“Probiotics are no longer just about digestive or immune health,” Gregory Bonfilio told NutraIngredients-USA. “More health specific areas have recently been clinically validated, and those areas are gaining momentum with US consumers.
“Because probiotics are also perceived as a general health benefit by the consumer, we expect to see probiotic organisms making their way into a variety of formulated products such as daily multi-vitamins, daily kid and senior products, meal replacement and protein powders, etc.
“The future of probiotics is in delivery forms,” he said.
Pharmachem has a licensing agreement with Italy’s Probiotical for its Microbac microencapsulated probiotics, which has seen these strains be slowly introduced to the US market over the last few years, said Bonfilio.
“Between 10 and 20 distinct products are currently using Microbac encapsulated strains, with an impressive range of delivery forms,” he noted. “Stability of tablets, capsules and chewables has been improved in real time comparisons to products made with typical uncoated strains.
“And ‘pixi stix’ type powders, as well as instant drinks, meal replacement and sports beverages are now possible. When you consider the advantages of using a microencapsulated strain in a chewable tablet, for instance, where uncoated strains would typically be harmed by enzymes in the mouth and the digestive process, the choice becomes easy.”
“Keeping probiotics alive is a major problem resulting in short shelf-lives of products, requiring overages (costs) and limiting delivery forms (e.g. beverages) or potential combinations with other active ingredients that could be detrimental to the survival of the strains,” explained Bonfilio. “In addition, probiotic efficacy is limited by gastric survival as only 10-25% survive passage through stomach and upper intestine due to the harsh conditions caused by acidity, proteases, or bile acid.”
Microencapsulation protects strains during manufacturing of finished products, resulting in lower overages needed, reducing costs and increasing shelf life, he said. The Microbac encapsulation technology utilizes controlled-release gastro-resistant vegetable oils.
“Clinical studies have shown that the target release post stomach increases gastroduodenal survival rate and subsequent colonization in comparison to uncoated strains. Microbac encapsulated strains colonize at the same rate than uncoated strains using 5 times lower amounts of probiotics.” (del Piano et al., J. Clin Gastroenterol. 2010, Vol. 44, pp. S42-S46)
Flow cytometry enumeration
Bonfilio said that, as a result of Probiotical’s work with encapsulated strains, flow cytometry enumeration methods have been developed and validated, which allows for the rapid and reliable distinction between live, dead and injured bacteria, even in a mixed population of bacteria types.
“We believe this new technology will greatly increase the speed and accuracy of testing probiotic organisms,” he said. “Flow cytometry may solve problems faced by the entire probiotic industry, because standard plating of probiotic strains often prevents an accurate, repeatable count of bacteria in multi-ingredient products.”
The technique uses a combination of cell permeant and cell impermeant dyes to determine cell health (impermeant dyes are unable to permeate through the semipermeable cell membrane).
The cell impermeant dyes have a higher affinity for DNA than cell permeant dyes and, because injured or dead cells have ‘leaky’ membranes, the cell impermeant dye will enter the cell and bind to the DNA more so than the permeant dye.
This leads to three populations of cells in a two color plot: live cells colored by the cell permeant dye only, injured cells with a combination of the impermeant and permeant dyes, and dead cells with only impermeant dyes.
“Acceptance of the method is currently ongoing with different agencies,” said Bonfilio. “Due to the clear advantages over the standard plate count technique, we expect flow cytometry to quickly replace the old technology as standard analytical method.”
The probiotic sector is the fastest growing in supplements, said Bonfilio, and consumer awareness has increased dramatically over the last few years. Despite this, he said that he still thinks that probiotics have a way to go before they reach the consumer ‘saturation’ level of omega-3s.
“By 2018 the marketplace will be more developed,” he said, “and I would expect continued strong growth (above 20%?) for probiotic supplements until then.”