PMF Newsletter - January 2012

The latest edition of the indispensable PMF Newsletter (Pharmaceutical Microbiology Forum) has been issued.

The current edition features articles by Scott Sutton on the 'Qualification of a Contract Microbiology Laboratory' and 'Current Contract Manufacturing Operation (CMO) Regulatory Issues' by Barry Friedman.


In his article, Scott Sutton looks at qualification of a contract microbiology testing lab in the GMP environment. This is a critical consideration in today's environment, as many companies are out-sourcing technical testing activities and reducing in-house capabilities. In addition, as the pool of in-house subject matter experts is reduced through lay-offs, transfers or attrition the remaining technical experts must learn to become more efficient and relying on outside expertise is one way to do this.

Barry Friedman considers how policy changes and enforcement strategies implemented by the FDA have led to an increase in the number of inspection of both Contract Manufacturing Operations (CMOs) and in-house sterile manufacturers - especially injectables.  There has been a greater oversight and detailed review of all protocols and processes.  Any issue or potential issue must have a corrective action with a demonstrated positive outcome.  There no longer exists a prescribed sequence of events after a Form FDA 483 is issued .

The newsletter can be accessed here: PMFNewsletter


Posted by: Dr. Tim Sandle

FDA and EMA co-operate on GMP

The EMA and FDA and working more closely together on GMP inspections. A new initiative for the agencies to work together on inspections was launched at the start of 2012.  This is aimed at sharing work for inspections of pharmaceutical manufacturers within each other's areas.

The objective of the change is to make better use of resources. This is the latest step in increased collaboration between European authorities and the FDA.

A joint statement between the EMA and FDA reads:

"The most likely impact will consequently be in the area of routine post-authorisation/surveillance
inspections.  Pre-authorisation/pre-approval inspections will continue largely unchanged, as by
definition the sites are unknown or a specific inspection trigger has been identified.  Exceptions
nevertheless may be made on a case-by-case basis.

The following considerations will be taken into account:

• The inspection history of the site. 
• The nature of the product. 
• Quality defects associated with the site. 
• Variations or significant changes since the last inspection. 
• Outstanding inspection follow up. 
• Whether there is an urgent public health need to expedite regulatory decision-making e.g., product 
• shortage. "

To read the statement in full, visit: Regulatory Joint Working

Posted by: Dr. Tim Sandle

A fragrant new bio-fuel?

The search for new bio-fuels represents potential big business with the decline in available fossil fuels. A team of scientists, backed by the US government, have made significant progress in using bacteria to create a bio-diesel from glucose.

Bio-fuels are either derived from plants or from agricultural waste or bio-fuels are synthesised through microbes that digest the biomass and convert its sugars into fuel molecules. With the projected decline of fossil fuels, the search for cheap and quick to produce bio-fuels continues to apace within industry and at university laboratories.

EcoFriend reports tat a team of scientists believe that they have made considerable progress with methyl ketone derived bio-fuels, made from sugar and genetically modified bacteria.

Chemicals called methyl ketones are naturally occurring compounds common in tomatoes and other plants. Methyl ketones are used to provide scents in essential oils and flavoring in cheese and other dairy products.

Methyl ketones can also be manufactured from bacteria like E. coli. It is from this bacteria that researchers are looking at ways to manufacture a new class of bio-fuel. The researchers, based at the U.S. Department of Energy's Joint BioEnergy Institute (JBEI), have used glucose and a genetically modified E. coli bacterium to manufacture a methyl ketone bio-fuel which has a high cetane rating (a measurement of the combustion quality of diesel fuel, similar to the octane rating used to classify gasoline). The bacteria was genetically modified so that it contained special enzymes which allowed it create a diesel bio-fuel.

Harry Beller, a JBEI microbiologist who led this study, is quoted by the Lawrence Berkeley National Laboratory as saying:

“Our findings add to the list of naturally occurring chemical compounds that could serve as bio-fuels, which means more flexibility and options for the bio-fuels industry. We’re especially encouraged by our finding that it is possible to increase the methyl ketone titer production of E. coli more than 4,000-fold with a relatively small number of genetic modifications.”

