Automated Microbiology Testing is the Key to Laboratory Growth in the 21st Century

Modern microbiology laboratories demand efficiency, and that means constantly improving data quality and lab productivity while simultaneously shortening time to results, and lowering the manufacturing cycle. This works by lowering the required working capital investment through faster release of inventory. As microbiologists become swamped with higher and higher volumes of work, traditional paper-intensive manual plate methodologies are becoming a thing of the past, and automated technology now holds the key to future growth. 

Manual systems are tedious and labor intensive; they are susceptible to transcription errors, and require skilled lab technicians to spend excessive amounts of time performing simple, monotonous tasks. Twenty years ago, when automated microbiological testing systems were not available, this elongated time to results and plodding flow of information was a necessary evil; with the rapidity of technological advancement in the 21st century, this is no longer the case. Automated microbiological testing systems are equipped with flexible electronic designs, allowing lab personnel to more efficiently manage, review, store, and report the high volume of analytical information needed for microbiology quality assurance. Repetition of the same task again and again is often the source of inconsistent results, but automation significantly reduces the chances of human error as well as the risk of lost data, resulting in huge productivity gains for manufacturers and laboratories. In this way, automation can provide companies with a higher level of quality assurance while requiring far less labor, and this equates to a greater profit margin.

In today’s economic climate, automation is essential in order to streamline workflows and ensure a quality product. Expedition of lab work through automation is changing the way that modern microbiology labs function, and the companies that utilize these new microbiology instruments are becoming more productive than ever before. The BioLumix system brings automation to current microbiological testing, providing a new generation of microbiology labs with the tools they need to excel and overcome their challenges. The system operates by using optical sensors, which detect color and fluorescent changes in ready to use carbon dioxide sensing vials, eliminating the need to make, prepare, or quality control traditional media. Automated sample tracking and data management also allows for the safe and immediate transfer of information in a paperless format.

With many labs bereft of a large number of qualified technicians, and subject to a great deal of pressure to analyze a higher volume of samples and deliver rapid results, the BioLumix automated microbiological testing system does the tedious work for you, and can be easily operated by a non-microbiologist. All in all, this equates to a 60% savings in labor, and this newfound time can be utilized by laboratory personnel to perform higher value tasks, such as the interpretation of results and diagnoses of potential problems as early as possible. We have all heard the old adage “time is money,” and that is where the BioLumix rapid testing system truly shines. Simply put, faster detection of microbial contamination results in faster corrective action, and that saves money. A nominal investment in an automated testing system will save your lab valuable days in their quality control process, and can reduce time to results from 7-10 days to 1-2 days. If contaminants are found, products can be recalled much earlier, and manufacturers can quickly take action to substitute raw materials that do not meet specifications. They can also resume production of a high quality product much earlier, which means that a single incident of faster intervention can save more than the cost of an automated rapid testing system.  Speeding up microbiology testing results in the speeding up of production and the reduction of inventory. By implementing a rapid micro method, you can re-allocate new found floor space to increase manufacturing capacity, add more SKU’s to inventory, or reduce distribution center costs.

The BioLumix automated microbiology testing system is easy to use, and can be utilized by modern laboratories to ensure quality products, speed up production, reduce labor, and increase manufacturing capacity by enabling a rapid release of inventory. With a lower working capital investment, and a more efficacious use of microbiologists’ skills, automating with BioLumix will have a profound affect on any organization’s key profit drivers, and may very well be the key to their growth in the 21st century marketplace.

Simplified, Rapid, Automated Microbiology

Simplified, Rapid, Automated Microbiology – mybiolumix.com

Customer support and innovation are the cornerstones of the BioLumix business. Our mission is to assist our customers in their quality assurance by offering simplified, rapid, automated, reliable solutions for microbiological quality control and assurance. We’re always available to answer any inquiries regarding the system specifications, installation, validation, performance, benefits and cost savings. If you would like to speak with one of our experienced microbiologists or engineers, please contact us directly: (734) 984-3100

We are committed to delivering high value products, with a real technological difference, to satisfy our customer’s needs and help improve their quality assurance. We’re always available to answer any inquiries regarding the system specifications, installation, validation, performance, benefits and cost savings. If you would like to speak with one of our experienced microbiologists or engineers, please contact us directly: (734) 984-3100

Microbiology of Dietary Supplements

Official Methods for Dietary Supplements

The USP methods for dietary supplements include:

USP <2750> Manufacturing Practices for Dietary Supplements: Contains the official FDA ruling on dietary supplement cGMPs (21 CFR Part 111).

USP <2021> Microbial Enumeration Tests-Nutritional and Dietary Supplements: Enumeration of the total number of aerobic bacteria, enumeration of the total number of combined yeast and molds, and the presence of bile-tolerant gram negative bacteria present in all nutritional supplements, from raw materials to the finished product.

USP <2022> Microbiological Procedures for Absence of Specified Microorganisms-Nutritional and Dietary Supplements: Procedure for determining the absence of specified objectionable organisms. Good manufacturing practices require that objectionable organisms be absent from non-sterile nutritional and dietary products. Objectionable organisms are defined as microorganisms that may cause potential health hazards to the user of the product and would adversely affect the product safety.

USP <2023> Microbiological Attributes of Non-sterile Nutritional and Dietary Supplements: contains information about frequency of microbiological sampling and testing, recommended microbial limits for botanical ingredients and products, and finally, recommended microbial limits for dietary supplement ingredients and products. However, specifications are typically are developed by the manufacturer with guidance from USP <2023>.

USP <1223> Validation of Alternative Microbiological Methods:  Any alternative method must be validated against these UPS methods using the methodology described in USP <1223> including side-by-side testing, determination of the accuracy, specificity, limit of detection, robustness, ruggedness, etc.

Many methods validated for food products, are not necessarily validated or appropriate for dietary supplements. For example, many methods have been written for very specific products such as eggs, meat, milk, nuts, etc. These methods were not written with dietary supplements in mind. And “dietary supplements” encompass such a large scope and variety of products therefore, it is risky to simply assume that a method written for example for eggs will also work for supplements.  Therefore, a method having AOAC certification is not necessarily appropriate for dietary supplements.  AOAC had not certified any microbiological method for dietary supplements, all their methods are certified for food only.

Often, one method will work for one product but not for a different product. Petrifilm methodologies are excellent methodology for many food products. However, Petrifilm should not be used on dietary supplement samples without thorough validation. For instance, samples that are dark in color and many common dietary ingredients might make it difficult to see growth on the Petrifilm. Viscous products such as Psyllium begins to harden before an appropriate spread on a Petrifilm can be obtained.

Product Interference or Matrix Effect

Spirulina, turmeric, Psyllium, honey, Calcium carbonate, dyes, and oils: these are just a few of the many products that the Nutraceutical industry works with, and struggles with. Many botanicals, herbals, enzymes or protein powders create challenges for microbiological testing (Tackling those challenging Nutraceutical Products with Rapid Automated Microbiology Testing!, C. Ockerman 2013). Granules (which can often be mistaken for a bacterial colony), extreme pH, viscous products can all interfere with the plate count methodology.

Microbial Testing of Dietary Supplement

Microbiological testing in the dietary supplement industry consists according to the USP chapters, of monitoring levels of naturally occurring indicator organisms and spoilage organisms, as well as the absence of objectionable organisms (e.g., Salmonella, Staphylococcus aureus, and E. coli) in finished products.

The typical battery of microbiological tests for a dietary supplement includes:

• Total Plate Count

• Yeast and Molds

• Enterobacteriaceae (gram negative bile tolerant bacteria)
• E. coli
• S. aureus

• Salmonella

• Pseudomonas (for liquid products or products with high water activity, or topical/cosmetic products)

How to Test for Microbiology

Many companies outsource the microbiology testing to third party commercial testing labs instead of performing it in house.  Some companies fell that whenever products are sent out for testing quality results cannot be questioned, but time is sacrificed and a premium price is paid for the service. Therefore, companies tend to only bring their microbiological testing in-house when they reach a critical mass such that the cost of sending out is equal to or more than starting up a laboratory.

