scholarly journals Spiral Gradient Dilution, a Rapid Method for Determining Growth Responses and 50% Effective Concentration Values in Fungus-Fungicide Interactions

2004 ◽  
Vol 94 (2) ◽  
pp. 163-170 ◽  
Author(s):  
Helga Förster ◽  
Loukas Kanetis ◽  
James E. Adaskaveg
Keyword(s):  
Mycelial Growth ◽  
Radial Distance ◽  
Conidial Germination ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Growth Responses ◽  
Continuous Growth ◽  
Agar Dilution ◽  
Regression Slopes ◽  
A New Technique

A new technique, the spiral gradient dilution method, was evaluated for determining 50% effective concentrations (EC50 values) of fungicides for the inhibition of mycelial growth and conidial germination of various fungi. In this method, an agar medium is plated with a fungicide solution by means of a spiral plater, which applies the fungicide in a 2.5-log dilution in a continuous radial concentration gradient. Fungal inoculum is then placed along radial lines across the gradient. After incubation of the plates, distinct growth shapes were observed in different fungus-fungicide interactions. Mycelial growth responses to increasing fungicide concentrations ranged from abrupt to gradual transitions. Conidial germination responses were similar; in addition, distinct zones of confluent growth, nonconfluent growth, and outlier colonies were also identified, depending on the fungus-fungicide interaction. The fungicide concentration at the radial distance at which 50% reduction of growth or spore germination occurred, compared with growth or germination on unamended media, was calculated by a computer program. EC50 values were obtained for mycelial growth in 22 fungus-fungicide interactions and for conidial germination in five interactions. The fungi evaluated were members of the Zygomycota, Ascomycota, Basidiomycota, and Deuteromycota. Nine fungicides, belonging to six different chemical classes, were tested. EC50 values were compared with those obtained by the traditional agar dilution method. In linear regression analyses of the two methods, the models were highly significant (P < 0.01), and coefficients of determination (r2) were 0.92 for the mycelial growth assays and 0.94 for the conidial germination assays. Regression slopes were not significantly different from 1 (P > 0.05) with optimal program settings in the software. Estimated bias, coefficients of variation, and actual confidence intervals for the regression slope were 13.5%, 6.5%, and 1.14 ± 0.14 for the mycelial growth assays and 7.5%, 14.5%, and 1.08 ± 0.37 for the conidial germination assays. These analyses indicate that the spiral gradient dilution method is accurate and precise compared with the agar dilution method for calculating EC50 values of fungicides in continuous growth responses to fungicide concentration gradients.

scholarly journals Antifungal activity of essential oils on Aspergillus parasiticus isolated from peanuts

2016 ◽  
Vol 56 (2) ◽  
pp. 139-142 ◽  
Author(s):  
Mina M. Yooussef ◽  
Quyen Pham ◽  
Premila N. Achar ◽  
Marikunte Yanjarappa Sreenivasa
Keyword(s):  
Antifungal Activity ◽  
Essential Oils ◽  
Mycelial Growth ◽  
Aspergillus Parasiticus ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Practical Application ◽  
Cinnamon Oil ◽  
Antifungal Effect ◽  
Agar Dilution

AbstractAspergillus parasiticus is one of the most common fungi which contaminates peanuts by destroying peanut shells before they are harvested and the fungus produces aflatoxins. The aim of this study was to evaluate the antifungal activities of seventeen essential oils on the growth of the aflatoxigenic form of A. parasiticus in contaminated peanuts from commercial outlets in Georgia. The agar dilution method was used to test the antifungal activity of essential oils against this form of A. parasiticus at various concentrations: 500; 1,000; 1,500; 2,000; 2,500 ppm. Among the seventeen essential oils tested, the antifungal effect of cinnamon, lemongrass, clove and thyme resulted in complete inhibition of mycelial growth. Cinnamon oil inhibited mycelial growth at ≥ 1,000 ppm, lemongrass and clove oils at ≥ 1,500 ppm and thyme at 2,500 ppm. However, cedar wood, citronella, cumin and peppermint oils showed partial inhibition of mycelial growth. Eucalyptus oil, on the other hand, had less antifungal properties against growth of A. parasiticus, irrespective of its concentration. Our results indicate that the aflatoxigenic form of A. parasiticus is sensitive to selected essential oils, especially cinnamon. These findings clearly indicate that essential oils may find a practical application in controlling the growth of A. parasiticus in stored peanuts.

