Water testing- Heterotrophic bacteria, coliforms and E. coli- Why and how to test

Water Quality

Water QualityWater is used in a variety of different industries as well as products within various industries, including Nutraceutical and Dietary Supplement, Pharmaceutical, cosmetics, toiletry industries.  Water can be used as a product ingredient, for example, to create the capsules that contain the supplement.  In the manufacture of the capsules many companies use their own water to create and encapsulate their products.   Water is also used for the cleaning of certain equipment and contact surfaces.

According to USP 1231, although there are no absolute microbial standards for water (other than water intended to be sterile), the CGMP regulations require the establishment of appropriate specifications. The specification must take into account the intended use of the water; i.e., water used to formulate a product should contain no organisms capable of growing in the product. Action or alert limits should be established based upon validation data and must be set low enough to signal significant changes from normal operating conditions.

Control of the microbiological quality of water is important for many of its uses. All packaged forms of water are required to be sterile because some of their intended uses require this for health and safety reasons. The needed microbial specification for a given bulk water depends upon its use. Some applications may require even more careful microbial control to avoid the proliferation of microorganisms ubiquitous to water during the purification, storage, and distribution.

To ensure adherence to certain minimal microbiological quality standards, water used in the production of drug substances or as source or feed water for the preparation of the various types of purified waters must meet the requirements of the National Primary Drinking Water Regulations (NPDWR) (40 CFR 141) issued by the U.S. Environmental Protection Agency (EPA) or the drinking water regulations of the European Union or Japan, or the WHO drinking water guidelines. Microbiological requirements of drinking water ensure the absence of coliforms, which, if determined to be of fecal origin, may indicate the potential presence of other potentially pathogenic microorganisms and viruses of fecal origin. Meeting these microbiological requirements does not rule out the presence of other microorganisms, which could be considered undesirable if found in a drug substance or formulated product.

USP<1115> deals with bioburden of non-sterile drug substances and products, and the chapter states that the biggest manufacturing risk is water as an ingredient.  Process water is the single most important risk factor contributing to the contamination of nonsterile products.  The purified waters that are used in manufacturing are deionized and do not contain chlorine that helps control microbial growth.  Purified water is capable of supporting growth of gram negative rod shaped bacteria and many different molds.

Water TestingThe FDA also covers a wide range of different types of water that can be used for pharmaceutical uses and describes different sources for water contamination.  The FDA even states that microbial contamination of oral liquids and topical drug products are a significant problem that is usually caused by contaminated water.  Due to the potential health risks involved with the use of contaminated water, particular attention should be paid to the deionized (DI) water systems, especially at smaller manufacturers.

Chlorinated water may be appropriate for early stage cleaning and sanitization activities, but the uses are risky and should only be used on a case by case basis.  Microbial enumeration is an integral component of a water monitoring system to assess the microbial quality of the water.  Some systems use both high-nutrient (PCA) and low-nutrient (R2A) media to allow the isolation of both heterotrophic organisms and slower growing oligotrophic bacteria.

Water testing is also important when dealing with well water, tap water and even bottled water.  The EPA uses coliform as an indicator of possible fecal contamination.  Coliforms naturally found in the environment, and are usually non-pathogenic, but their presence may indicate fecal coliforms.

The Rapid Automated BioLumix System

BioLumix SystemBioLumix automated; all-in-one microbial testing system is an ideal system for in plant water testing.  The system is fast, simple and cost-effective.  A novel optical system sensing color and fluorescence in ready-to-use vials provides faster results, labor savings, automation, and connectivity. The BioLumix system is capable of testing water for heterotrophic bacteria, total aerobic bacteria, E. coli, coliforms, fecal coliforms and yeast and molds. Using the BioLumix system will quickly determine the microbial quality of the water.

Heterotrophic Vial: This vial can detect organisms requiring low-nutrient media (similar to (R2A) to allow the isolation of both heterotrophic organisms and slower growing oligotrophic bacteria. In a study, over 50 samples of multiple different water types were tested by the BioLumix method and the plate count method side-by-side.  The BioLumix vials were directly inoculated with 0.1 mL of the water sample, or a 1.0 mL of a 1:100 dilution, and a few samples were inoculated with heterotrophic bacteria.  The samples were monitored in the BioLumix instrument for 35 hours.  The results showed that the BioLumix system was roughly 13 hours faster than the plate count method using Stand Methods Agar.  These particular samples were tested at specified levels <10 cfu/ml and <100cfu/ml, but the BioLumix method can detect organisms at levels of <1 cfu/ml of water.

Bottled water for human consumption also needs to be tested for coliforms, which are indicators of possible contamination. The FDA requires either MPN or membrane filtration to check 100 ml of water for any contamination. The MPN method which requires at least nine tubes to perform the test and up to 96 hours of testing; while BioLumix can do the same analysis using just one vial in less than quarter of the time.  The filter method can also be applied using the BioLumix system by filtering the 100ml onto a membrane filter and placing the filter directly into the vial.

What are the advantages of the BioLumix system?

The system serves, as a platform to perform all required assays- using the BioLumix system will allow the users to test for coliforms, heterotrophic bacteria, E. coli and Yeast/Mold. The system can be used for water testing as well as for testing raw materials, in process and finished products.

Saving time- The BioLumix system can save time when testing water for Heterotrophic bacteria instead of taking three days using traditional plates, the BioLumix system will give the same results in 35 hours.

Economical cost of assays: Instead of running an MPN assay, which will require up to 5 days of testing as well as 9 tubes of LTB and up to 9 tubes of EC Media to wait for confirmation of a positive fecal coliform, the BioLumix system requires less than 24 hours and a single vial.

References:

http://www.fda.gov/ICECI/Inspections/InspectionGuides/InspectionTechnicalGuides/ucm072925.htm -Water for Pharmaceutical Use

http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm064948.htm  Enumeration of Escherichia coli and the Coliform Bacteria

USP <1115> Bioburden Control of Nonsterile Drug Substances and Products

USP <1231> Water Treatment Systems For Industrial & Commercial Use.

TIPS FOR SUCCESSFUL ENVIRONMENTAL MONITORING

Putting together an environmental monitoring plan requires a thoughtful, scientific approach establishing and considering the risks of contamination from each location along the manufacturing process.
environment monitor
WHY MONITOR THE ENVIRONMENT?

Environmental monitoring (EM) is a basic requirement across many regulated industries such as food, cosmetics, Nutraceutical, dietary supplements, and pharmaceuticals. It measures the degree of maintaining environmental control and, therefore, the safety of the manufactured products. Maintaining environmental control can prevent product contamination.

The FDA evaluates the environment of manufacturing facilities against the regulatory code, and the potential of the product allowing for growth of organisms. They expect manufacturers to be in control of the environmental conditions in their facility. There is evidence that a relationship exists between the level of environmental control and the final quality of the product. In product safety, the EM program serves a critical role that the environment is under appropriate control.

In manufacturing facilities, there is a need to demonstrate that production equipment is sufficiently clean thereby the next production lots are not contaminated from the material of the previous lot.

An effective sampling plan needs to be established, the purpose of a microbial EM program is to:
• Provide crucial information on the quality of the work process environment during manufacturing
• Prevent future microbial contamination by detecting and reacting to adverse trends
• Prevent the release of a potentially contaminated batch if the appropriate standards are not met
• Prevent the risk of contamination of the product
• Ensure there are environmental controls in the production areas
• Provide a profile of of the microbial cleanliness of the manufacturing environment.

SITE SELECTION FOR DATA COLLECTION

The site’s selection for data collection should show the effectiveness of cleaning. Such as:
• Site where the product is exposed to people and equipment
• Critical sites that can change product integrity if compromised
• Areas and processes steps where microbial contamination must not happen, such as the final filling of the product into its containers.

In establishing an EM plan, the number and location for sampling sites should be established. When choosing the sampling sites, there needs to be a good representation of risk-based sites where the environment can affect the product quality. Examples of such location include areas that are difficult to clean, or areas close to critical operations.

For sterile products, the manufacturer needs to keep the bioburden of the pre-sterilization as low as possible. There is a need to monitor the air, water, personnel, and surfaces as they all can contribute to the bioburden.
For non-sterile, low bioburden products, there are two main reasons for monitoring the environment:
1. To keep product bioburden under control
2. To know if the environmental isolates could contain objectionable organisms.

Other important questions to consider include:
? What is the antimicrobial effectiveness of the product?
? Will the product promote microbial growth?
? If so, which organisms can grow and get around the inhibitors build into the products?

ESTABLISH A BASELINE

Before initiating an EM program, it is recommended to get a baseline of total aerobic count, Yeast and mold count, perhaps coliform or Enterobacteriaceae, and in some instances lack of objectionable organisms on surfaces and the air. Establishing a baseline also allows for assessments of the types of organisms present, and helps in developing a scientifically sound disinfection program to address chemical kill and physical removal of such contaminants. The data subsequently collected during microbiological performance qualification, and routine monitoring helps to validate the efficacy of disinfection and cleaning procedures.

CHOOSE SUITABLE METHOD FOR EM

method for monitoring - swabsMost EM is done by plate counting of colonies on agar media, which is simple and inexpensive. However, plate count methods are slow, requiring two to seven days to complete, thereby causing a delay in the detection of contamination, which can lead to an increase in product loss, plant downtime and expensive cleanup. Delays can cause increasing inventory holding cost. The delay in obtaining results impacts reaction to contamination issues and can make investigations very difficult. The plate count methodology is also labor-intensive and requires manual data entry and documentation. Such documentation is prone to human errors and compliance issues.
Methods are available to measure total particles in the air, including Total Organic Carbon (TOC), and Adenosine Tri-phosphate (ATP).These methods are very fast to perform but do not correlate well with total bacterial count or any specific group of organisms, and do not measure viable organisms (Carricket al., 2001; Easter, 2010). Therefore, these results do not measure viable organisms in the environment or on production lines.

