Example of Bacterial Classification

Bifidobacterium bifidum Malyoth Strain

Genus
Bifidobacterium
Different from Lactobacillus genus, which it was originally classified before reclassification occurred in 1974
Species
Bifidum
Different from other species of Bifidobacteria such as: longum, infantis, and adolescentis
Strain
Malyoth Strain
Different from other strains of the same biovar or biotype. Each strain has different properties; the Malyoth strain is an obligate anaerobe and one of the most difficult to produce
Biotype or Biovar
Biovar A
Sub grouping of similar strains, such as biovar A which are found specifically in adults (biovar B strains of B. bifidum are found specifically in infants)
*Biovar is specific for the bifidobacteria genus only

NOTE: Lactobacillus bifidus is an archaic term for the entire genus of bifidobacteria. As microscopes became stronger, bifidobacteria were found to have a different shape (Y-shaped) than lactobacilli and given their own separate genus that contains many species. Re-classification occurred in 1974 and was published in the 8th Edition of Bergey’s Manual of Determinative Bacteriology. Avoid any products that list Lactobacillus bifidus, Bifidobacteria bifidus, or bifidus as these are misleading terms.

Selecting the Most Beneficial Strains for Human Use
How to Trust the Right Manufacturer

One of the most important factors to ensure implantation of friendly bacteria is to make certain that the selected strains are host specific, or normal inhabitants, of the human intestinal tract. Strains of friendly microorganisms must be host specific to colonize the intestines. A strain that is suitable for a pig will not adhere to the intestinal wall of a chicken or other animals.

Beneficial bacteria reside naturally along your intestinal walls. However, these "residents" can and will be frequently displaced. Since the beneficial bacteria do not make an invasive attachment to the wall (they do not anchor themselves), they can lose their parking spaces due to factors such as: stress, aging, new cell replacement on the microvilli, too much roughage in the diet (which will scrape them off), as well as other factors that have not been clearly identified. This is why even resident bacteria must be replenished on a daily basis with a selection of the proper strain.

Choosing the right strain
Careful attention is given to strain selection. Our organisms come from culture banks. Culture banks are institutes where qualified researchers or scientists deposit microorganisms for safekeeping and for the use of other scientists or manufacturers. Unfortunately, most culture banks do not challenge the identification of the microorganisms they have received but rely on the depositor’s word as to the genus, species, and strain designation. This has led to disappointments for companies who did not verify these original designations. Without proper verification, a company may have no idea what they are selling. Therefore, our policy has been to verify the microorganisms with world-renowned authorities by using internationally acceptable procedures. Once in our possession, each purified and identified culture strain of our products is stored in cryoprotective vials in a deep freeze where the temperature is held at -170° F. This procedure is necessary to prevent strain mutation and ensure integrity until it is used in production.

Examples of Specific Probiotic Properties for Natren’s Strain Selection

• Production of large amounts of natural antibiotic and antimicrobial substances, i.e., H₂O₂, when compared to other strains of the same species
• Health benefits observable in human studies or in laboratory tests such as cholesterol reduction
• Better ability to adhere to intestinal walls and inhibit pathogens
• Aiding in the absorption of nutrients and processes of digestion

We try to select strains that are not widely available in other commercial products. We maximize each organism’s potential for health-giving benefits by specific growth media selection. Natren has had species verifications performed by Dr. Virginia Holdeman Moore at Virginia Polytechnic Institute, except for B. bifidum, which has been verified by the DMS in Germany using DNA technology. Dr. Sharon Hillier, University of Washington, confirmed that Natren’s DDS-1 is a H₂O₂ positive acidophilus. Silliker Laboratories and Dr. Moore confirmed that Natren’s DDS-1 strain conforms to published peer-reviewed literature. No other company, researcher, or basic manufacturer has ever provided independent verification that the strain they claim to be DDS-1 to be present in a finished product is truly that organism. Natren is the only company that had a final validation of its strain by Dr. Lars Peterson, chairman of the Probiotic Interest Group/Scientific Committee.


Why are Particular Genus and Species
More Important than Others for Human Use?

Lactobacillus acidophilus is the most prominent friendly bacteria in the small intestine and is considered part of the indigenous flora. Lactobacillus bulgaricus (the premier bacteria found in yogurt) are very beneficial transient bacteria. It is especially helpful in supporting healthy food digestion and peristaltic action. L. bulgaricus assists L. acidophilus and bifidobacteria to successfully compete with other microorganisms for attachment sites.