In house Laboratories can perform using the USP methodology.  In preparing its own media and reagents the lab needs to pay attention to USP <1117> (Good Microbiological Laboratory Practices).  Because in an internal laboratory the quality of the results depends on the quality of the culture media prepared.  This includes accurate weighing of dehydrated components, the use of high quality (USP Purified) water, completely dissolving the dehydrated media or individual ingredients, and the need to control the heating of the media to avoid damaging heat-labile components of the media. The quality control of each batch of the media is a critical concern.

It is important to take production timelines into consideration when determining what methods fit each company individual needs.  A major issue with microbiological testing is the hidden costs in labor and inventory hold time.  The one assay that often keeps a quality control manager up at night is their yeast and mold testing—which takes five days to obtain a result.

Why Automated Rapid Microbiology?

Internalizing the microbiology testing and especially adopting rapid microbiological methods (RMM) can speed up the time to results from 7-10 days to 24-48 hours. The time saved has a direct and REAL impact on many key operational profit drivers.  The Advantages of RMM can include: Greater accuracy, better sensitivity, and increased sample throughput, automated data capturing and allowing easier data handling, and reduced cost for product release.  Rapid methods allow test results to be obtained faster. More importantly, if a product is above a desired microbiological specification, the results are obtained a lot sooner, and corrective action can be taken days before traditional methodology would allow.

All analytical methods need to be validated prior to their introduction into routine use, and this is especially true for novel technology platforms, such as rapid microbiological methods (RMMs). Because many RMM technologies consist of a combination of instrumentation, software, consumables and reagents, in addition to specific detection, quantitative or identification methodologies, it is important to develop a comprehensive and holistic approach to the validation process to ensure that the entire RMM system is suitable for its intended use.

The BioLumix Advantage
The BioLumix System is a system that fits a need in the Dietary supplement industry in terms of speed, working with all product matrices, and automatically documenting the results in real-time, thereby reducing the labor requirement for microbiological testing.

The BioLumix system was extensively validated for dietary products and compared to the methodology described in USP <2021> and <2022>.  The validation methodology followed USP <1223> “Validation of Alternative Microbiological Methods”.  This included performance criteria such as: side-by-side comparison to the USP methodology, accuracy and precision, specificity,  limits of detection and quantification, along with ruggedness and robustness, reproducibility of the results, false positivity rate and false negative rate.

A full validation package including Installation Qualification (IQ), Operational qualifications (OQ) and Performance qualification (PQ) is included with the BioLumix system
The BioLumix Advantages include:

  • The only system with all assays on a single platform
  • Real-time communication – early warning of contamination
  • 48-hour Automated Certificate of Analysis
  • Simplified testing with ready-to-use vials
  • Each vial comes with a certificate of analysis, negating the need for QC of the vials
  • Validated for dietary supplement products – IQ, OQ, PQ
  • No product interference
  • 21 CFR Part 11–compliant software
  • Remote-access customer support 24/7/365

BioLumix RMM platform featured in Latest Edition of NutraCos Journal

Introduction

The BioLumix platforms Rapid Microbiological Method (RMM) was recently featured in NutraCos journal for its ability to offer a multitude of unprecedented advantages to companies, as well as its versatility across various industries. NutraCos is an international journal that features articles on everything from nutraceuticals and probiotics, to regulations and policy.1 Given the rapidity of technological advancement, and the complex structural nature of the cosmetic, OTC and nutraceutical industries, many companies and individuals utilize NutraCos as a way to stay current with the latest developments throughout the world. In the journals January/April 2013 edition, our President, Dr. Ruth Eden, and Senior Research Microbiologist, Roger Brideau, explain the technology, as well as the benefits of the BioLumix automated microbial testing system to an international audience.2

 

Why Use Rapid Microbiological Methods?
Rapid microbiological methods are technologies that allow their users to get microbiological test results faster compared to traditional USP methodologies, which use plates and require a minimum of two to seven days in order to achieve complete results.  The BioLumix system can report results within hours, and all final results are available within 48 hours. This can dramatically impact key profit drivers by eliminating the wait for microbial results that can tie up working capital and often results in storage expenses and delays in supplying products to the market. Essentially, inventory stays on the floor instead of going out the door to the customer/patient population.  A shortening of the length of the manufacturing cycle means less required working capital investments, and simplifies rapid microbiology.

RMM Using BIOLUMIX
The systems three components consist of a modular instrument, disposable vials, and a 21 CFR Part 11 compliant computerized software package, the latter of which provides the user with an audit trail, operator identification, a data management system, and multiple customizable report formats. This process functions by continually monitoring the changes of a broth medium in which target organisms grow. The BioLumix system uses a CO2 sensor at the top of each disposable vial to detect released CO2, which is produced by all microorganisms, as well as reagents at the bottom of each vial, which measure changes in color/fluorescence of the broth medium by way of light intensity units in response to microbial metabolism.

 

The BioLumix system is extremely versatile and is capable of performing most microbiological assays tests, such as those for microbial content (microbial enumeration of total counts and yeast and molds), objectionable organisms, suitability testing and preservative efficacy testing.

A suitability test, to show usage of an adequate neutralizer inactivates the preservative in the product being tested. This demonstrates that if organisms are in fact present they will be able to grow and be detected. The suitability test can be performed very simply and quickly with the BioLumix system.

Testing of any cosmetic, pharmaceutical or personal care product should include a survey for contamination often referred to as testing for Microbial Content, which uses assays designed to measure the growth of bacteria, yeasts and molds to flag products for the presence of contaminants.3, 4 For most cosmeceutical products, regulations state that total aerobic count should be <100 cfu/g, and that yeast and mold counts should be <10 cfu/g. The workflow of the testing of a personal care sample for the presence of microbial contamination is very simple and requires a fraction of the time and labor needed to perform the same tests using the USP methodology.  Each product can also be assayed specifically for the detection of E coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella species.

Upon determination that a product has been properly neutralized and has very low levels of contamination, the user can conduct a Preservative Efficacy Test (PET), sometimes referred to as the Antimicrobial Efficacy Tests. This test is required for all cosmetic and personal care products in the United States and follows the guidelines set forth by USP 51.5 Briefly put, the product must be challenged within individual containers, each going through this process separately using one of the five required organisms, which include ATCC established strains of E. coli, S. aureus, P. auruginosa, C. albicans, and A. brasiliensis. The PET assay measures the reduction of a high inoculum (106-107 cfu/g) in the presence of a product containing preservative over 28 days, which also tests the ability of the preservative to prevent a rebound of an organism (regrowth). After these tests the BioLumix software produces a certificate of analysis with a succinct report of the microbiological properties of each product for use at your convenience. This equates to proactive detection of contaminated products, and ensures that minimal resources are spent on production and subsequent recall of said products if they are found to be contaminated.

Conclusion

The BioLumix Rapid Microbiology System can save multiple days of time to reach the assay endpoint versus traditional plate methods.  This time saving could dramatically impact key profit drivers, such as elimination of idle time with raw materials, work in progress, and speed to finished goods. Faster detection of microbial contamination also results in faster corrective action in the event that material is above the targeted specification level. Earlier awareness of contamination problems in production runs can result in less production of defective final finished goods. By significantly decreasing the manufacturing cycle, one can lower their required working capital investment.

The BioLumix instrument is a straightforward and easy to use system that allows for rapid and accurate results.  The system is fully automated including data archiving, data maintenance, and report generation. It is paperless and efficient; saving on disposables, time and space. In short, BioLumix streamlines microbiological testing, delivering a simpler, faster, and automated solution saving your company significant time, labor and money.

