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Drigalski agar

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Drigalski agar is a selective and differential culture medium used for the isolation and identification of Gram-negative bacteria, particularly members of the Enterobacteriaceae family. It contains bile salts and crystal violet, which inhibit the growth of Gram-positive bacteria, while allowing the growth of Gram-negative bacteria. The medium also contains lactose, which allows for the differentiation of lactose-fermenting and non-lactose-fermenting bacteria based on colony color.

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Lab products found in correlation

Chocolate agar, by bioMérieux (2 mentions) Chromid esbl agar, by bioMérieux (2 mentions) Bact alert 3d system, by bioMérieux (1 mentions) Columbia agar, by bioMérieux (1 mentions) Mh agar, by Bio-Rad (1 mentions) Blood agar, by bioMérieux (1 mentions)

6 protocols using drigalski agar

1

Monitoring Neonatal Bacterial Colonization

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Blood cultures were processed for the diagnosis of bacteremia with automated microbial detection systems BacT/Alert 3D system (bioMérieux SA, Marcy l’Etoile, France). To determine the evolution of HAIs in neonatal patients and colonized babies, all newborns admitted were routinely screened for bacterial colonization and received a nasopharynx and rectum examination on their arrival in the unit and on a weekly basis following the admission. Rectal and cavum swabs collected from patients and surfaces were inoculated on Drigalski agar (bioMérieux SA, Marcy l’Etoile, France). All inoculated samples were incubated at 36°C for 48 h. The isolates recovered were routinely identified using MALDI-TOF MS.
Hernandez-Alonso E., Bourgeois-Nicolaos N., Lepainteur M., Derouin V., Barreault S., Waalkes A., Augusto L.A., Gera S., Gleizes O., Tissieres P., Salipante S.J., de Luca D, & Doucet-Populaire F. (2022). Contaminated Incubators: Source of a Multispecies Enterobacter Outbreak of Neonatal Sepsis. Microbiology Spectrum, 10(4), e00964-22.
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2

Fecal Sampling and Enterobacterales Enumeration

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During the CEREMI trial, faecal samples from all volunteers (Fig. 1) were stored at 4 °C after emission and transmitted to the bacteriology laboratory after blinding. One hundred milligrams of faeces were suspended in 1 mL of the brain–heart infusion broth containing 30% glycerol and stored at – 80 °C. Enterobacterales were counted by plating serial dilutions of broth on Drigalski agar (bioMérieux, Marcy-l’Etoile, France).
d’Humières C., Delavy M., Alla L., Ichou F., Gauliard E., Ghozlane A., Levenez F., Galleron N., Quinquis B., Pons N., Mullaert J., Bridier-Nahmias A., Condamine B., Touchon M., Rainteau D., Lamazière A., Lesnik P., Ponnaiah M., Lhomme M., Sertour N., Devente S., Docquier J.D., Bougnoux M.E., Tenaillon O., Magnan M., Ruppé E., Grall N., Duval X., Ehrlich D., Mentré F., Denamur E., Rocha E.P., Le Chatelier E, & Burdet C. (2024). Perturbation and resilience of the gut microbiome up to 3 months after β-lactams exposure in healthy volunteers suggest an important role of microbial β-lactamases. Microbiome, 12, 50.
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3

Fungal Spike in Patient Sputa Analysis

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Six sputa from patients (P1, P2, P3, P4, P5, and P6) hospitalized in Henri Mondor University Hospital (Créteil, France) were randomly selected from sputa addressed to the microbiological laboratory after assessment of their quality according to Murray and Washington criteria [23 (link)]. As part of routine bacterial analysis, they were inoculated onto Chocolate agar + PolyViteX, Columbia agar + 5% horse blood, Columbia ANC agar + 5% sheep blood, Trypicase Soy Agar and Drigalski agar (bioMérieux, Marcy L’Etoile, France) for 48h to 5 days at 35°C. After inclusion in our study, two sputa (P1 and P2) were spiked with 105 conidia of Aspergillus section Fumigati and 105 conidia of Aspergillus section Nigri. Ten microliters of the two spiked-sputa (P1 and P2) were inoculated onto two Sabouraud-Chloramphenicol-Gentamicin slants (Bio-rad, Marnes-la-Coquette, France) and onto one BBLTM CHROMagarTMCandida plate (Beckton Dickinson, Le Pont de Claix, France). Sabouraud slants were incubated for 21 days at 30°C or 35°C and CHROMagarTMCandida plate for 72h at 35°C. All sputa were divided into aliquots of 250 mg each and stored at -20°C for DNA extraction (Fig 1). This work was part of MucoBacMyco project, which was approved by an ad hoc Ethics Committee named Comité de Protection des Personnes d’Ile de France IX (N° CPP-IDF IX-12-011).
Angebault C., Payen M., Woerther P.L., Rodriguez C, & Botterel F. (2020). Combined bacterial and fungal targeted amplicon sequencing of respiratory samples: Does the DNA extraction method matter?. PLoS ONE, 15(4), e0232215.
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4

