KBI OSU DNA Sequence Study Analysis Report

By Dr. Chris Curtin & Keisha-Rose Harrison of Oregon State University & Hannah Crum of Kombucha Brewers International & Kombucha Kamp Thank you to everyone who participated in the first round of the KBI & Oregon State University Kombucha SCOBY DNA Sequencing study. With your help we sequenced nearly 100 samples provided by over 70 […]

Tuesday, 6th March 2018

By Dr. Chris Curtin & Keisha-Rose Harrison of Oregon State University & Hannah Crum of Kombucha Brewers International & Kombucha Kamp

Thank you to everyone who participated in the first round of the KBI & Oregon State University Kombucha SCOBY DNA Sequencing study. With your help we sequenced nearly 100 samples provided by over 70 different companies from 26 states and 9 different countries around the world. The largest SCOBY DNA Sequencing Study to date! Previous studies that sought to improve our knowledge of SCOBY microbial populations have been more limited in scope. We now have more data points which can provide a clearer understanding of which microbes are common across commercial SCOBYs, and an opportunity to learn which strains may be responsible for different fermentation and flavour outcomes.

DNA Sequencing Analysis Process

For each sample received, we blended a standardized amount of SCOBY under controlled conditions, and then extracted the DNA from all microbes present in the sample. Regions of DNA that can be interpreted as ‘barcodes’ were amplified from each sample and sequenced using Illumina Miseq technology. The sequences of these ‘barcodes’ were compared to large databases of fungi and bacteria, and assigned to Operational Taxonomic Units (OTUs). In this study OTUs were defined at a standard cutoff of 97% similarity. In other words, if two ‘barcodes’ are only 96% similar in DNA sequence they would not be grouped into the same OTU.

Is an OTU representative of a species? Sometimes yes, but often no – within many genera (particularly for bacteria) individual species cannot be reliably differentiated using current DNA ‘barcodes’. This approach is highly robust when used to provide resolution at the genus level, meaning that all OTUs for one type of organisms are grouped together. For example, if a SCOBY contained both Lactobacillus casei and Lactobacillus plantarum their DNA ‘barcodes’ would be grouped together as Lactobacillus.

After the samples are matched at the genus level, each ‘barcode’ detected is counted to provide the frequency with which it appears in the sample to determine the proportion at which each genus of the whole bacterial or fungal population exists. These are the numbers of the sample represented as as follows: Dekkera 0.67 means that 67% of the fungal population in your sample belongs to the Dekkera genus.

Interpreting Your Results

Each sequencing report provided participants the bacteria and fungi profile of the sampled SCOBY. Below is an example of a report of bacteria detected. The genus level is read as “g_GENUS”. The number provided indicates how much of bacteria was present in the sample. For instance, 94.8% of this example sample is Gluconoacetobacter. Some other OTU groupings could not be reliably defined at genus level, in which case they will be named at family level but assigned as ‘Other’ or ‘g__’ at genus level. In this example, 2.7% of the bacterial population is made up of bacteria within family Acetobacteraceae that could not be assigned to a genus.

Using the values provided we can create a visual of the bacteria composition.

Please note that the taxonomic name of organisms can change. For example, Dekkera is now known only as Brettanomyces (previously the names were interchangeable), while Gluconoacteobacter xylinum and Acetobacter xylinum have been reassigned to Komagataeibacter xylinum. We’re sorry, but this is just as confusing for seasoned microbiologists!

Q: “My report says “unknown” for some of the organisms. What does that mean?”
A: Sometimes ‘barcodes’ are not able to be definitely assigned at the genus level due to a lack of resolution or because the sample may contain a species that has not yet been identified by the microbiology community. These OTUs are then grouped at the family level and genus may be given as ‘undefined’, ‘unknown’ or ‘other’.

How do your results compare to the “average” SCOBY

Each company received an individual report listing the respective percentages of each type of bacteria and yeast found in their sample.