To read the whole article complete with direct comparison tables and figures, visit our scientific presentations page. (http://www.mybiolumix.com/scientific-presentations/)

 

References

  1. Lee, C. “NutraCos,” Vitafoods Europe. London, UK: Informa Exhibitions, 2013.
  2. Mori, F. L. “NutraCos,” Via M. Donati 6. Milano, Italy: B5 s.r.l., 2013.
  3. United States Pharmacopeia, Chapter <61>, 2009.
  4. United States Pharmacopeia, Chapter <62>, 2009.
  5. Moser, C.L.; Meyer B.K. AAPS PharmSciTech. 2009, 12 (1), 222-6

Case Study of a New Growth-Based Rapid Microbiological Method that Detects the Presence of Specific Organisms and Provides an Estimation of Viable Cell Count

Published In “Encyclopedia of Rapid Microbiological Methods”, vol 4. Editor Michael Miller 2013, PDA Bethesda, MD. DHI Publishing River Grove. IL.

 

Chapter  Highlights

The BioLumix Rapid Microbiological System is capable of performing testing with a similar outcome to USP <61> (USP, 2009a) and <62> (USP, 2009b) simultane­ously on a single platform faster than the conventional USP methodology. The automated growth-based system detects microbial growth, provides an estimation of viable cell counts (total aerobic count, yeast and mold, bile tolerant Gram-negatives), and identifies the absence of specified groups of microorganisms. The novel BioLumix Optical System using ready-to-use vials provides faster results, labor savings, automation, and connectivity. The streamlined testing design and rapid results lead to reduced material-holding time for faster product and raw materials release. The BioLumix System is capable of analyzing tablets, capsules, powder, liquid and viscous material without any product interference. BioLumix offers a comprehensive range of micro­biological tests for raw materials, in-process and finished products, as well as process water. Environmental monitoring (e.g., detecting micro-organisms on surfaces) is also performed easily by inserting the swab directly into the assay vial. Early warning of contaminated samples, as well as sample release informa­tion, could be automatically communicated through your intranet, significantly improving your company’s efficiencies. The software is 21 CFR Part 11 compliant and a Drug Master File (DMF) is on file with the FDA.

Industry Needs

Effective microbiological monitoring of pharmaceutical manufacturing processes is a necessary step to meet industry standards of product quality.  From a microbiology perspective, the industry should apply Quality Risk Management (QRM) principles in order to design a process to prevent contamination of product while investigating ways that correct contamination events.  QRM is an important part of science-based decisions that are essential for the quality management of pharmaceutical manufacturing.

Similarly, the FDA’s cGMPs for the 21st Century: A Risk-Based Approach, states “that using a scientific framework to find ways of mitigating risk while facilitating continuous improvement and innovation in pharmaceutical manufacturing, is a key public health objective, and that a new risk-based pharmaceutical quality assessment system will encourage the development of new technologies, such as Process Analytical Technology (PAT), to facilitate continuous manufacturing improvements via implementation of an effective quality system”.

In 2003 the FDA introduced the PAT initiative to “encourage the voluntary development and implementation of innovative pharmaceutical manufacturing and quality assurance.” PAT is comprised of four components: data analysis, process analytical tools, process monitoring, and continuous feedback. The quality-by-design aspect of PAT should reduce cycle time thus reducing the wastes of waiting and excess inventory already identified by lean manufacturing.

BioLumix Can Help Meet Industry Needs

Standard microbiological methods require up to seven days to complete.  At every point where microbial testing is carried out (raw materials, work-in-progress, or finished products) batches may be held in quarantine before they are pronounced ready to move to the next product stage.  Waiting for microbial results can tie up working capital and often results in storage expenses and delays in supplying products to the market. Essentially, inventory stays on the floor instead of going out the door to the customer and/or patient population.

The BioLumix Instrument is an example of a new generation Rapid Microbiological Method (RMM).  The BioLumix System is based upon detection of changes in color or fluores­cence due to microbial metabolism in liquid medium within a novel two-zone test vial. An optical sensor monitors these changes within the vial’s reading zone, which is physically separated from the incubation zone. This two-zone approach prevents masking of the optical pathway by product or microbial turbidity and, therefore, eliminates product interference.

The BioLumix System can be used to perform testing for total aerobic count, yeast and molds, and Gram-negative bile-tolerant bacteria as described in USP <61> (USP, 2009a).  The system can also detect objectionable organisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Salmonella according to USP <62> (USP, 2009b). Each of these assays can be simultaneously performed using the same instrumentation. The sensitivity of the BioLumix System is a single viable cell per sample vial; when a single cell replicates to a specific detection threshold level, a positive response is recorded (detection time (DT)). The threshold level is ~100,000 cells/mL for bacteria and ~10,000 cells/mL for both yeast and molds. BioLumix uses the dilute-to-spec protocol, which requires diluting the sample to product release specifications or in-process action levels. If growth is detected, the sample fails; if there is no detection, the sample passes (i.e., the counts are below the specification limit).

 

Summary of Data from the Chapter

  1. Side by side growth comparison to USP<61> was shown for Total Aerobic Counts, Yeast & Molds, and for Gram Negative Bile-Tolerant organisms.
  2. Specificity using Inclusivity and Exclusivity testing was demonstrated for the BioLumix system.
  3. The Limit of Detection was demonstrated using target organisms.
  4. Robustness of the system was demonstrated using the following parameters:  effect of instrument temperature, effect of sample size, and effect of media volume in BioLumix vials.
  5. Ruggedness of the system was demonstrated using the following parameters:  effect of the analyst, effect of reagent lots, and effect of different instruments.

 

References

  1. United States Pharmacopeia (USP) (2009a), Chapter <61>, Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests, The National Formulary, Rockville, MD, The United States Pharmaceopeial Convention.
  2. United States Pharmacopeia (USP) (2009b), Chapter <62>, Microbiological  Examination of Nonsterile Products: Tests for Specified Microorganisms, The national Formulary, Rockville, MD, The United States Pharmaceo­peial Convention.

BioLumix Partnership Program Highlighted During Supply Side Marketplace 2013

Be sure to stop by Booth # 216 during SupplySide Marketplace and learn about the benefits of joining the BioLumix Partnership Program.

The BioLumix Rapid Microbiology System can save multiple days versus current USP testing methods, and dramatically impact key profit drivers, such as elimination of idle time with raw materials, work in progress (WIP), and finished goods.
Here are just a few of the many benefits of the BioLumix Partnership Program:New rapid automated microbial testing methods, such as the BioLumix System, can speed up time to results from 7-10 days to 1-2 days.  The time saved has a direct impact on many operational profits drivers. . . and is THE reason over 400 BioLumix Systems are actively being used by many of the largest manufacturers in the industry.

  • Ready-to-use vials  with specialized media which do not require QC on your end
  • Very Rapid Results—C of A within 48 hours  for expedited product releases
  • Simplified, automated microbiology testing
    • Paperless laboratory
    •   60% savings in labor
    •   Operated by non-microbiologist
  • ALL assays available within one automated system
  • Validated Method in compliance with USP <2021>, <2022>, <2023> and USP <61> and <62>
  • Remote access/support for your system via internet and interactive phone calls to facilitate optimal instrument performance
  • Product reports tailored to specific applications
  • Dedicated microbiologist assigned to support your facility
  • Real-time continuous support for reviewing microbiology assays. . .  and more

BioLumix takes pride in delivering the highest level of customer support for our BioLumix Partners.  Did you know that in a recent customer satisfaction survey, ALL of our customers would recommend us to their vendors, suppliers and customers?

With the BioLumix System, we can help your company internalize microbiology testing.  If you already have an internal laboratory, we can help you save time and money by automating your testing!  Many of our customers have gone through audits and we have received great feedback from them.

