Screening of Antibacterial, Anti-Biofilm, Cell Proliferation Inhibition, and Synergistic Effects of Biogenic Synthesized Silver Nanostructures Using Trillium govanianum with Antibiotics

2020 ◽  
Vol 42 (1) ◽  
pp. 120-120
Author(s):  
Nazia Gulzar Nazia Gulzar ◽  
Saiqa Andleeb Saiqa Andleeb ◽  
Shaukat Ali Shaukat Ali ◽  
Sadia Nazer Sadia Nazer ◽  
Tariq Iqbal Muhammad Abdul Rauf Khan and Abida Raza Tariq Iqbal Muhammad Abdul Rauf Khan and Abida Raza

The aim of study is to develop the economic plant-based antibacterial agents to improve the treatment strategies for bacterial infections at nano-scale. In current research silver nanostructures synthesis was carried out using Trillium govanianum aqueous extract. Characterizations were done using UV–Visible spectrophotometer, scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. Antibacterial efficacy of green-synthesized silver nanostructures and aqueous extract was evaluated via agar well diffusion method, crystal violet assay (antibiofilm analysis), tetrazolium salt assay (cell viability assay), Direct agar overlay technique, and TLC- spot screening against clinical bacterial pathogens. Synergistic effect of synthesized nanostructures, aqueous extract along with antibiotics was determined by agar disc diffusion method. Minimum inhibitory concentration was also evaluated using agar well diffusion method. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) method was used for screening of antioxidant potential effect at various concentrations (0.5, 1 and 1.5 and#181;l). The indication of major phytochemical constituents was determined using both qualitative methods and Thin Layer Chromatography (TLC). Absorbance in UV region (295 nm) corresponds to the out of plane quadrupole Plasmon-resonance while at 350 nm corresponds to in-plane dipole resonance. SEM images shows prepared the morphology of Ag nanostructures TGAgNSs is not exactly spherical while XRD analysis shows that highly crystalline TGAgNSs have been prepared with average crystallite size of 27.94 nm. The FTIR spectrum represents sharp peaks of aldehyde, amide I, aromatic rings, and polysaccharides. Minimum inhibitory concentration showed the increased bactericidal effect with increase in concentration of silver nanostructures. Maximum inhibition of E. coli, S. epidermidis, S. pyogenes, and K. pneumonia was recorded at 0.12 mg/ml concentration of TGAgNSs with 7.6.0and#177;0.0 mm, 7.6and#177;0.0 mm, 6.6and#177;0.0 mm, and 7.0and#177;0.0 mm zones of inhibition. Similarly, TGAgNSs showed maximum inhibition of E. coli, S. pyogenes, and S. marcesscens at 0.14 mg/ml concentration (8.3and#177;0.0 mm, 7.0and#177;0.0 mm, 7.0and#177;0.0 mm), respectively. Cell viability, antibiofilm assay, TLC-spot screening, and TLC-bio-autography proves the potential antibacterial effect of TGAgNSs. Synergistic effect showed greater antibacterial effect. Glycosides, saponins, flavonoids, proteins, terpenoids, alkaloids, phenols, and tannins were detected. The scavenging potential of TGAgNSs was calculated as 74% at 1.5 and#181;l. It was concluded that green synthesized nanostructures have efficient antibacterial potential and would be used in medical devices to cure infections in near future.

2019 ◽  
Author(s):  
Sriraam Sankar ◽  
Ramasamy Thangamalai ◽  
Sriram Padmanaban ◽  
Porteen Kannan ◽  
M R Srinivasan ◽  
...  