In the aggregate, it was found that the organisms occurred in these relative percentages:

TABLE OF ORGANISMS FROM GREATEST TO LEAST FOR YEAST & BACTERIA

Making Sense of the Study

DOMINANT YEAST AND BACTERIA

As presented at KombuchaKon18, results from the study confirm that there is variability in SCOBY samples collected from different brewers. However we were able to identify core microbes that are present in most SCOBYs. On average, the most common fungi were Brettanomyces/Dekkera and Starmerella, while the most common bacteria were Gluconacetobacter, Gluconobacter, and Lactobacillus.

The near constant fluctuation of organism name changes presents its own kind of challenge. In previous SCOBY & Kombucha DNA Sequencing studies, for example, there are no instances of Starmerella being detected, however, when doing cursory research, it turns out this is a newer nomenclature for many species that used to be known as different types of Candida spp. which have been identified in other studies. However, without going to the species level, it is difficult to ascertain which stains correspond to those found in previous studies versus which may be novel to Kombucha.

HOW DOES THIS INFLUENCE FLAVOR AND FERMENTATION?

Identifying the prevalent organisms within the SCOBY is the first step towards answering this question. We do know that different types of yeast convert sugar to alcohol at different rates. It is also known that different types of bacteria produce varying kinds of organic acids in similar fermentation models, i.e. beer, wine (see table below). There is likely some overlap in how these microbes influence kombucha.

For the next study, we aim to determine how different microbial profiles influence your finished raw product. Continued participation in these studies will go a long way to addressing these questions.

In the meantime, some of the organisms found in Kombucha are also found in wine & beer. Here are some links to charts that outline the flavor profile and characteristics of some of those organisms.

UC Davis Wine Server Database

Our yeast and bacteria are considered spoilage for beer & wine!

ARE SCOBYs MORE VARIABLE IN THEIR BACTERIA OR YEAST POPULATIONS?

On the whole, there seems to be more variability in the bacteria profiles of SCOBY samples. Bacteria are responsible for converting most of the ethanol into organic acids and for building the SCOBY. We will continue to study how different types of bacteria influence the secondary fermentation.

VARIATION BETWEEN SCOBY AND STARTER FLUID MICROBIAL POPULATIONS

For this study we only examined the SCOBY and not the starter fluid. For the second sequencing study we will look at both! As mentioned in the presentation at Kombuchakon, acetic acid bacteria build the the SCOBY. They do not have to live in the SCOBY to produce acids (e.g. vinegar is made without a SCOBY), just as yeast do not have to live in the SCOBY to degrade sugars. The SCOBY does, however, enable repeated fermentations to be started with multiple species present, something that would be difficult to achieve otherwise. The SCOBY effectively buffers microbes against what would otherwise be a fluctuating environment as the sweet tea goes through alcoholic fermentation and acetification. .

METADATA ANALYSIS

At the time of sample collection, a couple of questions were asked in order to analyze the data according to a couple of variables – namely age of the culture sampled and location. We then sent out a follow up survey for additional metadata including age of the culture sampled, type & quantity of tea; type & quantity of sugar; and batch size to see if the data would show any trends or patterns based on these variables. Not all participants answered this part of the survey and we hope to have a more thorough Metadata Analysis available in future studies. This will greatly enhance the applicability of study results to the KBI community.

REGIONAL VARIATION

Preliminary analyses suggest there may be some region-to-region variances in the bacteria and fungal composition of the SCOBY, though it should be noted that sample numbers and the amount of metadata provided were uneven. Future studies will request additional metadata in order to determine the influence of types & quantities of tea, sugar and other variables.

NEXT STEPS

KBI & OSU will be partnering on a new DNA sequencing study and will also be conducting an analyte study. We hope to receive at least as many samples as last time if not more so that our knowledge base will continue to deepen. Stay tuned for more details coming soon!