We welcome all of our customers to stop by and say hello!  If you are not yet a customer, we welcome the opportunity to work with you.  Our jobs center around satisfying our customers and providing unparalleled support.  We would be happy to give you a demonstration of the system including testing your samples for free.  After all, we wouldn’t want to sell you a microbiology instrument that doesn’t work for you.  Still not convinced?  We would be more than happy to provide you a long list of customer referrals.

Allow us the opportunity to demonstrate the many benefits of the BioLumix Partnership Program.

See you at Booth #216!

BioLumix Exhibits at 2013 PDA Annual Meeting in Orlando, FL

This year’s Annual PDA Meeting –will be held in sunny Orlando at The Peabody Hotel, April 15-17th.  This meeting focuses mainly on present-day manufacturing, challenges in the industry, and those we will face in the future.  The meeting is separated into three tracks so each attendee may pick and choose different areas to learn more about.  Additionally, several planned networking and social activities will bring all kinds of professionals together to ask questions, learn and get to know one another.

FDA Approval - Drug Master FileAttendees will have an opportunity to visit the BioLumix booth – # 404.  The BioLumix System is a rapid, automated microbiological method that is the only system capable of concurrently testing both USP <61> and <62>.  The system provides a single platform for testing raw materials, in-process, finished product, environmental swabbing and processing water.  BioLumix provides a complete screening solution with real-time communication for early warning of contaminated samples (in hours rather than days), results for bacteria in 12-24 hours and Yeast & Mold in less than 48 hours.  Once all testing is complete, an automated Certificate of Analysis is available and can be printed via the printer or connected to a LIMS system.

The BioLumix system, as described in the “Encyclopedia of Microbiological Methods Volume 4” edited by Dr. Michael Miller’s, in Chapter 7, simultaneously detects microbial growth, provides an estimation of viable cell counts and identifies the presence of specified microorganisms.  Additionally, BioLumix has contributed Chapter 14 of “Environmental Monitoring,” edited by Jeanne Moldenhauer, “Validation of a Rapid System for Environmental Monitoring and Water Testing.”  Both books will be available to review at the BioLumix booth #404.

By encompassing both USP chapters (61 and 62), the BioLumix is the all-in-one microbial testing solution for companies looking to save significant time, labor and resources.  The system will bring automation to the current microbial testing! The system is based on optical sensors that detect color of fluorescent changes in ready to use carbon dioxide sensing vials.  These vials eliminate the need to make, prepare and QC traditional media.

BioLumix has a DMF (Drug Master File) on file with the FDA for prescription pharmaceuticals.  For OTC (Over the Counter) products, the system is fully Validatable.  BioLumix installers will generate the IQ (Installation Qualification) and OQ (Operation Qualification) on-site during the 2 ½ day training session.  We will also work with your team to complete the side-by-side testing portion of the PQ (Performance Qualification), leaving your company with a complete Validation package.

Please stop by Booth #404 to speak with a BioLumix expert, view our recent publications, ask questions, get answers and join the growing customer base of BioLumix users! Learn how BioLumix Rapid Microbiology can deliver significant reduction in time to results, reduced staff involvement, leading to faster product release. . . and how these can impact your key “operational profit drivers.” We look forward to earning another satisfied customer.  See you soon at the PDA Annual Meeting!

Microbiological Testing of Enzymes

What are Enzymes?

Enzymes are specialized proteins that accelerate the rates of chemical reactions by lowering the activation energy; the substrates are converted into products.  There are numerous different types of commercially available enzymes such as protease, amylase, cellulose and lipase.

Where are they used?

Many different industries use enzymes in the production of their products.  They range from animal nutrition, brewing industry, food processing, detergents, dairy industry, paper industry, biofuels, contact lens cleaners, photographic industry, carbohydrate processing and biochemicals.  Enzymes are also an integral part of the pharmaceutical and Nutraceutical industry.  In laundry detergents enzymes that degrade proteins causing stains, such as those found in grass stains, red wine and soil can be used.

Lipases are another useful class of enzymes that can be used to dissolve fat stains and clean grease traps or other fat-based cleaning applications.  Nutritional enzymes have risen to prominence during the past several years, driven by customers that would like to improve their digestive health. The biggest category for supplemental enzymes is still human digestion, an area that has become increasingly more important for maintaining optimum health. Supplementation of single enzymes, such as lactose to treat lactose intolerance, is probably the most familiar area of enzyme use. However, general dietary supplementation with a broad range of digestive enzymes is becoming increasingly more common. Supplemental enzymes can also be used to enhance the effect of herbs and botanicals. The active component of herbs and other botanicals can remain trapped within the cell wall matrix or may be in a form that is not easily broken down and absorbed. Select enzymes are added to these products to break open certain bonds to free the bioactive compounds.  The application of enzymes technology to pharmaceutical products is also a growing field.

How do they Work?

Amylases in the detergents can help by aiding in the breakdown of starch stains.  Also in the biofuel industry cellulases are used to break down cellulose into sugars that can be fermented, and then further processed to ethanol.  Trypsin is used in baby food to predigest the food for the babies.   Proteases can also be used in contact lens cleaners to remove proteins on the lens.  The figure below show a schematic diagram of how enzymes work:

Most enzymes require microbiological testing similar to other industrial products.   BioLumix has an easy method to complete this goal.  The versatility of the BioLumix system allows for a wide range of enzymes to be tested for a broad array of microorganisms; such as Total Aerobic Count, E. coli, Salmonella, Staphylococcus aureus, Pseudomonas aeruginosa, Lactic Acid Bacteria, Coliforms, Enterobacteriaceae, and Yeast and Molds. Liquid products allow a direct sample to be pipetted directly into the vial, while solid samples require a dilution step first.

The BioLumix Rapid Automated Method:

Enzymes traditionally can be very difficult to test due to their unique properties.  The two most common difficulties when working with enzymes is solubility and pH levels of the product.  When using the BioLumix system, pH is adjusted once the product has first been diluted at 1:10 in TSB or buffer during the preparation before the sample is placed into the vial.  Solubility issues can be solved in a few different ways. One way is by allowing the product to sit in solution and give it time to breakdown, and another way it to warm the product in a water bath or an incubator.

During enzyme testing at BioLumix, on plates the enzyme created a precipitate that looked very similar to colonies, which could lead to difficulties in reading the plate.  The plate needed to be  sub-cultured to determine if it was growth or just the precipitate.  The precipitate did not interfere with the BioLumix vials and provided a clear answer.  Additionally, the results were obtained two days sooner.  An oil based enzyme can also cause difficulties when using traditional plating methods versus the BioLumix system.  In the BioLumix system the oil does not have an effect on the result and can give a definitive answer.  While on plates, the oils can sometimes appear on top of the agar and give the appearance of colonies.

Another less common issue is gas production by the product when it is mixed with a liquid.  If the product is known to release gas the easy solution is to allow the product to off gas before adding it to the appropriate vial.  Then after an hour of being in the liquid, the sample can be stomached or mixed again to release the remaining gas.  While in traditional methods, the gas production can give false positive results in tests with Durham tubes, which would then require additional days to complete.

BioLumix Advantage:

The BioLumix system is designed to accelerate product release with a simplified, automated approach. This yields fast, accurate, real-time results while reducing costs. The BioLumix offers real-time results of contaminated samples saving hours, possibly days. Completed Total Aerobic Count, Coliform or Enterobacteriaceae results are obtained within 18-22 hours; Yeast and Mold results are obtained within 48 hours; the objectionable organisms can be detected within 24 hours after pre-enrichment. Also these assays are simpler to perform than the standard methods saving time, labor and money. The system is unaffected by product interference, delivering accurate results with faster product release.  When referring to batches of product that will be released days early due to the BioLumix system, which allows the storage and production less time to run and hastens the turnaround time of the product, the overall process of enzyme production will decrease.

