Abstract:The development of a specific agar diffusion bioassay for the quantitative determination of fluconazole formulated in capsules was carried out using a strain of Candida albicans ATCC 18804 as the test organism. A prospective validation of the method showed adequate linearity (r 2 =0.9995), precision (R.S.D. = 4.0% for intra-day and 4.5% for inter-day precision) and accuracy (mean recovery = 102.9%). High performance liquid chromatography was chosen as a comparison method for the fluconazole determination. The … Show more
“…(53). Moreover, a microbiological assay was a straightforward, robust, and inexpensive method compared to the routinely used approaches such as spectrophotometric detection and High pressure liquid chromatography (HPLC) and the results obtained are reproducible (58).…”
Background: Contamination of natural niches with pharmaceutical residues has emerged out as a serious concern. Disposal of untreated effluents from the pharmaceutical, hospital, and domestic settings has been identified as a significant source of such a massive spread of antibiotics. The unnecessary persistence of pharmaceutical residues including antibiotics has been related to the increased risk of resistance selection among pathogenic and non-pathogenic microorganisms. To date, several methods have been devised to eliminate such pollutants from wastewater, but their implication on larger scales is not feasible due to complexities and high costs of the processes, especially in developing and underdeveloped countries. This study aimed to isolate and characterize bacterial strains from domestic and pharmaceutical effluents having biotransformation potential towards most persistent antibiotics. Results: Antibiotic resistance screening and MIC determination experiments indicated highest resistivity of three bacterial isolates against two antibiotics Erythromycin and Sulfamethoxazole-trimethoprim, evincing extensive usage of these antibiotics in our healthcare settings. These isolates were identified as Comamonas jiangduensis, Aeromonas caviae and Aeromonas hydrophila by 16S rDNA sequencing. Growth conditions including incubation temperature, initial pH and inoculum size were optimized for these strains. Successful biotransformation of Erythromycin and Sulfamethoxazole-trimethoprim was achieved within 92 h under optimum growth conditions. Conclusions: Aeromonas and Comononas species were found to be potent degraders of antibiotics tested, presenting these strains as potential candidates to be utilized in the remediation processes.
“…(53). Moreover, a microbiological assay was a straightforward, robust, and inexpensive method compared to the routinely used approaches such as spectrophotometric detection and High pressure liquid chromatography (HPLC) and the results obtained are reproducible (58).…”
Background: Contamination of natural niches with pharmaceutical residues has emerged out as a serious concern. Disposal of untreated effluents from the pharmaceutical, hospital, and domestic settings has been identified as a significant source of such a massive spread of antibiotics. The unnecessary persistence of pharmaceutical residues including antibiotics has been related to the increased risk of resistance selection among pathogenic and non-pathogenic microorganisms. To date, several methods have been devised to eliminate such pollutants from wastewater, but their implication on larger scales is not feasible due to complexities and high costs of the processes, especially in developing and underdeveloped countries. This study aimed to isolate and characterize bacterial strains from domestic and pharmaceutical effluents having biotransformation potential towards most persistent antibiotics. Results: Antibiotic resistance screening and MIC determination experiments indicated highest resistivity of three bacterial isolates against two antibiotics Erythromycin and Sulfamethoxazole-trimethoprim, evincing extensive usage of these antibiotics in our healthcare settings. These isolates were identified as Comamonas jiangduensis, Aeromonas caviae and Aeromonas hydrophila by 16S rDNA sequencing. Growth conditions including incubation temperature, initial pH and inoculum size were optimized for these strains. Successful biotransformation of Erythromycin and Sulfamethoxazole-trimethoprim was achieved within 92 h under optimum growth conditions. Conclusions: Aeromonas and Comononas species were found to be potent degraders of antibiotics tested, presenting these strains as potential candidates to be utilized in the remediation processes.
“…When comparing the microbiological assay and chemical assay, each exhibits several advantages and deficiencies [51] . Both microbiological and HPLC methods are used to evaluate the potency of antibiotics [26] , [27] and to determine drug concentration in body fluids [52] and in therapeutic drug monitoring [53] .…”
Section: Comparison Of Microbiological Assay and Hplc Methodsmentioning
confidence: 99%
“…In 5×1 assay design, five Petri dishes were used for each assay in order to test the reference concentration concomitantly with each standard or sample concentration ( Fig. 1 C) [1] , [27] . Fig.…”
Section: Different Assay Designs For Agar Diffusion Assaymentioning
Antibiotics are the chemotherapeutic agents that kill or inhibit the pathogenic microorganisms. Resistance of microorganism to antibiotics is a growing problem around the world due to indiscriminate and irrational use of antibiotics. In order to overcome the resistance problem and to safely use antibiotics, the correct measurement of potency and bioactivity of antibiotics is essential. Microbiological assay and high performance liquid chromatography (HPLC) method are used to quantify the potency of antibiotics. HPLC method is commonly used for the quantification of potency of antibiotics, but unable to determine the bioactivity; whereas microbiological assay estimates both potency and bioactivity of antibiotics. Additionally, bioassay is used to estimate the effective dose against antibiotic resistant microbes.Simultaneously, microbiological assay addresses the several parameters such as minimal inhibitory concentration (MIC), minimum bactericidal concentration (MBC), mutation prevention concentration (MPC) and critical concentration (Ccr) which are used to describe the potency in a more informative way. Microbiological assay is a simple, sensitive, precise and cost effective method which gives reproducible results similar to HPLC. However, the HPLC cannot be a complete substitute for microbiological assay and both methods have their own significance to obtain more realistic and precise results.
“…The inhibition zone of growth of Candida albicans was measured after 24 hours, the mean inhibition zone was then calculated. The standard deviation (SD) was also calculated (Queiroz et al, 2009;Hurtado et al, 2008).…”
Section: Effect Of Storage At Room Temperature On Drug Content Hardnmentioning
Fluconazole is a synthetic triazole antifungal drug. It is available as tablets, capsules, oral suspension and injection for treatment of oral, esophageal and vaginal candidiasis. The present work was envisaged to develop a novel and aesthetic bioadhesive vaginal delivery system with improved efficacy. The compatibility between the drug, mucoadhesive polymers and tablet excipients (carbopol, HPMC, Xanthan gum and EC) was studied using Fourier transform infrared (FTIR) spectroscopy and differential scanning calorimeter (DSC). Fluconazole bioadhesive vaginal tablets (FBVTs) were prepared using mixtures of polymers that gave the optimal physico-mechanical properties and also showed the most delayed in-vitro release results. The in-vitro susceptibility profile of Candida albicans to Fluconazole was evaluated using agar cup method. The inhibition zones were calculated and these values were taken as the antifungal effect of the drug. The bioadhesive tablets were clinically evaluated in volunteers suffering from vaginal candidiasis and compared with commercial Fluconazole vaginal tablets (Diflucan ® 150 mg).
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