KBI Official Guidance – Added Sugars & Nutrition Panel Changes

Based on the rules put forth by the FDA effective July 26, 2016, commercial Kombucha producers in the United States are required to indicate the sugar they add to the fermentation process as “Added Sugars.” However, the amount required to put on the label is AFTER fermentation has occurred. To be in compliance with this […]

Thursday, 21st December 2017

To be in compliance with this rule, it is advised that Kombucha producers in the United States submit their products to a third party lab for sugar testing.

Clear records and scientific documentation that demonstrate the accurate amount of sugar grams in the final products along with a narrative of why the amount is different than the starting sugar are required to be kept on file.

Any Kombucha producer that adds juice concentrate, sugar or sugar substitutes to their final product would need to also include that information as part of the “Added Sugars” amount on the label.

RECAP:

The amount of sugars present must be declared as both TOTAL and ADDED sugars (21 CFR 101.9(c)(6)(iii)).
Added sugars may not exceed the Total sugars amount listed on the label.
Added sugars = the amount of sugar remaining in the product AFTER fermentation.
Clear documentation and narrative are kept on file and provided upon request.
Any other sugars added in the flavoring stage are also to be included as ADDED SUGAR
- sugars (free, mono- and disaccharides) (this would included fructose, sucrose, and glucose)
- sugars from syrups and honey
- sugars from concentrated fruit or vegetable juices that are in excess of what would be expected from the same volume of 100 percent fruit or vegetable juice of the same type

EXAMPLE:

15g of sugar are measured in the sweet tea substrate before fermentation
The product is tested after fermentation and has reduced to 12g of sugar
The label now reads:
- Total Sugar 12g
- Added Sugar 12g

The new rules put forth by the FDA regarding added sugars are listed here for reference (important details bolded by KBI):

Requiring the declaration of the gram amount of “added sugars” in a serving of a product, establishing a Daily Reference Value (DRV), and requiring the percent Daily Value (DV) declaration for added sugars;

Changing “Sugars” to “Total Sugars” and requiring that “Includes `X’ g Added Sugars” be indented and declared directly below “Total Sugars” on the label;

The rule requires a manufacturer with sugars added before and during the fermentation process to make and keep records of added sugars necessary to determine the amount of added sugars present in the finished food. The rule requires manufacturers of such foods to make and keep records of all relevant scientific data and information relied upon by the manufacturer that demonstrates the amount of added sugars in the food after fermentation and a narrative explaining why the data and information are sufficient to demonstrate the amount of added sugars declared in the finished food, provided the data and information used is specific to the type of fermented food manufactured.

Establishing a compliance date of 2 years after the final rule’s effective date (7/26/16), except that manufacturers with less than $10 million in annual food sales have a compliance date of 3 years after the final rule’s effective date. (For more details, see part III.)

ALL BRANDS WILL NEED TO BE IN COMPLIANCE WITH THIS LABEL CHANGE

January 1, 2020, for Manufacturers with $10 Million in sales and January 1, 2021, for Manufacturers with less than $10 Million in sales.

Example of where Added Sugars to be noted on label:

RECOMMENDED NARRATIVE TO KEEP WITH SCIENTIFIC RECORDS

Kombucha is a traditionally fermented beverage made of tea, sugar and SCOBY. Throughout the fermentation process, the sugar added at the beginning is consumed by the microorganisms in the SCOBY and is reduced over time in two ways. Cane sugar is sucrose, a disaccharide. Initially, the yeast split the sugar molecule into its monosaccharide components, fructose and glucose. Primarily, this results in a lower glycemic impact to the consumer because the complex carbohydrates are broken down into simpler sugars. Secondarily, the amount of sugar remaining is also reduced the longer the fermentation continues. The byproducts of this process are converted into organic acids giving Kombucha is unique flavor and properties.