AbstractThe colossal rise in antimicrobial resistance has led to treatment failures and so mastitis has become cumbersome to treat. The objective of this study was to evaluate the antibacterial effect of non-antibiotic drug, atorvastatin in combination with antimicrobial, ampicillin against two commonly isolated bacterial species Staphylococcus spp and E. coli from bovine mastitis. Milk samples were collected from mastitis cows, visiting Veterinary Clinical Complex. Bacterial isolation was performed using Eosin Methylene Blue (EMB) agar and Mannitol Salt Agar (MSA), followed by characterization and identification by biochemical tests and gram staining. Genotypic confirmation was done by Polymerase Chain Reaction (PCR) with subsequent screening for resistant genes-mec A, blaTEM. Antibiotic Sensitivity Test (ABST) of the isolates against 12 different antimicrobials, atorvastatin only, and combination of atorvastatin with ampicillin were performed using Kirby-Bauer disc diffusion method. Minimum Inhibitory Concentration (MIC) of ampicillin alone and ampicillin in combination with atorvastatin were determined by modified microdilution method. Staphylococcus spp (77.5%) and E.coli (35%) were the two major pathogens isolated in the current study and multi-drug resistance was observed. Among the antimicrobials, the ampicillin showed 100% resistance against Staphylococcus spp and 85.71% resistance against E. coli. Atorvastatin did not display antibacterial effect as a sole agent but displayed synergistic antibacterial activity with ampicillin. There was an average increase in Minimum Inhibitory Concentration of ampicillin for E.coli and Staphylococcus spp isolates and atorvastatin decreased the Minimum Inhibitory Concentration of ampicillin in combination. The ampicillin shows more resistance against both Staphylococcus spp and E.coli, while atorvastatin improves the effect of ampicillin in-vitro. So, atorvastatin may be combined with ampicillin for the treatment of Gram-positive and Gram-negative infections. However, further studies are required to ascertain the exact mechanism of action of atorvastatin with respect to their antibacterial effect for them to be redeployed as an antimicrobial drug in the future.


2018 ◽  
Vol 20 (87) ◽  
pp. 65-69
Author(s):  
R.A. Peleno

The data of the influence of active substances of anthelmintic and antiprotozoal preparations on the growth of L. casei IMB B-7280 and E. coli 055K59 are provided in the article. Their minimal inhibitory concentrations were determined for these strains of microorganisms and the active substances with which possible simultaneous application of probiotic strain L. casei IMB В-7280 is established. With this aim, the effect on the growth of L. casei IMB B-7280 and E. coli 055K59 and the minimum inhibitory concentration of fenbendazole, levamisole and ivermectin, which are part of the anthelmintic preparations and amprolium, tylosin, sodium sulfadimexone and sodium sulfatyazole, which are active substances of antiprotozoal drugs, were investigated. The determination of the minimum inhibitory concentration of the active substances of antiparasitic agents against these strains of microorganisms was carried out in in vitro experiments by serial dilutions in a dense MRS environment and MPA, and a study of the effect on the growth by diffusion method, followed by measurement of growth retardation zones in millimeters. It is established that among active substances of anthelmintic preparations only phenbendazole caused growth retardation and only relative to L. casei IMB B-7280. Among the active substances of antiprotozoal drugs, sodium sulfatyazole was the most active, which inhibited growth as L. casei IMB-7280 and E. coli 055K59 № 3912/41. Thylosin was effective only in relation to L. casei IMB B-7280 and at the highest concentration of 0.03%, the growth retardation zone was 23.4 ± 0.92 mm. Sodium sulfadimetoxin caused the growth retardation of L. сasei IMB В-7280 only at the highest concentration. The minimum inhibitory concentration of active substances of anti-parasitic drugs was different for strains L. casei IMB B-7280 and E. coli 055K59 № 3912/4. The strongest inhibitory effect was shown by tylosin, which stopped the growth of L. casei IMB B-7280 and E. coli 055K59 № 3912/41 respectively at concentrations of 0.00125 and 50.0 mg/ml. Active substances such as amprolium, levamisole and ivermectin did not significantly inhibit the growth of L. casei, IMB B-7280 and E. coli 055K59 № 3912/41, since their minimal inhibitory concentration was in the range of 4000 to 6000 mg ml.


