scholarly journals Russia-made Mueller–Hinton agar: compliance with contemporary requirements

2019 ◽  
Vol 9 (2) ◽  
pp. 409-416
Author(s):  
L. V. Domotenko ◽  
I. S. Kosilova ◽  
A. P. Shepelin

At present, a rise of antimicrobial resistance requires that susceptibility of infectious agents to antimicrobial agents could be accurately evaluated as related errors may lead to selecting improper therapeutics provoking spread of drug resistance. Pathogen sensitivity to antimicrobial agents is commonly determined by a disc diffusion method. A quality of nutrient medium used in assays plays a crucial role influencing final results. In Russia, it turned out that regulatory documents such as the nationwide guidelines and clinical recommendations outlining methodology for antimicrobial susceptibility testing underlay availability in domestic market few nutrient media, including Mueller–Hinton Agar, AGV medium etc. exhibiting sometimes unsatisfactory quality. To harmonize such methodology with international requirements, theStateResearchCenterfor Applied Microbiology and Biotechnology has developed a technology and promoted manufacture of Russia-made Mueller–Hinton agar satisfying requirements of EUCAST documents, clinical guidelines, and ISO/TS 16782:2016. The main objective of this study was to compare quality of new agar product with five similar foreign media while examining 11 test strains by disc diffusion method. As a result, some of nutrient media available to the Russian market turned out to be off-standard: not all of them satisfy to the EUCAST requirements and clinical guidelines since diameter distribution for growth inhibition recommended by EUCAST for quality control does not fit into permissible range. Moreover, susceptibility of P. aeruginosa ATCC 27853 to aminoglycosides, fluoroquinolones, Meropenem, as well as S. aureus ATSS 25923 and E. faecalis ATCC 29212 to tigecycline was assessed with certain mistakes. The data obtained by us were analyzed in accordance to the new document ISO/TS 16782:2016 “Clinical laboratory testing — criterion for acceptable lots of dehydrated Mueller–Hinton agar and broth for antimicrobial susceptibility testing”, not approved yet In Russia. To determine potential reason for deviation of data from reference range, we measured concentration of bivalent metals in all nutrient media examined by atomic emission spectrometry with inductively coupled plasma. We determined new patterns affecting reliability of results on microbial antibiotic susceptibility. A need to check intralaboratory quality control of nutrient media was emphasized.  

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Anwar Ali Chandio ◽  
Ayaz Ali Memon ◽  
Shahabuddin Memon ◽  
Fakhar N. Memon ◽  
Qadeer Khan Panhwar ◽  
...  

Present study deals with the synthesis of the p-tert-butylcalix[4]arene diamide derivative as ligand (L) and its Fe3+ complex, followed by its characterization using TLC and FT-IR, while UV-Vis and Job’s plot study were performed for complex formation. Antimicrobial activity of the derivative (L) and its metal complex was carried out by the disc diffusion method against bacteria (Escherichia coli and Staphylococcus albus) and fungi (R. stolonifer). Different concentrations of the derivative (L) (6, 3, 1.5, 0.75, and 0.37 μg/mL) and its Fe3+ complex were prepared, and Mueller–Hinton agar was used as the medium for the growth of microorganisms. Six successive dilutions of the derivative (L) and Fe3+ complex were used against microorganisms. Two successive dilutions (6 and 3 μg/mL) of the derivative (L) showed antibacterial action against both Gram-positive and Gram-negative bacteria. In addition, three successive dilutions (6, 3, and 1.5 μg/mL) of the derivative (L) showed antifungal activity. However, all of six dilutions of the Fe3+ complex showed antimicrobial activity. Derivative (L) showed 3 and 1.5 μg/mL minimum inhibitory concentrations (MIC) against bacteria and fungi, respectively. On the contrary, its Fe3+ complex showed 0.37 μg/mL value of MIC against bacteria and fungi. Hence, Fe3+ complex of the derivative (L) was found to be a more effective antimicrobial agent against selected bacteria and fungi than the diamide derivative (L).


2017 ◽  
Vol 62 (No. 7) ◽  
pp. 410-415 ◽  
Author(s):  
J. Bzdil ◽  
O. Holy ◽  
J. Toporcak

The aim of this study was to describe two case studies of reptile disease, in which the primary pathogen was apparently Salmonella enterica subspecies enterica serovar Paratyphi B. Pathological examinations, sampling from different organs and cloacal swabs was performed on a dead savannah monitor (Varanus exanthematicus) and a sick green tree python (Chondropython viridis). This material was subjected to culture examination, including selective enrichment using standard methods. Typing was performed using MALDI-TOF and strains were also serologically typified. The utilisation of d-tartrate was confirmed biochemically and also using PCR. Antibiotic susceptibility was determined by the standard disc diffusion method using Mueller-Hinton agar without blood and antibiotic discs. In both cases the detected Salmonella enterica subspecies enterica serovar Paratyphi B was positive for d-tartrate and exquisitely susceptible to chloramphenicol, tetracycline, ampicillin, amoxycillin/clavulanic acid, gentamicin, ceftazidime, enrofloxacin and piperacillin.