CHANGES TO THE NUTRITION FACTS LABEL

In an effort to provide additional time to comply with the new Nutrition Facts Panel and Supplement Facts Panel regulations, FDA has announced that, for the first six months following the January 1, 2020 compliance date, FDA plans to work cooperatively with manufacturers to meet the new requirements and will not focus on enforcement actions regarding these requirements. For more information, go to https://www.fda.gov/food/food-labeling-nutrition/industry-resources-changes-nutrition-facts-label#Compliance.

REFERENCES:

https://www.federalregister.gov/articles/2016/05/27/2016-11867/food-labeling-revision-of-the-nutrition-and-supplement-facts-labels

https://www.fda.gov/media/102614/download

https://www.fda.gov/food/food-labeling-nutrition/industry-resources-changes-nutrition-facts-label#AddedSugars

https://www.fda.gov/food/food-labeling-nutrition/changes-nutrition-facts-label

https://www.fda.gov/food/food-labeling-nutrition/industry-resources-changes-nutrition-facts-label#Compliance

Help Your Patients Use the Nutrition Facts Label to Cut Down on Added Sugars

REVISIONS:

Rev.120617 Approved KBI Board December 2017

Updated 022218 to clarify types of sugars that need to be classified as “Added Sugar”

Updated 080719 to list example and clarify Added Sugars may not exceed Total Sugars on label

Update 102919 to add Nutrition Facts Panel and Supplement Facts Panel regulations

KBI Position Paper – Municipal Sugar Tax & Kombucha

Municipalities are eager to find new ways to fund their operations and programs. Lately it has been popular to enact a “sugar tax” on non-alcoholic beverages that exceed whatever threshold the municipality has deemed as acceptable. There is a long history of enacting “sin taxes” on products that are perceived or proven to have a […]

Friday, 27th October 2017

Kombucha is a traditional fermented beverage made from tea and sugar. During the fermentation process, most of the sugars are consumed by the microorganisms present in the SCOBY (Symbiotic Culture of Bacteria and Yeast) and converted into trace amounts of ethanol and organic acids. Each producer has their own fermentation process and method as well as their own flavor profile – some of which may skew sweeter in order to attract a certain type of consumer.

However, despite having some sugar present, the fermentation process materially changes the structure of the sugars – the disaccharide sucrose (table sugar) is broken into its monosaccharide components thus creating a lower glycemic impact. Also, the living organisms and nutrition in a living form (trace amounts of B vitamins, vitamin C, amino acids, etc) contribute a net positive effect to the consumer unlike comparable non-alcoholic beverages – i.e. sodas, energy drinks, juices, etc.

Moreover, many of these statutes include exemptions for products such as diet soda which, while their sugar content may be low, do contain known carcinogens. The unintended consequence is that products that are healthier end up taxed while less healthy options cost less and could influence consumer behavior in a negative way.

While KBI has been active in supporting its members who are dealing with these local statutes, ultimately, the decision whether or not to tax Kombucha falls to the cities themselves. Any taxes required are charged at the discretion of those locales and are not in any way under the purview of the Kombucha producer.

REFERENCES

Jayabalan, Rasu, et al. “A review on kombucha tea—microbiology, composition, fermentation, beneficial effects, toxicity, and tea fungus.” Comprehensive Reviews in Food Science and Food Safety 13.4 (2014): 538-550.

Rev.090617 Adopted by KBI Board Oct 2017

KBI Official Guidance – Cold Storage for Raw, Unpasteurized Kombucha

Kombucha is a traditionally fermented, low alcohol, non-intoxicating beverage that is most commonly consumed raw, meaning unpasteurized, to protect the probiotics and nutrients in a living form from being damaged. Unpasteurized means not subject to pasteurization via heat or chemical means. As a result of remaining a raw product, it can experience slight shifts in […]

Friday, 27th October 2017

Unpasteurized means not subject to pasteurization via heat or chemical means. As a result of remaining a raw product, it can experience slight shifts in ethanol as the fermentation process continues in the bottle.

