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====Killer Wine Yeast====
Many wine yeast strains are known to be "killer" yeast strains that produce toxins that kill nearby yeast cells in order to give the killer yeast strains a competitive edge over sensitive strains (the term for these toxins has been changed to "zymocides" in science <ref name="Stewart_2018">[https://link.springer.com/chapter/10.1007/978-3-319-69126-8_10 "Killer (Zymocidal) Yeasts." Brewing and Distilling Yeasts. Graham G. Stewart. 2018.]</ref>, but is also sometimes called "mycocins" or "zymocins" <ref name="Boynton_2019">[https://onlinelibrary.wiley.com/doi/pdf/10.1002/yea.3398 The ecology of killer yeasts: interference competition in natural habitats. Primrose J. Boynton. 2019. DOI: https://doi.org/10.1002/yea.3398.]</ref>). Unwanted killer strains of ''Saccharomyces'' have been suspected to cause stuck wine fermentations by killing off the sensitive wine strain that was pitched into the wine by the winemaker <ref name="Bajaj_2017">[https://link.springer.com/chapter/10.1007/978-981-10-2621-8_7 Biology of Killer Yeast and Technological Implications. Bijender Kumar Bajaj, Satbir Singh. 2017.]</ref>. In ''Saccharomyces'', killer strains are genetically determined to secrete toxins (in the form of extracellular proteins or glycoproteins) that kill sensitive strains (there is no evidence that these toxins affect humans). These killer strains are immune to their own toxin. The mycocin toxins can act on sensitive strains in a number of ways: by blocking DNA synthesis and preventing cell division, inhibiting the synthesis of beta-glucans (β-1,6-glucan) that a part of their cell wall formation, and by causing ions to leak through the cell wall. In low dosages, which is typical in the natural environment, toxin triggers active cell death ([http://www.biology-pages.info/A/Apoptosis.html apoptosis]), while large dosages cause necrotic cell killing ([https://en.wikipedia.org/wiki/Necrosis necrosis]). One study in wine found that the use of killer strains to outcompete sensitive strains resulted in off-flavors from yeast autolysis <ref>[http://www.nature.com/nrmicro/journal/v4/n3/full/nrmicro1347.html Yeast viral killer toxins: lethality and self-protection. Manfred J. Schmitt & Frank Breinig. 2006.]</ref><ref name="Hatoum2012">[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3525881/ Rima Hatoum, Steve Labrie, and Ismail Fliss. 2012.]</ref>. Another study found that a lager strain that was genetically modified to secrete killer toxins was able to eliminate all cells of a sensitive ale strain within 24 hours of beer fermentation at a pitching rate of 99% sensitive ale strain to 1% killer lager strain, indicating that even a small amount of killer cells is can be enough to kill a larger population of sensitive cells <ref name="Stewart_2018" />. Ratios of killer cells to sensitive cells that have shown to completely eliminate or nearly eliminate the sensitive population on lab media includes 1:1, 1;10, and 1:100. In a starter of a sensitive strain that had 10% cells of a killer strain introduced, the viability of the sensitive strain was greatly reduced <ref name="Bajaj_2017" />. Neutral strains do not produce toxins, nor are they killed by them <ref>[https://books.google.com/books?hl=en&lr=&id=mvORN6OXHh4C&oi=fnd&pg=PA93&dq=Bussey,+H.+1981.+Physiology+of+killer+factor+in+yeast.+Adv.+Microb.+Physiol.+22:93-121&ots=jUY4T9NpgB&sig=aw-Y1um0KsDnGe6rRe5PTWIDYdI#v=onepage&q&f=false Advances in Microbial Physiology, Volume 22. Academic Press, Sep 15, 1981. Pg 94-95.]</ref>. Almost all domesticated ale and lager strains are sensitive to the toxins produced by killer strains <ref>[https://www.facebook.com/groups/MilkTheFunk/permalink/1179271825434363/?comment_id=1179424538752425&offset=0&total_comments=5&comment_tracking=%7B%22tn%22%3A%22R%22%7D Conversation with Bryan of Sui Generis Blog on MTF on Killer Factor for Saccharomyces. 11/16/2015.]</ref><ref>[http://onlinelibrary.wiley.com/doi/10.1002/j.2050-0416.1973.tb03515.x/pdf Strains of Yeast Lethal to Brewery Yeasts. A.P. Maule and P.D. Thomas. 1972.]</ref>.
In ''Saccharomyces cerevisiae'', four toxins have been identified: K1, K2, K28, and Klus, the first three of which can only kill other strains/species of ''Saccharomyces''. The Klus toxin has been found to kill all strains of ''S. cerevisiae'' (including those that produce the previous three toxins), as well as yeast from other genera, such as ''Hanseniaspora spp.'', ''Kluyveromyces lactis'', ''Candida albicans'', ''Candida dubliniensis'', ''Candida kefir'' and ''Candida tropicalis''. Rodriguez et al. (2011) reported that out of 1,114 strains of ''S. cerevisiae'' isolated from spontaneous wine fermentations, 38% of them were killer positive with most producing K2. Only 7% of produced the Klus toxin (no commercial wine yeast strains have been reported to produce the Klus toxin that we know of) <ref name="Rodriguez">[http://aem.asm.org/content/77/5/1822.long A New Wine Saccharomyces cerevisiae Killer Toxin (Klus), Encoded by a Double-Stranded RNA Virus, with Broad Antifungal Activity Is Evolutionarily Related to a Chromosomal Host Gene. Nieves Rodríguez-Cousiño, Matilde Maqueda, Jesús Ambrona, Emiliano Zamora, Rosa Esteban and Manuel Ramírez. 2011]</ref>. The K1 toxin is most active between a pH of 4.6 and 4.8, while K2 and Klus are active around a pH of 4.0 to 4.3 <ref name="Rodriguez"></ref>. The activity of the toxin is greatest during the log phase of growth, and decays during the stationary phase of fermentation <ref name="Buyuksirit"></ref>. Generally, none of the toxins secreted by killer strains of ''Saccharomyces'' have been found to kill ''Brettanomyces'' <ref>[http://www.scielo.org.za/scielo.php?pid=S2224-79042015000100010&script=sci_arttext&tlng=pt Non-Saccharomyces killer toxins: Possible biocontrol agents against Brettanomyces in wine? S. Afr. J. Enol. Vitic. vol.36 n.1 Stellenbosch. 2015.]</ref>. One study from India reported that a wild ''S. cerevisiae'' strain caught from flowers killed another wild caught strain of ''Brettanoyces anomulus'', however, their methodology was not explicit and potentially not scientifically rigorous enough <ref>[http://nopr.niscair.res.in/handle/123456789/7735 Production and effect of killer toxin by Saccharomyces cerevisiae and Pichia kluyveri on sensitive yeasts and fungal pathogens. Dabhole, Madhusudan P, Joishy, K N. 2005.]</ref>. For example, this study did not use DNA fingerprinting to identify the wild yeast strains used in the study and instead relied on morphology and media selection, and they did not identify the type of toxin produced by the killer strain of wild ''S. cerevisiae''. They also reported that the ''B. anamulus'' strain did not ferment glucose, which is not typical for this species and indicates that it might have been misidentified.