The research was published in the following journal:

E.-B. Goh, E. E. K. Baidoo, J. D. Keasling, H. R. Beller. Engineering of Bacterial Methyl Ketone Synthesis for Bio-fuels. Applied and Environmental Microbiology, 2011; 78 (1): 70

Bio-diesels are the fastest growing class of bio-fuels and it is being produced at increasing rates across the U.S. from primarily soy-bean oil, but also canola, peanut, sunflower, mustard and even algae. Posted by: Dr. Tim Sandle

Dengue-resistance spreads in mosquitoes

Researchers based in Australia have modified a strain of bacteria (Wolbachia pipientis), which has been  used to infect mosquitoes carrying the dengue virus. The researchers have shown that the bacterium can be passed rapidly throughout a local population of mosquitoes in the wild.

The bacterium can halt the reproduction cycle of the dengue virus.  While the exact mechanism is unknown, the bacteria likely compete for limited sub-cellular resources required by the virus for replication.

This was originally reported in Nature.

Posted by: Dr. Tim Sandle

EMA Guidance on Genetic Variability

The EMA has published a guideline that addresses the influence of patients’ genetic variability on drug pharmacokinetics. The guideline on the use of pharmacogenetic methodologies in the pharmacokinetic evaluation of medicinal products clarifies the requirements for drug developers to analyse the effect of genetic variability on their medicines. Adopted by the EMA’s Committee for Medicinal Products for Human Use in January following a public consultation, the guideline is expected to come into effect on 1 August 2012.
Genetic variation can affect the body’s absorption, distribution, metabolism and excretion of medicines, which can subsequently impact a medicine’s benefits and risks. Although pharmacogenetics are not equally important for every drug, the guideline recommends that prospective banking of DNA for genotype analyses be conducted in all clinical phases of development. Even if there is no obvious indication of a genetic influence on pharmacokinetics, effects may be identified at later stages of development or in postmarketing, which was the case for tamoxifen and clopidogrel where activation by polymorphic enzymes was identified during pharmacovigilance monitoring.

According to the guidance, studies of the effect of pharmacogeneitcs will usually be required when the magnitude of interindividual variation in drug exposure is high enough to potentially affect a medicine’s safety and/or efficacy. The guideline provides recommendations on where pharmacogenetics should be implemented into the drug development process and applies mainly to small-molecule drugs, as the genetic effects on the pharmacokinetics of biological drugs are not yet as well understood.
The guideline also addresses:

• recommendations and requirements for study designs, subject selection and sampling
• how to evaluate the clinical impact of genetic differences between patients and recommendations for further studies
• possible consequences of differences and the impact on treatment recommendations and labelling
• consideration of drug–drug interactions
effects of impaired or immature organ functions on pharmacogenetics related pharmacokinetic studies.

The full guideline can be read on the EMA website.

Posted by: Dr. Tim Sandle

Natural Killer Cells

Here is an interesting video from the Cancer Foundation.




Natural Killer Cells are the most aggressive white cells in the immune system. They make up about 5% to 15% of the total lymphocyte circulating population. They target tumor cell and protect against a wide variety of infectious microbes. Natural Killer Cells are a very important factor in the fight against cancer. Immune Stimulation is the key to keeping the white blood cell count high and giving the Natural Killer Cells a chance to fight cancer and other diseases.

Posted by: Dr. Tim Sandle

Cleanroom Cleaning and Disinfection: Eight Steps for Success

Cleanrooms in healthcare and pharmaceutical facilities must be kept in a state of microbiological control. This is achieved in a number of ways, including the physical operation of Heating, Ventilation, and Air Conditioning (HVAC) systems, control of materials, properly gowned and trained personnel, and through the use of defined cleaning techniques, together with the application of detergents and disinfectants.

The object of cleaning and disinfection is to achieve appropriate microbiological cleanliness levels for the class of cleanroom for an appropriate period of time. Thus the cleaning and disinfection of cleanrooms is an important part of contamination control.