To arrest that process, raw Kombucha must be kept cold at all times to maintain the integrity of the product and to prevent over fermentation in the bottle.

Cold storage ought to be maintained at 34-40º F /1.1-4.4ºC to slow the fermentation of the Kombucha throughout the supply chain.

Shelf life of Kombucha is determined by how long the product can remain in cold storage before the ethanol level goes above the prescribed legal limit (will vary based on location).

Products that claim to be both shelf-stable and raw are not currently recognized as stable without further evidence vis a vis shelf life testing by a third party lab to verify that the ethanol level remains within the legal limit for that location. No such products currently exist in the US marketplace as all raw Kombucha is subject to refrigeration.

Rev.090517 Approved KBI Board October 2017

SCOBY DNA Sequencing Study

KBI is excited to announce a partnership with Oregon State University to study the organisms present in Kombucha cultures from around the United States. The purpose of the study is to try sequence numerous samples from a wide variety of places to answer the question of “What is a SCOBY?” After sequencing all of the […]

Monday, 18th September 2017

After sequencing all of the samples, the data will be analyzed to see which organisms are common to all cultures, which are not, which are unique to specific locations. This information is the first step to gaining a deeper understanding of how the microorganisms contribute to flavor, alcohol content and more.

HOW TO PARTICIPATE IN THE STUDY

We are aiming for 200+ total samples to be analyzed in order to have a sufficient pool to draw conclusions. We invite you to take advantage of this opportunity to learn what is in your culture while also contributing to the deeper body of knowledge about Kombucha as a whole. Each participant will be provided a detailed report with all of the organisms present in their culture.

If you are a current KBI member, the cost is only $100 per sample. Part of the cost goes directly to the university to cover the sequencing, part of it is to cover shipping of kits and the purchase of the kit supplies and the rest covers administrative costs.

Non-KBI members are also invited to participate!. The cost is $250 per sample for non-members. Or join KBI today to receive the member pricing.

DNA sequencing typically costs several hundred up to thousands of dollars per sample, so this is a significant savings for valuable information. The data will only ever be presented in an aggregate format to protect confidentiality for all participants.

We have a tight timeline (all samples need to be received by ~~October 15th~~ extended to October 30th!), so please sign up today.

We look forward to your participation and hope to see you at KombuchaKon18 – Defining Our Culture where the results will be presented.

Expert Review Panel on Ethanol Testing for Kombucha – AOAC Annual Meeting

Join us at the Expert Review Panel on Ethanol Testing for Kombucha at the AOAC’s 131’st Annual Meeting taking place at the Atlanta Marriott on Sunday September 24th from 1-4pm. The meeting is free for all to attend. KBI President, Hannah Crum, will be on the panel. KBI Secretary, Melanie Wade (Golda Kombucha), along with […]

Wednesday, 9th August 2017

KBI President, Hannah Crum, will be on the panel. KBI Secretary, Melanie Wade (Golda Kombucha), along with Claire Irie of Lupa’s Kitchen and Adina Martinez of Mighty Bucha will also be in attendance and will be bringing Kombucha to share with the panel.

We invite you to join us! Please RSVP by sending an email to admin@kombuchabrewers.org

Hill Climb Success!

(from L to R: Elixir, Funky Fresh, Golda, Humm, Craft, High Country, KBI, Health-Ade, Bootleg Bucha) On Wednesday, March 22nd, KBI members from around the United States converged in Washington DC to educate lawmakers about our growing industry and regulatory […]

Wednesday, 12th April 2017

Image may contain: 13 people, people smiling, people standing

(from L to R: Elixir, Funky Fresh, Golda, Humm, Craft, High Country, KBI, Health-Ade, Bootleg Bucha)

On Wednesday, March 22nd, KBI members from around the United States converged in Washington DC to educate lawmakers about our growing industry and regulatory needs. In total, 15 member brand attended 28 meetings with members of Congress and the Senate. After a long day on the Hill talking about the KOMBUCHA Act, everyone was invited to attend a Kombucha tasting reception on the Hill. We had a fantastic turn out as over 100 different Hill staffers came out to nosh and enjoy some booch (some were seen squirreling bottles away as they left!).