WHY BIOLUMIX

why BioLumixThe BioLumix system is useful for rapid and simple monitoring of the manufacturing environment. A large validation study was performed (Eden and Brideau 2012) to show that is correlates well with the plate count method. The BioLumix system was validated as an alternative to the plate count method for EM. The study involved 549 surface coupons representing five diverse types of material. These five surfaces represent those encountered in manufacturing, including metal, plastics and rubber. Some of the coupons were inoculated with bacteria, yeast, or mold. There was 100% agreement between BioLumix assay and the plate count assay for the 260 coupons that were determined to be below the specified level by the plate count method. There was an overall agreement of 97.2%between the two methods when swabs containing counts above the specified level were used. In general, discrepancies in swab results between the BioLumix vial method and the traditional plate count method reflected marginal samples that were very close to the specified testing level, and thus were variable.

REFERENCES

Carrick, K, Barney M, Navarro, A. and Ryder D. (2001). The Comparison of Four Bioluminometers and Their Swab Kits for Instant Hygiene Monitoring and Detection of Microorganisms in the Brewery. J. Institute of Brewing 107, 32-37
Easter M. (2010) A comparison of commercial ATP hygiene monitoring systems. Next Generation Food issue 9, 2010.
Eden, R. and Brideau, R. (2012). Validation of a Rapid System for Environmental Monitoring and Water Testing. In Environmental Monitoring Volume 6, Jeanne Moldenhauer, Ed

Come and See us at the IAFP in Indianapolis, IN August 3-6, 2014; Booth 610

The 2014 Annual International Association of Food Protection (IAFP): The IAFP meeting will provide attendees with information on current and emerging food safety issues, the latest science, innovative solutions to new and recurring problems. It also provides an opportunity to network with thousands of food safety professionals from around the globe. This meeting has grown over the years to become the leading food safety conference worldwide.
More than 2,800 scientists, from six continents, attend the IAFP Meeting. This event owes its reputation and success to the quantity, quality and diversity of each year’s program; the quality and relevance of exhibits sharing the latest in available technologies; leading experts speaking on a variety of timely topics.
All of us at BioLumix want to take this opportunity to invite you to our exhibit (booth #610) and show you what’s new at BioLumix. We continue to innovate, growing our assay repertoire and capabilities – allowing you to perform all required assays on raw ingredients and finished products, as well as test environmental samples and process water.
During the IAFP show, we will feature:
New Listeria vial: The vial helps detect Listeria in environmental swabs. The Listeria vial capitalizes on the ability of Listeria to hydrolyze esculin, which in the presence of ferric ions, results in the yellow broth turning black and a reduction of the absorbance. A Listeria detection obtained with the BioLumix system can be further verified using a simple and quick Rapid Listeria Immunostrip Test. There was 100% agreement between the BioLumix system and plate methodology when product swabs contained Listeria.
Enhanced B. cereus vial: Recently the inhibitory system of the B. cereus vial was enhanced to eliminate the detection of non-target organisms. It uses the CO2 sensor at the bottom of the vial. The new BC vial recovers well vegetative cells as well as spores. The method used is very simple; just pipette the diluted sample into the vial. The results are available in 24 hours or less.
Direct inoculation of products into vials: Many products can be introduced directly into the BioLumix test vial without the need of a dilution step. These include yogurts and sour cream, as well as milks. Complete coliform test results are obtained within 12 hours with one vial substituting for nine or more MPN (Most Probable Number) test tubes. Yeast and mold results are obtained in 48 hours rather than 5-7 days using plate methods.

The yogurt can be directly added (Figure 1) into BioLumix vials to measure growth of Coliforms or Yeast and Molds. Specialized high pH BioLumix vials can also be used for yogurt samples. When a low pH yogurt sample is added to the BioLumix high pH Coliform vial (CC), the pH conditions become near neutral. This ensures the yogurt manufacturer to be able to correctly test for coliforms using a direct (without dilution) sample of product. As much as 1 gram of product can be directly tested in each BioLumix test vial.

With automated monitoring of ready-to-use assay vials, along with automated data processing and archiving, the microbiologist’s job gets a lot easier with the same accurate results in less than half the time. Our innovative vial design prevents product interference even when directly testing products, such as yogurt and salad dressing. Come to our booth (#610) or contact us directly (734-984-3100) to learn more about the exciting new developments we are featuring at this year’s IAFP.

The BioLumix Advantage
• Vial design prevents product interference
• Automation and connectivity allows faster product release
• Real-time communication for immediate action
• Expedited results: most results in 12-18 hours; Yeast and mold assay in 48 hours
• Automated data archiving and audit trail
• Streamlined testing increases laboratory efficiencies
• Paperless laboratory: centralized test data automatically stored and protected
• Barcode capability for automated sample entry
• Environmental testing made easy

Free Product Trial
Give us your most difficult samples and we will test them for free. We will provide you with a detailed report which includes a side-by-side comparison to your current manual methodology. The data generated is strictly confidential and is only used to show the high correlation of results should your company decide to purchase.

We look forward to working with you and earning another satisfied customer!

The versatility of the BioLumix System – One system for all your microbial needs.

Why go to an automated system?

There has been a growing awareness over the years that utilizing the current methodology for determining microbial contamination is not cost effective due to the slow turnaround.  Internalizing the microbiology testing and especially adopting rapid microbiological methods (RMM’s) can speed up significantly the time to results from 7-10 days to 15-48 hours.  An automated detection system provides contamination information earlier than the conventional method, products can be released faster, manufacturing at risk is minimized, and investigations can be conducted and concluded closer to the time of the actual contamination.  The obvious benefit of an automated system is the quicker release of finished products, where the shorter ‘time to result’ may generate considerable cost savings. Some of the other advantages of RMM can include greater accuracy, better sensitivity, increased sample throughput, and automated data capture allowing easier data handling and paperless laboratory.

Why You Should Invest in the BioLumix System?

BioLumix SystemTime is money.  For a nominal investment in automation, the BioLumix Rapid Microbiology System will save valuable days in the QC process: optimize operations; increase throughput; and directly impact company profitability!  The BioLumix system allows for all of your microbiology needs to be met with one automated system.  It can be used to test Raw, In-process, and Finished Materials along with environmental and water samples.  The BioLumix system allows the user to test as frequently and as broadly as desired without significant cost or delays. The results are increased capacity along with cost reduction, improvement of logistics, and overall better efficiencies! With shorter sample preparation time and automated data entry, data archiving, report generation, and product release, the BioLumix system simplifies and automates your laboratory procedures. Tests can be also be performed by non-microbiologists, providing significant savings in laboratory labor.

Advantages of the BioLumix System

1)      All assays performed on a single system.  This allows laboratories to purchase one system but offer and perform many different tests, thus eliminating the problem of finding space for multiple systems in the lab and training personnel on multiple instruments.

a)      Microbial results can be generated in 24-48 hours in assays such as Total Aerobic Count, Yeast and Mold, Escherichia coli, Salmonella, Staphylococcus aureus, and Pseudomonas aeruginosa while utilizing one system.  The system is unaffected by product interference and delivers accurate results. BioLumix is dedicated to producing affordable and easily performable assays that laboratories of all sizes can integrate into their daily work flow and yearly budget.

b)      BioLumix has improved the way microbiology can be performed in the cosmetic, toiletry and pharmaceutical industries.BioLumix has improved the way microbiology can be performed in the cosmetic, toiletry and pharmaceutical industries.  BioLumix offers the first automated PET and Microbial Limits system on a single platform, without any product interference! Preservative Efficacy Testing or Challenge Testing (USP<51>), Microbial Enumeration Testing (USP<61>), and Testing for Specified Microorganisms (USP<62>) can be easily performed without the hassle of preparing and counting multiple plates that would be expected using the standard methodology yielding faster results and being more cost effective.

c)      Environmental Monitoring.  In manufacturing facilities there is a need to demonstrate that the production equipment and environment are sufficiently clean so that the next production lot will not be contaminated by the material from the previous lot, removing the potential of cross contamination. One needs to prove with high degree of certainty that the cleaning process was effective. To do so an effective sampling plan needs to be established.  The BioLumix system can be utilized for rapid and simple monitoring of the manufacturing environment.

d)    Water Testing. Water is widely used as a raw material, ingredient, and a solvent in the processing, formulation, and manufacture of pharmaceutical products, active pharmaceutical ingredients and intermediates.Water Testing As such, all water purification systems must be monitored regularly to verify the quality of the water produced. Monitoring of water for microbiological quality may include testing for total heterotrophic plate count, coliforms/E. coli, or by checking for the presence of other organisms suspected to be present in a water sample. The relevant standards relating to pharmaceutical grade water are USP <1231> Water for Pharmaceutical purposes.

2)      Scarcity of skilled personnel: Microbiology laboratories face major challenges such as the growing scarcity of skilled laboratory workers and the burden of the ever-increasing workload.  The BioLumix system requires limited training to successfully set up the tests.  The system is fully automated including archiving of data, data maintenance and report generation, and it can be used to create a paperless laboratory.

3)      In house testing allows a laboratory to be more cost effective.  Increased turnaround time may be one of the most appealing features of an automated system.  Contamination can be caught quicker and actions can be taken immediately so that products can be released faster into the marketplace.