2002 ◽  
Author(s):  
◽  
Mbuso Mabuza

The aim of this in vitro microbial study was to evaluate the efficacy of Calendula officinalis tincture 60% (v/v) ethanol as an antibacterial on in vitro Pseudomonas aeruginosa. The standardised disc - diffusion method was employed. Seven pairs of Mueller - Hinton agar plates were used.


2016 ◽  
Vol 78 (11-2) ◽  
Author(s):  
Fatimatul Akmal Sulaiman ◽  
Nurfarahin Fuad ◽  
Farawahida Rahman ◽  
Anwar Iqbal ◽  
Deny Susanti Darnis

Apart from being the primary source of food to other living things, plants also have medicinal value to treat various kinds of diseases. In recent years, it has been proposed that the extract from plants may be used as natural antioxidants which can help to prevent the generation of carcinogens in human body. In addition, plants also have antimicrobial agents to inhibit the growth of pathogenic microbes. This study was intended to investigate the antioxidant properties and antimicrobial activity of methanolic extract of Tinospora crispa stems extracted using soxhlet extraction method. The antimicrobial properties of T. crispa stems extract were tested using disc diffusion method against Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 11778, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Candida albicans IMR C S23/11 A and Saccharomyces cerevisiae IMR S 617/068. The antioxidant properties of the extract were investigated by using Total Phenolics Content (TPC), Total Flavonoids Content (TFC), DPPH free radical scavenging and b–carotene bleaching assays.  The TPC value was 6.12 g GAE/100 g of dried extract while the TFC value was 55.58 g QE/100 g of dried extract. The IC50 of DPPH scavenging assay for the extract and ascorbic acid were 0.21 and 0.04 mg/mL, respectively. The average percentage of b–carotene bleaching assay was 38.3 % as compared to BHT, which was 45.1%.  The disc diffusion method showed no inhibition zone against all the strains of microorganisms at all concentrations of the extracts (0.5, 1.0, 2.5 and 5.0 mg/disc). 


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Comfort Nkogwe ◽  
Juliah Raletobana ◽  
Alva Stewart-Johnson ◽  
Sharianne Suepaul ◽  
Abiodun Adesiyun

The study was conducted to determine the frequency of isolation ofSalmonella,CampylobacterandE. coliO157 in the faecal samples of rats trapped across the regional corporations in Trinidad and to assess their resistance to antimicrobial agents. A total of 204 rats were trapped for the detection of selected bacteria. Standard methods were used to isolateSalmonella,CampylobacterandE. coliO157. Characterization ofE. coliwas done on sorbitol MacConkey agar to determine non-sorbitol fermentation, blood agar to determine haemolytic and mucoid colonies and by usingE. coliO157 antiserum to determine O157 strain. The disc diffusion method was used to determine resistance to nine antimicrobial agents. Of the 204 rats, 4 (2.0%), 7 (3.4%) and 171 (83.8%) were positive forSalmonellaspp.,Campylobacterspp. andE. coli, respectively. Of the 171 isolates ofE. colitested 0 (0.0%), 25 (14.6%) and 19 (11.1%) were haemolytic, mucoid and non-sorbitol fermenters, respectively. All isolates were negative for the O157 strain. The frequency of resistance to the 9 antimicrobial agents tested was 75% (3 of 4) forSalmonella, 85.7% (6 of 7) ofCampylobacterspp. and 36.3% (62 of 171) forE. coli(;χ2).


2021 ◽  
Vol 12 (6) ◽  
pp. 87-90
Author(s):  
Virendra Vaishnav ◽  
Debasish Sahoo ◽  
Tanushree Chatterjee

Medicinal Plants are the good source of natural antimicrobial agents. The main aim of present study was to evaluate the antibacterial activity of stem and root of Rauwolfia serpentina against six microorganism, Powdered stem and root of plant were extracted with acetone, chloroform and methanol and streptomycin used as positive control. The antibacterial activity of Rauwolfia serpentine was detected by using disc diffusion method and agar well diffusion method on the following bacteria- Bacillus cereus, Staphylococcus aureus, Bacillus fusiformis, Escherichia coli, Pseudomonas aeruginosa and P. luminescens. The experiment reported that R. serpentina Root methanol extract shown 14.86 ± 1.11 highest antibacterial activity against Pseudomonas aeruginosa through well diffusion method. Whereas root chloroform recorded 13.46 ± 1.28 highest antibacterial activity against E. coli through disc diffusion method, maximum zone of inhibition 22.66±0.52 mm was found for the positive control, streptomycin through well diffusion method. Further studies should be undertaken to reveal the correct mechanism of action of antimicrobial effect to identify the active ingredients which can be used in drug development program.


