This year Probiota Americas took microbiome stakeholders to Miami to learn about new prebiotics and probiotics, about the shift of the market to internet, what next-generation analytical tools will drive the future, and much more. Special focuses described the potential for selected probiotics in sports performance, vaginal health, cardiovascular health and mental health.
1) Prebiotic workshop
Nutrasource Dr. Anna De Boer exposed work on the modes of action behind prebiotics (a prebiotic is a substrate that is selectively utilized by host microorganisms to confer a health benefit – new definition by ISAPP 2017). The consumption of prebiotics is recommended at 5 g/day but most people get from food only half of that. Based on clinical research, prebiotics can decrease plasma total cholesterol, LDL cholesterol, triglycerides and increase HDL cholesterol, increase self-reported satiety in healthy adults, decrease postprandial glucose and insulin, and 14 studies on xylo-oligosaccharides (XOS) showed it to increase Bifidobacteria, shor chain fatty acids (SCFA) and decrease the pH.
Liliana Cardemil, Pr. in Chile, introduced two molecules deriving from aloe vera, acemannan and fructans, and her lab showed that they present better prebiotic effects than commercial fructo-oligosaccharides (FOS), based on Bifidobacteria growth and SCFA production. Curiously, glucomannan purified from water-depleted stressed plants increased the butyric acid several-fold, so the way to grow the aloe vera could be tailored depending on the function looked for (healing effect on the skin or prebiotic effect).
Monika Fleshner from Univ. Colorado Boulder prolonged these ideas to the impact on wellbeing. Feeding mice with chow enriched with galacto-oligosaccharides (GOS), polydextrose and/or lactoferrin, her lab managed to increase the mice’s robustness to stress. In these experiments, prebiotics protect the mice against disturbed sleep after intense acute stress (tailshocks) and chronic stress exposure (switching the circadian rhythms). The mechanisms behind this reduction of the impact of stressors on the mind and body include the upregulation of serotonin receptors in the brain and improved immune regulation. The military appears interested in these findings!
Johanna Maukonen from Dupont presented other works on 2-fucosyllactose (2’FL), the main human milk oligosaccharide (HMO). While non of the test pathogens are able to grow on 2’FL, B. longum subsp. infantis and B. bifidum could metabolize it. As highly bifidogenic, it is an important prebiotic for the newborn, and could also be beneficial for adults since the increase in Bifidobacteria in the gut is a generally positive feature.
The first day was closed with an exciting overview on the microbiome revolution in precision medicine, by Jack Gilbert, co-author of Dirt is Good, Pr. of surgery at the Univ. of Chicago and and Head of the Microbiome Center. “Medicine can no longer be practiced separately from ecology” he says, with a profusion of examples showing that usually the disease is not due to the pathogen but to the disturbance in the balance which allows it to overgrow. Precision medicine takes into account that C. diff sits calmly in most of us without degenerating into potentially lethal diarrhea, that antibiotic treatment in the year prior to chemotherapy correlates with higher mortality, that colorectal cancer is associated with Fusobacterium nucleatum, that the Amish (traditional farmers) have 7 times less asthma than the Hutterites (industrial farmers)... It was also great to learn that every person releases about 36 million cfu worth of microorganisms every hour!
2) Market trends and IPA news
Ewa Hudson from William Reed reported as in Probiota Europe on the major study undergone screening probiotic products in 20 countries over the 4 categories dietary supplements, derma, juices and kombucha. It turns out that for probiotics sold online “reviews are the new currency” (already envisioned in a Black Mirror show!). Ewa highlighted that the personalization trend helps consumers to find more targeted products while it provides the brands with more space to advertise the products, and allows to receive more reviews and obtain a higher ranking on Google and online retailers.
In terms of dosage form, convenience is key, with sticks and sachets the leading formats and drops representing 19% of the products for infants. The main health trends are 1. Digestion, 2. Immune and 3. Women health (vaginal and urinary alike). Major trends also include delivery technologies such as microencapsulation, and free-from claims.
