The aim of broth and agar dilution methods is to determine the lowest concentration of the assayed antimicrobial agent (minimal inhibitory concentration, MIC) that, under defined test conditions, inhibits the visible growth of the bacterium being investigated. MIC values are used to determine susceptibilities of bacteria to drugs and also to evaluate the activity of new antimicrobial agents. Agar dilution involves the incorporation of different concentrations of the antimicrobial substance into a nutrient agar medium followed by the application of a standardized number of cells to the surface of the agar plate. For broth dilution, often determined in 96-well microtiter plate format, bacteria are inoculated into a liquid growth medium in the presence of different concentrations of an antimicrobial agent. Growth is assessed after incubation for a defined period of time (16-20 h) and the MIC value is read. This protocol applies only to aerobic bacteria and can be completed in 3 d.

Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances

In the last decade we have witnessed a dramatic increase in the proportion and absolute number of bacterial pathogens resistant to multiple antibacterial agents. Multidrug-resistant bacteria are currently considered as an emergent global disease and a major public health problem. The B-Debate meeting brought together renowned experts representing the main stakeholders (i.e. policy makers, public health authorities, regulatory agencies, pharmaceutical companies and the scientific community at large) to review the global threat of antibiotic resistance and come up with a coordinated set of strategies to fight antimicrobial resistance in a multifaceted approach. We summarize the views of the B-Debate participants regarding the current situation of antimicrobial resistance in animals and the food chain, within the community and the healthcare setting as well as the role of the environment and the development of novel diagnostic and therapeutic strategies, providing expert recommendations to tackle the global threat of antimicrobial resistance.

https://wrap.warwick.ac.uk/67444/1/WRAP_main.pdf

The global threat of antimicrobial resistance: science for intervention

In this Opinion article, Baquero and colleagues propose a hierarchical system for estimating the risks associated with genes present in environmental resistomes, by evaluating the likelihood of their introduction into human pathogens, and the consequences of such introduction events for the treatment of bacterial infections.

What is a resistance gene? Ranking risk in resistomes.

The changes that have been made to the previous version of the recommendations (version 6) are as follows: medium and incubation condition for testing Acinetobacter spp. (Tables 1 and 6); use of cefoxitin as an indicator antibiotic for detecting methicillin/oxacillin/cefoxitin resistance in coagulase-negative staphylococci (Tables 1, 6 and 11); MIC breakpoint for co-trimoxazole based on the trimethoprim concentration in a 1:19 combination with sulfamethoxazole (Tables 7, 10, 11, 12, 15, 16 and 19); advice on the use of azithromycin for the treatment of infections with Salmonella typhi (footnote to Table 7); amendment to the recommendation for cefuroxime for the treatment of infections with Proteus mirabilis (footnote Table 7); MIC and zone diameter breakpoints for Stenotrophomonas maltophilia only (Table 10); MIC breakpoints for daptomycin (Tables 11 and 15); clarification for staphylococci that the neomycin zone diameter breakpoints are for topical use only and differentiate the isolates outside the 'wild-type' population in Table 11; clarification for beta-haemolytic streptococci that the linezolid zone diameter breakpoints relate to an MIC breakpoint of 2 mg/L as no data for the intermediate category are currently available (Table 15); clarification that strains with reduced susceptibility to fluoroquinolones give no zone of inhibition with a 30 microg nalidixic acid disc (Tables 16 and 21); erythromycin is no longer used for therapy of Neisseria gonorrhoeae, but may be tested for epidemiological purposes (Table 17); clarification that the ciprofloxacin zone diameter breakpoint for Neisseria meningitidis relates to the MIC breakpoint of 0.03 mg/L as no data for the intermediate category are currently available; clarification that the ciprofloxacin zone diameter breakpoints for Campylobacter spp. relate to an MIC breakpoint of 0.5 mg/L as no data for the intermediate category are currently available; clarification that for ciprofloxacin and vancomycin zone diameter breakpoints for coryneform organisms relate to an MIC breakpoint of 0.5 and 4 mg/L, respectively, as no data for the intermediate category are currently available; MIC and zone diameter breakpoints for Gram-negative rods isolated from urinary tract infections have been expanded to include Klebsiella spp.; and a definition of coliforms is also included (Table 26).

BSAC standardized disc susceptibility testing method (version 7)

https://www.clinicalmicrobiologyandinfection.com/article/S1198-743X(14)60298-6/pdf

Development of the EUCAST disk diffusion antimicrobial susceptibility testing method and its implementation in routine microbiology laboratories

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/pdf/european-harmonization-of-mic-breakpoints-for-antimicrobial-58xps52fqc.pdf

European harmonization of MIC breakpoints for antimicrobial susceptibility testing of bacteria

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European harmonization of MIC breakpoints for antimicrobial susceptibility testing of bacteria