Author(s):  
Gaurab Risal ◽  
Aayush Shrestha ◽  
Saroj Kunwar ◽  
Gajal Paudel ◽  
Rameshwor Dhital ◽  
...  

Background: In urinary tract infections, an important role is played by bacterial biofilms which are responsible for persistence infections together with the antimicrobial resistance. Higher resistance can be seen in biofilm forming uropathogens in comparison with free-floating bacteria. So, the present study was performed with a goal to find the prevalence of biofilm formation and also the antimicrobial resistant pattern of uropathogens.Methods: A descriptive method was conducted at Modern Technical College, Sanepa, Lalitpur in samples isolated from UTI suspected patients. The overall duration of this study was approximately 3 months. Total of 50 isolated E. coli was tested for biofilm formation and antimicrobial susceptibility testing was done by Kirby-Bauer disc diffusion method on Mueller Hinton agar as per CLSI guidelines.Results: From the 50 isolates of E. coli, 32 were biofilm producers (3 strong and 29 moderate) and 18 were weak/non-biofilm producers. Among the biofilm producers, cefotaxime was more resistant in 20 of the isolates followed by ceftriaxone in 16 and amoxyclav in 13, whereas amikacin was least resistant in 2 of the isolates.Conclusions: Among the isolated E. coli, biofilm-forming isolates showed higher antimicrobial resistance as compared to the non-biofilm producer. Thus, uropathogen should be routinely screened for biofilm formation. 


2021 ◽  
Vol 30 (1) ◽  
pp. 101-106
Author(s):  
K. F. Chah ◽  
S. C. Okafor ◽  
S. I. Oboegbulem

This study was carried out to determine resistance profiles of Escherichia coli strains isolated from clinically healthy chickens in Nsukka, southeast Nigeria. A total of 324 E. coli strains isolated from cloaca swabs from 390 chickens were tested against 16 antimicrobial agents using the disc diffusion method. The antibiotics used in the study were: ampicillin (25µg), amoxycillin-clavulanic acid (30µg), gentamicin (10µg), Streptomycin (30µg). cefuroxime (20µg), cephalexin (10µg), nalidixic acid (30µg), ciprofloxacin (5µg), norfloxacin (10µg), ofloxacin (5µg), pefloxacin (5µg), tetracycline (30µg), chloramphenicol (10µg), cotrimoxazole (50µg), colistin (25µg) and nitrofurantoin (100µg).The strains demonstrated high rates of resistance (34.6%  66.1%) to ampicillin, tetracycline, nitrofurantoin, cefuroxime and cotrimoxazole. None of the isolates was resistant to colistin, ofloxacin and pefloxacin. For each antimicrobial agent (except cephalexin), strains from the intensively reared chickens (layers and broilers) displayed higher resistance frequencies than those from the local birds. A total of 49 resistant patterns were recorded for the 228 strains resistant to at least one antimicrobial drug, with AmTeCoS and AmTeCfN being the predominant patterns. Because of the great variation in the drug resistance patterns of the Escherichia coli strains, use of antimicrobial agents in the management of E. coli infections in the study area should be based on results of sensitivity tests.


2019 ◽  
Vol 64 (6) ◽  
pp. 360-367
Author(s):  
I. S. Kosilova ◽  
L. V. Domotenko ◽  
N. K. Fursova ◽  
S. V. Dentovskaya ◽  
M. G. Ershova ◽  
...  

The results of the comparative tests of the «Agar Muller-Hinton II - Obolensk» nutrient medium developed in SRCAMB, Obolensk, and the control nutrient medium imported «Mueller Hinton II Agar» are presented in the study. The susceptibility of bacterial clinical strains to antimicrobial agents (AMP) was determined by the disc diffusion method and the method of gradient diffusion (E-test). The carbapenemase activity of the strains carrying the carbapenemase genes was determined by CIM-test. Total 173 characterized bacterial strains of species Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, Serratia marcescens, Enterobacter aerogenes, Escherichia coli; Photorhabdus spp., Staphylococcus aureus, Enterococcus spp. were used in the study, including producers of OXA- and NDM-types carbapenemases for gram negative bacteria. A high degree of coincidence of the results obtained on both nutrient media was shown. The consistency index of the strain sensitivity categories to AMPs (S, I, and R) was 98.2% for the disc diffusion method, and 94.4-100% - for E-test and CIM-test methods. Thus, within the framework of the Import Substitution Program, the domestic nutrient medium «MHA II-Obolensk» has been successfully developed. The nutrient medium meets the requirements of GOST R ISO 20776-2-2010 «Clinical laboratory testing and in vitro diagnostic test systems - Susceptibility testing of infectious agents and evaluation of performance of antimicrobial susceptibility test devices».


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