no better identification methods are available for detection of biofilm producing gram negative bacteria. MATERIALS AND METHODS: In this research work, multi drug resistant strains of Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) were identified based on the specific antibiotics and third generation cephalosporin discs by disc diffusion assay. Subsequently, biofilm forming ability of selected pathogens were identified tissue culture plate and tube test. Based on the multi-drug resistant ability and biofilm production, the molecular identification of P. aeruginosa and K. pneumoniae were confirmed by PCR using universal primers. RESULTS AND CONCLUSIONS: No zone of inhibition present around the discs of muller hinton agar plates were confirm, selected P. aeruginosa and K. pneumoniae strains were multi drug resistant pathogens. Performed third generation cephalosporin antibiotics were also highly sensitive to selected pathogens of P. aeruginosa and K. pneumoniae. Further, biofilm forming ability of selected P. aeruginosa and K. pneumoniae was confirmed by tissue culture plate and tube methods. Finally, molecular identification of P. aeruginosa and K. pneumoniae was named as P. aeruginosa and K. pneumoniae. Our result was conclude, selected P. aeruginosa and K. pneumoniae as biofilm producing pathogens and also highly resistant to current antibiotics.

Identification of carbapenems resistant genes on biofilm forming K. pneumoniae from urinary tract infection.

The multi-drug resistant effect of the Gram negative bacteria K. pneumoniae was identified by disc diffusion method using specific UTI panel discs of Kleb 1 HX077 and Kleb 2 HX090 HEXA. Among the multi-drug resistant bacteria, the carbapenem resistant (CR) effect of the K. pneumoniae was screened by specific carbapenem detection antibiotics of HEXA HX066 and HX0103 HEXA by disc diffusion method. In addition, the effective antibiotics were further performed against K. pneumoniae by minimum inhibition concentration method. Further, the carbapenemase genes of VIM 1 and IMP 1 were detected from the isolated strains by multiplex PCR method. Furthermore, the biofilm forming ability of selected carbapenem resistant K. pneumoniae was initially identified by tissue culture plate method and confirmed by exopolysaccharide arrest ability of congo red agar assay. Finally, our result was proved that the identified K. pneumoniae is carbapenemase producing strain, and its virulence was extended with strong biofilm formation.

Phytochemical screening and anti-oxidant activity of Sargassum wightii enhances the anti-bacterial activity against Pseudomonas aeruginosa.

In this study, the phytochemical, phenolic, flavonoid and bioactive compounds were successfully screened from crude extract of Sargassum wightii by LC-MS analysis after NIST interpretation. Bacterial growth inhibition study result was shown with 24 mm zone inhibition at 200 µg/mL concentration against P. aeruginosa. The increased phenolic content was much closed to gallic acid and the range was observed at 250 µg/mL concentration. In addition, flavonoid contents of the algae extract was indicated more significant with rutin at 200 µg/mL. In result, both the phenolic and flavonoid contents of the extract were more correlated with gallic acid and rutin. Further, the total anti-oxidant and DPPH radical scavenging activities were shown increased activity at 200 µg/mL concentrations. Furthermore, the excellent anti-bacterial alteration result was observed at 200 µg/mL concentration by minimum inhibition concentration. Therefore, the result was revealed that the marine algae Sargassum wightii has excellent phytochemical and anti-oxidant activities, and it has improved anti-bacterial activity against P. aeruginosa.

Biosynthesized silver nanoparticles using Caulerpa taxifolia against A549 lung cancer cell line through cytotoxicity effect/morphological damage.