Ewa got the audience headspinning and confused over the highest dosages found worldwide, with Chogenryoky from Japan declaring 22,5 trillion cfu! While the technical and economic feasibilities are questioned, the impact of a delivery of 22 trillion bacteria in one dose in the microbiome of a host that contains a total of 38 trillion bacteria according to the updated estimates (Ron Sender, 2016) should be evaluated.
The International Probiotics Association (IPA), following its mission to promote safe and efficacious probiotics around the world, came on stage to present the guidelines to produce probiotics - or “Quality Book”, undergoing last review among IPA members in July before its officialization, which is expected to raise the level of probiotics production standards worldwide. These guidelines were summed-up in a one-pager infographic focusing on declaring dosage, the expression of quantity in cfu (as opposed to mg), strain specificity, quantity till expiration, and storage conditions. Notably, analogous guidelines detailing high quality standards for probiotics have existed in Italy since 2013, published by the Ministry of Health, giving Italian actors a head start.
The IPA is also working on the Codex initiative, in order to help regulators from all over the world as they struggle with the specificities of live microorganisms.
At the regulatory panel discussion, the lack of harmonization was again pointed at, and while there is movement in the LATAM regulations, the positive list of claims authorized for probiotics in Chile and Colombia are applicable in food – but not in supplements. These authorized claims are general (ie. “balance of intestinal flora” or “regulation of intestinal transit”) and there is still need for a system allowing specific health claims, when the benefits are demonstrated.
3) Analytical tools from the future
One of the culminating moments of the event was the experts panel discussion on next generation analytical tools.
Marco Pane from Probiotical described flow cytometry as a cell enumeration method allowing to discriminate between different bacterial groups: total fluorescent units or tfu count the total of cells in a sample, while active fluorescent units or afu count the number of live cells in the sample, which include viable but not culturable (VBNC) cells, as opposed to plate counting. Flow cytometry is a universal, accurate and fast method to enumerate live microorganisms made official by the ISO standards published in 2015, which have allowed this method to rapidly blossom in the industry. Strikingly, the release of a White Paper by Martin Wilkinson from Univ. of Limerick on the potential of flow cytometry as a tool for measuring bacterial viability in probiotic products was announced on the same day (https://www.sciencedirect.com/science/article/pii/S0924224417307793?via%3Dihub).
Buffy Stahl from Dupont described digital PCR as the analytical tool of the future, presenting the advantage of strain identification and enumeration simultaneously, including in a mixed population.
David Erickson, Nutrasource, stood behind whole genome sequencing for species identification – with a need to be transparent with regards to which reference is used. Scott Jackson, NIST, also backs sequencing, including as a cell counting tool. Metagenomics sequencing can read unexpected bacteria in a sample, which may be due to contamination, sequencing errors or computer errors. “I can tell you the operator who ran the analysis based on the contamination level” he revealed. The sources of variability are many and data can hardly be compared from lab to lab. However data showing relative changes using the same assumptions and analytical tools are deemed interpretable. Leading microbiome sequencing platform uBiome R&D director Sara Bird assures that the lab’s tests are repeatible and reproduceable and that two samples from the same toilet wipe cluster well together compared to others. A number of scientific papers are coming from the company, that works with the biggest data bank worldwide. A recent achievement from uBiome is the development of the first clinical healthy range… and we know in the field what a challenge it is to define healthy.
To the final question from Marco Van Es, Winclove: “if in 5 years you want to test the number of live bacteria in each product on the market, what method would you use?” all panelists came up with a different answer but all agreed with Marco Pane: one method would not give the necessary precision, a stand-alone approach doesn’t allow to identify different aspects, the future should combine methodologies. So get ready to juggle with more complexity and more information in the future!