KBI hopes that this buzz will turn into additional bipartisan co-sponsor signatures for the bills in the House and the Senate. Since it deals specifically with updating the tax code, the goal is to include it in any bills that might be up for a vote on the issue of tax reform.

Thank you to those who attended the Hill Climb:

Aqua ViTea Kombucha (VT)

BAO Food & Drink (NJ)

Blue Ridge Bucha (VA)

Bootleg Bucha (NY)

Craft Kombucha (DC)

Elixir Kombucha (KY)

Funky Fresh (PA)

Golda Kombucha (GA)

GT’s Kombucha (CA)

Health-Ade Kombucha (CA)

High Country Kombucha (CO)

Holy Kombucha (TX)

Humm Kombucha (OR)

Ninja Kombucha (VA)

Sole Kombucha (PA)

WANT TO GET INVOLVED? HERE’S HOW:

Send your Congressperson and Senator an email requesting their support.
- Not sure who that is – use this handy online tool to find their info.
Invite your Congressperson/Senator to visit your facility when they are next in-state.
Traveling to DC or your state capitol? Set up a meeting with your Rep or Senator. Email KBI to get a copy of the talking points.
Stay tuned to the KBI newsletter for updates on how else you can be involved!

KBI Presentation to AOAC SPSFAM Panel – Working Group to develop new testing method for ethanol in kombucha

Sunday, 25th October 2015

On Sunday, September 27th, KBI president Hannah Crum and and Heath-Ade CEO Daina Trout, head of LGO committee (Special Projects Team) presented to the Stakeholder Panel on Strategic Food Analytical Methods (SPSFAM) at the annual conference of the Association of Official Analytical Chemists (AOAC) to establish a new Working Group with the aim of developing a new testing standard for the low levels of ethanol in kombucha. Experts across various industries specializing in manufacturing, food chemistry, and laboratories were present as well as representatives from government organizations like the TTB.

erik konings nestle daina trout healthade hannah crum kombucha kamp 9.27.15 — Erik Konings of Nestle presiding over the SPSFAM with presenters Daina Trout of Health-Ade & Hannah Crum of KBI

Hannah and Daina shared with those in attendance the explosive growth in the kombucha industry while enumerating the difficulties with the testing methods in use by regulators in the US today. The message was clear: currently, an accurate, standard method of testing that takes into consideration the complexities of kombucha such as the strands of living culture, yeast bodies, organic acids, low pH and other factors simply does not exist. The Q & A was lively with questions, support and enthusiasm from the audience for this fascinating scientific problem and very much echoed that current methodologies need to be revised.

The stakeholder panel eagerly took up the process of formulating a Fitness for Purpose (FOP) statement. The FOP is ultimately the parameters that the Working Group will establish before sending out a request for methods to the international membership.

Fitness for Purpose statement: Methods need to accurately and precisely measure the ethanol concentrations to comply with alcohol and non-alcohol declarations in Kombucha in-process and finished products.

The Working Group will now proceed under co-chairs Hannah Crum and Sam Labonia of Cornerstone Labs. The goal is to hone in on the issues that will be necessary to take into consideration for determining accurate testing methods. KBI anticipates presenting the Standard Method Performance Requirements (SMPR®) to the SPSFAM Panel at the AOAC Midyear Meeting in March 2016 in Maryland. Since AOAC standards are in use by governments around the world, including the US, the TTB (Tax & Trade Bureau) has also been invited to participate in the process and are members of the Stakeholder Panel that will vote to determine the ultimate methodology that is selected.