The Caulerpa taxifolia is excellent marine green algae, which produced enormous bioactive compounds with more biological activities. Also, it is an excellent source for synthesis of Ag NPs with increased bioactivity against various infections. In our study, the marine algae marine algae Caulerpa taxifolia mediated Ag NPs was synthesized effectively. The synthesized Ag NPs was characterized well using UV-spectrometer and X-ray powder diffraction (XRD) and confirmed as synthesized particle was Ag NPs. The available structure of the Ag NPs was morphologically identified by scanning electron microscope (SEM), and exact minimum size, polydispersive spherical shape of the entire Ag NPs structure was confirmed by Transmission electron microscope (TEM). Further, the anti-cancer efficiency of biosynthesized Ag NPs against A549 lung cancer cells was found at 40 µg/mL concentration by cytotoxicity experiment. In addition, the phase contrast images of the result were supported the Ag NPs, which damaged the A549 morphologically clearly. Finally, florescence microscopic images were effectively proved the anti-cancerous effect against A549 lung cancer cells due to the condensed morphology of increased death cells. All the confirmed in-vitro results were clearly stated that the Caulerpa taxifolia mediated Ag NPs has superior anti-cancer agent against A549 lung cancer cells.

Anti-cancer activity of biosynthesized silver nanoparticles using Avicennia marina against A549 lung cancer cells through ROS/mitochondrial damages.

The biosynthesized Ag NPs was synthesized by using marine mangrove plant extract Avicennia marina. The synthesized Ag NPs was confirmed by various physiochemical characterization including UV-spectrometer and XRD analysis. In addition, the shape and of the synthesized Ag NPs was morphologically identified by SEM initially and TEM finally. After confirmation, the anti-cancer property of synthesized Ag NPs was confirmed at 50 µg/mL concentration against A549 lung cancer cells by MTT assay. Further, the ability to stimulate the ROS generation and mitochondrial membrane at the IC50 concentration of Ag NPs was confirmed by fluorescence microscopy using DCFH-DA and rhodamine 123 dyes respectively. Finally, the result was concluded that the synthesized Ag NPs has improved anti-cancer activity against A549 cells at lowest concentration.

Preparative HPLC fraction of Hibiscus rosa-sinensis essential oil against biofilm forming Klebsiella pneumoniae.

Recent years Klebsiella pneumoniae (K. pneumoniae) biofilm formation (BF) is emerging thread worldwide. For tackling this problem, we have chosen Hibiscus rosa-. pneumoniae. The HPLC purified essential oils (EOs sinensis (H. rosa-sinensis) (HRS) to inhibit the BF K) of H. rosa-sinensis was performed against BF K. pneumoniae and showed concentration dependent biofilm inhibition. At the MBIC of EOs (90 µg/ml), the biofilm inhibition was showed at 92% against selected BF K. Pneumoniae. The biofilm metabolic assay, exopolysaccharide quantification and hydrophobicity index variation results exhibited with 88%, 92% and 89% reduction at 90 µg/mL was observed respectively. In addition, the morphological modification of MBIC treated K. pneumoniae was clearly viewed by scanning electron microscope (SEM). Overall, all the invitro experiments result were confirmed that the MBIC of H. rosa-sinensis EOs was very effective against BF K. pneumonia.

Enhanced anti-cancer activity of chitosan loaded Morinda citrifolia essential oil against A549 human lung cancer cells.

In the present study, the chemical composition of Morinda citrifolia essential oils was determined by gas chromatography-mass spectrometry and was found to contain several anti-cancer compounds including L-scopoletin, nordamnacanthal, ß-morindone, α-copaene, 9-H-pyrido[3,4-b]indole, ß-thujene and terpinolene. The physico-chemical characterization of chitosan, chitosan nanoparticles and Morinda citrifolia essential oils loaded chitosan nanoparticles combination was carried out by Fourier transform infrared spectroscopy, powder X-ray diffraction and dynamic light scattering coupled with zeta potential. The morphological observation obtained by scanning electron microscopy and transmission electron microscopy provided clear indication that the immobile chitosan polymer formed a coating onto the Morinda citrifolia essential oils surface. The cytotoxic effect of Morinda citrifolia essential oils loaded chitosan nanoparticles against A549 cells were investigated, resulting in 54% inhibition at 40 µg/ml-1. Information about in vitro morphological modification, nucleus damages, ROS generation and cell cycle arrest was obtained by fluorescence microscopy and flow cytometer analysis. The toxicity evaluation against human red blood cells suggested that the Morinda citrifolia essential oils loaded chitosan nanoparticles possess minimum cytotoxicity. Altogether, the present study suggests that these Morinda citrifolia essential oils loaded chitosan nanoparticles are valuable biomaterials owing to their ability to fight against A549 cancer cells.