Bifidobacteria are considered the most important indigenous bacteria of the large intestine. Bifidobacterium bifidum is one of the most researched species of bifidobacteria and is essential to a healthy intestinal tract. Natren sells a super strain of B. bifidum that is specific for the large intestine of adults. Bifidobacteria are also considered to be the most important organisms for infants. To accommodate this need, Natren makes a supplement specifically for infants, which contains a super strain of Bifidobacterium infantis. Recent evidence has led Japanese scientists to consider bifidobacteria more important than lactobacilli in human health from infants to adults. Natren, however, believes that proper supplementation should be taken as a system. By using specific strains of L. acidophilus, B. bifidum, and L. bulgaricus, each type of beneficial bacteria has its own specific function, creating a system to optimize the health of the gastrointestinal tract.

Recently, an exhaustive study was presented to the FDA providing concrete evidence (in excess of 600 citations) that the lactobacillus and bifidobacteria genera are entirely safe for human consumption.

Producing Products of Optimum Quality

Early probiotic supplements were simply yogurt tablets with very low potency, as only about 1% of fresh yogurt products are actually viable bacteria. To make stronger probiotic products, after culturing the beneficial bacteria, most manufacturers concentrate the percentage of bacteria in their products by ultra-filtration or centrifugation. Production methods such as ultra-filtration and centrifugation concentrate the cell biomass by reducing the volume of the original growth medium, thereby increasing and concentrating the percentage of cells. With centrifugation, the strong centrifugal forces damage the structure of the cells. Injured cells may have less resistance to unfavorable conditions in the gastrointestinal tract. The original use for concentrated centrifuged cells was to make a bulk starter for the dairy industry. In the manufacture of fermented foods such as cheese or yogurt, microorganisms may be able to partially repair the damage during the culturing process in the bulk starter. However, this process is not applicable to the probiotics industry. In freeze-dried supplements made from centrifuged cells, there is no opportunity to repair the damage.

The main drawback of both centrifugation and ultra-filtration for the production of probiotic health supplements is: when you discard the liquid culturing medium (supernatant), you lose an essential part of the product. The supernatant, which may be as important as the live bacteria itself, contains antimicrobial by-products, enzymes and other beneficial factors produced by the microorganisms. Most probiotic manufacturers avoid retaining the supernatant when they freeze-dry their products because it adds substantially their production costs. After a product is centrifuged, only 3% of the cell biomass concentrate remains. Manufacturers will blend the 3% biomass with 97% of arbitrary filler, allowing them to sell these "probiotics" for a substantially lower price.

Natren’s specific production technology employs a very gentle technique to concentrate cells. The major advantage to this method is the retention of the supernatant, which produces a complete supplement and maximizes the probiotic benefits for the consumer.

The cells are freeze dried in a complete state. When they are packaged in dark glass bottles and refrigerated, they are in a state of arrested growth. The beneficial bacteria are activated when ingested with tepid, filtered water.

Reported potency refers to the total number of single cells, pairs, short chains, and/or long chains present in the product. Laboratory reports refer to these chains as colony forming units (CFU). Lactobacillus acidophilus occurs naturally in long or short chains (similar in shape to sausage links). These chains normally break apart to form new CFU when they mature. Occasionally they do not break apart and continue growing in one long clumped chain. This phenomenon is known as agglomeration. Centrifugation and ultra-filtration causes forced cell division of these chains resulting in higher potencies. Breaking these chains prematurely may boost the potency but the quality of the product is sacrificed. The full culture method, which Natren employs exclusively for all our current products, does not promote premature chain separation, allowing the bacteria to divide naturally into new colony forming units. This method ensures the optimum levels of both potency and quality for the consumer.

Chemical preservatives can be added to prolong shelf life. Depending on the ingredient, these processing agents can be used in products, ranging from 1% to 5% by weight, without being listed on the label. MSG and BHT are among the favored agents as they shield the cells from rapid die-off. Natren does not use these unlisted ingredients in our products.

Three quality control assays are run for potency and contaminants, including a screen for individual pathogens, which must test negative before each product lot is released for sale. Periodic re-verification of species is no longer required on Natren products since our cultures have been DNA fingerprinted. Periodic random shelf life testing is performed on products during their guaranteed shelf life. This shelf life testing has been continually performed over the past 18 years.

Rounding Out the Cycle: Packaging

Natren relies on specific production, special freeze drying techniques, and packaging to maintain the arrested growth state of friendly microorganisms. Bacteria are kept in an environment that is 18% drier than the Sahara desert and there are no plastic containers that can support this type of environment. Natren uses dark amber glass bottles to provide the best moisture and light barrier to protect the cells from premature die-off.