Armen Mirzoian Cathy Halverson of TTB Daina Trout Hannah Crum — Armen Mirzoian, Chief Science Officer, and Cathy Halverson, Head of Alcohol Testing Lab, of the TTB attend the AOAC Stakeholder Panel and continue to participate in the Working Group spearheaded by KBI, Health-Ade and other KBI members

The reaction from not only the stakeholders, but the general AOAC attendees was overwhelmingly positive and enthusiastic about the direction of this research. Not only was the presentation very well received, the questions asked after were informed and thought-provoking. Overall, KBI is thrilled to get the opportunity to work with the AOAC to find a method of testing ethanol in kombucha that produces accurate, consistent, repeatable results. “The AOAC is pleased to be working with the kombucha industry, regulatory bodies, and other stakeholders to develop such a standard,” stated E. James Bradford, Ph.D., Executive Director of AOAC International.

The working group meets regularly (biweekly) to refine definitions and identify criteria for the SMPR®

Get the latest updates on the Working Group in the Alcohol – Testing & Control Group on the KBI Member Forum.

Current Testing Methods for Kombucha

Wednesday, 14th October 2015

Kombucha is a complex, living product with an enormous range of styles and flavors that delight the senses and enliven the body. Mixed with juice, spices or brewed with other beverages; it is most often consumed raw but can also be filtered, force carbonated or in some cases pasteurized; and it can be fermented with a variety of substrates yielding new and exotic combinations. This amazing diversity of expression is part of the mystique of kombucha but also presents difficulty when testing for ethanol due to its inherent biodiversity and complexity, as well as the wide spectrum of serving possibilities.

Kombucha companies currently rely on individual labs to test for ethanol. The KBI Special Projects Team (sub-committee of LGO) contacted over a dozen labs with experience working with kombucha and discovered that each lab has its own method for testing for ethanol. Three main methods are being utilized for testing ethanol in kombucha – alcolyzer (refraction), densitometry and gas chromatography. Moreover, each lab will apply their own tweaks and adjustments to the process in order to account for kombucha’s unique characteristics. The inconsistency in methodology is concerning because kombucha has so many quirks that need to be accounted for and numerous decisions in the testing process can have a significant effect on the final result. With so many changing the methods ever so slightly, it is little wonder that consistent results are simply not achievable.

Oftentimes, the sample is simply run through a beer alcolyzer system designed to effortlessly detect the much higher, and therefore easier to measure, levels of ethanol. Kombucha, being far more complex than beer, contains organic acids, fermented sugars, and live culture strands that are simply not present in most commercial beers. Moreover, the unique kombucha culture is a symbiosis and the bacteria consume the ethanol created by the yeast into organic acids that may have a similar density to ethanol thereby creating false positives.

Just a handful of labs are currently using more sophisticated techniques, like headspace gas chromatography to suss out the nuances. This method shows a lot of promise as it opts to use far lower detection limits than is recommended in the AOAC method for testing beer, consistent with the lower levels of ethanol observed in kombucha. Another difference in sample handling is to not centrifuge it to avoid adulterating the analyte.

Still others utilize AOAC 983.13 method for testing wine that relies on centrifuging the kombucha sample to remove the high levels of sediment present in kombucha and then later factors in the removed sediment into the final mass percentage. This method shows similar results to the modified 984.14 method, and shows promise when adjusted for kombucha’s low detection levels. What is abundantly clear, however, is that there is no consensus available for the appropriate method to use when testing kombucha.

In one lab, the same bottle of kombucha was tested using the beer alcolyzer and a modified AOAC 984.14 method for testing beer. The beer alcolyzer system claimed that the kombucha contained levels of ethanol higher than 0.5%, the level above which a beverage must be marketed as alcoholic. When analyzed using the modified AOAC method, a gas chromatography method, the kombucha was tested to be well under 0.5% ethanol. Because the limit for alcohol content in non-alcoholic kombucha is so low at 0.5%, it is imperative that the testing is done accurately and precisely. When an unmarked bottle of Bud Light was tested, both the beer alcolyzer system and the gas chromatography method were highly precise, possibly because its higher alcohol content is easier to detect with higher detection limits.