On this important subject, Tim Sandle has written an article for Controlled Environments magazine entitled "Cleanroom Cleaning and Disinfection: Eight Steps for Success".

Here is an extract:

When selecting a disinfectant, points to consider are:


a) To satisfy GMP regulations, two disinfectants should be used in rotation. While scientifically this may not be necessary, many regulatory agencies expect to see two different disinfectants in place. For this, the two agents selected should have different modes of activity.⁴ It may be prudent for one of the disinfectants to be sporicidal.
b) The disinfectant should have a wide spectrum of activity. The spectrum of activity refers to the properties of a disinfectant being effective against a wide range of vegetative microorganisms including Gram-negative and Gram-positive bacteria.
c) Ideally the disinfectant should have a fairly rapid action. The speed of action depends upon the contact time required for the disinfectant to destroy a microbial population. The contact time is the period of contact when the surface to which the disinfectant is applied must remain wet.
d) Residues from organic materials or detergent residues should not interfere with the disinfectant.
e) Disinfectants used in higher grade cleanrooms (like ISO 14644 classes 5 and 7) must be supplied sterile or be sterile filtered by the cleanroom operators.
f) The disinfectant should be able to be used at the temperature at which the cleanroom operates. If a cleanroom is a cold store then it needs to be checked whether the disinfectant will work at that temperature.
g) The disinfectant should not damage the material to which it is applied or some other measures should be taken. Many sporicidal disinfectants are chlorine based and will damage material like stainless steel unless the residue is wiped away after use.
h) The disinfectant should be safe for operators to use and meet local health and safety laws.
i) The disinfectant should be cost effective and be available in the required formats like trigger spray bottles or ready-to-dilute concentrates.

The article can be viewed and printed on-line, go to: Controlled Environments

Please reference the article:

Sandle, T. (2012). Cleanroom Cleaning and Disinfection: Eight Steps for Success, Controlled Environments Magazine, March 2012 

Posted by: Dr. Tim Sandle

Cleanrooms and the Cold War

There is an interesting article on the the role of cleanrooms in the Cold War relating to the manufacture of spy satellites and components.

Here is an extract:



"For more than a decade they toiled in the strange, boxy-looking building on the
hill above the municipal airport, the building with no windows (except in the cafeteria), the building
filled with secrets.

They wore protective white jumpsuits, and had to walk through air-shower chambers before entering
the sanitized “cleanroom” where the equipment was stored.

They spoke in code."

The article is written by Helen O'Neill and a pdf can be found here: cleanrooms.

Posted by: Dr. Tim Sandle

Contamination Control in the Life Sciences

The latest issue of Life Sciences Industry Insights features an article by Gordon Farquharson, an expert in cleanroom design and standards, on contamination control practices in the Life Science industry with a focus on bio-containment and cleanroom applications.   Don Thornburg, a Director of R&D for HVAC products and Chair of the 52.2 committee, reviews the history and current state of ASHRAE testing standards.

There is also an article by Jan Andersson, Deputy Managing Director for the Nordic countries and Chairman of PG4B within Eurovent, updates readers on EN779 2011, and its mission of classifying air filters based on their lowest filtration efficiency (ME).  Andersson also discusses the increasing influence of "ZEBs" (zero-energy buildings). Other contributors include Tomm Frungillo, who writes on dust collection within the Latin American pharma sector, and Sean O'Reilly, Global Director, Cleanroom & Bio-Pharma Segment, who offers a broad perspective on the changes within the industry over the last decade.

To view the digital publication, go to: Life Sciences 

Posted by: Dr. Tim Sandle

FDA to protect important class of antimicrobial drugs for treating human illness

The U.S. Food and Drug Administration (FDA) have announced that certain uses of the cephalosporin class of antimicrobial drugs in cattle, swine, chickens and turkeys will be prohibited as of April 5, 2012 (prohibition FDA-2008-N-0326).

Cephalosporins are commonly used in humans to treat pneumonia as well as to treat skin and soft tissue infections.