4) Focuses on specific roles for probiotics in health
Robert Schiestl from UCLA and CEO of Microbiopharma presented impressive data on strain L. johnsonii 456: able to survive at different pH and to exposure to bile acids, it also shows exceptional capacity to persevere and reproduce in the gut for over 60 days. With its strong anti-inflammatory capacity (decreases pro-inflammatory markers and increases anti-inflammatory ones), it could be of great benefits for inflammatory diseases ranging from cancer to obesity, IBS, depression, Parkinson, etc.
Harvard PhD and CEO of Fitbiomics Jonathan Scheiman presented, as in Probiota Barcelona, his exciting discovery resulting from metagenomics studies from marathoners: a special bug spikes during endurance. This special bug is able to break down lactic acid, and convert it to anti-inflammatory SCFA propionate. With sports nutrition a market of 115 billion $ in 2015 and probiotics a 58 billion $ market, and with probiotics representing just 1% of the sports nutrition market, Jonathan argues there’s a place to take, and engages with the industry, the athletes, and the athletes-to-be to step in.
Clinical reminders on the syptoms of the major vaginal infections were given by Pr. of Internal Medicine Jürgen Schrezenmeir. He showed evidence from several clinical trials that probiotics, even when delivered orally, presented better cure rates than the current basic treatments: antibiotics against Bacterial Vaginosis (BV) cured about 65% of the women after 3 months, and Gregor Reid strains cured 88% after 1 month – vs. 40% in the placebo group. Studies on oral administration of a blend of L. crispatus, L. gasseri, L. jensenii and L. rhamnosus isolated from healthy pregnant women showed the probiotics had a positive influence on the vaginal microbiota in patients with breast cancer, cured BV better than the placebo and halved the incidence of placental insufficiency and fetal complications in pregnant women with herpes virus infection.
UAS Labs’ CSO Greg Leyer presented the capacity of L. reuteri LRCTM to reduce total cholesterol, LDL and inflammation (CRP) by the deconjugation of bile acids while improving vitamin D status in supplemented subjects – maybe because vitamin D and cholesterol have a common precursor, he says. More science is in the making.
Gut brain axis
The gut brain axis session was opened by Caroline Wallace from Queen’s University Canada with the same presentation as in Probiota Barcelona. The pilot study on 10 adults with Major Depressive Disorder (MDD) showed significant improvement on mood, anhedonia, anxiety and sleep quality after 4 weeks on Lallemand’s Probio’Stick. These are important results because of low compliance and serious side effects of antidepressants while 350 million people worldwide suffer from depression. A further study is planned.
Rebecca Slykerman from University of Oakland, New Zealand, gave an excellent talk on pregnant women and postnatal depression. A study on 423 pregnant women showed the probiotic L. rhamnosus HN001 to halve the risk of clinically relevant anxiety compared to placebo. Following a cohort of 871 children born in the 90s to their teenagehood, she also found that 70% had been treated with antibiotics in their first year of life, a treatment which consistently correlated with an increase in behavioral and emotional difficulties, attention-deficit hyperactivity disorder (ADHD) as well as self-reported symptoms of depression at 11 years old.
As for Stefanie Malan-Müller, post-doc in psychiatry in South Africa, she focused on post-traumatic stress disorder (PTSD) and the understanding of what factors determine robustness and resilience to trauma and whether we can we have an effect on these. Fecal analyses of PTSD patients were compared to those of healthy subjects who had also experienced trauma and showed PTSD sufferers tended to have decreased levels of Actinobacteria (the group including Bifidobacteria), Verrucomicrobia and Akkermansia. Therefore, vulnerability could come from higher levels of inflammation in the gut and the loss of “Old Friends”. A larger cohort study is under course to confirm these findings.
5) Panel on colonization, diversity and next generation probiotics
This experts panel finally addressed the major question “when it comes to probiotics, is colonization important?” and the experts all agreed that colonization is not conditioning the efficacy of probiotics. As Jordi Espadaler, AB-Biotics CSO, rightly put it: “do drugs need to remain permanently in the body to have an effect? NO!”. What matters is the clinical effect.