Quantifying Gram-Negative Enteric Bacilli

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Total counts of Gram‐negative enteric bacilli (GNB) were determined on all samples by plating serial dilutions of fresh stools on Drigalski agar (bioMérieux, Marcy l'Etoile, France). Third generation resistant (3GC) GNB were detected and counted on ChromID ESBL agar (bioMérieux) and biplate ESBL agar (AES Chemunex; 37°C for 48 h). Full details are available in the previous results.18 For each sample, 3GC‐resistant GNB counts were designed as the smallest value obtained from these two agar media.
Guk J., Bridier‐Nahmias A., Magnan M., Grall N., Duval X., Clermont O., Ruppé E., d'Humières C., Tenaillon O., Denamur E., Mentré F., Guedj J, & Burdet C. (2022). Modeling the bacterial dynamics in the gut microbiota following an antibiotic‐induced perturbation. CPT: Pharmacometrics & Systems Pharmacology, 11(7), 906-918.
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5

Evaluation of Culture Media for Isolates

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Eleven representative isolates were grown on 5 commonly used culture media: Mueller-Hinton (MH) agar (Biorad, Marnes la Coquette, France), URISelect 4 (Uri-4; Biorad, Marnes la Coquette, France), Columbia agar plus 5% horse blood (COH; bioMérieux, Marcy l’Etoile, France), ChromID ESBL agar (bioMérieux, Marcy l’Etoile, France) and Drigalski agar (bioMérieux, Marcy l’Etoile, France).
Bernabeu S., Ratnam K.C., Boutal H., Gonzalez C., Vogel A., Devilliers K., Plaisance M., Oueslati S., Malhotra-Kumar S., Dortet L., Fortineau N., Simon S., Volland H, & Naas T. (2020). A Lateral Flow Immunoassay for the Rapid Identification of CTX-M-Producing Enterobacterales from Culture Plates and Positive Blood Cultures. Diagnostics, 10(10), 764.
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6

Bacterial Analysis of Tracheal Aspirates

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Tracheal aspirates (TA) were obtained from the mid-part of the trachea with sterile catheters. In the laboratory, serial dilution of liquefied TA in sterile saline solution (1/1000 and 1/100000) were cultured on the following media: blood agar (aerobic and anaerobic), chocolate agar in 5% CO2 and Drigalski agar to select Gram-negative bacteria (Biomérieux, Marcy-l'Etoile, France) [15] . Bacterial identification was performed by Maldi-TOF mass spectrometry (Microflex and biotyper Bruker Daltonique ® , Wissembourg, France). Bacteria were quantified and antibiotics susceptibility assayed only for potentially pathogenic bacteria: non-fermenting Gram-negative bacilli, Enterobacteriacea, Staphylococcus aureus, Haemophilus sp., Moraxella sp. and Streptococcus pneumoniae. Susceptibilities were determined by the disc diffusion method and interpreted according to the 2015 EUCAST/ CASFM recommendations (http://www.sfm-microbiologie.org). For each patient, overall susceptibility for all the bacteria isolated from each TA was determined according to natural and acquired resistances, identifying antibiotics that would have been effective if the patient required to be treated.
Lepainteur M., Ogna A., Clair B., Dinh A., Tarragon C., Prigent H., Davido B., Barbot F., Vaugier I., Afif M., Roux A.L., Rottman M., Orlikowski D., Herrmann J.L., Annane D, & Lawrence C. (2019). Risk factors for respiratory tract bacterial colonization in adults with neuromuscular or neurological disorders and chronic tracheostomy. Respiratory medicine, 152.
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