The FDA has taken this action to preserve the effectiveness of cephalosporin drugs for treating disease in humans. Prohibiting the uses in animals is intended to reduce the risk of cephalosporin resistance in certain bacterial pathogens.

To read more: FDA

Posted by: Dr. Tim Sandle

Norovirus: interesting video

Here is an interesting video about norovirus (first broadcast on CBS News).

Noroviruses are a group of related, single-stranded RNA, non-enveloped viruses that cause acute gastroenteritis in humans. The most common symptoms of acute gastroenteritis are diarrhea, vomiting, and stomach pain. Norovirus is the official genus name for the group of viruses previously described as “Norwalk-like viruses” (NLV).

Noroviruses spread from person to person, through contaminated food or water, and by touching contaminated surfaces. Norovirus is recognized as a leading cause of foodborne-disease outbreaks.


Posted by: Dr. Tim Sandle

Cleanroom humidity

PharmPro have an interesting white paper on their website. The paper relates to the monitoring of cleanrooms for humidity and it is written by Jim Tennermann of Vaisala.

Here is an extract:

“Every cleanroom has specifications for environmental control that define upper and lower limits for temperature and relative humidity (RH). Creating and maintaining these conditions in the cleanroom is no small feat, and this must be achieved while also controlling pressure, flow, and contamination. Devices used for measuring RH (variously referred to as sensors, transmitters, or instruments) play a small role in the overall picture, but problems with these devices can cause significant headaches for cleanroom operators. What follows are some observations that may be relevant to anyone in the cleanroom community who has struggled with humidity issues.”

To review the paper go to: PharmPro


Posted by: Dr. Tim Sandle

Cleaning cleanrooms

Posted by: Dr. Tim Sandle

New take on antibiotics

With antibiotic resistance on the rise in strains of pathogenic bacteria, innovative strategies are needed to discover ways of treating bacterial infections in both humans and in agriculture. Scientists have studied the molecular machine known as the 'type II bacterial secretion system', which is responsible for delivering potent toxins from bacteria such as enterotoxigenic E. coli and Vibrio cholerae into an infected individual.

The research was published in:

Shuang Gu, Saima Rehman, Xiaohui Wang, Vladimir E. Shevchik, Richard W. Pickersgill. Structural and Functional Insights into the Pilotin-Secretin Complex of the Type II Secretion System. PLoS Pathogens, 2012; 8 (2): e1002531 DOI: 10.1371/journal.ppat.1002531

According to Science Daily, Professor Richard Pickersgill, who led the research, said: "Bacterial secretion systems deliver disease causing toxins into host tissue. If we can understand how these machines work, then we can work out how it they might be stopped."

In order to infect, Gram negative bacteria have to export their toxins into their host across both an inner and outer membrane. Professor Pickersgill explains: "The pore in the outer membrane which the toxins pass through is formed from protein subunits which are guided into place by a protein pilot. The protein pilot interacts with the subunits that form the pore in the outer membrane; if the protein pilot is missing, then the pore forms in the inner membrane and not the outer membrane and secretion is stopped."

Professor Pickersgill adds: "If we can successfully interfere with this or with other interactions we are discovering then we might be able to halt the secretion system and prevent these harmful diseases."

The same type II secretion system that enables E.coli and cholera is also used by bacteria that cause substantial food spoilage, such as Dickeya dadantii. Crop spoilage by plant infecting bacteria is becoming an increasing problem in the UK due to the warmer and wetter summers caused by global climate change, and the team hope that the results of this study will be of interest to both agrichemical scientists seeking ways of preventing crop damage and pharmaceutical companies seeking new antibiotics.

Posted by: Dr. Tim Sandle

Clostridium toxin: possible treatment for asthma and arthritis

The conditions of rheumatoid arthritis, asthma, psoriasis and other diseases might be relieved by the toxins produced by the Clostridium botulinum bacterium (which is normally associated with a rare but severe form of food poisoning. The toxin is the most powerful neurotoxin ever discovered). This is according to a paper published in American Chemical Society (ACS) Journal of Biochemistry.