Extracts from many types of plants show several evidences of beneficial health effects in the living system. Calendula officinalis is a well known medicinal plant in our country. The purpose of this study was to examine the effectiveness of C. officinalis for control of growth and survival of microorganisms. Inhibition of growth was tested by the paper disc agar diffusion method. Minimum inhibitory concentration (MIC) was determined by the tube dilution method. C. officinalis leaf extract showed inhibition (MIC, inhibitory) to Escherichia coli, Klebsiella pneumoniae, and Bacillus subtilis and to the other bacteria tested. Antimicrobial effects of leaf extract of C. officinalis on some microorganisms including pathogens were investigated. The extract of C. officinalis leaf which is prepared in was tested on bacterial cultures such as B. subtilis, S. lutea, E. coli, K. pneumoniae and also investigated that minimum inhibitory concentration 4µg/ml of leaf extract of C. officinalis in Petroleum ether against K. pneumoniae and largest inhibitory zone are created by 512mg/ml chloroform extract against E. coli leaf extract of C. officinalis in Petroleum ether has proved better for antibacterial activity. This study demonstrates that the potentiality of C. officinalis as a source of antimicrobials that could be harness for use in the health care delivery process.


Extracts from many types of plants show several evidences of beneficial health effects in the living system. Calendula officinalis is a well known medicinal plant in our country. The purpose of this study was to examine the effectiveness of C. officinalis for control of the growth and survival of microorganisms. The inhibition of growth was tested by the paper disc agar diffusion method. Minimum inhibitory concentration (MIC) was determined by the tube dilution method. C. officinalis leaf extract showed inhibition (MIC, inhibitory) to Escherichia coli, Klebsiella pneumoniae, and Bacillus subtilis and to the other bacteria tested. Antimicrobial effects of leaf extract of C. officinalis on some microorganisms including pathogens were investigated. The extract of C. officinalis leaf which is prepared in was tested on bacterial cultures such as B. subtilis, S. lutea, E. coli, K. pneumoniae and also investigated that minimum inhibitory concentration 4µg/ml of leaf extract of C. officinalis in petroleum ether against K. pneumoniae and largest inhibitory zone are created by 512mg/ml chloroform extract against E. coli leaf extract of C. officinalis in petroleum ether has proved better for antibacterial activity. This study demonstrates that the potentiality of C. officinalis as a source of antimicrobials that could be harness for use in the health care delivery process.


Author(s):  
D. A. Awomukwu ◽  
C. K. Anumudu ◽  
A. J. Ogbolosingha

The research was undertaken to investigate the comparative phytochemical and in-vitro antibacterial activity of the single and combined strengths of the leaf extracts of Ocimum gratissimum Linn and Gongronema latifolium Benth. on some enteric bacterial isolates. The sensitivity test and minimum inhibitory concentration (MIC) were carried out using a modified agar-well diffusion method.  The enteric bacterial isolates tested included Escherichia coli, Shigella dysenteriae, Salmonella Typhi and Enterobacter aerogenes.  Standard methods were applied to obtain the ethanol and aqueous leaf extracts. The ethanol extracts of O. gratissimum and G. latifolium produced highly significant inhibitory activity against all the enteric bacterial isolates tested.  Comparatively, the ethanol plant extracts were more potent than the commercially available drug, Ciprofloxacin and the aqueous plant extracts. The isolates were sensitive at a minimum inhibitory concentration of 1.25 mg/ml for the ethanol extract but varied from 2.5 mg/ml - 5.0mg/ml in the aqueous extract.  It was also observed that the synergistic antibacterial effect of the medicinal plant extracts was greater than the singular antibacterial effect of the individual plant extracts in both the ethanol and aqueous extracts. The potency of the individual extracts and the combined effect may be due to the presence of flavonoids, alkaloids, phenols, tannins and saponins in the leaves of the plants.  This study partly validates the use of the plant extracts in the treatment of disease caused by the enteric bacterial isolates by multiple traditional medicine practitioners in Nigeria, however, strict adherence to dosage is recommended.  The leaf extract is a potential source of the new drug if the components are purified and enhanced for treating infections caused by these enteric pathogens.


2020 ◽  
Vol 7 (2) ◽  
pp. 75
Author(s):  
Amalia Khairunnisa ◽  
Nashrul Wathan ◽  
Mia Fitriana ◽  
Fadlilaturrahmah Fadlilaturrahmah ◽  
Nisriyati Fiddina

