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Caenorhabditis elegans is a model system widely used in fundamental research. Even though, nematodes are easy to maintain in the laboratory, obtaining large populations of worms require a lot of work and is time consuming. Furthermore, because C. elegans are mainly hermaphrodite it is even more complicated to obtain large amounts of males which make high-throughput experiments using C. elegans males very challenging.In order to overcome these limitations, we developed affordable and rapid methods to:(1) grow large synchronous worm populations(2) easily obtain large amounts of malesWe developed a culture method on plates to grow big synchronized worm populations with the standard incubators used on all worm labs. We also established an easy filtration method allowing to obtain large male populations in an hour. After filtering, the worm population contains more than 90% of adult males and no adult hermaphrodites since all the contaminants are larva and embryos.The culture and the filtering methods we developed are easy to implement and require a very limited investment in equipment and consumables beside the standard one present in worm labs. In addition, this filtering method could be applied to nematode's species similar in size to C. elegans.

Culture-based microdilution and disk diffusion tests are two commonly used reference methods for determining the susceptibility of causative bacteria to antibiotics. However, these methods are slow and laborious. Automated antimicrobial susceptibility test (AST) instruments are extensively used in clinical microbiology labs, replacing manual methods to perform gold standard microdilution or disk diffusion methods. These automated instruments require the use of isolated bacteria grown in pure culture against a fixed antimicrobial panel, and the susceptibility tests are based on measuring bacterial growth or turbidity changes over a range of pre-determined antimicrobial conditions. As a result, these automated technologies remain inherently inflexible to frequent adjustment of minimum inhibitory concentrations published by the Clinical and Laboratory Standards Institute and are limited by the detection methods that consumables were designed for. Here, we present a stripwell that is compatible with the 96-well format of most lab automation systems to provide a streamlined workflow to inoculate microorganisms for a customized or routine AST. The main goal of this method of stripwell preparation with various antibiotic conditions is to enable the utility of lab automation for phenotypic antibiotic response assays to address the reproducibility issues due to manual operation.• A standardized and scalable solution from inoculation to antimicrobial incubation• Microplates in stripwell format offer the advantage of greater flexibility in clinical microbiology and diagnostics• Customized antimicrobials and dilution ranges tailored to unique specifications for research and development

The ability to assess and eliminate the matrix effect in bioanalytical methods is critical for reproducibility, but sample preparation procedures necessary to address the matrix effect for microbiological methods could be significantly different if viable pathogens are required for downstream microbiological response analysis. A pure bacterial culture remains essential for virulence, antibiotic susceptibility, and phenotypic response studies in order to facilitate the understanding and treatment of caused diseases. Bacterial culture involves the collection, inoculation, incubation, growth, and detection of viable organisms while avoiding contamination throughout the entire process. The goal of this method is to concentrate viable pathogens directly from clinical specimens such as whole blood and urine while removing most interfering matrix components through pelleting in an enriched media, which is designed to facilitate the growth of clinically relevant microorganisms. Nonselective culture media with no inhibitors is used to permit the growth of most of the microorganisms present in the clinical samples studied. Most of the species implicated in clinical infections are mesophilic bacterial species, so the pelleting procedure is conducted at medium temperatures of 37°C to facilitate optimal growth. • Viable bacterial pelleting for phenotypic response analysis. • Concentration of bacteria by centrifugation and matrix component removal for direct-from-specimen molecular analysis. • Viable pathogen recovery directly from whole blood and urine.

The emergence and rapid spread of resistant bacteria has become a serious public health concern worldwide. Delayed antimicrobial therapy significantly increases mortality in high-risk infections with a particularly strong association with septic shock. Therefore, antimicrobial agents are often injudiciously used without any evidence-based microbiological confirmation. Antimicrobial consumption is strongly linked to the emergence and dissemination of antimicrobial-resistant bacteria strains in several epidemiological studies. According to CDC's recent publication, an estimated 30% of outpatient oral antimicrobial prescriptions may have been inappropriate. A compact and rapid pathogen identification (ID) and antimicrobial susceptibility testing (AST) can assist to address both the unnecessary use and overuse of antimicrobials, and therefore effectively reduce antimicrobial resistance. The overall goal of these AST protocols is to deliver a molecular diagnostic platform that is capable of profiling the antimicrobial susceptibility of causative pathogens in hours, not days. The presented AST utilizes an electrochemical sensor to quantify the microbial changes of 16S rRNA after exposure to various antimicrobial conditions either from clinical isolates or directly from unprocessed clinical specimens such as urine and blood. These protocols can be performed by our robotic lab automation systems or manual benchtop assays with associated reagent kits, AST stripwells and sensor chips. • A rapid, quantifiable antimicrobial efficacy profiling comparable to traditional AST reporting. • Customized antimicrobials and dilution ranges tailored to unique specifications for research and development. • Direct antimicrobial susceptibility of viable pathogen from whole blood, urine, or subculture.