Glass is the one of the most non-porous and non-toxic packaging material available commercially for health foods. It is environmentally safe and recyclable. Plastics are not impermeable, they break down and allow moisture and air to pass over time and may even give off objectionable gases. Plastics are made from petroleum and more difficult to recycle.

Even though bottle companies sanitize glass bottles prior to shipment, Natren employs our own unique procedures to sanitize the bottles a second time. Because our strict, sanitary guidelines and attention to ensuring high quality, we believe that we exceed the drug GMP certification standards.

Natren pioneered global overnight or second day delivery airfreighting shipments of probiotics. We pack our products in thermally controlled boxes, cooled with ice packs to deliver the freshest and most potent probiotics possible.
CAUTION: Once product leaves the manufacturer, there is no guarantee that the product will not be exposed to damaging heat or mishandled. There are no probiotics that do not require refrigeration. Controlled room or ambient temperature does not exist once the product leaves the manufacturer’s door.

What Does Our Guarantee Mean?

With Natren’s manufacturing standards, we are able to offer a potency guarantee through the printed expiration date. This type of guarantee is in accordance with the NNFA Probiotic Labeling Standard, which was adopted in March 1989. The NNFA is an American association of health food industry retailers, distributors, and manufacturers. Industry organizations adopted the standard to regulate themselves rather than allow the government to dictate terms. The criterion specifically calls for a "label statement of the minimum number of viable cells or colony-forming units (CFU) per unit of measure such as a capsule, tablespoon, gram, etc."

For the last 18 years, every lot of Natren products has been tested for potency and purity to ensure the highest quality through the printed expiration date. We have undergone rigorous consumer testing in the U.S. and U.K. Further testing has been done in Australia. We were among the first to enroll our products in the industry quality assurance program, called TruLabel. Natasha Trenev, president and founder of Natren developed the Probiotic Standards for the Natural Products Quality Assurance Alliance as part of good manufacturing practices guidelines, which were read into Congressional record. Natren consistently conforms to potency guarantee and label declaration guidelines.

Why Not Mix Various Bacteria in a Single Bottle?

Natren has always refused to mix bacteria in a single product. Each type of microorganism is suited for a specific niche in the intestinal environment. Numerous studies support the use of both L. acidophilus and L. bulgaricus or L. acidophilus and bifidobacteria species. However, storing different strains and species in the same bottle for extended periods of time may cause dramatic shifts in the proportions of microorganisms present. We do not know of any high potency products on the market that mix various bacteria offering extended potency guarantees for specific percentages of microorganisms through a printed expiration date. We assume that other manufacturers have not discovered a way to combine high potencies of various freeze-dried microorganisms and guarantee that ratios will not shift in the product. These types of specific potency guarantee statements are avoided so that less expensive strains can dominate the content of the product.

The only carrier we have found where the bacteria can be kept separated and micro enrobed is in our special oil-matrix system. Some companies explain that keeping the product extremely dry allows them to mix various bacteria in the same container or bottle. This feature was not designed to keep bacteria separate but instead designed to have a better shelf life. They cannot guarantee a product with high quality and potency without the right system to mix the bacteria.

We Have Never Wavered on Storage Conditions

Don’t be fooled by storage claims at "ambient temperature"
Ambient temperatures do not exist once the product leaves the manufacturer. Over thirty years of perishable shipping and distribution experience has shown that delivery truck temperatures can reach 120°F to 130°F. Merchandise that is not marked perishable or kept refrigerated is left for days on loading docks or doorsteps. Refrigeration is the only way to avoid product mishandling and give a potency guarantee through the printed expiration date. Refrigerated storage is mandatory to maintain high potencies of lactobacilli and bifidobacteria species. Extensive industry and NNFA testing has revealed that refrigerated storage is required for extended shelf life stability. Some manufacturers use microorganisms from the streptococci or enterococci group. One prime example, Enterococcus faecium, is heat stable and will survive much higher temperatures and adverse storage conditions than either lactobacilli or bifidobacteria. E. faecium may also be less sensitive to non-refrigerated storage conditions. Many products that add this microorganism to their microbial mix claim potency for the total count of all the bacteria present in their product. This practice is very misleading because it does not clearly state the potency of each individual beneficial bacterial strain.

We have not seen any guarantee of product potency stating specific numbers of each lactobacilli or bifidobacteria through a visible, verifiable expiration date in these mixed products that claim shelf life stability with or without refrigeration.