The Economics of Adding Automation and Speed to Microbiological Testing

Are you growing tired of waiting on QC in order to release thousands of dollars worth of finished products?       Do you work in the lab and are you receiving inquires from senior management looking for ways to speed up the QC process?

For a nominal investment in automation, the BioLumix Rapid Microbiology System will save valuable days in the QC process, enabling you to optimize operations, increase throughput —AND  directly impact company profitability!

New rapid automated methods, such as the BioLumix System, can speed up time to results from 7–10 days to 1–2 days.  The time saved has a REAL impact on key operation profit drivers, such as elimination of idle time with raw materials, work in progress (WIP), and finished goods.

TIME IS MONEY. . . a few of the advantages of the simplified automated microbiology system includes:

Faster detection of microbial contamination results in faster corrective action.  This will result in faster substitution of out of spec raw materials and faster resumption of production of high quality product.  Faster detection of problems can result in less production of defective products.  A single incident of faster intervention can save more than the cost of a new rapid detection system.

Speed up your microbiology testing and eliminate idol time thereby speeding up production and reducing inventory.  By significantly decreasing the manufacturing cycle, you can lower the required working capital investment.  Increase your manufacturing capacity through a faster release of inventory.  By implementing a rapid micro method, you can re-allocate new found floor space to increase manufacturing capacity, add more SKU’s to inventory, or reduce distribution center costs.

The BioLumix straightforward, streamlined testing design and rapid, accurate results lead to reduced material-holding time for faster product release.   Early warning of contaminated samples as well as sample release information could be automatically communicated through the internet, significantly improving your company’s efficiencies.

 

The BioLumix Advantages include:

  • Single-platform testing for all assays
  • Real-time communication—early warning of contamination
  • 48-hour Automated Certificate of Analysis
  • Simplified testing with ready-to-use vials
  • Validated for your product—IQ, OQ, PQ
  • No product interference
  • 21 CFR Part 11 compliant software
  • Technology based on traditional USP media
  • Remote-access customer support 24/7/365

BioLumix Exhibits at Engredea March 2013!

Join BioLumix at booth #224 in Anaheim March 8th – 10th, 2013 for Engredea 2013 at the Anaheim Convention Center.

Engredea encompasses innovation, cutting-edge technology and services making it a leader in the global nutrition tradeshow industry.Biolumix Exhibit  This year suppliers and manufacturers come together to offer hundreds of exhibits, networking events and educational opportunities for the industry.  Food, beverage, dietary supplements and nutricosmetic exhibits will be present.

The state of the art validated BioLumix system allows easy compliance with cGMP will be featured in Engredea 2013.  With over 400 instruments in action world-wide, the cost-effective BioLumix system revolutionizes your current testing methodology. The BioLumix System can save multiple days versus current testing methods, and dramatically impact key profit drivers, such as elimination of idle time with raw materials, work in progress, and finished goods. It will streamline your microbiological testing – saving significant time, labor and money. It is fully automated including data archiving, data maintenance, and report generation. It is paperless and efficient-saving on disposables, time and space. The system eliminates product interference, delivering accurate results.

Are cGMP Regulations Driving Up Your Costs?

Learn how we can save you time and money!  Streamline your microbiological testing, reduce cost, increase efficiency and see rapid return on investment with the BioLumix system.  BioLumix is fully automated, validated against USP methodology, 21 CFR Part 11 Compliant that allows for complete microbiology testing results within 24-48 hours, allowing for faster product release time, shorter sample prep time – saving time, money and other resources.  Faster detection of microbial contamination results in faster corrective action.

Why Visit us at Booth #224?

Be sure to stop by the BioLumix booth (#224) to view a product demo, meet some of our customers, learn the benefits that the system brought to them and inquire about our show special pricing.  Be sure to examine our numerous product literature and scientific white papers.

The BioLumix System is completely modular, able to handle the microbiological testing needs of both large and small companies.  In many cases a Return-On-Investment can be proved within 6 months.  We would like to show you how the BioLumix System will make your company more efficient and add profits to your bottom line!  Don’t just take our word for it, speak to our customers directly.

Learn how:

Faster detection of microbial contamination results in faster corrective action

Speeding up your microbiology testing can eliminate idol time thereby speeding up production and reducing inventory

Reduce Warehouse space when you obtain micro results in half the time.

We look forward to the opportunity to earn another satisfied customer!

Tackling those challenging Nutraceutical Products with Rapid Automated Microbiology Testing!

Spirulina, turmeric, Psyllium, honey, Calcium carbonate, dyes, and oils.  These are just a few of the many products that the Nutraceutical industry works with, and struggles with.  However, microbiology quality assurance requires any such product to be tested.  Nutraceutical products come in a wide variety of colors and compositions that can make it difficult to process the samples and count colonies on plates.  Some products cause the coloring on plates to be so dark that one cannot see the actual colonies, other samples have particulate matter that resembles colonies or hides them altogether making counting plates time consuming and inaccurate.  The BioLumix vials not only streamline microbiological testing, but also make the challenging samples easier to work with due to the fact that color, particulate matter, and sticky samples do not interfere with the ability of the system to detect growth of organisms. Below are a few examples of samples that many struggle with:


Psyllium is the husk of the seed of a Plantain and is used to improve digestion and cleanse the digestive system; it is an excellent dietary supplement.  However, before it reaches the shelves for sale, the product needs to be tested in the laboratory, to be sure it is free of harmful microorganisms and is safe for consumption.  This particular product is a spongy fiber, and when it is added to an enrichment broth or diluent, it begins to swell, and harden into a sticky substance that is very difficult to pipette and mix with the agar on the plate. Figure 1 shows the lack of mixing of 1.0 ml of a 1:10 dilution of the Psyllium with the agar.

To avoid this sticky situation, necessary dilutions can be performed quickly and added to the appropriate BioLumix vials before the product has a chance to harden.  Even though, the product will become a bit harder after being added to the BioLumix vial, the growth of organisms will not be inhibited.
Spirulina and turmeric – both colorful raw materials (and different dyes used to color capsules) may make it difficult or even impossible to see and count colonies on pour plates as can be seen in figure 2.  Because the BioLumix CO2 sensor is impermeable to liquids and colors, no product interference due to color of the product will impact the results.

Particulate matter is always a concern during sample preparation, and rightfully so.  As shown in Figure 3, it is sometimes impossible to tell the difference between product and colony.  The BioLumix vial is not affected by particulate matter because only CO2 gas can penetrate the CO2 sensor. As a result, there is no product interference from particulate matter present in the sample.

The Problem: The inability to achieve homogenous mixtures, excess color, and particulate matter are three major causes of inaccurate plate count results when testing Nutraceutical and dietary supplement products for microbiology.

The Solution: The BioLumix vial is able to support growth of organisms in a favorable broth environment, and these problems do not interfere with results.   The BioLumix System is an automated microbial testing system designed to accelerate product release with a simplified, approach.  This yields fast, accurate results while reducing costs.  No more waiting days for completed assays.  The BioLumix System brings automated microbiological testing in house with most assay results achieved over night.  It is simple enough to be operated by non-microbiologists, providing accurate data of raw materials, environmental samples, and finished products.

Way to Increase profits with faster automated microbiology

Looking For Ways to Increase Profits in 2013?
Unlock the QC bottleneck?

New rapid automated methods, such as the BioLumix system, can speed up time to results from 7-10 days to 1-2 days. The time saved has a direct impact on key operational drivers.Way to Increase profits with faster automated microbiology

TIME IS MONEY…a few of the advantages of the simplified automated microbiological system includes:

The BioLumix Rapid Microbiology System can save multiple days versus current testing methods, and dramatically impact key profit drivers, such as elimination of idle time with raw materials, work in progress (WIP), and finished goods.