Nymphaea pubescens Willd telah diketahui mempunyai efek antibakteri, terutama pada biji dan daunnya. Tetapi sampai saat ini bagian bunga dari tanaman tersebut belum dilakukan pengujian aktivitas antibakteri. Tujuan dari penelitian ini untuk melakukan skrining fitokimia, uji aktivitas antibakteri dan penentuan konsentrasi hambat minimum (KHM) dari ekstrak metanol bunga N. pubescens terhadap S. aureus dan E. coli. Proses ekstraksi bunga N. pubescens dimaserasi menggunakan pelarut metanol dengan perbandingan 1: 4 b/v. Metode pengujian yang digunakan ada dua yaitu metode difusi untuk pengujian aktivitas antibakteri dan metode dilusi untuk pengukuran konsentrasi hambat minimum (KHM). Hasil skrining fitokimia didapatkan bahwa ekstrak metanol bunga N. pubescens mengandung senyawa golongan fenolik, saponin dan flavonoid. Hasil pengujian aktivitas antibakteri menunjukkan bahwa ekstrak metanol bunga N.pubescens mampu menghambat S. aureus (diameter hambat 10 ± 0,29 mm) dan E.coli (diameter hambat 10,2 ± 0,50 mm). Konsentrasi hambat minimum dari ekstrak metanol bunga N. pubescens terhadap S.aureus sebesar 12,5% dan terhadap E.coli sebesar 25%. Dapat disimpulkan bahwa ekstrak metanol bunga N. pubescens memiliki aktivitas sebagai antibakteri. Nymphaea pubescens Willd has known to have antibacterial effects, especially on the seeds and leaves. However, until now the flower of the plant has not been tested for antibacterial activity. The purpose of this study was to perform phytochemical screening, antibacterial activity test and determine the minimum inhibitory concentration (MIC) of the methanol extract of N. pubescens flowers against S. aureus and E. coli. The process of extracting N. pubescens flowers is macerated using methanol as a solvent with a ratio of 1: 4 w / v. There are two test methods used, namely the diffusion method for testing antibacterial activity and the dilution method for measuring the minimum inhibitory concentration (MIC). The results of phytochemical screening showed that the methanol extract of N. pubescens flowers contained phenolic compounds, saponins, and flavonoids. The results of the antibacterial activity test showed that the methanol extract of N.pubescens flowers was able to inhibit S. aureus (inhibition diameter 10 ± 0.29 mm) and E. coli (inhibitory diameter 10.2 ± 0.50 mm). The minimum inhibitory concentration of the methanol extract of N. pubescens flowers against S.aureus was 12.5% and against E. coli was 25%. It can be concluded that the methanol extract of N. pubescens flowers has antibacterial activityKeywords: Nymphaea pubescens, lotus flower, methanol extract, antibacterial


2016 ◽  
Vol 5 (04) ◽  
pp. 4512
Author(s):  
Jackie K. Obey ◽  
Anthoney Swamy T* ◽  
Lasiti Timothy ◽  
Makani Rachel

The determination of the antibacterial activity (zone of inhibition) and minimum inhibitory concentration of medicinal plants a crucial step in drug development. In this study, the antibacterial activity and minimum inhibitory concentration of the ethanol extract of Myrsine africana were determined for Escherichia coli, Bacillus cereus, Staphylococcus epidermidis and Streptococcus pneumoniae. The zones of inhibition (mm±S.E) of 500mg/ml of M. africana ethanol extract were 22.00± 0.00 for E. coli,20.33 ±0.33 for B. cereus,25.00± 0.00 for S. epidermidis and 18. 17±0.17 for S. pneumoniae. The minimum inhibitory concentration(MIC) is the minimum dose required to inhibit growth a microorganism. Upon further double dilution of the 500mg/ml of M. africana extract, MIC was obtained for each organism. The MIC for E. coli, B. cereus, S. epidermidis and S. pneumoniae were 7.81mg/ml, 7.81mg/ml, 15.63mg/ml and 15.63mg/ml respectively. Crude extracts are considered active when they inhibit microorganisms with zones of inhibition of 8mm and above. Therefore, this study has shown that the ethanol extract of M. africana can control the growth of the four organisms tested.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Abdulkader Masri ◽  
Naveed Ahmed Khan ◽  
Muhammad Zarul Hanifah Md Zoqratt ◽  
Qasim Ayub ◽  
Ayaz Anwar ◽  
...  