4)      The Validated BioLumix System.  Since June 25, 2010, all dietary supplement manufacturers have been required to comply with the Food and Drug Administration’s (FDA) current good manufacturing practices (cGMP’s) according to the guidance outlined in USP chapters <2021>, <2022>, and <2023>.  The BioLumix system has software that is 21 CFR Part 111 compliant and BioLumix generates a customized validation book for each of its customers that include:

a)      Installation qualification (IQ):  Identification and validation of the system components; validation of the environmental conditions; electrical requirements; computer qualification; verifying that all installation steps were followed; and documentation of instrument calibration.

b)     Operational qualification (OQ):  Verifying that the equipment is properly installed calibrated and is operational.  It includes a unique SOP for all products and assay combinations to be performed on the instrument; software characteristics and the verification that the software is 21 CFR part 11 compliant; verification that all the instrument functions operate as expected; Verification of the instrument temperature accuracy; and training records.

c)      Performance qualification (PQ): is the most extensive part of the BioLumix validation book. It shows equivalency with USP methodology when following USP <1223> “Validation of alternative microbiological methods”.

 

 

References:

1)       United States Pharmacopeia Chapter  <51> Antimicrobial Effectiveness Testing

2)       United States Pharmacopeia Chapter  <61> Microbiological Examination of Nonsterile Products:  Microbial Enumeration Tests

3)       United States Pharmacopeia Chapter  <62> Microbiological Examination of Nonsterile Products:  Tests for Specified Microorganisms

4)       United States Pharmacopeia Chapter <1231> Water for Pharmaceutical Purpose

5)       United States Pharmacopeia Chapter <2021> Microbial Enumeration Tests – Nutritional and Dietary Supplements

6)       United States Pharmacopeia Chapter <2022> Microbial Procedures for Absence of Specified Microorganisms – Nutritional and Dietary Supplements

7)       United States Pharmacopeia Chapter <2023> Microbial Attributes of Non-Sterile Nutritional and Dietary Supplements

8)       United States Pharmacopeia Chapter <1223> Validation of Alternative Microbiological Methods

PRESERVATIVE EFFICACY TESTING: GUIDELINES TO AN AUTOMATED SIMPLIFIED TESTING SYSTEM

Introduction

The ultimate purpose of the Preservative Efficacy Test (PET) is to determine the effectiveness of the preservative(s) present in a cosmetic, toiletry, or pharmaceutical product.  USP Chapter <51> ANTIMICROBIAL EFFECTIVENESS TESTING describes the type of products to be tested (categories), the specified microorganisms to be used for testing, and the inoculum and log reduction amounts required per category of product.  The product to be tested is inoculated with a high number of bacteria, yeast, and mold, and the reduction in the initial inoculum amount is calculated over a 28 day period.

Current Methodology

Until now, the only way to perform PET was by the plate count method.  Typically, a product is inoculated with a high number of organisms (usually 105 - 107), and after 7, 14, 21, and 28 days, samples of that inoculated product are tested to determine the log reduction that occurred to the organisms when subjected to the preservative system.  Because it is not known how effective the preservative system is, serial dilutions have to be plated to determine the number of organisms remaining in one gram of product.  As a result every organism require a number of plates, dilution bottles and tips as shown in figure 1.

Advantages of BioLumix Simplified Automated System

The main advantages the customer gains using BioLumix are savings on time, labor, and materialsPreservative Efficacy Test

The BioLumix assay (Figure 2) takes about 75% less hands-on labor as compared to the standard plate count method, and the simplicity of the BioLumix method is unparalleled.  Ease of use, less materials (as seen in comparing Fig1 to Fig2), and less dilution reduces the chance of error.  After inoculation of product with pure cultures, the BioLumix method allows the operator to perform testing in 4 easy steps:

  1. Weigh 1.0 gram of inoculated product to a sterile sample bag.
  2. Add 9.0 mL neutralizing broth and allow to sit for up to 45 minutes.
  3. Add 1.0 mL to the BioLumix vial.
  4. Enter the vial into the BioLumix Instrument and begin the automated testing.

Pre-programmed calibration curves as shown in Figure 3 are used to generate the colony forming unit count per gram of product based on the detection time of the curve.  This eliminates tedious counting of multiple plates and interpretation of results when working with thick or waxy products, or products containing materials that may resemble bacterial or yeast colonies.  The BioLumix method also saves time in obtaining results.  For each day of sampling, BioLumix reduces the time-to-results for bacteria from 48 hours to 24 hours and from 5-7 days for yeasts and mold to 48 hours.  The BioLumix system uses a single vial to replace a number of plates, dilution bottles and pipet tips, thereby reducing disposable costs.calibration curves

Example of Results Obtained

The example, show the results of PET obtained with eye drops with and without benzalkonium chloride.  The results obtained by both the BioLumix system (BL) and the plate count methodology are shown in the two table below.

results obtained by both the BioLumix system (BL) and the plate count methodology

results obtained by both the BioLumix system (BL) and the plate count methodologyThe data by both methods shows that in the product with the inhibitor benzalkonium chloride (Table1) after 7 days all bacteria are reduced by more than 5 log cycles, as are the counts of Candida. It took 14 days for Aspergillus to be reduced by 5 log cycles.

When the product did not contain benzalkonium chloride (Table 2) and as a result was not properly preserved, only P. aeruginosa was reduced by 5 logs after 7 days. E. coli took 14 days to reduce the numbers by 5 logs. The other organism had a very slow reduction over time.  The counts of Aspergillus were reduced by less that 2 log cycles.  The results show that the two methods yield very similar results.

Many other products were tested with the BioLumix PET methodology including scrubs, creams, lotions, Shampoos, hand and facial cleansers to name a few, and equivalency with the plate count methodology and reproducibility of results was demonstrated for all products.

The BioLumix system allows the operator to save time, materials, and money by drastically cutting hands-on labor and time-to-results.  If offers labor reduction by 75-80% of the labor required for the Petri dish method, with much less disposable used, faster time to results and good correlation with current methodology.  It allows for the creation of a paperless laboratory.  The system automation provides automated data achieving, and automated reporting including log reduction calculations.

Growth Promotion and Negative Controls – Does Shipping Affect the Results?

Why Growth Promotion?

USP Chapter <61>, Microbiological Examination of Non-sterile Products: Microbial Enumeration Tests ,there is a requirement for the testing of Growth Promotion (GP) for each batch of medium as well as testing for Negative Controls (NC).  Growth Promotion must be tested for each new batch of nutritive broth.  The purpose of the GP Test is to determine the suitability of the culture media for the growth of target organisms.  The medium is challenged with a small number (not more than 100 cfu) of microorganisms to assure its nutritive properties.

USP Chapter 61USP Chapter <61> lists the following acceptance criteria for liquid medium: “Liquid media are suitable if clearly visible growth of the microorganisms comparable to that previously obtained with a previously tested and approved batch of medium occurs.” It is recommended to use parallel testing; in parallel testing the new medium and the previously approved medium are inoculated with the same microorganism suspension, by the same technician, using the same method and same environmental conditions.  The only variable is the medium. GPT is performed by inoculating the product with ? 100 CFU of microorganisms, defined by the pharmacopoeia.

BioLumix Quality Assurance and Certificate of Analysis

For every batch of vials prepared by BioLumix a thorough testing, including growth promotion and negative controls, is done.  Growth promotion is tested performed, for each organism, by inoculating 4 vials of the new lot and comparing the detection time (DT), and amplitude of the curve to a lot that previously passed the PG. At least 2 different strains of organisms are used in each media.  Additionally at least one negative control organisms is tested and compared to the reference.  In the growth control assay the DT of the new lot has to be within a range of 50-200% from the reference vial.  A certificate of analysis is issued with each lot of vials.

Is there a need to redo GP and NC after shipping the vials to the end users?

In other words, does the shipping of the vials to the end users influence the GP and NC of the vials? The study described below was undertaken to answer this question.  BioLumix vials are always shipped UPS second day air.  The furthest destination in the USA is California.  The study was conducted by double shipping second day air to California and back.

Summary of Validation Data

Methodology-Quadruplicate vials that were shipped to California and were returned from there where compared to vials that stayed in the lab. This test was performed on three different lots of media, using three different media types (Total Aerobic Count, Enterobacteriaceae, and Yeast and Mold Vials).

Results-The data shows that the shipped vials and the vials that stayed in the laboratory results of the test show identical growth curveshad identical growth curves, falling close to each other.   There is no difference in growth promotion or negative control between vials that were shipped (actually double shipped) and vials that stayed on the shelf for any of these lots, as can be seen from the attached example.  The example compares the curves of total aerobic count vials (TAC) inoculated with Bacillus spizizenii var subtilis ATCC 6633, shipped to vials that remained in the laboratory (Key: Shipped vials: Purple, Yellow, Beige and Dark Green; vials that remained in the lab: Dark Blue, light Green, Light Blue and Red lines). As can be seen from this example the Curves are indistinguishable

Similar results were obtained for all three lots of TAC inoculated with Bacillus spizizenii var subtilis ATCC 6633; Escherichia coli ATCC 8739; and Staphylococcus aureus ATCC 6538;

A buffer was used as a negative control for the TAC vials, and no difference was seen for all three lots between shipped and vials that remained in the lab.

Similar results were obtained with combined yeast and mold vials (YM) inoculated with Aspergillus brasiliensis ATCC 16404; Candida albicans ATCC 10231; Saccharomyces cerevisiae ATCC 9763; and Staphylococcus aureus ATCC 6538 (SAT1) as negative control.  There is no difference in growth promotion or negative control between vials that were shipped and vials that stayed.

The shipping experiment was also conducted with Bile-Tolerant Gram-Negative Bacteria vials (ENT) inoculated with Escherichia coli ATCC 8739; Salmonella enteritidis subsp. enterica serovar enteritidis ATCC 13076; Citrobacter freundii ATCC 8090; and Staphylococcus aureus ATCC 6538 as a negative control. There was no difference between the data generated by the system for shipped vials, as compared to vials that remained in the laboratory.