Faster detection of microbial contamination results in faster corrective action. This will result in faster substitution of out of spec raw material and faster resumption of production of high quality product. Faster detection of problems can result in less production of defective products. A single incident of faster intervention can save more than the cost of a new rapid detection system.

Speed up your microbiology testing and eliminate idol time thereby speeding up production and reducing inventory. By significantly decreasing the manufacturing cycle, you can lower the required working capital investment.
Increase your manufacturing capacity through a faster release of inventory. By implementing a rapid micro method, you can re-allocate new found floor space to increase manufacturing capacity, add more SKU’s to inventory, or reduce distribution center costs.

Reduce Warehouse space when you obtain micro results in half the time. Waiting for micro testing is expensive. When considering an investment in rapid micro testing, it is critical to weigh the nominal incremental testing cost versus the significant financial benefits that accrue to operations.

The BioLumix straightforward, streamlined testing design and rapid, accurate results lead to reduced material-holding time for faster product release. Early warning of contaminated samples as well as sample release information could be automatically communicated through your intranet, significantly improving your company’s efficiencies.

The BioLumix Advantages include:
• Single-platform testing for all assays
• Real-time communication – early warning of contamination
• 48-hour Automated Certificate of Analysis
• Simplified testing with ready-to-use vials
• Validated for your product – IQ, OQ, PQ
• No product interference
• 21 CFR Part 11–compliant software
• Technology based on traditional media
• Remote-access customer support 24/7/365

Biolumix Automation Drives Profits

BioLumix Automation Drives Profits!

No longer look at micro testing of raw materials and finished products as a “necessary evil” in remaining cGMP compliant.   Rather, with BioLumix Rapid, Automated Microbiology, you can view microbiology product testing as a position of company strength and competitive advantage!

TIME IS MONEY. . .  we have detailed financial models prepared by our current customers that demonstrate the dramatic impact BioLumix is having when you apply Multiple Days Saved to real profit drivers, such as:  

Biolumix System

  •   Savings as a result of early detection of contamination
  •   Lower cost of inventory
  •   Increase capacity of existing factory and lab
  •   Reduce cost of storage
  •   Lower cost of test related overhead
  •   Savings on cost of capital
  •   Potential improvement in account receivables. . .  all of which directly impact profits!

Automation Drives Profits
The enacted cGMP regulations require manufacturers to ensure that products are free of contamination by objectionable organisms, and that appropriate microbiological testing is conducted.

BioLumix Rapid Microbiology offers a high degree of automation, significant reduction in time to results, reduced staff involvement, and faster product release. . .  all which impact key “operational profit drivers,” such as:

  • Savings as a result of early detection of contamination
  • Lower cost of inventory
  • Increase capacity of existing factory and lab
  • Reduce cost of storage
  • Lower cost of test related overhead
  • Savings on cost of capital
  • Potential improvement in account receivables

 

Contact Us TODAY To Learn More About Our FREE DEMO PROGRAM!

Thank you for your Continued Loyalty to BioLumix!

As we near the close of our sixth year in business, we would like to take this opportunity to express our appreciation to all of our valued customers. 2012 was a great year for us at BioLumix, as we experienced tremendous growth. During this year more and more companies have adopted our novel streamline technology and enjoy accelerated product release with our simplified, automated microbiology.
BioLumix is proud to have almost 400 systems in place throughout the world, as more and more companies are adopting our rapid microbiology testing system. Thank you for choosing BioLumix as your partner in microbiological testing!
For those that are looking to add new building blocks for improved company profitability in the coming year, we hope that you will allow us the opportunity to demonstrate how the BioLumix automated method for rapid micro testing can positively impact your company’s bottom line.
We would like to invite you to take a tour of our system that allows you to test raw materials, finished products, including powders, oils, enzymes, botanical material, tablets, capsules, and liquids easily and without product interference. With automated monitoring of ready-to-use assay vials and automated data processing and archiving (paperless), the microbiologist‘s job got a lot easier with the same accurate results in less than half the time.
Our entire organization wishes you a wonderful holiday season, a prosperous New Year and we look forward to earning many more satisfied customers in the coming year. Happy Holidays and Best Wishes for a bright and successful year ahead, filled with joy and treasured moments.

Significant Tax Benefits if the BioLumix System is Purchased Prior to the End of Year!

The BioLumix Advantage

BioLumix can help internalize the microbiology testing or make the existing microbial testing simpler, faster and automated- saving you significant time, labor and money. With BioLumix you can test raw materials, in-process and finished products as well as processed water and environmental testing.  The system can analyze tablets, capsules, powders, liquids and viscous material without any product interference and automatically generate a certificate of analysis.

While enjoying all these advantages you can take advantage of the IRS Section 179 deduction, if Agreements are reached to purchase a BioLumix System prior to the end of this year.

Most people think the Section 179 deduction is some mysterious or complicated tax code.  It really isn’t, as you will see below. Essentially, Section 179 of the IRS tax code allows businesses to deduct the full purchase price of qualifying equipment and/or software purchased or financed during the tax year.  That means that if you buy (or lease) a piece of qualifying equipment, you can deduct the FULL PURCHASE PRICE from your gross income.  It’s an incentive created by the U.S. government to encourage businesses to buy equipment and invest in their company.

2012 Limit on equipment purchases = $560,000.
This is the maximum amount that can be spent on equipment before the Section 179 Deduction available to your company begins to be reduced.

 

Bonus Depreciation = 50%
This is taken after the $560k limit in capital equipment purchases is reached. Note: Bonus Depreciation is available for new equipment only. Bonus Depreciation can also be taken by businesses that will have net operating losses in 2012.

 

Now is the Time

The first-year bonus depreciation, which applies to new business equipment, has been set at 50 percent through 2012, and will drop to zero percent starting January 1, 2013. Lastly, the current cap on total capital equipment purchases is $560,000, and will decrease to $200,000 in 2013.

Here is an example of Section 179 at work:

 

 

Who Qualifies for Section 179?

All businesses that purchase, finance, and/or lease less than $560,000 in new or used business equipment during tax year 2012 should qualify for the Section 179 Deduction. If a business is unprofitable in 2012, and has no taxable income to use the deduction, that business can elect to use 50% Bonus Depreciation and carry-forward to a year when the business is profitable.  The deduction begins to phase out if more than $560,000 of equipment is purchased – in fact, the deduction decreases on a dollar for dollar scale after that, making Section 179 a deduction specifically for small and medium-sized businesses. However, as noted above, large businesses can expense all qualifying capital expenditures with 50% Bonus Depreciation for the 2012 tax year.

 

What’s the difference between Section 179 and Bonus Depreciation?
The most important difference is both new and used equipment qualifies for the Section 179 Deduction (as long as the used equipment is “new to you”), while Bonus Depreciation covers new equipment only. Bonus Depreciation is useful to very large businesses spending more than $560,000 on new capital equipment in 2012. Also, businesses with a net loss in 2012 qualify to deduct some of the cost of new equipment and carry-forward the loss. When applying these provisions, Section 179 is generally taken first, followed by Bonus Depreciation – unless the business has no taxable profit in 2012 because the unprofitable business is allowed to carry the loss forward to future years.

This is not legal, tax or financial advice.  You should consult with your tax advisor for the specific impact to your business.  For additional information, please visit www.irs.gov.  The information above is an overall, “simplified” view of the Section 179 Deduction for 2012. For more details on limits and qualifying equipment, please consult your tax attorney or financial advisor.

 

The debate: Coliforms, Fecal coliforms, and Enterobacteriaceae as Indicator Organisms

Which one Should you Use?

What are Indicator Organisms and why use them?

Indicator organisms are organisms used as a sign of quality or hygienic status in food, Dietary supplements, water, or the environment. The initial goal in finding a safety indicator was to find a group of bacteria that could indicate the presence of fecal material and serve as a surrogate for Salmonella, but was easier and simpler to detect. Such a group may signify the potential presence of pathogens, a lapse in sanitation as required in good manufacturing practices (GMPs), or a process failure.

