scholarly journals Expanding the Use of a Fluorogenic Method to Determine Activity and Mode of Action ofBacillus thuringiensisBacteriocins Against Gram-Positive and Gram-Negative Bacteria

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Norma M. de la Fuente-Salcido ◽  
J. Eleazar Barboza-Corona ◽  
A. N. Espino Monzón ◽  
R. D. Pacheco Cano ◽  
N. Balagurusamy ◽  
...  

Previously we described a rapid fluorogenic method to measure the activity of five bacteriocins produced by Mexican strains ofBacillus thuringiensisagainstB. cereus183. Here we standardize this method to efficiently determine the activity of bacteriocins against both Gram-positive and Gram-negative bacteria. It was determined that the crucial parameter required to obtain reproducible results was the number of cells used in the assay, that is,~4 × 108 cell/mL and~7 × 108 cell/mL, respectively, for target Gram-positive and Gram-negative bacteria. Comparative analyses of the fluorogenic and traditional well-diffusion assays showed correlation coefficients of 0.88 to 0.99 and 0.83 to 0.99, respectively, for Gram-positive and Gram-negative bacteria. The fluorogenic method demonstrated that the five bacteriocins ofB. thuringiensishave bacteriolytic and bacteriostatic activities against all microorganisms tested, including clinically significant bacteria such asListeria monocytogenes, Proteus vulgaris,andShigella flexnerireported previously to be resistant to the antimicrobials as determined using the well-diffusion protocol. These results demonstrate that the fluorogenic assay is a more sensitive, reliable, and rapid method when compared with the well-diffusion method and can easily be adapted in screening protocols for bacteriocin production by other microorganisms.

Author(s):  
Sushma Vashisht ◽  
Manish Pal Singh ◽  
Viney Chawla

The methanolic extract of the resin of Shorea robusta was subjected to investigate its antioxidant and antibacterial properties its utility in free radical mediated diseases including diabetic, cardiovascular, cancer etc. The methanol extract of the resin was tested for antioxidant activity using scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazil) radical method, reducing power by FeCl3 and antibacterial activity against gram positive and gram negative bacteria using disc diffusion method. The phytochemical screening considered the presence of triterpenoids, tannins and flavoniods. Overall, the plant extract is a source of natural antioxidants which might be helpful in preventing the progress of various oxidative stress mediated diseases including aging. The half inhibition concentration (IC50) of resin extract of Shorea robusta and ascorbic acid were 35.60 µg/ml and 31.91 µg/ml respectively. The resin extract exhibit a significant dose dependent inhibition of DPPH activity. Antibacterial activity was observed against gram positive and gram negative bacteria in dose dependent manner.Key Words: Shorea robusta, antioxidant, antibacterial, Disc-diffusion, DPPH.


Author(s):  
Elaf Ayad Kadhem ◽  
Miaad Hamzah Zghair ◽  
Sarah , Hussam H. Tizkam, Shoeb Alahmad Salih Mahdi ◽  
Hussam H. Tizkam ◽  
Shoeb Alahmad

magnesium oxide nanoparticles (MgO NPs) were prepared by simple wet chemical method using different calcination temperatures. The prepared NPs were characterized by Electrostatic Discharge (ESD), Scanning Electron Microscope (SEM) and X-ray Diffraction (XRD). It demonstrates sharp intensive peak with the increase of crystallinty and increase of the size with varying morphologies with respect to increase of calcination temperature. Antibacterial studies were done on gram negative bacteria (E.coli) and gram positive bacteria (S.aureus) by agar disc diffusion method. The zones of inhibitions were found larger for gram positive bacteria than gram negative bacteria, this mean, antibacterial MgO NPs activity more active on gram positive bacteria than gram negative bacteria because of the structural differences. It was found that antibacterial activity of MgO NPs was found it has directly proportional with their concentration.