scholarly journals Ammoides pusilla Essential Oil: A Potent Inhibitor of the Growth of Fusarium avenaceum and Its Enniatin Production

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6906
Author(s):  
Yasmine Chakroun ◽  
Souheib Oueslati ◽  
Vessela Atanasova ◽  
Florence Richard-Forget ◽  
Manef Abderrabba ◽  
...  
Keyword(s):  
Essential Oil ◽  
Mycelial Growth ◽  
Fusarium Species ◽  
Fusarium Avenaceum ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Agricultural Commodities ◽  
Fungistatic Activity ◽  
Antifungal Potential ◽  
Agar Dilution

Enniatins are mycotoxins produced by Fusarium species contaminating cereals and various agricultural commodities. The co-occurrence of these mycotoxins in large quantities with other mycotoxins such as trichothecenes and the possible synergies in toxicity could lead to serious food safety problems. Using the agar dilution method, Ammoides pusilla was selected among eight Tunisian plants for the antifungal potential of its essential oil (EO) on Fusarium avenaceum mycelial growth and its production of enniatins. Two EO batches were produced and analyzed by GC/MS-MS. Their activities were measured using both contact assays and fumigant tests (estimated IC50 were 0.1 µL·mL−1 and 7.6 µL·L−1, respectively). The A. pusilla EOs and their volatiles inhibited the germination of spores and the mycelial growth, showing a fungistatic but not fungicidal activity. The accumulation of enniatins was also significantly reduced (estimated IC50 were 0.05 µL·mL−1 for the contact assays and 4.2 µL·L−1 for the fumigation assays). The most active batch of EO was richer in thymol, the main volatile compound found. Thymol used as fumigant showed a potent fungistatic activity but not a significant antimycotoxigenic activity. Overall, our data demonstrated the bioactivity of A. pusilla EO and its high potential to control F. avenaceum and its enniatins production in agricultural commodities.

scholarly journals Evaluation of the Antimicrobial Activity of Some Components of the Essential Oils of Plants Used in the Traditional Medicine of the Tehuacán-Cuicatlán Valley, Puebla, México

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 295
Author(s):  
Sebastián Candelaria-Dueñas ◽  
Rocío Serrano-Parrales ◽  
Marisol Ávila-Romero ◽  
Samuel Meraz-Martínez ◽  
Julieta Orozco-Martínez ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Essential Oils ◽  
Qualitative Evaluation ◽  
Bactericidal Effect ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
E Coli ◽  
Gram Positive Bacteria ◽  
Diffusion Technique ◽  
Agar Dilution

In Tehuacán-Cuicatlán valley (Mexico), studies have been carried out on the essential oils of medicinal plants with antimicrobial activity and it was found that they present compounds in common such as: α-pinene, β-pinene, carvacrol, eugenol, limonene, myrcene, ocimene, cineole, methyl salicylate, farnesene, and thymol. The goal of this study was to assess the antimicrobial activity of essential oils’ compounds. The qualitative evaluation was carried out by the Kirby Baüer agar diffusion technique in Gram-positive bacteria (11 strains), Gram-negative bacteria (18 strains), and yeasts (8 strains). For the determination of the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), the agar dilution method was used. All the evaluated compounds presented antimicrobial activity. The compounds eugenol and carvacrol showed the largest inhibition zones. Regarding yeasts, the compounds ocimene, cineole, and farnesene did not show any activity. The compounds eugenol, carvacrol, and thymol presented the lowest MIC; bactericidal effect was observed at MIC level for S. aureus 75MR, E. coli 128 MR, and C albicans CUSI, for different compounds, eugenol, carvacrol, and thymol. Finally, this study shows that the essential oils of plants used by the population of Tehuacán-Cuicatlán valley share compounds and some of them have antibacterial and fungicidal activity.

scholarly journals Antibiotic sensitivities of Neisseria meningitidis isolates from patients and carriers in Greece