 Conclusion

The data shows that the shipped vials and the vials that stayed in the laboratory had identical growth curves, falling on top of each other.   There is no impact of shipping on growth promotion or negative control.  The results validate that there is no reason to perform again the growth promotion and negative controls after shipping to the end user. The C of A supplied with the BioLumix vials is indicative of Growth Promotion and Negative Control followed the shipping.

BioLumix Simplified, Rapid, Automated Microbiology on Display at Ingredient Marketplace 2014 in New York

BioLumix Simplified, Rapid, Automated Microbiology on Display at Ingredient Marketplace 2014 in New York

May 5, 2014 by Linda Schmaler & Paul Dudley
Since its beginnings in 2006, BioLumix, Inc. has been on the leading edge of technology in the world of microbiology.  On June 2 – 3, 2014, that technology will be on display at Ingredient Marketplace 2014 at the Jacob Javits Center in New York City.  This year, BioLumix has added a new assay to its already robust line of test vials: Listeria

Ingredient MarketplaceSupplySide Marketplace has changed its name to Ingredient Marketplace, and is built upon the theme, “What’s Inside Matters.”  With consumers seeking safer, healthier products, manufacturers today are more focused than ever on providing their products to satisfy their demands while at the same time coming up with effective ways to improve both their products and the means by which they are brought to market.  This is where BioLumix can make a difference!

The BioLumix System monitors changes in broth medium in which target organisms grow.  Microbial metabolism causes the reagents on the proprietary vial to change their color or fluorescence, and real-time monitoring of these changes by optical sensors provide immediate feedback.  This means that a sample that tests above spec is known right away.  The optical sensor monitors vials in the testing unit 10 times per hour.

BioLumix provides test vials for a wide range of assays:

  • Total Aerobic Count
  • Coliform
  • Enterobacteriaceae
  • Yeast and mold count
  • Gram negative bacteria
  • Heterotrophic bacteria
  • Sterility testing
  • Preservative efficacy test
  • Microbial limits
  • Suitability testing
  • Pseudomonas aeruginosa
  • Staphylococcus aureus
  • Escherichia coli
  • Listeria
  • Bacillus cereus
  • Lactic acid bacteria

Listeria VialThe impact of the BioLumix automated rapid microbiology system can be realized in a wide variety of benefits:

  • Ready-to-use vials come with a Certificate of Analysis
  • All confirmation steps are done directly from the vial
  • Certificate of Analysis (C of A) for all assays produced within 48 hours
  • Customized Validation Package in compliance with USP <2021>, <2022>, <2023>, or USP <61> and <62>
  • Automated real-time monitoring for rapid results
  • Ease-of-use – the system can be operated by non-microbiologists, which can allow microbiologists to perform other tasks
  • Centralized data processing and archiving for easy management of results
  • Same-day Total Aerobic Counts and Coliform results
  • Yeast and Mold results within 48 hours
  • Objectionable organisms can be tested on the same platform

These benefits translate into significant business advantages, such as early detection of problems, faster product release, product validation, improvements in factory and lab capacity, lower cost of inventory, and reduced cost of capital.  In addition, all installations include real-time, 24/7 support by a dedicated BioLumix microbiologist.

Please join us at Ingredient Marketplace 2014, and be sure to stop by our booth (417).  For more information, visit http://www.mybiolumix.com.

Why is it important to test Pet Food for microbiology?

Pet Food microbiologyThe pet food industry is nearly a $22.2 Billion dollar a year industry and projected to almost double by the year 2017.  Nearly 101 million homes have at least one pet in the household, and there are nearly 170 million cats and dogs owned in the United States alone.  New trends in pet food are emerging as consumers want to give their pets the freshest food possible and make sure that it is healthier for them as well.  In the past few years there have been multiple outbreaks related to pet food affecting the health of both pets and humans.  Most people associate Salmonella as a bacterium linked to food borne illness in people food, but in recent years there have been quite a few outbreaks of Salmonella in pet food that has also affected humans.  The most concerning aspect is that it primarily caused illness in small children.

Several recalls of pet food due to Salmonella happened in the recent past as shown in the examples that follows. On February 5, 2014 – Pro-Pet LLC, has initiated a voluntary recall of a limited number of Dry Dog and Cat Foods for possible Salmonella contamination. A single field test indicated products manufactured during a two-day period, on a single production line might have the potential for Salmonella contamination1.  On January 25, 2014 – PMI Nutrition, LLC (PMI), has initiated a voluntary recall of its 20 lb. bags of Red Flannel® Cat Formula cat food for possible Salmonella contamination2. On November 4, 2013 – Bailey’s Choice LLC, had recalled its 5 oz. packages of chicken treats because they have the potential to be contaminated with Salmonella, an organism which can cause serious and sometimes fatal infections in young children, frail or elderly people, and others with weakened immune systems3.

The CDC also added an information page on keeping people and pet healthy and safe from Salmonella4.  There was also a pet food recall based on an aflatoxin contamination.  The Center for Disease Control (CDC) categorizes aflatoxin as a naturally occurring fungal toxin that contaminates maize and other types of crops during production, harvest storage or processing5.  The aflatoxin outbreak was linked to the death of over a hundred pets.  In the past year Kroger stores recalled a wide variety of pet foods due to a possible contamination caused by aflatoxin4.

Microbiology Testing of Pet Foods

Why test for indicator organisms? It is more effective to test for indicator organisms rather than to test for pathogens such as Salmonella.  Indicator organisms are used to measure potential fecal contamination of environmental samples. The presence of coliform bacteria, such as E. coli, is a common indicator of fecal contamination. Indicator organisms are typically used to demonstrate the potential presence or absence of groups of pathogens. The use of indicators is attractive because it reduces the complexity and cost of analyzing. Indicator bacteria are selected for the following reasons:

1) They are initially abundant in the matrix to be assayed.

2) A relatively rapid, accurate, and cost effective analytical method for enumerating the indicator exists or can be readily developed.

3) A reasonably strong correlation exists between the presence/absence of the indicator and a particular pathogen or group of pathogens. The strength of the correlation will determine the effectiveness and accuracy of the indicator as a measure of pathogen occurrence.

4) Indicator organisms can be used to pet food manufacturing to cleanliness and sanitary issues within the facility.

Assays Performed on pet foods: in pet food, testing is conducted for Enterobacteriaceae or fecal coliform as indicator of fecal contamination and yeast and mold as indicators for general quality and aflaxoins.

What are the advantages of the BioLumix system?

The system serves, as a platform to perform all required assays- using the BioLumix system will allow the pet food manufacturers to test their products not only for Salmonella and yeast/molds, but also for indicator organisms such as coliforms, fecal coliforms, Enterobacteriaceae and more.

Saving time- The BioLumix system can save time when testing pet food products for Yeast and Mold, instead of taking five days using traditional plates, the BioLumix system will give the same results in under 48 hours.  This can help the manufacturers to avoid a potential aflatoxin contamination by knowing if their product contains any amount of mold.

Economical cost of assays: Instead of running an MPN assay, which will require up to 5 days of testing as well as 9 tubes of LTB and up to 9 tubes of EC Media to wait for confirmation of a positive fecal coliform, the BioLumix system requires less than 24 hours and a single vial.  Finally, the last confirmation step is to streak the positive EC Media to L-EMB agar plates; the BioLumix system instead requires one test vial and 1ml of the sample in order to detect a level as low at <10 cfu/gram, and can give results in under 24 hours.  Similarly, the Enterobacteriaceae test in BioLumix requires one vial instead of multiple MPN tubes required by the European method.

Screening Products: BioLumix Rapid Microbiology Testing can also be helpful in screening products to determine what the next steps are.  Some manufacturers sample the product from the line and test for total aerobic count. If the level is below a certain number, then the product can be sent out to the market, if it is above the specification level then it has to go through a special sterilization procedure which costs more money as well as a delay in the product reaching the customer.

BioLumix Pet Food Study

BioLumix originally conducted a study of different store bought pet foods, ranging from dry dog food samples to wet (oil based) samples.  All samples matched the results for Yeast/Mold, Enterobacteriaceae, Total Aerobic Count, E. coli and fecal coliforms when comparing between the BioLumix System and traditional plating methods.  The products were processed and tested using FDA-BAM methods7.

Total Aerobic Count: There was 100% agreement between the two methods for all samples tested. Fourteen samples were below the specified level by both methods.  One sample was above the specified level by both methods.  One sample was inoculated to show the ability of the system to detect positive samples.

Yeast and Mold Count: There was 100% agreement between the two methods. Fifteen samples were below the specified level by both methods and two samples were above the specified level by both methods. One samples was inoculated with yeast or mold to show the ability of the system to detect positive samples.

Enterobacteriaceae: There was 100% agreement between the two methods. Thirteen samples were below the specified level by both methods and two samples were above the specified level by both methods.

E. coli: Fifteen products were tested for E. coli at a level of Absence in 10 grams.  There was 100% agreement between the two methods. Fifteen samples were below the specified level by both methods. One sample was inoculated with E. coli and were detected as containing E. coli by both methods

Salmonella: Ten products were tested for Salmonella at a level of Absence in 25 grams.  There was 100% agreement between the two methods. Ten samples were below the specified level by both methods after a confirmation step. One sample was inoculated with Salmonella and was detected as containing Salmonella.

BioLumix has also conducted a study using fresh pet food, which is an emerging product in the marketplace.  The study yielded similar results as the initial BioLumix study, except Lactic Acid Bacteria was also tested.

The BioLumix System showed a high correlation between the instrument results and the BAM methodology.  It simplified the microbiological testing, offers a significant reduction in time to obtain results and reduces hands-on labor due to its automation and simplicity of use.  The time to results for bacteria was hours rather than days while yeast and mold required only 48 hours instead of 5 days.