2012 ◽  
Vol 28 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Md Atiar Rahman ◽  
Rumana Sharmin ◽  
Md Nazim Uddin ◽  
Md Sohel Rana ◽  
Nazim Uddin Ahmed

Antibacterial effect of Crinum asiaticum bulb extract (1mg/disc) was tested on four Gram- positive and six Gram-negative bacteria by disc diffusion method using kanamycin (30 ìg/disc) as standard antibiotic disc. The bulb extract (250-1000mg/disc) showed significant zone of inhibition against all Gram-positive and Gram-negative bacteria ranging from 12-14 mm in diameter. Antioxidant potential of the same extract was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging method. The extract showed remarkable free radical scavenging effect (95.96%) providing the IC50 value of 5.62 for the bulb extract and 5.46 for ascorbic acid (standard antioxidant) at the concentration of 1000 ìg/ml. The bulb extract was found to be (LC50 value 94.06 ?g/ml) in Brine-Shrimp lethality test. DOI: http://dx.doi.org/10.3329/bjm.v28i1.11801 Bangladesh J Microbiol, Volume 28, Number 1, June 2011, pp 1-5


2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S386-S386
Author(s):  
Susan M Novak-Weekley ◽  
Aye Aye Khine ◽  
Tino Alavie ◽  
Namidha Fernandez ◽  
Laxman Pandey ◽  
...  

Abstract Background Conventional antimicrobial susceptibility testing (AST) of microorganisms from positive blood cultures (PBC) can take ≥ 2 days. In order to improve the turnaround time for AST on a PBC, CLSI and EUCAST have made efforts to standardize procedures for disk diffusion (DD) direct from a PBC. Qvella Corporation (Richmond Hill, ON, Canada) has recently developed FAST-Prep, an automated centrifugal sample preparation system that rapidly delivers a Liquid Colony consisting of a purified, concentrated, viable cell suspension directly from a PBC. This study was performed to investigate the feasibility of DD AST off of a PBC using a FAST-Prep Liquid Colony. Methods Contrived PBC samples were prepared by spiking 6 species of Gram-positive and 4 species of Gram-negative bacteria (3-5 strains per species) into FA® Plus bottles and incubating in the BACT/ALERT® VIRTUO® System (bioMerieux, Durham, NC). After positivity, 3 mL of PBC was added to the FAST-Prep cartridge. After 20 minutes of processing in the FAST-Prep instrument, the Liquid Colony was removed from the cartridge and a 0.5 McFarland sample was prepared for DD AST. In parallel, the DD AST from a PBC was performed using 4 drops of PBC (CLSI direct method). Both methods were compared to conventional colony-based DD AST. After 16-18 hours of incubation zone diameters and S/I/R interpretations were determined. Categorical agreement (CA) and errors for both DD AST methods were calculated. In addition, colony plate counting was performed on 0.5 McFarland suspensions of Liquid Colony and the plate colony to determine biomass recovery and sample purity. Results CA for a FAST-Prep DD AST for Gram-positive and Gram-negative bacteria was 95.6% and 98.6%, respectively, compared to CA for CLSI DD AST of 77.2% and 81.9%, respectively. Biomass in the Liquid Colony was 7.2x108 and 1.2x109 CFU for Gram-positive and Gram-negative bacteria, respectively. Cell concentration in the 0.5 McFarland suspension of the Liquid Colony was 3.7x107 and 5.9x107 CFU/mL for Gram-positive and Gram-negative bacteria, respectively, which was similar to the concentration for the reference colony suspension. Conclusion The results support the potential role of FAST-Prep in providing a Liquid Colony for use in rapid AST. Disclosures Susan M. Novak-Weekley, PhD, D(ABMM), Qvella (Employee, Shareholder) Aye Aye Khine, PhD, Qvella (Employee, Shareholder) Tino Alavie, PhD, Qvella (Employee) Namidha Fernandez, MS, Qvella (Employee) Laxman Pandey, MS, Qvella (Employee) Abdossamad Talebpour, PhD, Qvella (Employee, Shareholder)


2021 ◽  
Vol 2 (2) ◽  
Author(s):  
Eti Nurwening Sholikhah ◽  
Maulina Diah ◽  
Mustofa ◽  
Masriani ◽  
Susi Iravati ◽  
...  