The longest used indicator organism was the coliform group that was recommended for use in the early 1900s for water testing. Fecal coliforms and E. coli followed as more specific indicators of potential presence of pathogens. The Pasteurized Milk Ordinance includes a requirement of coliform testing of pasteurized for milk and milk products.

Many different types of safety indicators have been proposed for use in particular applications. A thorough review of the indicator organisms is given in Tortorello (2003).

Definitions

Coliforms

Coliforms are gram negative, oxidase negative, non spore-forming, aerobic or facultative anaerobic rod shaped bacteria. The coliform group is not a distinct valid taxonomic group, but is defined functionally as organisms that ferment lactose with both gas and acid production at 35°C. The coliform members include Citrobacter, Enterobacter, Escherichia, and Klebsiella. Some also add to the group Serratia and Hafnia to the coliform group. Many of these bacteria are found naturally in the intestines of humans and animals, and some are even found naturally in soil and water. However, of the 1% of coliforms found naturally in the human gut, E. coli represents the majority and is found exclusively in the intestines of humans and animals. It is important to note that many of the coliforms can be found also in plants and the environment, thus, a positive coliform test does not necessarily indicate fecal contamination.

Enterobacteriaceae

The family Enterobacteriaceae encompasses approximately 20 genera, including E. coli and all members of the coliform group; in addition it includes foodborne pathogens Salmonella, Shigella, and Yersinia. The family was originally proposed as an indicator alternative to the coliform group because testing for the entire family would be more inclusive for the pathogenic bacteria. The Enterobacteriaceae may be superior to coliforms as indicators of sanitation GMPs because they have collectively greater resistance to the environment than the coliforms. This group is more widely used as indicators in Europe than in the United States. The determining factor separating coliforms from Enterobacteriaceae is the ability of coliform to ferment lactose, while the Enterobacteriaceae family ferments glucose.

Fecal Coliform

These organisms are a subset of the total coliform group. The fecal coliforms have the same properties as the coliform group, except that the fermentation is able to proceed at 44.5°–45.5°C. They are considered a better indicator of fecal contamination than the coliform group.

E. coli

E. coli is present in all mammalian feces at high concentrations; it does not multiply appreciably, but can survive in water for weeks, and so it is useful as an indicator of fecal pollution of drinking water systems. E. coli meets all the criteria used for the definition of both total coliforms and fecal coliforms. In addition, the organism can be distinguished from other fecal coliforms by the lack of urease and the presence of B-glucuronidase enzymes.

When to test and how to test

There are some regulations in various industries that require testing of one or several of these organisms. In some industries, product manufacturers use these indicators to assure that there is no lapse in sanitation or process failure.

Coliform

This test is required in the dairy industry, bottled water and drinking water. Many producers in the food industry also utilize the coliform test, especially in the USA. The BioLumix Coliform vial (CC-C) is useful for water testing, for food, and in some cases also in dietary supplements. The coliform vial contains lactose as the sole carbon source and selective ingredient such as bile salts. The assay is simple to perform – just add the appropriate amount of the liquefied sample to the ready-to-use vial and run it in the instrument at 35°C for 16-18 hours.

Enterobacteriaceae

USP(ENUMERATION TESTS—NUTRITIONAL AND DIETARY SUPPLEMENTS) recommends the testing of Enterobacterial Count (Bile-Tolerant Gram-Negative Bacteria) in dietary supplements. Most European producers prefer to use this test instead of the coliform test for foods. The BioLumix Enterobacterial vial (ENT-C) is very similar to the Coliform vial, except the medium contains glucose in addition to lactose. It is mainly used for dietary supplement industry.

Fecal coliforms

The fecal coliform test is used instead of the coliform test in industries where it is considered to be more directly associated with fecal contamination from warm-blooded vertebrates than are other members of the coliforms, such as in seafood, nuts, etc. The BioLumix coliform vial is used for this application however is incubated at an elevated temperature of 44.0°C.

E. coli

E. coli testing is required in drinking water systems. Also, USPsuggests the testing of Dietary supplement for the absence of E. coli in 10 grams of product. In the meat industry there are regulations relating to the presence of generic E. coli on carcasses.

The BioLumix E. coli vial (EC) contains a highly selective medium and includes MUG that is the most commonly used fluorogenic substrate for the detection of E. coli. It detects the activity of glucuronidase by E. coli. For tests with a specified level of 10 cfu/g of product, the liquefied sample can be added directly into the EC vial and the vial is run in the instrument at 35°C for 18-20 hours. For tests with a specified level of absent in 10 grams, the samples are diluted 1:10 in TSB for 18-24 hours followed by the addition of 0.1 mL of sample into the EC vial.

BioLumix has them all!

The BioLumix system is ideal for testing of any of the indicator organisms. With the BioLumix automated microbiological system, users have control over their testing, leading to accurate results and accelerated product release while reducing costs. The BioLumix Advantage:

  • All assays can be performed on one Automated Microbial testing system
  • Fast, automated results of all assays
  • Can be operated by non-Microbiologist
  • Complies with FDA cGMP Regulations
  • System is validated during system installation
  • Designed to accelerate product release
  • Fully automated data archiving
References:
  • Ockerman, C. “Rapid Microbiological Testing of E. coli with the BioLumix Vial” (2012). BioLumix Blog.
  • Tortorello, M. “Indicator Orgnanisms for Safety and Quality – Uses and Methods for Detection: Minireview.” Journal of AOAC International 86 (2003): 1208-1217.
  • United State PharmacopeiaENUMERATION TESTS—NUTRITIONAL AND DIETARY SUPPLEMENTS (2005). The National Formulatory . Rockville MD.
  • United State PharmacopeiaMICROBIOLOGICAL ATTRIBUTES OF NONSTERILE NUTRITIONAL AND DIETARY SUPPLEMENTS. (2005). The National Formulatory . Rockville MD.

Pseudomonads and Their Rapid Detection

Description of Pseudomonad Organisms

Pseudomonas bacteria (Pseudomonads) encompass gram negative, motile, non-fermenting rods. This genus is ubiquitous in nature and these organisms can impact a number of environments and patient populations. The Pseudomonads may be found in soil, on plant material, in water, and can be isolated from various tissues and body fluids from mammals. In human health, some of these organisms, primarily Pseudomonas aeruginosa, can be an opportunist pathogen and cause serious health problems. If allowed to reach unsafe levels, this organism may cause several health problems including skin rash and other skin infections, ear infection, urinary tract infection, and in rare instances, pneumonia. Other Pseudomonads, for example, P. stutzeri can be isolated from wounds but are generally not associated with human disease. Many Pseudomonads found in the soil can damage plant materials by causing spoilage.

Who tests for Pseudomonas and why?

Water Testing: Pseudomonas aeruginosa is a bacterium commonly found in purified water systems. Pseudomonas grows in water. It thrives at warm temperatures, which is why it is so often associated with spas. It can also grow in purified water systems.

Pharmaceutical and Cosmetic Products: Analysis of FDA product recall data for 134 non-sterile pharmaceutical products from 1998 to September 2006 demonstrated that 48% of recalls were due to contamination by either Burkholderia cepacia, or Pseudomonas spp (Jimenez L. 2007). In cosmetic products, P. aeruginosa was recovered from contaminated mascara material and was identified as the agent responsible for corneal ulcers in the 1970s (Ortho 2009). Pseudomonads can survive and grow in DI water—Contaminated DI water may be the source of microbial contamination if it is used for the final rinse of equipment that has been cleaned and sanitized, and it may be the source of contamination for finished products in these industries.

