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Agitation can be an important factor for any species of microbe (yeast and bacteria). Gentle stirring on a stir plate or orbital shaker, or frequent gentle manual agitation leads to faster growth and a higher number of organisms. Agitation keeps the microbes in solution. It also maximizes the microbes' access to nutrients and disperses waste evenly. In a non-agitated starter, the microbes are limited to the diffusion rate of nutrients, leading to a slower and more stressful growth <ref name="BryanHeit_starters">[https://www.facebook.com/groups/MilkTheFunk/permalink/1168024059892473/?comment_id=1174865305875015&reply_comment_id=1176092372418975&total_comments=1&comment_tracking=%7B%22tn%22%3A%22R9%22%7D Conversation with Bryan of Sui Generis Blog about starters and agitation. 11/09/2015.]</ref>. Allowing for at least some oxygen to enter the vessel will help ''Brettanomyces'' grow, and limiting oxygen will help any surviving ''Saccharomyces'' to grow. For more information regarding aeration and agitation effects on ''Brettanomyces'' growth, see [[Brettanomyces_Propagation_Experiment|Mark Trent's Brettanomyces Propagation Experiment]] and [[Mixed_Cultures#Effects_of_Mixed_Cultures_on_Growth|Effects of Mixed Cultures on Growth]].
The starter should be stored at room temperature for at least 5-7 days and sometimes longer if growth does not occur within that time. It can be stored at a higher temperature if bacteria is the main microbe that the brewer is after, although avoid going above 86°F/30°C. Monitor the starter for activity during this time to ensure that there are viable microbes. Visual activity may not always be present, may be brief, or minimal. Visual indicators of successful growth include turbidity and continuously produced bubbles on the surface of the liquid. Monitoring the pH and the gravity can assist the brewer with deciding whether or not the commercial sour beer has any viable microorganisms still alive in it. If mold grows in the starter, it should be discarded and not used (see [[Mold]]). If the starter krausens within 2-3 days then it might have viable ''Saccharomyces'', otherwise strains of ''Saccharomyces'' might be dead (especially if the beer was not freshly bottled and was a low pH sour beer; see [[Commercial_Beer_Dregs_Inoculation#The_Dreaded_Killer_Champagne.2FWine_Yeast|"Dreaded Killer Champagne/Wine Yeast" below]]). ''Brettanomyces'' may not show signs of fermentation for up to 3-7 days (in some cases and depending on the viability of the microbes, signs of growth may not appear until up to 2 weeks). Once the starter starts showing signs of fermentation, it should be aged for another 3-7 days to finish fermentation. After the fermentation starts to slow, create a another 1 liter starter of DME wort, and add the original 100 mL dregs starter to it. Repeat the process above, giving the 1 liter starter another 5-7 days of growth <ref name="preiss"></ref>. Pitch the 1 liter starter into a ~20 liter batch, or continue to step up the starter for larger batch sizes. Avoid storing the starter for extended amounts of time because a low pH could result in lower viability over time in an acidic environment; see [[Mixed_Fermentation#Storing_a_Yeast_Cake_or_Sample|Storing a Yeast Cake]] and [[Mixed_Cultures#Starters_and_Other_Manufacturer_Tips|Mixed Culture Starters]] for more information.
===Supplementing another fermentation===