Studying sulfur dioxide effects with better DNA technology suggests we may not need much of it

Fast: In a new study using better-than-ever microbiology, 25 mg/L SO2 added after pressing was enough to “stabilize” yeast and bacterial growth during fermentation, and higher concentrations actually seemed to slow fermentation. Inoculating the must with commercial S. cerevisiae had a very similar effect, even without adding SO2, which looks really, really good for no-added-sulfur wines. BUT (and this is a big but) the study only included one wine (a California chardonnay) made in one way, in smallish (19 L) lab volumes. Goodness only knows if their results will generalize, but let’s hope this encourages someone to look.

More: Sulfur dioxide is the single most commonly used winemaking chemical worldwide. That familiarity probably has something to do with our not understanding it better: we know it’s safe, we know how to use it, and so we don’t have much reason to study it.

In all fairness we do understand SO2 well, but microbiology keeps changing. The publishing dynamo* of Nicholas Bokulich and David Mills – responsible for really excellent recent research on how microbes are spread around a working winery over space and time – plus UC Davis wine microbiologist Linda Bisson (and another Davis student and a Japanese collaborator have published a new American Journal of Enology and Viticulture article on how SO2 affects bacteria and yeast populations in fermenting wine.

The question isn’t new, but the technology they’re using is. Short story: better DNA detection techniques let them pick up on the presence of a bigger range of both bacteria and yeast than previous strategies.

Longer story: Microbes in wine (and elsewhere, for that matter) can be “viable but nonculturable” (VBNC), a new idea ten years ago when microbiologists could still think that agar and Petri dishes were a reasonable way of identifying bugs in a sample. Until better DNA technology made clear a serious issue: yeast and bacteria might be stressed out enough by environments (like wine) to not grow on command but still be alive and able to multiply and cause problems, aka VBNC. (The unculturables who won’t grow in dishes at all are trouble, too.) The details of the high-throughput DNA sequencing they used to ensure that VBNC bugs weren’t left out of their survey aren’t important except to note that it lets them detect more microbes than previous studies.

The other great = new element of this study is its looking at multiple SO2 concentrations, from 150 mg/L down to nil. More work for them; more data for everyone. They also included ferments inoculated with commercial S. cerevisiae and not, which ended up being important.

Their results say that the most important factor in determining what grows in fermenting wine seems to be the degree to which a single strain has the opportunity to take over. One way of encouraging dominance is inoculating with commercial yeast: it more or less takes over and overall microbe diversity declines. But another way is adding SO2, which knocks down some microbes and gives tolerant ones (S. cerevisiae strains included) an opening. Adding SO2 and inoculating S. cerevisiae even without SO2 had similar effects on overall microbial diversity. And, moreover, 25 mg/L was enough SO2 to “stabilize” the ferment. In other words, sulfur-free wines may be less risky than winemakers are generally inclined to believe if they inoculate (which plenty of people inclined not to use sulfur are also inclined to avoid).

The obvious problem: one wine, one vintage, one set of processing techniques, and 19 L volumes. All of these are major reasons to question whether these results will hold for any other set of circumstances. pH and a slew of sulfur-binding compounds affect SO2 efficacy. Fermentation temperature, oxygen, clarification, means of harvesting…the list of processing steps important to microbial diversity is too long to list. And it’s well-known that fermentation volume is important to microbial kinetics.

In short? This article is almost certainly more important to wine microbiologists as a methods paper than to winemakers. (It’s not incidental that the methods section abbreviates the winemaking protocol — “grapes were harvested, crushed, and pressed according to standard winemaking procedures,” whatever that means – and uses nearly a full page of text to describe the DNA sequencing technique.) Nevertheless, it may well serve as impetus for more experimentation with low- and no-sulfur wines, and a good reminder that we always have more to learn about SO2.

Even more: find the full paper, with many more details on which specific yeast and bacteria species were detected and when they peaked (unfortunately behind the AJEV‘s lovable paywall) here. Read the full paper if you can; it contains plenty of potentially idea-generating details that I’ve not even attempted to summarize here.

*Hackneyed maybe, but, seriously, what else do you call them? Bokulich’s CV, as a PhD student, could put to shame plenty of tenured professors. When I’m not just feeling horribly inadequate, I’m wondering where this guy will end up post-graduation. Barring his speaking French and fancying living overseas – or starting a lucrative consulting firm – he’s probably lined up to make tenure at Davis in record time. Heck, he probably already qualifies for tenure.

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