  1. http://www.fda.gov/Safety/Recalls/ucm384876.htm
  2. http://www.fda.gov/Safety/Recalls/ucm374043.htm
  3. http://www.cdc.gov/features/salmonelladrypetfood/
  4. http://www.cdc.gov/features/salmonelladrypetfood/
  5. http://www.cdc.gov/nceh/hsb/aflatoxin/
  6. http://www.prnewswire.com/news-releases/kroger-recalls-pet-foods-due-to-possible-health-risk-112125284.htm
  7. http://www.fda.gov/food/foodscienceresearch/laboratorymethods/ucm2006949.htm

Are You Ready for an FDA Inspection? Surviving the Audit

When it comes to facility audits, tensions run high, even for the most prepared.  When your facility is being audited, the documents used for evaluation come from the Code of Federal Regulations, 21 CFR 111 (Current Good Manufacturing Practice in Manufacturing, Packaging, Labeling, or Holding Operations for Dietary Supplements).  These documents review specifications, responsibilities to meet those specifications, written procedures, and laboratory operations.  Here, we focus on the microbial aspect of these requirements.

Microbiology Lab Ready for FDAWhat specifications must you establish?

For dietary supplements, specification limits must be set on contaminants that may adulterate (change) or lead to adulteration of the in process or finished batch.  Documentation as to why these specifications will help to ensure contaminants will not lead to adulteration should be in place.  It is your responsibility to ensure the established specifications are met and properly documented.

What steps should be taken to make sure specifications are met?

Appropriate testing should be in place to be sure specifications are within compliance.  You may also rely on a certificate of analysis (C of A) from the supplier of the component that you receive provided you first qualify that supplier’s certificate of analysis through testing.  If you rely on the suppliers C of A, it should include a description of the testing performed, specifications, and actual results of the test.  Periodic auditing of C of A results is required, and should be part of the microbial testing.  To ensure the finished product specifications are met, the in-process procedure should be in place to monitored and make sure that  unanticipated contamination does not occur at any stage.

Requirements for Laboratory Operations

Written procedures (Standard operating Procedures-SOP), for laboratory operations for tests and examination of product must be in place.  Lab operations should ensure that all lab material specifications and dietary supplement specifications are met.  Test methods should be well documented and validated, and sample-collecting plans should also be in place and followed.

Out of Specification Operations

If specifications are not met, a procedure (SOP) must be present and followed to correct for any error made during processing or testing.  A modified method of testing may be necessary.

Microbiology Lab RequirementsThe Validated BioLumix System

BioLumix generates a customized validation book for each of its customers that include:

Installation qualification (IQ):  Identification and validation of the system components; validation of the environmental conditions; electrical requirements; computer qualification; verifying that all installation steps were followed;  and documentation of instrument calibration.

Operational qualification (OQ):  Verifying that the equipment is properly installed, calibrated and is operational.  It includes a unique SOP for all products and assay combinations to be performed on the instrument; software characteristics and the verification that the software is 21 CFR part 11 compliant; verification that all the instrument functions operate as expected; Verification of the instrument temperature accuracy; and training records.

Performance qualification (PQ): is the most extensive part of the BioLumix validation book. It shows equivalency with USP methodology when following USP <1223> “Validation of alternative microbiological methods”.

BioLumix has some quick tips to be sure that your BioLumix system passes inspection without any fuss.

How to Use the BioLumix Validation Book?

Your customized validation book is a major component in your system, just like the instruments and vials themselves.  Make sure the person in charge of validation knows exactly where the validation book is.  Also, be sure that all pages of the book requiring signatures, are signed by the appropriate personnel, and the operator has a good grasp on the meaning of each attachment.  The training log of attachment 9 should show that appropriate training was provided to all operators.  All SOPs used in product testing must also be included in the book.

Side-by-Side Testing

Attachment 16 in the validation book shows your company’s own product testing and the data generated not only with the BioLumix system, but with the USP plate count method as well.  It generally starts with the report that was generated when samples were first sent to BioLumix for testing.  Since then, your company should have added to this data to show continuity between the BioLumix and plate count methods.  Attachment 18 is much more in depth, showing a wide variety of products, vial types, and specification limits.  This data verifies the comparability between the two methods on a broad spectrum.

Vials and Reagents

Make sure that the vials and reagents in use are within their expire dates.  Also be sure the Certificate of Analysis matches the vials.  It may seem like common sense, but when days get busy, some things may be overlooked.  Be sure all reagents are stored at the proper temperature conditions as specified by BioLumix.

Optical Calibration

It is important that your instruments be calibrated optically at least every 6 months, and full temperature and optical calibration be performed annually.  These documents should be held in Attachment 8 of your validation book.  This will show that care was taken to keep the instruments performing at optimal levels.

By following these guidelines, you should be ready for your audit.  As always, BioLumix is just a phone call away with any questions or concerns you or the auditor may have.

It is important for all to know that BioLumix customers pass FDA microbiological audits without any problems.

BioLumix Exhibits at Engredea March 2014!

Join BioLumix at booth #616 in Anaheim March 7th – 9th, 2014 for Engredea 2014 at the Anaheim Convention Center.

Engredea 2014: Anaheim, CAEngredea brings together the community of leading suppliers and manufacturers of new ingredients, packaging, technologies, equipment, and services in the global nutrition industry. Engredea, co-located with Natural Products Expo West, is the only trade show that brings together the full food chain of supply to shelf.

The BioLumix Rapid Microbiology System  delivers significant reduction in time to results, reduced staff involvement,  and faster product release.  BioLumix can save multiple days versus current testing methods and dramatically impact key profit drivers. The validated BioLumix system allows for easy compliance with cGMP. BioLumix will streamline your microbiological testing saving significant time labor and money.  BioLumix delivers shorter sample preparation, automated real-time communication, fast product release and early warning of contamination.  It is paperless and efficient, saving on disposables, time and space.  The system eliminates product interference, delivering accurate results. Engredea 2014: Anaheim, CA

Driving Profit Ability!
With BioLumix Rapid, Automated Microbiology you can view microbiology product testing from a position of company strength and competitive advantage!  Streamline your microbiological testing, reduce cost, increase efficiency and show rapid return on investment.  The BioLumix system is fully automated, validated against USP methodology and 21 CFR Part 11 Compliant.  The system delivers complete microbiology testing results within 24-48 hours, allowing for faster product release time, shorter sample prep time – saving time, money and other resources.

Why Visit us at Booth #616?

Be sure to stop by the BioLumix booth (#616) to view a product demo and inquire about our special show pricing.  Be sure to examine our numerous product literature and scientific white papers.

We would like to show you how the BioLumix System will make your company more efficient and add profits to your bottom line!  For a nominal investment in automation, the BioLumix Rapid Microbiology System will:

Save valuable days in the Microbiology Quality Control Process

Optimize Operations

Increase Throughput    AND…..

Directly Impact Company Profitability!

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

BioLumix Validation Book for Dietary Supplements

According to cGMP all dietary supplement products should to be tested in accordance with the methods described in USP <2021> and <2022>. The required assays might include Total Aerobic Microbial Count, Total Combined Mold and Yeast, Bile-Tolerant-Gram-Negative bacteria, and objectionable organisms (absent in 10 grams of organism such as Salmonella, E. coli, and sometimes S. aureus). To do these tests companies either send product samples to contract laboratories for microbiological testing or test their samples internally.  Many outside laboratories utilize BAM and AOAC methods that were never validated against the USP methodology for Dietary Supplement products.

BioLumix Validation BookFDA’s GMP inspection program for dietary supplements has evolved since its debut a few years ago, with the agency increasingly strict in its interpretations of the regulation and requirements.  Validation of any new microbiological method is required prior to entering its use in the commercial arena. The validation assures equivalency of the new method to the reference method. This means that the new technique or device is giving us “real results” that are reproducible and that can be trusted. USP <1223> Validation of alternative microbiological methods and ISO 16140 defines the general principle and the technical protocol for the validation of alternative methods in the field of microbiological analysis.

BioLumix generates a customized validation book for each of its customers. The book includes:

Installation qualification (IQ):  Identification and validation of the system components; validation of the environmental conditions; electrical requirements; computer qualification; verifying that all installation steps were followed;  and documentation of instrument calibration.

Operational qualification (OQ):  Verifying that the equipment is properly installed, calibrated and is operational.  It includes a unique SOP for all products and assay combinations to be performed on the instrument; software characteristics and the verification that the software is 21 CFR part 11 compliant; verification that all the instrument functions operate as expected; Verification of the instrument temperature accuracy; and training records.

Performance Qualification (PQ):

Performance qualification is the most extensive part of the BioLumix validation book. It shows equivalency with USP methodology when following USP <1223> “Validation of alternative microbiological methods”.

