Rapid Automated Testing of Probiotic Organisms

Definition and Health benefits of Probiotics: The World Health Organization’s 2001 definition of probiotics is “live micro-organisms which, when administered in adequate amounts, confer health benefits on the host”.[1] This definition, although widely adopted, is not acceptable to the European Food Safety Authority because it embeds a health claim which is not measurable.[2] Etymologically, the term appears to be a composite of the Latin preposition pro (“for”) and the Greek βιωτικος(biotic), the latter deriving from the noun βιος (bios, “life”) [3].

Health benefits: Some digestive disease specialists are recommending the use of probiotic organisms to help in the treatment of disorders that frustrate conventional medicine, such as irritable bowel syndrome. Since the mid-1990s, clinical studies have established that probiotic therapy can help in the treatment of several gastrointestinal ills, may delay the development of allergies in children, and both treat and prevent vaginal and urinary infections in women.

Examples of Probiotic Organisms: There are hundreds of strains of probiotic bacteria. The most commonly used organisms include Lactobacillus sp. (such as L. acidophilus, L. casei, L. fermentum , L. rhamnosus) Bifidobacterium sp, (such as B. Bifidum, B. lactis and B. longum), Streptococcus thermophilus, Bacillus coagulans,and Enterococcus faecium.
Potency Testing: Probiotics offer a broad range of health benefits. As with any supplement, the efficacy of a probiotic depends on dosage. Essentially the titer of live organisms is the critical part in determining potency. Recommending an adequate dose for an individual patient requires clear knowledge of the potency of a product. Probiotic potency is specified as the numbers of viable cells of the beneficiary organism. Confidence in the accuracy of this number is essential for successful and consistent clinical results.
Enumeration of bacteria has been a routine practice in microbiology for over 100 years. The gold-standard method used to determine the titer of organisms is known as the viable plate count that is used to generate a count referred to as colony forming units (CFU). On probiotic product labels, results are expressed in CFU per serving. Since probiotic cells are sensitive to their environment, potency is subject to change. Therefore, potency must be determined after manufacturing, shipping and storage. Thus the supplement industry needs a rapid, accurate, and reliable method for testing of probiotic organisms is therefore needed. BioLumix offers a novel rapid method for enumeration of Probiotic organisms.

BioLumix Methodology for Potency Testing:
A calibration curve is generated to easily relate the number of colony forming units determined using the plate count method to the detection times (DT) in the BioLumix instrument. These calibration curves are embedded into the instrument software and are used to access the number of probiotic organisms present in the product sample for individual organisms. An example is shown in the Graph for the Lactobacillus acidophilus. Currently, individual calibration curves are available for the following organisms: L. casei, L. acidophilus, L. rhamnosus, L. bulgarus, B. coagulans, B. longum, B. bifidum, E. feacalis, and S. thermophilus. The procedure used to test sample cultures involves a single 1:10,000 dilution of the sample followed by the addition of 0.1 ml to the appropriate test vial. Organism growth may occur rapidly, often in less than 24 hr, and the BioLumix instrument generates an estimate of the cfu per gram of sample. This is a much more rapid method than the traditional plate methods that often takes 3-7 days for Lactobacillus species. Using the BioLumix rapid method can be much less expensive than traditional plate methods for Lactobacillus species as these organisms often require specialty media under conditions of low oxygen (candle jars).

Microbial contamination: Good manufacturing Practices must be applied in the manufacture of probiotic containing products. Contamination of probiotic products with undesirable microorganisms is possible in uncontrolled fermentation and during handling. Therefore, most probiotic batches need to be tested for indicator organisms such as coliforms and to also show the absence of potentially harmful organisms such as E. coli, Staphylococcus and Salmonella.

BioLumix Methodology for Microbial Contamination: The BioLumix simplified automated system can detect indicator organisms and objectionable organisms, if present, in a fraction of the time of traditional methods, with significantly less hands-on time. The system offers a wide variety of rapid assays for samples, including assays to detect yeast & molds, coliforms, E. coli, Staphylococcus, Pseudomonas and Salmonella.

References
1. Schlundt, Jorgen. “Health and Nutritional Properties of Probiotics in Food including Powder Milk with Live Lactic Acid Bacteria”. Report of a Joint FAO/WHO Expert Consultation on Evaluation of Health and Nutritional Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria. FAO / WHO.
2. Rijkers, Ger T.; De Vos, Willem M.; Brummer, Robert-Jan; Morelli, Lorenzo; Corthier, Gerard; Marteau, Philippe (2011). “Health benefits and health claims of probiotics: Bridging science and marketing”. British Journal of Nutrition 106 (9): 1291–6.
3. Hamilton-Miller, J. M. T.; Gibson, G. R.; Bruck, W. “Some insights into the derivation and early uses of the word ‘probiotic’”. British Journal of Nutrition 2003 (90): 845.