CHROMagar™ Vibrio

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-DO NOT HEAT TO MORE THAN 100 °C. DO NOT AUTOCLAVE AT 121 °C. - If using an autoclave, do so without pressure. - 2: In case of samples with a high presence of Aeromonas, 50 mg of cefsulodin can be added to the mix once cooled down at 45-50 °C (50 mg/L). -FINAL MEDIA pH 9.0 +/- 0.2	-Plates can be destroyed by autoclaving at 121 °C for at least 20 minutes.

Protocol tips
-DO NOT HEAT TO MORE THAN 100 °C. DO NOT AUTOCLAVE AT 121 °C. - If using an autoclave, do so without pressure. - 2: In case of samples with a high presence of Aeromonas, 50 mg of cefsulodin can be added to the mix once cooled down at 45-50 °C (50 mg/L). -FINAL MEDIA pH 9.0 +/- 0.2
Downstream tips
-Plates can be destroyed by autoclaving at 121 °C for at least 20 minutes.

Publication protocol

Although developed several decades ago, traditional culturing methods continue to be improved with the result of more reliable detection of V. cholerae from environmental samples. The process involves either directly plating the study sample (water, plankton, sediment, shellfish) onto TCBS, TTGA, or CHROMAgar™ Vibrio or pre-enrichment of the sample in alkaline peptone water followed by streaking for isolation onto one or a combination of these three agars. For environmental samples, it is highly recommended to use pre-enrichment step to improve detection or isolation. Following an incubation period, presumptive V. cholerae can be isolated from these media and confirmed by either biochemical analyses (Huq, Grim et al. 2006) or immediately by PCR. Isolates can then be serogrouped as O1, O139, or non-O1/non-O139 by a slide agglutination assay using antisera for the O1 and O139 antigens or by PCR using primers developed to target O1 and O139 coding regions of the genomic DNA. These V. cholerae isolates can then be archived with replicates in nutrient agar containing 0.5% NaCl overlaid with sterile mineral oil or as a glycerol stock at −70°C. Stock cultures of V. cholerae should be periodically re-streaked for isolation to ensure their viability and purity.

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Manufacturer protocol

Download the product protocol from ChroMagar for CHROMagar™ Vibrio below.

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