1992 ◽  
Vol 108 (3) ◽  
pp. 449-455 ◽  
Author(s):  
G. Tzanakaki ◽  
C. C. Blackwell ◽  
J. Kremastinou ◽  
C. Kallergi ◽  
G. Kouppari ◽  
...  
Keyword(s):  
Neisseria Meningitidis ◽  
School Children ◽  
Penicillin G ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Southern Europe ◽  
Agar Dilution ◽  
Of Greece

SUMMARYUsage of antibiotics in southern Europe is less well regulated than in northern countries. The proportion (48%) of meningococci in Spain insensitive to penicillin (MIC ≥ 0·1 mg/l) prompted this investigation of antibiotic sensitivities of isolates from Greek patients with meningitis (31) and carriers (47 school-children and 472 recruits). The agar dilution method was used to determine MIC to penicillin G (PN), sulphamethoxazole (SU), rifampicin (RF), cefaclor (CF) and ciprofloxacin (CP).The proportion of isolates insensitive to PN was 48% for isolates from patients, 19% from school-children and 36·6% from recruits. Resistance to SU (MIC ≥ 16 mg/l) was found in 16% of those from patients, 10·6% from children and 40% from recruits. None of the isolates from patients was resistant to RF (≥ 1 mg/l) but 6% of those from carriers were. Resistance to CF (≥ 4 mg/l) was found in 9·2% of patient isolates, 6·4% from children and 23·7% from recruits. All isolates except one were sensitive to CP (MIC range < 0·0015–0·125 mg/l).Resistances to PN, SU and RF were analysed by serogroup, serotype and subtype of the bacteria. The proportion of resistant isolates showed some variation between different areas of Greece, but it was not statistically significant.

scholarly journals Prevalence and characterization of extended spectrum beta-lactamase-producing Enterobacter cloacae strains in Algeria

2013 ◽  
Vol 7 (11) ◽  
pp. 804-811 ◽  
Author(s):  
Sabrina Nedjai ◽  
Abouddihaj Barguigua ◽  
Nassima Djahmi ◽  
Loubna Jamali ◽  
Khalid Zerouali ◽  
...  
Keyword(s):  
Enterobacter Cloacae ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Extended Spectrum Beta Lactamase ◽  
Agar Dilution ◽  
Synergy Test ◽  
High Level ◽  
Genomic Typing ◽  
Microbiological Data

Introduction: Expended spectrum β-lactamase (ESBL)-producing Enterobacter cloacae is an important nosocomial pathogen. In this study, the prevalence and the molecular epidemiology of ESBL producing E. cloacae strains isolated from various hospitals in Annaba, Algeria were investigated. Methodology: The study involved 63 isolates of E. cloacae obtained during 2009 at the four hospitals in Annaba. The detection of ESBL was performed using the double-disk synergy test and the combined disk test. Minimum inhibitory concentrations (MICs) were determined using the agar dilution method. The presence of blaCTX-M, blaSHV, blaTEM, and blaDHA β-lactamase genes was evaluated by PCR, and genomic typing was determined by pulsed-field gel electrophoresis (PFGE) analysis. The clinical and microbiological data were entered into the EpiI Info database. Results: Thirty isolates (47.6%) had an ESBL phenotype. BlaCTX-M group1 (76%); blaTEM (70%) were the most prevalent, followed by blaDHA (16.6%) and blaSHV (10%). Eighteen strains expressed at least two bla genes. MICs revealed a high level of resistance to cefotaxime, ceftazidime, and cefepime.  PFGE revealed an epidemic clonal dissemination of these isolates. Various risk factors associated with the occurrence of ESBL-producing E. cloacae were detected. Conclusions: A higher frequency of ESBL-producing isolates and a diversity of β-lactamases were detected among ESBL-producing E. cloacae; these resulted from an epidemic clonal dissemination and high transference of ESBL genes between bacteria in hospital settings. Strict measures will be required to control the further spread of these pathogens in hospital settings.

scholarly journals Antifungal and Antibacterial Metabolites from a French Poplar Type Propolis

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Séverine Boisard ◽  
Anne-Marie Le Ray ◽  
Anne Landreau ◽  
Marie Kempf ◽  
Viviane Cassisa ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Antifungal Activity ◽  
Antimicrobial Effect ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Gram Positive ◽  
Weak Activity ◽  
Agar Dilution

