How do you control biofilms in your GMP facility?

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Biofilms are complex communities of microorganisms that attach to surfaces and form a protective matrix. They can pose serious risks to the quality, safety, and efficacy of your products, as well as to the integrity and performance of your equipment and facilities. Therefore, controlling biofilms is a crucial aspect of good manufacturing practice (GMP) in any industry that involves biological processes or products. In this article, you will learn what biofilms are, how they form and grow, what factors influence their development, and how you can prevent and remove them effectively in your GMP facility.

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Brent Lloyd, RN

Innovation through transformative disruption
Tasnim Adel

Quality Assurance Engineer | ???? Food Safety awareness | Food Science & Technology | Nutrition " ??????????? ???…
Huda Abbasi

Life scientist R&D

1 What are biofilms?

Biofilms are not just random collections of microbes. They are highly organized and dynamic structures that can adapt to different environmental conditions and resist external stresses. Biofilms consist of microbial cells embedded in a self-produced matrix of extracellular polymeric substances (EPS), which can include polysaccharides, proteins, DNA, and other molecules. The EPS matrix provides physical and chemical protection to the biofilm cells, as well as a scaffold for attachment and communication. Biofilms can also contain water channels, pores, and gradients of nutrients, oxygen, and pH, creating heterogeneous and diverse microenvironments within the biofilm.

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Tasnim Adel

Quality Assurance Engineer | ???? Food Safety awareness | Food Science & Technology | Nutrition " ??????????? ??? ??????????? ??? ??? ?????? ???? ?? ????????? ??? ???? ??? ?????? ????? ??? ???????.."
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??Biofilms are communities of microorganisms that stick to each other & to surfaces.? ? ??They are all around us, from the slime on rocks in streams, to washing machines, to our own bodies. ? ??The biofilm matrix consists of extracellular polymeric substances (EPS), which are a complex mix of substances like proteins, polysaccharides, & eDNA.? ? ??This matrix provides a protective barrier that protects them from environmental stresses, including antimicrobial treatments. ? ??In food processing environments, biofilms can form on many surfaces, including equipment, pipes, & containers, leading to food contamination & spoilage. ? ?? So, managing biofilms is crucial to food safety to prevent microbial contamination & maintain product quality.

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2 How do biofilms form and grow?

Biofilms can form on any surface that is exposed to moisture and nutrients, such as pipes, tanks, filters, valves, or product contact surfaces. The biofilm formation process can be divided into four main stages: attachment, growth, maturation, and dispersal. In the attachment stage, free-floating microbial cells (planktonic cells) adhere to the surface by using various mechanisms, such as flagella, pili, or EPS. In the growth stage, the attached cells multiply and secrete more EPS, forming microcolonies and increasing the biofilm thickness and complexity. In the maturation stage, the biofilm reaches a stable and mature state, where the cells communicate with each other through chemical signals (quorum sensing) and regulate their gene expression and metabolism. In the dispersal stage, some biofilm cells detach from the biofilm and return to the planktonic state, either spontaneously or in response to environmental cues, such as nutrient depletion, shear stress, or antimicrobial agents. The dispersed cells can then colonize new surfaces or rejoin existing biofilms.

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Tasnim Adel

Quality Assurance Engineer | ???? Food Safety awareness | Food Science & Technology | Nutrition " ??????????? ??? ??????????? ??? ??? ?????? ???? ?? ????????? ??? ???? ??? ?????? ????? ??? ???????.."
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???Biofilms are communities of microorganisms that stick to each other & to surfaces.? ???Their matrix consists of extracellular polymeric substances (EPS), which are a complex mix of substances like proteins, polysaccharides, & eDNA.? ???Biofilms form & grow through a series of steps, including: 1?? Individual microorganisms attach to a surface. 2?? Once attached, they start to multiply & produce extracellular polymeric substances (EPS), creating a matrix that holds the biofilm together. 3?? The biofilm matures as more microorganisms join the community & the EPS matrix grows. 4?? The structure becomes more organized, with distinct layers of microorganisms. This structure provides protection & resilience to the biofilm community.

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3 What factors influence biofilm development?

Biofilm development is influenced by many factors, both biotic and abiotic. Biotic factors include the type, number, and diversity of microbial species involved in the biofilm, as well as their interactions and synergies. Abiotic factors include the characteristics of the surface, such as its material, texture, roughness, hydrophobicity, and charge, as well as the properties of the fluid, such as its flow rate, temperature, pH, oxygen level, and nutrient concentration. These factors can affect the initial attachment, growth rate, structure, composition, and resistance of the biofilm. For example, a rough and hydrophobic surface can facilitate biofilm formation by providing more surface area and hiding places for microbes. A high flow rate can prevent biofilm formation by removing planktonic cells and EPS, but it can also enhance biofilm formation by providing more nutrients and oxygen. A low pH can inhibit biofilm formation by damaging microbial cells and EPS, but it can also promote biofilm formation by reducing the solubility of minerals that can coat the surface.