Why We Prefer Freeze-Dried Powders

L. acidophilus, B. bifidum, and L. bulgaricus powders, which are freeze-dried rather than spray-dried, are the internationally preferred form for the optimum survival of beneficial microorganisms in health food probiotic supplements. Natren’s freeze-dried powders are exceptionally superior because they include the all-important supernatant, which acts as a buffering agent and food source to facilitate better survival for the bacteria. Our powder forms are also desirable because they can impact our entire digestive/intestinal tract beginning in the mouth.

Natasha Trenev has been involved with the manufacture of probiotics for over 30 years. Her experience has shown that liquid acidophilus supplements can support high viable cell counts for no more than several weeks (at best), depending on the additives used with continuous refrigeration. The beneficial effect that consumers derive from liquid acidophilus is due primarily to the supernatant or the growth medium where the beneficial by-products of L. acidophilus metabolism have been deposited.

Encapsulation of probiotic products in two-piece hard gelatin capsules is problematic because the capsules may contain a considerable amount (approximately 20%) of moisture. This moisture, or water activity, jeopardizes the survival of probiotics during the shelf life under the current market conditions. Pure L. acidophilus capsules did not have a ‘potency guaranteed through an extended expiration date’ before we came out with our Natradophilus non-dairy capsules (called Megadophilus in U.S.). Natradophilus is encapsulated in a unique capsule that minimizes exposure to moisture allowing us to ensure a potency guarantee through a printed expiration date.

Some manufacturers use enteric coating, which protects the contents of the gelatin capsule for up to an hour against the acid found in the stomach. This was developed for pharmaceutical drugs and is not appropriate for protecting probiotic products. Natren’s capsules are not enteric coated.

Testing – What is the Suitable Criteria?

From University of Washington, U.S.A.: "We have recently completed testing of your ‘Megadophilus DDS-1’ product [marketed under the brand name Natradophilus in Australia] (Lot number N133, expiration 7-30-92). We found it to contain 109 [billions] colony-forming units per gram of L. acidophilus, which was H₂O₂ [hydrogen peroxide] positive." Sharon L. Hillier, Ph.D., Director, Research Microbiology Laboratory, Department of Obstetrics and Gynecology.

From U.S. National Nutritional Foods Association, Natradophilus shelf-life testing results: "These viable cell counts compare favorably with your label claim of ‘billions per gram.’ Species identification was also performed, using the API method. The organism present was identified as Lactobacillus acidophilus." Burton Kallman, Ph.D., Director of Science and Technology, NNFA.
NOTE: This was the initial test run at one month and included species identification. Testing after nine months found that the product met labeled claims for potency when refrigerated as required.

From Silliker Laboratories of California, Inc., addressed to the president of Natren, Inc., Natasha Trenev: "This letter is in response to your inquiry of 15 February. You were concerned about possible causes for misidentification [of] the L. acidophilus in your Megadophilus [Natradophilus] by other institutions. As with any bacterial identification, there could be a host of reasons for misidentifying an organism." Daryl W. Osato, M.S., Laboratory Director.
NOTE: An earlier letter from Mr. Osato confirmed culture identification for Megadophilus [Natradophilus]: "Most of these cultures have been identified by this laboratory, with definitive verification being done by a distinguished professor at Virginia Polytechnic Institute…Definitive confirmation for L. acidophilus took place in late 1987…"

It is very difficult to test probiotic organisms accurately. Many species have similar attributes and are hard to distinguish without expensive phenotypic characterization or DNA testing. We have employed such testing to verify the species used in Natradophilus as L. acidophilus. This is the definitive verification mentioned above by the Virginia Polytechnic Institute.

Other institutions may not recognize that our products require testing procedures that are different from those developed for other high potency acidophilus products. The unique production processes we employ to manufacture superior products necessitates the use of very specific testing protocol. Thus, erroneous lab reports are generated which ultimately embarrass the issuer.

Our industry has a testing program called TruLabel in the United States where Natradophilus powder and Natradophilus non-dairy capsules is manufactured. Natural product manufacturers voluntarily enroll and pay for independent verification of their label claims. As part of this program, a committee of probiotic manufacturers has been formed to agree upon testing procedures for the TruLabel program. As products are found to meet label claims, retailers are notified. This program gives customers, retailers, and health care practitioners a simple way to know which brands to trust and stop the proliferation of inaccurate tests, which hurt the industry’s credibility.

Over the past 18 years, recognized independent laboratories have run thousands of quality control tests on Natren’s products. Natren’s products have been tested in three different consumer quality assurance programs, including other countries, and have passed with flying colors. You can count on the superior quality of Natren’s products, which is recognized as the leader in the probiotics industry worldwide.