Abstract Backgrounds Escherichia coli K1 causes neonatal meningitis. Transcriptome studies are indispensable to comprehend the pathology and biology of these bacteria. Recently, we showed that nanoparticles loaded with Hesperidin are potential novel antibacterial agents against E. coli K1. Here, bacteria were treated with and without Hesperidin conjugated with silver nanoparticles, and silver alone, and 50% minimum inhibitory concentration was determined. Differential gene expression analysis using RNA-seq, was performed using Degust software and a set of genes involved in cell stress response and metabolism were selected for the study. Results 50% minimum inhibitory concentration with silver-conjugated Hesperidin was achieved with 0.5 μg/ml of Hesperidin conjugated with silver nanoparticles at 1 h. Differential genetic analysis revealed the expression of 122 genes (≥ 2-log FC, P< 0.01) in both E. coli K1 treated with Hesperidin conjugated silver nanoparticles and E. coli K1 treated with silver alone, compared to untreated E. coli K1. Of note, the expression levels of cation efflux genes (cusA and copA) and translocation of ions, across the membrane genes (rsxB) were found to increase 2.6, 3.1, and 3.3- log FC, respectively. Significant regulation was observed for metabolic genes and several genes involved in the coordination of flagella. Conclusions The antibacterial mechanism of nanoparticles maybe due to disruption of the cell membrane, oxidative stress, and metabolism in E. coli K1. Further studies will lead to a better understanding of the genetic mechanisms underlying treatment with nanoparticles and identification of much needed novel antimicrobial drug candidates.


2021 ◽  
Vol 50 (2) ◽  
pp. 219-226
Author(s):  
Abdul Haq ◽  
Alam Khan ◽  
Zulfiqar Ali Malik ◽  
Mushtaq Ahmed ◽  
Samiullah Khan ◽  
...  

Antimicrobial activities of deoiled seed kernel (mechanically pressed), fruit coat and seed coat of Jatropha curcas Linn. collected from two regions (Bannu and Peshawar) of Pakistan were investigated. The antimicrobial activities were carried out against Klebsiella pneumoniae (ATCC 43816), Escherichia coli (ATCC 10536), Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6633) and two clinical fungal isolates Aspergillus fumigatus and Candida albicans using agar well diffusion method. The antibacterial activities of Peshawar sample were found to be higher than Bannu, against selected strains. While antifungal activities of both samples were similar. Highest zone of inhibition 31.5 ± 0.7 mm was exhibited by n-hexane extract of deoiled seed kernel of Peshawar sample against Bacillus subtilis (ATCC 6633). The minimum inhibitory concentration of ethanolic extracts of deoiled seed kernel and seed coat of Peshawar sample was 31.25 - 25 mg/ml. Whereas, minimum inhibitory concentration of ethanolic and n-hexane extracts of Bannu sample was 62.5 - 125 mg/ml. The results suggested that antimicrobial potential of J. curcas Linn. varied with geographical distribution. The investigation of different varieties of medicinal plants belonging to the same species will greatly enhance the chances of best pharmaceuticals discovery. Bangladesh J. Bot. 50(2): 219-226, 2021 (June)


2021 ◽  
Author(s):  
li li jiang ◽  
Su Xu ◽  
Haitao Yu ◽  
Qi Cui ◽  
Rui Cao

Abstract In this study, graphene oxide (GO) was first prepared by the modified Hummer method. Then, the GO/trichloroisocyanuric acid (TCCA) composite was prepared by loading TCCA into GO with the blending method. X-ray diffraction, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy and atomic force microscopy were used to characterize the composite. The results showed that TCCA was successfully loaded on the surface of GO or intercalated among GO layers. Next, the antibacterial performance of the composite against Escherichia coli and Staphylococcus aureus was tested by the 96-well plate assay. A bactericidal kinetic curve, bacterial inhibition tests, and the mechanism of bacterial inhibition is discussed. The results showed that the minimum inhibitory concentration of the GO/TCCA composite (GO:TCCA ratio = 1:50) was 327.5 µg/mL against E. coli and 655 µg/mL against S. aureus. At the minimum inhibitory concentration, the inhibition rate of the GO/TCCA composite exceeded 99.46% against E. coli and 99.17% against S. aureus. The bactericidal kinetic curves indicate that the GO/TCCA composite has an excellent bactericidal effect against E. coli and S. aureus.


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