The toxins have been used for some time to treat nerve disorders and facial wrinkles ('botox'). For these treatments, small doses of the toxin is injected. The toxin blocks the release of the neurotransmitters, or chemical messengers, that transmit signals from one nerve cell to another by breaking down the protein within the nerve cells which functions to release neurotransmitter. This lack of neurone response disrupts the nerve signals that cause pain, muscle spasms and other symptoms in certain diseases.

Because similar proteins exist as receptors in other cells in the human body, the Clostridium toxin can theoretically be applied elsewhere. This has been the basis of research undertaken by Edwin Champan at the Howard Hughes Medical Institute.  Initial findings indicate that a re-engineered botulinum toxin blocks the release of a protein from immune cells linked to inflammation. Inflammation is part of the immune response and is the key to the way in which the human body reacts to many diseases.  These diseases include chronic inflammatory diseases including celiac disease, vasculitis, lupus, chronic obstructive pulmonary disease (COPD), irritable bowel disease, atherosclerosis, arthritis, and psoriasis.

Journal reference:

“Retargeted Clostridial Neurotoxins as Novel Agents for Treating Chronic Diseases” Biochemistry, 2011, 50 (48), pp 10419–10421. DOI: 10.1021/bi201490t

Posted by: Dr. Tim Sandle

Cleanroom gloves (survey)

The Cleanroom Forum has a feature on cleanroom gloves.

When working in cleanroom environments, in which strong measures are taken to control the level of contamination that occurs within the space, common supplies such as work gloves must be specially manufactured to generate minimal pollution. Cleanroom gloves are made from fibers and materials that are suitable for cleanrooms, providing workers and researchers with the dexterity and form fitting comfort necessary to perform precise and accurate work.

In the post there is a discussion of materials used for gloves. These include:

• Latex: Latex is a natural rubber and is highly flexible. It is made from the liquids secreted by the rubber plant and by virtue of its conformity, latex gloves are comfortably used in precision activities, such as carrying out surgical procedures and other lab operations.
• Nitrile: It is a copolymer fabric which contains propane nitrile in different measurements. Based on its propane nitrile content, its chemical resistance and stiffness varies. The higher the content, the more the stiffness and chemical resistance.
• Polyvinyl chloride (PVC): It is a stiff material that is made softer by adding plasticizers. PVC is the perfect option for protective clothing and gloves in a cleanroom.
• Polyurethane: It is an anti-abrasion material that is coated onto other fabrics and used to make PPE.
• Neoprene: It is a synthetic rubber and is superior to latex. It comes with chemical resistance features and is widely used in chemical operations in labs.

To read more visit: cleanroom gloves.

Posted by: Dr. Tim Sandle

Pharmaceutical Microbiology on Facebook

Pharmaceutical Microbiology's page on Facebook has a new look.

It can be found here: Pharma Micro

Posted by: Dr. Tim Sandle

Bacteria - Magnetic Swimmers

A bacteria species that uses a magnetic sulfide compound to swim towards food.

The Scientist has an interesting feature on so-called 'magnetic bacteria'. It relates to findings published in the following paper:

C. Lefèvre, et al, “A Cultured Greigite-Producing Magnetotactic Bacterium in a Novel Group of Sulfate-Reducing Bacteriam” Science, 334, pp. 1720-1723, 2011.


The Scientist surmises:

"Inside their cells, magnetotactic bacteria grow chains of 40- to 100-nanometer-long magnetic particles to help them align with the earth’s magnetic field. These tiny compasses allow the water-borne bacteria to swim back and forth in one direction, rather than wiggling around in three dimensions in a more random hunt for food."

In the paper, scientists state that they have been able to isolate and grow magnetotactic bacteria that produces greigite in pure culture. Here research has shown that magnetotactic bacteria are able to grow chains of 40- to 100-nanometer-long magnetic particles to help them align with the earth’s magnetic field.

To read more, go to: magnetic bacteria


Posted by: Dr. Tim Sandle