Pycnarrhena cauliflora (Miers.) Diels., local name sengkubak, is one of indigenous plants from West Kalimantan that has been used as natural flavor. Pycnorrhena cauliflora is one of species of Menispermaceae family which is rich in bisbenzylisoquinoline alkaloids. This alkaloids are known to have various biological activities including antiprotozoal, antiplasmodial, antifungal and antibacterial activities. This study aimed to investigate antimicrobial activity of  the P. cauliflora (Miers.) Diels. methanolic extracts against gram-positive and gram-negative bacteria. The methanolic extract of P. cauliflora (Miers.) Diels., root, leaf and stem were prepared by maceration. The disk-diffusion method was then used to determine the antimicrobial activity of the extracts against Streptococcus pyogenes, S. mutants, Staphylococcus aureus, S. epidermidis, Salmonella typhi, Shigella flexneri, Pseudomonas aeruginosa and Escherichia coli after 18-24 h incubation at 37 oC. Amoxicillin was used as positive control for gram-positive bacteria and ciprofloxacin was used as gram-negative bacteria. The inhibition zones were then measured in mm. Analysis were conducted in duplicates. The results showed in general the methanolic extracts of P. cauliflora (Miers.) Diels. root (inhibition zone diameter= 10-23 mm) were more active than that leaf (0-15 mm) and stem (0-17 mm) extracts against gram-positive bacteria. The zone inhibition diameter of amoxicillin as positive control was 8-42 mm. In addition, the methanolic extracts of P. cauliflora (Miers.) Diels. root (12-17 mm) were also more active than that leaf (0-12 mm) and stem (0-12 mm) extracts against gram-negative bacteria. The zone inhibition diameter of ciprofloxacin as positive control was 33-36 mm. In conclusion, the methanolic extract of P. caulifloria (Miers.) Diels. root is the most extract active against both gram-positive and gram-negative bacteria. Further study will be focused to isolate active compounds in the methanolic extract of the root.


2021 ◽  
Vol 13 (1) ◽  
pp. 106-112
Author(s):  
Sri Kasmiyati ◽  
Elizabeth Betty Elok Kristiani ◽  
Maria Marina Herawati ◽  
Andreas Binar Aji Sukmana

The medicinal plant-derived bioactive compounds have a potential for many biological activities, including antimicrobial activity. Artemisia cina is a medicinal plant from the Compositae family with the potential of having antitumor, antifungal, and antibacterial activity. This study aimed to determine the antibacterial activity and the flavonoid content of A. Cina’s ethyl acetate extract. Plants samples were extracted by ethyl acetate maceration method. Antibacterial activity was tested against Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus) by a disk diffusion method using 25, 50, and 100 mg/l extract concentrations. The flavonoid contents (quercetin and kaempferol) were measured using High-Performance Liquid Chromatography. The extracts of diploid and polyploid A. cina displayed some antibacterial activity, with the Gram-negative bacteria being more resistant than the Gram-positive counterpart. However, no significant difference was observed between the diploid and polyploid extracts. As for the flavonoid content, the highest quercetin content (0.5501 mg/ml) was found in the polyploid A. cina (J), while the highest kaempferol content (0.5818 mg/ml) was observed in the diploid A. cina (KJT). Although A. cina is widely grown in Indonesia, compared to other Artemisia species, A. cina has not been widely studied, especially its antibacterial  potential and in related to its flavonoid content and the use of ethyl acetate as the extraction solvent.  This study reveals the potential of A. cina as a natural antibacterial agent. 


2014 ◽  
Vol 8 (3) ◽  
pp. 40-45
Author(s):  
Zina Hashem Shehab ◽  
Huda Suhail Abid ◽  
Sumaya Fadhil Hamad ◽  
Sara Haitham

The study was conducted to evaluate the inhibitory activity of methanol extract of Gardenia jasminoides leaves compared with leaf crude extracts for some organic solvents namely Methanol, Ethanol, Petroleum ether, Asetone and Chloroform on growth of some pathogenic bacteria and yeast, which included four gram positive isolates Staphylococcus aureus, Enterococcus faecalis, Streptococcus pyogenes and Bacillus cereus and gram negative isolates Escherichia coli, Salmonella typhi, Proteus vulgaris and Pseudomonas aeruginosa and some yeasts Candida albicans and Saccharomyces boulardii, by using well diffusion method. The inhibitory activity of extracts in the tested bacterial strains and yeasts was varied according to the type of extracting solvents and are tested microorganisms. The methanol callus extract which grown on Murashige and Skoog (MS) media by using (Naphthalen acitic acid) NAA and (Benzyle adenine) BA as growth regulator highly effective as compared to the other extracts as for inhibition of three gram positive bacteria and three gram negative bacteria,which include Staphylococcus aureus and, Proteus vulgaris, followed by acetone and ethanolic extracts which include two gram positive bacteria and two gram negative bacteria. All extracts had highly effect in growth of Candida albicans while all crude extracts didn’t show any sensitivity against Saccharomyces boulardii, and when we’d done (High Performance Liquid Chromatography) HPLC test for detection of some active compound we found Quinic acid, Iridiods glycosides and Crocin which its rate in fresh callus was higher than fresh leaves.