The lack of consistency is evident even at the same lab. The same samples were sent with different labels to a lab and the test results for the exact same samples had a variance of +/- 1% ABV – that’s more than double the legal limit! So, how do we all get on the same page and find an accurate, repeatable, consistent test for ethanol in kombucha? Collaboration, of course! We are symbiotic after all.

The kombucha industry is reaching out to collaborate with the most educated body of scientists to establish a new testing standard for ethanol in kombucha. By partnering with AOAC, KBI participates in an ethical, scientific process for creating testing methods. Moreover, the TTB, FDA and other government, non-profit, academic and others will work together and decide through consensus on the appropriate method for accurate, consistent and repeatable results. The lack of definitive research in this field has frustrated kombucha brewers who are taking measures to brew compliant products and regulatory agencies. Working with industry experts, KBI hopes to reach a universal truth for testing kombucha to better serve and inform consumers.

HACCP Plan – What is it and why its important

What is a HACCP Plan? A Hazard Analysis and Critical Control Points or HACCP Plan is a systematic approach to the identification, evaluation, and control of food safety hazards (biological, chemical, and physical) throughout the production process to prevent the risk of foodborne illness or other safety concerns. HACCP is based on the following seven […]

Monday, 31st August 2015

What is a HACCP Plan?

A Hazard Analysis and Critical Control Points or HACCP Plan is a systematic approach to the identification, evaluation, and control of food safety hazards (biological, chemical, and physical) throughout the production process to prevent the risk of foodborne illness or other safety concerns. HACCP is based on the following seven key activities:

Conduct a hazard analysis
Determine critical control points (CCP)
Establish critical limits
Establish a monitoring system
Establish a procedure for corrective action for use when monitoring a CCP indicates a deviation from an established critical limit
Establish procedures for verification to confirm the effectiveness of the HACCP Plan
Establish documentation concerning all procedures and records appropriate to these activities and their applications

Which industries require it?

HACCP should be used in conjunction with other food protection programs and is mandatory for meat & poultry (USDA), seafood (FDA), and juice (FDA), and voluntary for all other food and beverage industries. Now that FSMA is being enforced for every size business, KBI highly recommends that all commercial Kombucha brewers have a HACCP Plan in place.

Why does KBI recommend it?

KBI’s newly established Best Practices list having an HACCP Plan in place as an important action item for commercial Kombucha brewers. These Best Practices have been voted into place by KBI’s own member organizations, which represent the majority views of some of the world’s leading Kombucha producers.

Furthermore, products produced by the Kombucha industry are often raw (unpasteurized) and are consumed by people around the world primarily for their health benefits. Therefore, it is in the best interest of public health to take as many precautionary measures as possible to produce healthful, safe beverages which meet established food safety standards. As this rapidly growing industry continues to expand, we feel that it is important to operate our brewing companies with integrity, safety, and quality in mind, while also keeping in mind the extra layer of protection a HACCP Plan can offer the public.

In addition, when local, state, and/or federal inspections occur, an established HACCP Plan highlights the company’s efforts to produce safe products and demonstrates/documents all of the key elements required. HACCP Plans also make it much easier to communicate with inspectors who may not be familiar with Kombucha products and/or the Kombucha industry as a whole.

How do I develop an HACCP Plan?

Developing an HACCP Plan can be very time consuming and rigorous . Fortunately, KBI has developed a hazard analysis template, which is available to brewery members at a discounted price (it is also available to non-members).

Not a KBI member yet?

Whether you’re just starting out or already have an established Kombucha company, we invite you to join KBI to take advantage of our many member’s only resources and benefits, including the HACCP Plan template mentioned above. To learn more about our member benefits, to view our membership requirements or to join KBI, please click here.

Updated: May 2022