  • Side-by-side comparison: Side-by-side results are comparable between the BioLumix method and the reference method when testing by the BioLumix Dilute-to-Specification Protocol.   A generic study was performed testing all types of Nutraceutical, botanicals and dietary supplements for Total aerobic count. A total of 252 samples with counts in the range of 100 cfu/g to 10,000,000 cfu/g   were analyzed, including   botanical powders, botanical extracts, infusions, alcohol extracts, raw materials, and finished products. There was a 99.6% agreement between the BioLumix method and the SP plate count method. Testing similar products for yeast and mold counts in the range of 10 cfu/g to 100,000 cfu/g there was a 98.1% agreement; and for Gram negative Bile Tolerant bacteria 98.7% agreement.  Similar results are shown for all objectionable groups of organisms such as E. coli, S. aureus, P. aeruginosa, and Salmonella.  Unique customer samples are also tested and compared to USP results.
  • Specificity (Inclusivity and Exclusivity): is the ability of the media in each vial to detect a wide range of organisms belonging to the target group, and not detect using non-target organisms, which demonstrates that the method is fit for its intended purpose.  The inclusivity and exclusivity of each of the media used by the system was extensively studied.  The data shows good inclusivity and exclusivity.
  • Limit of detection (LOD):  are the lowest microbial counts in a sample that can be detected under the conditions used.  The limit of detection was evaluated for Total Aerobic count, Yeast and Mold count and Gram negative Bile Tolerant bacteria.  The data shows that for all 3 media the limit of detection was close to 1-3 cfu/vial.
  • Precision and Repeatability:  is the degree of agreement among individual test results when repeatedly testing multiple samples of suspensions of microorganisms across the range of the test.  It is usually expressed as standard deviation (SD) or coefficient of variation (CV) of a series of measurements. Generally, a %CV of 15%-35% is acceptable. All the results obtained were well within the acceptable range.
  • Robustness: indicates the capacity of the device or technique to absorb small variations such as: incubation conditions, incubation temperature, media and sample volumes, etc.   The data demonstrates the robustness of the system with various media.
  • Ruggedness: is related to the reproducibility of the results considering different equipment (different instrument units), personnel, different lots of reagents, etc.  The BioLumix system has been demonstrated to be very rugged with high precision of test results obtained by analyzing the same microorganisms under a variety of different operating conditions including different analysts, different instrument units, and various  lots of media.
  • False negative rate: A false negative is a test in which samples that are inoculated or naturally contaminated generate a positive result in the reference method (plate count) but are negative in the test method (BioLumix). Naturally contaminated samples were tested at various specification levels, the BioLumix procedure detected the samples as being contaminated in all cases while the plate count method had a false negative result.
  • False positive rate: A false positive is a test in which a sample that does not contain the target organism generates a positive result in the BioLumix system but not in the Plate Count method. A false positive rate of 2.5% was seen in total aerobic count and no false positives were seen in the yeast and mold assay.

Conclusion

There is a need for rapid microbial detection technologies in order to improve the quality of products and their safety and speed up time to results. Their benefit could include significant reductions in time-to-result over conventional methods, improved sensitivity, specificity and accuracy, benefits of automation, reduced requirements for staff training, rapid product release, and lower inventory.  New rapid automated methods in microbiology need to be validated, and the BioLumix system comes with a validation book.

  • Unique validation book created for each customer.
  • Single platform testing for all assays
  • Real-time communication – early warning of contamination
  • Automated Certificate of Analysis in 48 hours
  • No product interference with ready-to-use vials with organism specific enrichment medium
  • Technology based on traditional media
  • Stackable compact instruments take up little bench-top space
  • Technical service available 24/7

References

Food  and  Drug  Administration  (FDA)  (2007),  21  CFR  Part  11,  Guidance  for Industry; Electronic Records; Electronic Signatures – Scope and Application. The National Formulary, Rockville, MD

United States Pharmacopeia (USP) (2008), Chapter <1223>, Validation of Alternative Microbiological Methods. The National Formulary, Rockville, MD

United States Pharmacopeia (USP) (2008), Chapter <2021>, Microbial enumeration tests-Nutritional and dietary supplements. The National Formulary, Rockville, MD

United States Pharmacopeia (USP) (2008), Chapter <2022> Microbiological procedures for Absence of Specified Microorganisms- Nutritional and dietary supplements. The National Formulary, Rockville, MD

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.

Thank you for your Continued Loyalty to BioLumix!

We would like to take this opportunity to express our appreciation to all of our valued customers: Thank you for choosing BioLumix as your partner in microbiological testing!  BioLumix is proud to have more than 500 systems in place throughout the world, as more companies adopt our rapid microbiology testing system.  During 2013 more and more companies have chosen our novel streamline technology and enjoy accelerated product release with our simplified, automated microbiology.

For those that are looking for improved company profitability in the coming year, we hope 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 which allows you to test raw materials and finished products.  Powders, oils, enzymes, botanical material, tablets, capsules, food products, dairy products, and beverages can be easily tested without product interference. Automated monitoring of ready-to-use assay vials, automated data processing and archiving (paperless), makes the microbiologist‘s job a lot easier with the same accurate results in less than half the time. Please start the New Year by contacting us for a free product trial.

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.

Detection of psuedomonads in dairy and water samples using a quantatative one-step testing protocol in just one day

By Roger Brideau*
*Presented in part at the International Association of Food Preservation
‘IAFP’ Conference in Charlotte NC USA

dairy microbiology detection of psuedomonads Introduction- Pseudomonad organisms are a major cause of bacterial spoilage of pasteurized milk and dairy products due to post process contamination.  Early detection of Pseudomonad’s can be a predictor of product shelf-life as they are the predominant psychotropic bacteria present.  BioLumix has developed a rapid method for the detection of Pseudomonad’s in dairy products and the method is also applicable to their detection in process water.

Purpose- To evaluate the ability of the BioLumix system to detect Pseudomonad’s in dairy products, determine the speed to results, sensitivity, selectivity and ability to predict shelf-life.

Methods- the BioLumix system is an optical system that detects growth of Pseudomonad’s using a CO2 sensor in selective growth media.  The BioLumix system was directly compared to the plate count methodology for milk samples stored at refrigerated temperatures and held overnight at room temperatures (enriched).  Testing of water was also accomplished in side by side studies to show the capability of the BioLumix system for quantitation of Pseudomonads.

Results:  Growth of Pseudomonad’s in the BioLumix vial
Table of quantitation of Pseudomonads
A growth comparison was made for detection of each Pseudomonad in the BioLumix system using PSE-B vials and on CFC (Pseudomonas agar) spread plates.  Table 1 summarizes the growth of freshly diluted samples of organisms that were enriched in TSB during the prior 18-24 hrs.  The PSE-B vials are selective, as shown by not allowing growth of unrelated gram positive and gram negative bacteria, yeast or mold.  Four different species of Pseudomonad’s grew in the PSE-B vial and on CFC plates.

Milk Sample Testing
Commercial milk samples were tested upon arrival in the laboratory.  Five of twenty were positive for the presence of Pseudomonad’s using both PSE-B vials and CFC spread agar plates.  After storage for 3-7 days, twelve of twenty samples were positive for Pseudomonad’s including after enrichment at RT for 16-18 hrs.  Thus, refrigerated milk samples have varying incidence of Pseudomonad flora.

Dairy Microbiology Calibration dataMilk Calibration Curve
Organisms from milk samples that grew in PSE-B vials and on CFC plates were used to generate the Calibration Curve shown in Figure 1. These data suggest that low numbers (~10) of Pseudomonad’s should detect within 24 hrs in the PSE-B vial. The Calibration Curve can be embedded into the BioLumix software on the instrument and used to generate a read-out of cfu per gram of milk.  This enables quantitation of the milk sample for the presence of Pseudomonad’s before 24 hr; a distinct advantage over plate methodology taking 48-72 hours.

Dairy Microbiology detection time distributionDistribution of Data
In the dairy settings the goal is to separate a “good” sample that has a potential to maintain quality over a product’s shelf-life from a “bad: sample that will have a shorter shelf life. Criteria for separation between a “good” and “bad” product based upon the Pseudomonad’s numbers can be established. If one selects a count of 1,000 cfu/ml as the separation point: the Histogram shown in Figure 4 indicated at 12.5 hrs all samples with higher counts (in red) detected, while all the samples below 1,000 cfu/ml did not.

Results: Testing of Process Water for the presence of Pseudomonads
Eight different types of process water samples were found to be free of Pseudomonad’s after testing using PSE-B vials and CFC spread plates (data not shown).  Clean process water samples were then inoculated with individual isolates of Pseudomonad’s were used to generate a calibration curves for water, similar to the milk calibration curve. Pseudomonad growth in inoculated process water was measured using PSE-B vials and PA spread plates and was used to generate the Calibration Curve. Detecting vials were confirmed to contain Pseudomonas by the Oxydase test.

Summary
The data presented show equivalency between the BioLumix PSE-B vial and CFC (Pseudomonas agar) plates for the detection of Pseudomonad’s found in commercial milk samples and in inoculated process water samples.  PSE-B vials detected as little as 1-3 organisms (data not shown).
The number of organisms in commercial milk was found to increase over time at refrigerated temperatures and this agreed with a previously published report (Burdova et al 2002) showing the affect of storage temperature on milk shelf-life.
The BioLumix assay is completed in 18 hours and offers an advantage over spread plate methods for time to results and ease of calculation of cfu per gram of milk or water.  A single vial is all that is needed and thus both time and material costs are reduced.  Calibration Curves were easily generated for both milk and water sampling and can be used to generate a cfu/ml of sample in less than 1 day to yield an estimate of cfu/gram.

REFERENCE:  Burdova, O. et al (2002).  Bulletin Vet Med. Poland. 46:325-329. Hygiene of Pasteurized Milk Depending on Psychrotrophic Microorganisms.

BioLumix Exhibits @ SupplySide West 2013 – Stop by Booth # 30038!

SupplySide WestSupplySide West brings together the suppliers and buyers that drive the dietary supplement, food, beverage, animal nutrition, personal care and cosmetic marketplace. Key professionals and thought leaders from executive management, R&D, QA/QC and marketing teams will meet in Las Vegas November 12-16.  The 2013 show floor is the largest yet, featuring over 1,700 booths, please make sure that you visit BioLumix at Booth #30038.

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 many operational profits drivers. . . and is The reason over 450 BioLumix Systems are actively being used by many of the largest manufacturers in the industry.

Here are just a few of the many benefits of the BioLumix Partnership Program:

Ready-to-use vials  with specialized media which do not require QC on your end

Very Fast 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.

biolumix automation drives profitsWe 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 product 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.