During this study, thein vitroantifungal and antibacterial activities of different extracts (aqueous and organic) obtained from a French propolis batch were evaluated. Antifungal activity was evaluated by broth microdilution on three pathogenic strains:Candida albicans, C. glabrata, andAspergillus fumigatus. Antibacterial activity was assayed using agar dilution method on 36 Gram-negative and Gram-positive strains includingStaphylococcus aureus. Organic extracts showed a significant antifungal activity againstC. albicansandC. glabrata(MIC80between 16 and 31 µg/mL) but only a weak activity towardsA. fumigatus(MIC80= 250 µg/mL). DCM based extracts exhibited a selective Gram-positive antibacterial activity, especially againstS. aureus(SA) and several of its methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains (MIC10030–97 µg/mL). A new and active derivative of catechin was also identified whereas a synergistic antimicrobial effect was noticed during this study.

scholarly journals Antimicrobial activity of honeys from two stingless honeybee species and Apis mellifera (Hymenoptera: Apidae) against pathogenic microorganisms

2014 ◽  
Vol 44 (2) ◽  
pp. 287-290 ◽  
Author(s):  
Carolinie Batista Nobre da Cruz ◽  
Fabio Alessandro Pieri ◽  
Gislene Almeida Carvalho-Zilse ◽  
Patrícia Puccinelli Orlandi ◽  
Carlos Gustavo Nunes-Silva ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Apis Mellifera ◽  
Antimicrobial Agents ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Therapeutic Approaches ◽  
E Coli ◽  
Agar Dilution ◽  
Amazonas State

Honeys are described possessing different properties including antimicrobial. Many studies have presented this activity of honeys produced by Apis mellifera bees, however studies including activities of stingless bees honeys are scarce. The aim of this study was to compare the antimicrobial activity of honeys collected in the Amazonas State from Melipona compressipes, Melipona seminigra and Apis mellifera against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, and Candida albicans. Minimum inhibitory concentrations were determined using the agar dilution method with Müller-Hinton agar (for bacteria) or Saboraud agar (for yeast). Staphylococcus aureus and E. faecalis were inhibited by all honeys at concentrations below 12%, while E. coli and C. violaceum were inhibited by stingless bee honeys at concentrations between 10 and 20%. A. mellifera honey inhibited E. coli at a concentration of 7% and Candida violaceum at 0.7%. C. albicans were inhibited only with honey concentrations between 30 and 40%. All examined honey had antimicrobial activity against the tested pathogens, thus serving as potential antimicrobial agents for several therapeutic approaches.

scholarly journals Drug resistance of yeasts isolated from oropharyngeal candidiasis in aids patients

1998 ◽  
Vol 29 (4) ◽  
pp. 271-275
Author(s):  
Maria do Rosário Rodrigues Silva ◽  
Claudete Rodrigues de Paula ◽  
Soraya Cristina Silva ◽  
Théo Rodrigues Costa ◽  
Márcio Rodrigues Costa
Keyword(s):  
Amphotericin B ◽  
High Frequency ◽  
Inhibitory Concentration ◽  
Serum Levels ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Oropharyngeal Candidiasis ◽  
Candida Spp ◽  
Aids Patients ◽  
Agar Dilution

Candida spp was isolated from 59 (68.60%) out of eighty six samples of oral mucosa of AIDS patients. The identification, based or the production of a germ tube and chlamydospores, and on the assimilation and fermentation of carbohydrates, revealed 52 strains (88.13%) of C. albicans, 4 (6.77%) of C. tropicalis and 3 (5.08%) of C. krusei. The susceptibility of these strains to amphotericin B, flucytosine, itraconazole, fluconazole and ketoconazole was determined using the agar dilution method. Comparing the minimum inhibitory concentration values found in the susceptibility test with the serum levels achieved by these drugs, only 8.47% and 5.08% of the yeasts strains proved to be resistant to amphotericin B and flucitosyne, respectively. A high frequency of strains resistant to azole derivatives (25.42%, to itraconazole, 45.76%, to ketoconazole and 66.10% to fluconazole) was observed.

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