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Huda Abbasi

Life scientist R&D
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In a water purification system, biofilms can develop on surfaces of pipes, membranes, filters, and inner walls of storage tanks. Biofilm formation negatively impacts the quality of purified water by Harboring pathogenic bacteria, and promoting the growth of undesirable microorganisms, leading to waterborne diseases. Trapping particles, organic matter, and other impurities and reducing the effectiveness of the purification process. Obstructing water flow through pipes, filters, and membranes, decreasing flow rates and increasing energy consumption. Fouling & clogging critical components, increasing maintenance costs and decreasing lifespan of equipment. Increasing resistance to chemical disinfectants and antimicrobial agents.

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4 How can you prevent biofilm formation?

To prevent biofilm formation, it is essential to design and maintain a GMP facility in a way that minimizes the conditions that favor biofilm development. This can be achieved by selecting materials such as stainless steel, glass, or plastic that are resistant to corrosion, biofouling, and biofilm formation. Additionally, it is important to smooth and polish surfaces to reduce their roughness and porosity, and avoid cracks, crevices, joints, or dead ends that can trap microbes and nutrients. Furthermore, filters, screens, or UV lamps should be installed to reduce the number of planktonic cells and EPS in the fluid. Additionally, controlling the temperature, pH, oxygen level, and nutrient concentration of the fluid can limit microbial growth and activity. Moreover, it is important to monitor the microbial load and biofilm presence on the surfaces by using methods such as swabbing, culturing, staining, microscopy, or molecular techniques. Finally, a regular and effective cleaning and sanitation program should be implemented to remove any organic and inorganic residues that can serve as substrates for biofilm formation.

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Fathima Doole

Membrane Biophysicist | Analytical Method Development | Dissolution Scientist
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Incubators, especially those used for cell culture, provide a warm, humid environment that is conducive to microbial growth. If not properly cleaned and maintained, these conditions can promote the formation of biofilms. Regularly clean and disinfect the incubator. Ensure all components, including shelves and water pans, are cleaned. Use only distilled/deionized water in the humidity pan, and change it frequently. Ensure a regular maintenance schedule and check for contamination. Educate lab personnel on the importance of aseptic techniques. Prevention is the best strategy. Once a biofilm has formed, it becomes significantly more challenging to ensure complete removal and can jeopardize the validity of cell culture experiments.

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Jonathan Reno

Quality Assurance and Laboratory Manager @ Losurdo Foods, Inc.
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Antimicrobial resistance is a real thing and sanitation procedures need to be top notch to keep biofilm and niche growth areas from developing. ATP swabbing can be a verification factor in evaluating your sanitation program is working. Additionally, manipulating the environment can help deter growth as some values are unfavorable and require more resistance to change.

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5 How can you remove biofilm contamination?

If biofilm contamination occurs in your GMP facility, it is important to act quickly and thoroughly to eliminate it and prevent it from recurring. Biofilms are difficult to remove due to their resistance to physical and chemical agents that usually kill planktonic cells. Therefore, a combination of mechanical, thermal, and chemical methods should be used to disrupt and remove the biofilm. Scrubbing, brushing, or scraping the surfaces can physically detach the biofilm and EPS. Flushing, jetting, or sonication can apply high-pressure or high-frequency water or air to dislodge the biofilm and EPS. Heating, steaming, or boiling can apply high temperatures to kill the biofilm cells and degrade the EPS. Sanitizing, disinfecting, or sterilizing with agents such as detergents, acids, bases, oxidizers, or biocides can dissolve the EPS and destroy the biofilm cells. However, it is important to choose the right agent, concentration, contact time, and frequency for each type of biofilm and surface while following safety and environmental regulations. Rinsing, drying, or storing can remove any residual biofilm or chemical agent from the surfaces and prevent biofilm reformation.

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Brent Lloyd, RN

Innovation through transformative disruption
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How to locate the biofilms are a major concern. There are new technologies using biofluorescence to identify components within the EPS of the biofilm. The color observed at certain wavelengths determine the potential for biofilms. ATP testing has been a method to measure the level of cleanliness with RLUs which show the presence of organisms-dead or alive-which can be covered by the biofilm. The problem is that ATP is a blind test. Certain surfaces are tested routinely while other surfaces are tested randomly. Biofluorescent technology quickly scans surfaces for biofilms which drive additional testing and or cleaning. The goal is to get the facility back into production mode safely.

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Huda Abbasi

Life scientist R&D
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Nanotechnology is the next logical step for biofilms prevention in water purification systems. Nanocoatings to prevent bacterial adhesion, nanofibers with bactericidal functionalization for sterilization. Nanoimaging for bacterial growth monitoring. Nanobubbles for film disruption... These are the few of many exciting and promising technologies which can be explored for biofilms treatment and prevention in water purification systems.

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6 Here’s what else to consider

This is a space to share examples, stories, or insights that don’t fit into any of the previous sections. What else would you like to add?

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How do you control biofilms in your GMP facility?

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1 What are biofilms?

2 How do biofilms form and grow?

3 What factors influence biofilm development?

4 How can you prevent biofilm formation?

5 How can you remove biofilm contamination?

6 Here’s what else to consider

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