Dealing with difficult to read plates or manual data transfer is a thing of the past. 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.  Real-time data communication: Early warning of contaminated samples as well as sample release information could be automatically communicated through your intranet, significantly improving your company’s efficiencies.  BioLumix system can be used to test powders, oils, enzymes, botanical material, tablets, capsules, powders and liquids easily and without product interference.

BioLumix trainers will provide each customer with a full and customized Validation book, during the two and a half days of on-site training and installation of the system, including installation qualification, Operation Qualification and Performance qualification.

With this internal automated microbiological system, customers have control over their testing, leading to accurate results and accelerated product release while reducing costs.

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, and finished goods.

International Dairy Show 2013

International Dairy Show 2013
November 3–6, 2013, McCormick Place • Chicago, Illinois

International Dairy Show 2013The International Dairy Show is back in Chicago and is bigger, and better than ever. Dairy, food and beverage processors will see technology and innovations in formulation, processing, packaging, distribution, food safety, and more. Operations, research, and sales and marketing teams will attend to learn trends, cost-cutting innovations, and ways to deliver more value to your customers. We at BioLumix wanted to take this opportunity to invite you to our exhibit (booth #452) and show you what’s new at BioLumix. We continue to innovate, growing our assay repertoire and capabilities.
BioLumix has developed a simple rapid microbiological method for the detection of various groups of bacteria, yeasts and molds. The system can make the microbiology testing simpler, faster and automated, saving you time, labor and money. BioLumix has a comprehensive range of microbiological tests, including specific vials for Coliforms and Pseudomonas. The system is designed to accelerate product release in a simplified automated approach. Complete coliform test results are obtained within 12 hours with one vial substituting for nine or more MPN (Most Probable Number) test tubes. Yeast and mold results are obtained in 48 hours rather than 5-7 days using plate methods. Many products can be introduced directly into the BioLumix test vial without the need of a dilution step. These include yogurts and sour cream, as well as milks.
Dairy product Testing for Yeast and Mold and Coliform
The yogurt can be directly added (Figure 1) into BioLumix vials to measure growth of Coliforms or Yeast and Molds. Specialized high pH BioLumix vials can also be used for yogurt samples. When a low pH yogurt sample is added to the BioLumix high pH Coliform vial (CC), the pH conditions become near neutral. This ensures the yogurt manufacturer to be able to correctly test for coliforms using a direct (without dilution) sample of product. As much as 1 gram of product can be directly tested in each BioLumix test vial.

A clear advantage of the BioLumix vial is that it can be used on the same work-day to yield data about the status of any contamination in dairy samples and thus enable the manufacturer to make a decision on the acceptability of the lot of product for sale and consumption.

Rapid Detection of Pseudomonads in Dairy Products as an Indicator of Product Shelf-Life The data presented in this study suggested that the BioLumix Pseudomonas vials are capable of early detection of Pseudomonads in dairy products and in process water. The system offered a reduction in time to results as compared to the plate methodology and eliminates any product interference. It allows for rapid assessment of any post processing associated problems.

The BioLumix Advantage
• Vial design prevents product interference
• Automation and connectivity allows faster product release
• Real-time communication for immediate action
• Expedited results: most results in 12-18 hours; Yeast and mold assay in 48 hours
• Automated data archiving and audit trail
• Streamlined testing increases laboratory efficiencies
• Paperless laboratory: centralized test data automatically stored and protected
• Barcode capability for automated sample entry
• Environmental testing made easy

Free Product Trial
Give us your most difficult samples and we will test them for free. We will provide you with a detailed report which includes a side-by-side comparison to your current manual methodology. The data generated is strictly confidential and is only used to show the high correlation of results should your company decide to purchase.

We look forward to working with you and earning another satisfied customer!

PDA’s 8th Annual Global Conference on Pharmaceutical Microbiology

October 21-24, 2013 | Bethesda North Marriott Hotel | Bethesda, Maryland; Exhibition: October 21-22

PDA’s 8th Annual Global Conference on Pharmaceutical Microbiology brings together all levels of industry professionals to network and benefit from a program that reveals the essential science of microbiology and seeks to solve the problems that the Pharmaceutical industry faces on a daily basis. During the conference, PDA will host an exhibition of leading bio/pharmaceutical companies who will showcase new technologies and trends for pharmaceutical microbiology strategies, providing invaluable sessions dedicated to pharmaceutical microbiology.  Visit the BioLumix booth (#13) to learn about the most advanced rapid microbiological testing system.

advanced rapid microbiological testing system
Rapid Automated USP <61> and <62> on a single Platform
BioLumix Rapid Microbiological System is capable of performing testing equivalent to both USP <61>and <62> simultaneously on a single platform. The novel automated growth-based system simultaneously detects microbial growth, provides an estimation of viable cell counts, and identifies the presence of specified micro-organisms. By encompassing both USP types of testing, the BioLumix is a complete screening solution making the existing microbial testing simpler, faster, and automated – saving your company significant time, labor and money. BioLumix has a DMF on file with the FDA, and the system is easy to validate due to growth based format using traditional USP media.

Adding Speed and Automation to Microbiological Testing – The Automated BioLumix System will accelerate your microbiological testing resulting in faster product release and decreased costs. The streamlined testing design reduces labor, time and supplies. The BioLumix software is 21 CFR Part 11 compliant with log-in, log-out and audit trail. 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, and finished goods. 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 manufacturers.

Here are just a few of the many benefits of the BioLumix System:

  •  Simplified, automated microbiology testing
  •   Paperless laboratory
  •   60% savings in labor
  •   Real-time continuous support for reviewing microbiology assays.

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 on 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 product that doesn’t work for you.  Still not convinced?  We would be more than happy to provide you a long list of customer referrals.

The Pharmaceutical Industry is Growing; the BioLumix System Can Help Your Lab Grow With it

According to the United States National Institute of Health, there are nearly 140,000 clinical studies currently underway involving potential pharmaceutical products. Pharma is a huge industry, and companies both big and small are investing roughly $50 billion a year to get their products to market.Pharmaceutical Microbiological Testing This fervor spans some 200 countries, and with all this in mind it is no wonder that research has increased by an average of 28% each year since 2000. Still, whenever there are huge sums of money involved, people and companies can be enticed to hastily release their products, and this potential for microbial contamination in pharmaceutical products can have serious and far-reaching repercussions. The goal of the BioLumix Microbiological Testing System is to promote an expedient, automated, and efficient laboratory environment, allowing companies to increase their bottom line and ensuring that only safe products make it to the shelves.

Pharmaceutical microbiology is essential to the quality control process of any drug or supplement. Companies can spend immeasurable sums of time and money during this stage, and current plate count methods are incapable of providing labs with an avenue for advancement. BioLumix Rapid Microbiology Methods offer innovative new testing solutions that are designed specifically for the challenges of the pharmaceutical industry, including a multitude of microbial tests for raw materials, in-process and finished products, as well as the capacity to perform environmental testing. This simplifies, expedites, and reduces the cost of performing microbiological assays.

Biolumix Microbiology SystemThe key to the BioLumix System is its use of patented two-zone vials to monitor changes in a broth medium. At the top of each vial, an incubation zone allows a microbiologist to insert samples for testing, and at the bottom, a reading zone is utilized for the automated monitoring of up to 32 specimens by a single unit. Changes in color or fluorescence, which may be a sign of microbial growth, are expressed as light intensity units, which are detected by the systems optical sensor and recorded in its computer. Sample volumes of up to 2 ml can be used in each vial. Additionally, each vial slot features a dual-color Light Emitting Diode (LED) and a fluorescent tube that covers the full spectrum of colors and ultraviolet light. These two-zone vials eliminate the masking of the optical pathway by the products microbial turbidity, and because changes in color and fluorescence are monitored in the reading zone, results cannot be influenced by the sample or any of the microorganisms.

A few of the applications of the BioLumix System include total counts, yeast and mold, Bile tolerant gram negatives, and objectionable organisms. Results are easy to interpret and can be automatically transmitted to other locations, such as a company warehouse, to rapidly ship safe products. Perhaps one of the most convenient aspects of the BioLumix System is the fact that it can be easily operated non-microbiologists. This lessens the amount of tedious work for skilled lab-technicians, and allows them to shift their focus to higher value tasks.

In short, the BioLumix System allows pharmaceutical companies to perform as many tests as are needed to maintain a safe and beneficial product, without sacrificing time or money. Instead, for a one-time investment in BioLumix rapid microbiological method technology, your lab will be able to expedite its quality control process and decrease its overhead, allowing your company to expand in the growing market.  The BioLumix rapid microbiology system will allow for increase capacity, logistical improvements and better efficiencies.

Recently Dr. Miller presented a web seminar about a case study of the implementation of the BioLumix system.  To listen to this seminar click on the link http://cnpg.comparenetworks.com/144579-Implementation-of-New-RMM-BioLumix-Case-Study/

Rapid Microbiology Testing of Dairy Products using the BioLumix Instrument

Background

Testing Dairy Products using Rapid Microbiology with the BioLumix InstrumentCommercial Dairy Products need to be tested for microbial contaminants.  Dairy products include: cultured dairy products (kefir, cultured buttermilk, sour cream, and yogurt), cheeses (soft cheese products, semi-hard, hard, and extra hard cheeses), processed cheeses, butters, butter creams, dried milks, and ice creams.

Manufacturers require a simple, cost effective and rapid microbiological method to assess samples for the presence of specific organisms as an indication of the sanitary conditions.  The most common dairy microbiology tests include: Aerobic plate count, Yeast and Molds, Coliforms, and could also include some indicator of cold spoilage such as Pseudomonads.  Rapid and early testing for microorganisms can reduce product quarantine time, allows for faster response to contamination, decreases inventory holding and cold warehouse costs, and aids in prediction of product shelf-life for manufacturers.

