Table 1.

Target organism, media, inoculation technique, and incubation used to identify specific microbial populations in spirulina samples.

Target Organisms	Plating Medium	Inoculation Technique	Incubation
Mesophilic microorganisms & Bacillus spores	Plate count agar (PCA) 1	Surface spread, quarter plate, 20 μL	30 °C, 72 h
Clostridium & Clostridium spores	Reinforced Clostridial Agar (RCA) 1	Surface spread, quarter plate, 20 μL	30 °C, 72 h
Lactic acid bacteria	De Man, Rogosa and Sharpe (MRSA)1	Surface spread, quarter plate, 20 μL	30 °C, 72 h, candle jars
Thermotolerant enterococci & streptococci	KF streptococcus medium (KF) 2	Surface spread, quarter plate, 20 μL	37 °C, 4 h then 44 °C, 44 h.
Staphylococcus aureus	Mannitol Salt Agar (MSA) 1	Surface spread, quarter plate, 20 μL	37 °C, 48 h
Yeasts and moulds	Yeast glucose chloramphenicol agar (YGC) 3	Surface spread, 100 μL	25 °C, 3–5 days
Listeria spp.	Oxford agar 1	Surface spread, 500 μL	30 °C, 48 h
Coliforms	Violet red bile agar (VRBA) 1	Pour plate, 1 mL, with overlay	37 °C, 48 h
Pseudomonas spp.	Cetrimide Agar medium 4 + 10 mL glycerol 3	Surface spread, quarter plate, 20 μL	37 °C, 48 h
Vibrio spp.	-Alkaline Peptone water 5 (Enrichment broth) -Thiosulfate Citrate Bile Salts Sucrose Agar 3 (TCBS)	10 g of sample in 90 mL   Plates were streaked with isolation technique	37 °C, 24 h   37 °C, 24 h
Salmonella spp.	-Skimmed Milk Powder 5 (Pre-enrichment broth) -Tetrathionate Broth & Selenite broth 5 (Enrichment broths) -Xylose Lysine Desoxycholate Agar (XLD) & Salmonella-Shigella Agar 5 (SS) (Isolation on selective agars)	10 g of sample in 90 mL pre-enrichment broth   1 mL of pre-enrichment broth in 10 mL enrichment broths Plates were streaked with isolation technique	37 °C, 24 h   37 °C, 24 h   37 °C, 24 h

¹ Lab M, Heywood, UK. ² Panreac Quimica SA, Barcelona, Spain. ³ Merck, Darmstadt, Germany ⁴ Scharlau Chemie SA, Barcelona, Spain ⁵ Oxoid, Basingstoke, UK.