Molecular identification and structural characterization of marine endophytic actinomycetes Nocardiopsis sp. GRG 2 (KT 235641) and its antibacterial efficacy against isolated ESBL producing bacteria.

The present study was designed to identify the potential bioactive compound from endophytic actinomycetes (EA) Nocardiopsis sp. GRG 2 (KT 235641) against selected extended spectrum beta lactamase (ESBL) producing Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae). Initially, the multi drug resistance (MDR) effect of selected uropathogens was confirmed by respective UTI panel of Hexa antibiotics disc methods. The zone of inhibition ≤22 mm for ceftazidime, ≤ 27 mm for cefotaxime and ≤8 mm zone of MIC stripe against both the uropathogens of phenotypic methods confirmed, the selected strains were ESBL producer. Among the various EA extracts, GRG 2 extract showed excellent antibacterial activity against both ESBL producing P. aeruginosa and K. pneumonia by agar well diffution method. The molecular identification of selected GRG 2 strain was named as Nocardiopsis sp. GRG 2 (KT235641). The antibacterial metabolites present in the TLC elution was exhibited at 274 nm by UV visible spectrometer. The partial purification of preparative HPLC fraction 3 showed 14, 16 mm against P. aeruginosa and K. pneumoniae, respectively. Based on the antibacterial effect, the FT-IR, GC-MS and LC-MS analysis of fraction 3 was confirmed as 1, 4-diaza-2, 5-dioxo-3-isobutyl bicyclo[4.3.0]nonane (DDIBN). Further, the dose dependent inhibition of DDIBN against both ESBL producing pathogens was observed at 75 µg/mL by minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC). The increased cell death and disrupted cell membrane integrity were observed at MIC of DDIBN by confocal laser scanning electron microscope (CLSM) and scanning electron microscope (SEM). The results were proved that the DDIBN has potential antibacterial metabolites against ESBL producing pathogens and it can be applied for various other biomedical fields.

Biologically synthesized copper oxide nanoparticles enhanced intracellular damage in ciprofloxacin resistant ESBL producing bacteria.

Copper oxide nanoparticles (CuO NPs) were synthesized biologically using leaf extract of Camilla japonica. The typical UV-visible spectral peak of CuO NPs was observed at a wavelength of ∼290 nm, which confirmed their successful synthesis. From scanning electron microscope (SEM) and transmission electron microscope (TEM) analyses, the synthesized CuO NPs were found to possess spherical shape. Energy dispersive X-ray analyzer (EDX) results revealed that the CuO NPs are almost pure with atomic percentages of 50.92 for Cu and 49.08 for O. Fourier transform infrared (FTIR) confirmed the presence of an absorption peak located at a wavenumber position of ∼480 cm-1 typical for highly pure CuO NPs. TEM images displayed that the particles are relatively uniform in size ∼15-25 nm. The P. aeruginosa and K. pneumonia showed complete resistance against Hexa 077 antibiotic discs. The result of ≤22 ceftazidime and ≤27 cefotaxime confirmed that both the uropathogens were ESBL producers. The ≥8 mm of the MIC stripe further confirmed that both the uropathogens were ESBL producers. Furthermore, the antibacterial activity of CuO NPs against selected ESBL producing P. aeruginosa and K. pneumoniae at minimum inhibition concentration (MIC) of 100 µg/mL. The decreased cell viability and damaged membrane construction of both the uropathogens were observed by confocal laser scanning microscope (CLSM) using AO/EB stains at desired MIC dose. The morphological damage of the bacterial cells was demonstrated by SEM analysis. Hence, based on the above in vitro findings, the results suggested that the CuO NPs are efficient antibacterial compounds against ESBL producing bacteria, and that the plant leaf mediated CuO NPs can be considered as novel and promising material to act against various infectious bacteria.