Dairy and Food: The predominant microorganisms limiting the shelf life of processed fluid milk at 4°C are Pseudomonas spp. these species are able to grow to high numbers during refrigerated storage. Pseudomonas species accounted for79% of the psychotropic isolates that spoiled pasteurized milk (Dogan and Boor 2003). Important characteristics of Pseudomonads include their abilities to grow at low temperatures (3–7?C) and to hydrolyze and use large molecules of proteins and lipids for growth.

Biolumix Offers Two Options for Detecting Pseudomonads

For certain industries it is important to detect Pseudomonas aeruginosa, while for others it is important to detect all Pseudomonas spp, including the closely related Burkholderia cepacia. As a result BioLumix offers two different types of vials: the PSE vial for the detection of P. aeruginosa; and the PSB vial for the detection of all strains of Pseudomonas and for B. cepacia.

Detection of Pseudomonas aeruginosa (PSE Vial)

For the Pharma (OTC), Cosmetic and Nutraceutical Industries the primary cause for concern is the absence or presence of Pseudomonas aeruginosa. P. aeruginosa is common and is able to become an opportunistic pathogen in people and may cause severe disease (Hugh and Gilardi 1974). The ability to detect P. aeruginosa is critical in the non sterile Pharmaceutical products, Cosmetic and Nutraceutical Industries to ensure the product material is safe. BioLumix offers a highly selective media in the form of a test vial (PSE) that primarily only allows for the growth of Pseudomonas aeruginosa organisms. Confirmation of the presence of this organism is accomplished using the simple Oxidase reaction on vial contents. The test sample is merely enriched in TSB (Tryptic Soy Broth) per USP instructions and then tested directly in the BioLumix PSE vial. Other common Pseudomonads and closely related organisms, including B. cepacia and P putida, as examples, are excluded from growth due to the use of antibiotic supplements in the BioLumix PSE vial. P. aeruginosa is typically more antibiotic resistant than other Pseudomonas organisms (Blazevic, DL et al 1973). Figure 1 illustrates the growth curve of Pseudomonas aeruginosa ATCC 9027 in the BioLumix PSE vial.

KEY:Dark Blue Curve- P. aeruginosa Green Curve- Negative Control

Detection of other Pseudomonads (PSB Vial)

For many industries including the dairy industry and manufacturers using water, there is a need to test for all Pseudomonads as they impact these industries economically. Other Pseudomonads may include P. fluorescens, P. putida, and P. stutzeri. Burkholderia cepacia, can also be detected using the BioLumix PSE-B vial. Specific to the use of water in manufacturing: Pseudomonas bacteria can be found naturally in the ground and within drinking water sources such as aquifers. Contamination of either dairy products or water systems by Pseudomonads is something to avoid and early detection of goods using a rapid microbiological detection system such as the BioLumix Instrument System, would offer an advantage to the manufacturer. Figure 2 illustrates the growth of many types of Pseudomonads and Burkholderia cepacia in the BioLumix PSE-B vial.

KEY: Dark Blue Curve- B. cepacia; Green Curve -P aeruginosa; Light Blue Curve – P. putida; and Red Curve– P. fluorescens growth

REFERENCES

Blazevic, DJ, Koecke, M.H., and Mastsen J.M. (1973). Incidence and identification of Pseudomonas fluorescens and Pseudomonas putida in the clinical laboratory. Applied Microbiology 25: (1)

Dogan, B. and Boor, K J. (2003). Genetic diversity and spoilage potentials among Pseudomonas ssp. isolated from fluid milk products and dairy processing plants. Appl. Microbiol.,69: 130-138.

Hugh, R. and Gilardi, G. (1974) In “Manual of Clinical Microbiology” Edited by Spaulding, Lennette, Spaulding and Truant. Chapter 23 Pseudomonas.

Jimenez L.(2007). Microbial diversity in pharmaceutical product recalls and environments. Review. PDA J Pharm Sci Technol. 2007 Sep-Oct;61(5):383-99.

Ortho D. (2009). Insight into Cosmetic Microbiology, Chapter 8 263-267

Rapid Microbiological Testing of Osmophilic Yeast with the BioLumix Vial

Osmophilic or osmotolerant yeasts are able to grow in products containing high concentrations of organic solutes, particularly sugars. These organisms do not require a high water activity value and tolerate drier environments than non-osmotolerant organisms. Osmophilic yeast can cause spoilage of honey, corn syrup, jam, molasses,flavored syrups and toppings, and concentrated fruit juices. Many of the yeasts causing this spoilage belong to the genus Zygosaccharomyces. It is of great importance in the food industry to test for these organisms. Although no osmophilic organisms are highly pathogenic, they may cause food poisoning and opportunistic infections, especially in people with weakened immune systems.

Current Methodology

Although there are many techniques used to test and enumerate Osmophilic yeast, a standard method has not been attained. Different types of growth media have been suggested, but like the techniques, not one type of agar has been adopted as the standard. Using the plate count method, samples are blended with sterile phosphate buffer containing high concentration of glucose. Petri dishes are then inoculated with the appropriate dilution of the product, and agar medium that would promote growth of the Osmophilic yeast is added, depending on the type of product. Plates are incubated for 5-7 days. Membrane filtration technique can also be utilized to test the Osmophilic yeast. Typically, a 25-gram sample is added to the flask and mixed well with sterile distilled water. After filtration, provided that the product can be filtered, the walls of the flask and funnel are rinsed with the distilled water, and the filter placed on a desired growth medium and incubated for 5-7 days. Both the plate count and membrane filtration techniques pose threats to damaged organisms from general processing procedures, temperature of agar, and temperature, pressure and duration of incubation. Colonies may also be very small and difficult to count.

The BioLumix Osmophilic Yeast Vial

The BioLumix system provides Osmophilic Yeast results 3 days faster than the aforementioned methodologies. The Osmophilic Yeast vial has a CO2 sensor that is monitored by the yellow LED light in the BioLumix Instrument. This particular vial contains much more sugar than a typical Yeast and Mold vial, to allow only Osmophilic yeast to grow. The vial also contains Chloramphenicol to prevent the growth of bacteria.

The procedure to test for Osmophilic yeast is very simple; just add 10 grams of product to 90mL Tryptic Soy Broth or buffer, and add the appropriate amount to ready-to-use Osmophilic vials. For some product the sample can be added directly into the vial. From this 1:10 dilution, other dilutions can be made if necessary. From the appropriate dilution, 0.1 to 1.0 mL is added to the BioLumix vial. compared to the plate method. Figure 1 shows curves generated by the Zygosaccharomyces rouxii ATCC 34890 (blue) and 28253 (green). organisms while preventing product interference. It offers a reduced work load, faster time-to-results, and fewer costs associated with testing. The BioLumix system can make the microbiological testing simpler, faster and automated. The test runs for 48 hours, cutting the time-to-results by three days as The BioLumix system allows for selectivity of organisms while preventing product interference. It offers a reduced work load, faster time-to-results, and fewer costs associated with testing. The BioLumix system can make the microbiological testing simpler, faster and automated.

References:

Downes, Frances P., and Keith Ito. Microbiological Examination of Foods. 4th ed. Washington, DC: American Public Health Association, 2001.

Why New Rapid Automated Methods Are Desired?

Newer microbiological methods can improve company’s quality of testing and result in significant financial savings.  Such methods can reduce cycle times resulting in a leaner and more responsive supply chain.  At every point where microbial testing is done (raw materials, work-in-progress, or finished products) batches may be held in quarantine for up to seven days before they are pronounced ready to move to the next stage.  Waiting for microbial results can tie up working capital and results in storage expenses and a delay in supplying products to the market.  The faster results can identify a contamination and enable implementation of corrective action, and cost savings.  Consequently, an increasing number of pharmaceutical companies are becoming interested in adopting RMM’s.  By utilizing rapid methods for microbiology, manufacturers can dramatically slash the amount of time products must be held for microbiology testing and drive new efficiencies with less capital tied up in finished goods inventory, and reduced warehouse space requirements.