Troublesome spoilage microorganisms include aerobic Psychrotrophic bacteria, yeasts, molds, heterofermentative lactobacilli, and spore-forming bacteria (Ledenbach and Marshall 2009).  Psychrotrophic bacteria can produce large amounts of extracellular hydrolytic enzymes, and the extent of recontamination of pasteurized fluid milk products with these bacteria is a major determinant of the milk products shelf life. Fungal spoilage of dairy foods is manifested by the presence of a wide variety of metabolic by-products, causing off-odors and flavors, in addition to visible changes in either color or texture and may also cause gassing.

BioLumix has developed a simple rapid microbiological method for the detection of various groups of bacteria, yeasts and molds.  The system can make the microbiology testing simpler, faster and automated, saving manufacturers time, labor and money. BioLumix has a comprehensive range of microbiological tests, including specific vials for Coliforms and Pseudomonas.  The system is designed to accelerate product release in a simplified automated approach.  Complete coliform test results are obtained within 12 hours with one vial substituting for nine or more MPN (Most Probable Number) test tubes.  Yeast and mold results are obtained in 48 hours rather than 5-7 days using plate methods.  Many products can be introduced directly into the BioLumix test vial without the need of a dilution step.  These include yogurts and sour cream, as well as milks.

BioLumix Applications

Yogurt Testing

Yogurt is a dairy product, which is made by blending fermented milk with various ingredients that provide flavor and color.  Manufacturers have responded to the growth in the yogurt market by introducing many different types of yogurt including low fat and no-fat, creamy, bio-yogurt, organic, baby, and frozen.  BioLumix has developed assay methods for measuring contaminating microorganisms in dairy products.

The yogurt can be directly added (Figure 1) into BioLumix vials to measure growth of Coliforms or Yeast and Molds.  Specialized high pH BioLumix vials can also be used for yogurt samples.  When a low pH yogurt sample is added to the BioLumix high pH Coliform vial (CC), the pH conditions become near neutral.  This ensures the yogurt manufacturer to be able to correctly test for coliforms using a direct (without dilution) sample of product.  As much as 1 gram of product can be directly tested in each BioLumix test vial.

Coliform organisms generally include four key groups of enteric bacteria.  They comprise of the following species: Escherichia, Citrobacter, Klebsiella and Enterobacter.  Each of these organisms can grow in the BioLumix high pH CC vial.  Figure 2 illustrates the growth of a coliform (Citrobacter) in the BioLumix CC vial (green) while the Dark Blue curve shows a clean sample.

A clear advantage of the BioLumix vial is that it can be used on the same work-day to yield data about the status of any contamination in dairy samples and thus enable the manufacturer to make a decision on the acceptability of the lot of product for sale and consumption.

Fermented Dairy Products

Yeast and Mold Testing: An example of the growth of a mold in cream cheese is shown below. Figure 3 illustrates the clean test result, using BioLumix Yeast & Mold vials, for the cream cheese product as shown in the Green curve.  The Dark Blue curve illustrates the growth curve in BioLumix YM vials when cream cheese was inoculated with the mold organism Geotrichum candidum.

Coliforms Testing:Similarly, testing of sour cream showed the lack of Coliforms in the product as illustrated in the Dark Blue curve in Figure 4.  The sour cream product sample was found to have other flora as shown by growth in the BioLumix TAC vial which detected total aerobic bacterial counts.  Thus, the BioLumix high pH CC vial was used to show selectivity for the presence or absence of Coliform organisms.

Rapid Detection of Pseudomonas in Dairy Products as an Indicator of Product Shelf-Life
Pseudomonad organisms are a major cause of bacterial spoilage of pasteurized milk and dairy products due to post process contamination.  Early detection of Pseudomonas in can be a predictor of product shelf-life as it is the predominant psychotropic bacteria present.  BioLumix has developed a rapid method for the detection of Pseudomonads in dairy products; the method is also applicable to detection of Pseudomonads in process water.

The BioLumix system was directly compared to the plate count methodology for milk samples stored at refrigerated temperatures and held overnight at room temperatures and to detected Pseudomonads in process water and other dairy products.

Commercial milk products were used to measure the presence of Pseudomonads during refrigerated and elevated temperature storage.  Pseudomonads were present at varying levels in dairy samples and were detected within 16-24 hours using BioLumix vials.  All process water samples tested were free of Pseudomonads by both methods.  Process water samples inoculated with different Pseudomonads strains were found to detect in the BioLumix system.  The vial were selective enough not allowing for growth of unrelated gram positive and gram negative bacteria, mold or yeast.  The combination of the Pseudomonas vial with overnight pre-incubation could serve as an indicator of shelf-life of products.

The data in this study suggests that the BioLumix Pseudomonas vials are capable of early detection of Pseudomonads in dairy products and in processing water. The system offers a reduction in time to results as compared to the plate methodology and eliminates any product interference.  It allows for rapid assessment of problems associated post processing problems.

BioLumix has developed a vial specific for testing of Pseudomonads that are known to commonly contaminate milk and also survive pasteurization, albeit in low numbers. These Pseudomonads may include P aeruginosa, P fluorescens, P putida, and P stutzeri.  Each of these species of pseudomonas was found to grow readily in the BioLumix PSE-B vials.

Summary

The BioLumix System is designed to accelerate product release with a simplified, automated approach.  This yields fast, accurate, real-time results while reducing costs and eliminates the time required for the assays to be completed.  The system offers real-time results of contaminated samples saving hours, possibly days.  The Coliform or Enterobacteriaceae results can be available within 12 hours; Yeast and Mold results within 48 hours.  Thus the BioLumix system allows for rapid detection of Coliforms and Yeast and mold.  For low coliform numbers, one vial can substitute for 9+ MPN tubes.  BioLumix will streamline and simplify the microbiological procedures, save labor and create a paperless laboratory, while generating results that correlate well with plate count methodology.

Reference
Ledenbach L. H. and R.T. Marshall. 2009 In “Compendium of the Microbiological Spoilage of Foods and Beverages” . Chapter Microbiological Spoilage of Dairy Products.

BioLumix System Saves Valuable Time in Yeast and Mold Testing

Rapid Microbiology Yeast and Mold TestingIf you are a microbiologist, or work in close relation with a laboratory, you probably already know that yeast and mold testing is one of the longest quality control processes. Both the plate count and the membrane filtration techniques used by many microbiology labs often take between 5-7 days to complete, and this ends up costing you and your organization valuable time and money in the mean time. The BioLumix rapid microbiology system is based on cutting edge knowledge of the microbial metabolic process, and is the most versatile method around when it comes to the expedient detection of microorganisms. For the nominal cost of our automated testing systems, your company could shave several days off of their production and distribution time, and secure a catalyst that will increase profits for years to come.

Yeasts and molds constitute a large and diverse group of microscopic foodborne fungi, totaling several hundred different species. One of the main reasons that these organisms are so successful in attacking foods and cosmetics is their versatility, having very broad temperature (5-60 ?C) and acid/alkaline (pH 2-pH 9+) requirements for growth. This means yeast and mold are capable of invading crops, such as those producing grains, nuts, beans, and fruits both before harvesting and during storage, and can also make their way into our processed food. In many cases Yeasts and molds are not immediately visible, however these fungi can cause varying degrees of deterioration and decomposition in products, with the manufacturer only discovering their presence when it’s too late.

Several different kinds of molds and yeasts can be hazardous to humans and animals due to their ability to produce toxic metabolites called mycotoxins. These metabolites are stable compounds, and the performed toxins are usually not destroyed during food processing, or even when cooking at home. Other yeasts and molds can also cause allergic reactions or infections, especially in immunocompromised demographics, such as the elderly, the young, the debilitated, or those undergoing antibiotic and chemotherapy treatment. To make matters worse, yeasts and molds often cause products to spoil after they have left the processing facility, posing potentially a health hazards and product quality issues to the consumers. For this reason, a wide range of industries require that companies test their products for yeast and mold before they go on sale. In a nutshell, the quicker you can produce and quality test products, the quicker you can get your inventory to market, and the more profitable you will be.

This value proposition is the premise upon which our rapid microbiological testing system was founded. The BioLumix system is committed to simplifying, expediting, and reducing the cost of performing microbiological assays so that companies can bolster production with a fraction of their current working capital. Our ready to use vials are capable of detecting yeast and mold in 48 hours, saving 3-5 days when compared to plate count and membrane filtration techniques and keeping more money in your pocket.

optical technology for yeast and mold testingThe keys to our systems celerity are its patented CO­­­2 sensors and optical technology, which continuously monitor growth media in each vial. Because yeasts and molds are obligate aerobes, they must take in oxygen and release Carbon Dioxide (CO­­­2) in order to grow. This cellular respiration is part of the Krebs cycle, and is used by all aerobic organisms to generate energy. When the sensor detects the presence of CO­­­2 it is a sign that microbial growth is present. The optical sensor simultaneously takes readings of the vials at 6 minutes intervals, detecting changes in color, and allowing for rapid detection of growth. Results are easy to interpret and can be automatically transmitted via a multitude of communication channels to its needed location, such as a warehouse or production facility to expedite shipping of a safe product. Our fully automated system can also be operated by a non-microbiologist, reducing labor by up to 60%, and allowing skilled scientists to spend their time in the lab more efficaciously. With less tedious work, microbiologists can turn their attention to higher value tasks, such as interpretation of data and product safety analysis, making your lab far more efficient.

With the BioLumix rapid microbiology system, labs can revolutionize the way that one conducts their quality control process. Our platform makes production times shorter, products safer, and recalls less frequent and far less costly. Adding value and increasing lab productivity with BioLumix will allow your organization to thrive, and will put you in a position to be successful for years to come.