2018 ◽  
Vol 54 (4) ◽  
pp. 294
Author(s):  
S Zuhaira ◽  
Noorhaniz Mohd Nizam ◽  
PM Ridzuan

Antibiotic is a type of medication that helps in fighting bacterial infection. Treating bacterial infections in clinical setting become more complicated and costly due to drug resistance. This study was conducted to determine the antibacterial potential of Psidium guajava Linn leaf extracts against Gram-positive and Gram-negative bacteria. P. guajava Linn leaf was obtained from Research Orchards at University Putra Malaysia (UPM). Leaves were extracted using three types of extracts; hot, cold and methanol extract. Freeze dried was used in this study and temperature was set at -104°C for 24 hours. Antibacterial testing was determined using disc diffusion and well diffusion method. Antibacterial activity was evaluated by measuring the inhibitory zone of the tested bacteria. Phytochemical analysis was conducted by adding few drop of diluted NaOH (Sodium Hydroxide) solution to detect the active flavanoid from leaves extract. Antimicrobial activity showed all extracts were effective against Gram-positive and Gram-negative bacteria. Phytochemical analysis of P. guajava Linn leaf extracts showed the methanol extract indicating the presence of tannins, phenols, flavonoids, terpenoids, glycosides and saponins. Results showed that P. guajava Linn leaf was effective against Gram-positive and Gram-negative bacteria, and pythochemical analysis also exhibited a few active compounds that were determined. P. guajava Linn leaf had potential natural product that may be used for further analysis in the future research.


2020 ◽  
Vol 9 (6) ◽  
pp. 238-244
Author(s):  
Meharrar Nadjet ◽  
Bekada Ahmed Mohamed Ali ◽  
Dris Ibrahim ◽  
Jacques Dommes ◽  
Arabi Abed ◽  
...  

The genera Cistus present groups of about twenty species of shrubs belon- ging to the Cistaceae family are very common throughout the Mediterranean region. Cistus salviifolius, known as Melliya is a species of this genus that occurs in the forest region of Mount Ouarsenis (northwest Algeria), exactly in Thniet El Had Na onal Park in the Wilaya of Tissemsilt. This study was un- dertaken to evaluate the an bacterial effects of C. salviifolius and its chemical composition in addition the effect of shade drying me on essen al oil yield in order to op mize this parameter which is most o en not controlled for producers. The op mal yield recorded is about 0.07± 0.02%. Its characte- riza on by gas chromatography coupled with mass spectrometry shows a dominance of hydrocarbon compounds. The minimum inhibitory concentration (MIC) is established using 6 strains of Gram-positive and negative bacteria. The antibiotic susceptibility test is performed using the giliose- environment diffusion method, a bacterial suspension is prepared and subsequently well homogenized. Opacity must be adjusted to 0.5 McF (McFarland), from a young exponen al culture (18 to 24 hours). The an mi- crobial ac vity of C. salviifolius EO was tested against three Gram-posi ve and three Gram-nega ve bacteria. In fact, a strong inhibitory and bactericidal effect has been demonstrated against all Gram-positive and Gram- negative bacteria tested. It appears from this study that the EO of C. salviifo- lius has a very high antimicrobial activity, it shows a wide spectrum covering Gram positive and Gram negative bacteria. Taking into account the results of this study, new an bacterial agents could be developed and the use of C.salviifolius should be encouraged in the pharmaceutical and perfumery industries.


2012 ◽  
Vol 7 (5) ◽  
pp. 1934578X1200700 ◽  
Author(s):  
Alexis Peña ◽  
Luis Rojas ◽  
Rosa Aparicio ◽  
Libia Alarcón ◽  
José Gregorio Baptista ◽  
...  

The essential oil of the leaves of Espeletia nana Cuatrec, obtained by hydrodistillation, was analyzed by GC-MS, which allowed the identification of 24 components, which made up 99.9% of the oil. The most abundant compounds were α-pinene (38.1%), β-pinene (17.2%), myrcene (15.0%), spathulenol (4.2%), bicyclogermacrene (4.0%), α-zingiberene (4.0%), and γhimachalene (3.7%). Antibacterial activity was tested against Gram-positive and Gram-negative bacteria using the agar disk diffusion method. Activity was observed only against Gram-positive bacteria. MIC values were determined for Staphylococcus aureus ATCC 25923(200 μg/mL) and Enterococcus faecalis ATCC 29212 (600 μg/mL).


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