Shellfish toxins

Domoic acid is a toxin naturally produced by species of algae belonging to thePseudo-nitzschia genus, particularly affecting shellfish. Scientists at the University of California, Santa Cruz, USA, investigated the toxin’s persistence in the marine life, long after the bloom had disappeared, and found that it had found its way into the muscle tissue of some commercial species of fish. Although the toxicity levels were well below regulated limits, being present in muscle tissue means it stays in the food web for much longer than if it was simply in the fish’s digestive system. These results could have implications in the seafood trade, and researchers will need to continue to monitor and predict when prolonged algal blooms occur.

See:

• ScienceDaily

 Posted by Dr. Tim Sandle

Protein purification workflow

Identifying and establishing an efficient protein recovery process can be a time-consuming endeavour. In addition, a case study compares the traditional sequential chromatography to a multidimensional workflow, demonstrating that both methods yield proteins of equivalent purity.

To assist with this, Pharmaceutical Process has issues a new e-book. The e–book shares best practices for downstream purification, and shows how continuous purification methods can offer increased productivity and greater process reliability and reproducibility in the laboratory and when scaling protein purification processes for the production of commercial quantities.

In addition, a case study compares the traditional sequential chromatography to a multidimensional workflow, demonstrating that both methods yield proteins of equivalent purity.

For details see: PharmaPro



 Posted by Dr. Tim Sandle

Microbe Menaces Emerging Infectious Diseases

Infection Control Today have a free digital issue on the subject “Microbe Menaces Emerging Infectious Diseases, Hospital Pathogens Remain an Ongoing Challenge”.

The article can be accessed here.

This publication explores disease caused by emerging infectious threats as well as hospital pathogens, and the interventions – such as hand hygiene, contact precautions and environmental cleaning – that can be used to control and prevent infection transmission.



 Posted by Dr. Tim Sandle

Lessons Learned in the Cleanroom

Cleanroom components: Install anti-contamination flooring to minimize particulate carry-over, both foot-borne and from cart wheels. Find a qualified third party expert to annually certify the acceptable performance of the air handling systems inside the controlled areas (HEPA filters, BSC, LFH, isolators). Integrate automation/robotics when feasible to limit interventions by personnel.

In relation to this by Deborah Hoffer has written an article for Controlled Environments magazine. The piece can be accessed here.

Posted by Dr. Tim Sandle

What is the impact of disinfectants on environmental testing?

Posted by Dr. Tim Sandle

Sporicides – As Part Of Your Transfer Process in Practice

Do you monitor, run or work in an aspectic facility?  If yes – you should be attending this event!

Please see below (or contact Pharmig):

Update on Sporicides as Part of Your Transfer Process - And Booking From

Posted by Dr. Tim Sandle

Environmental Monitoring Program: Hot topics in Microbiology & Best Practices

The manufacture of sterile pharmaceutical product requires specific care Regarding environment, premises and practice of the personnel. Regulations in place to be used by the Pharmaceutical Industry give requirements on the environmental monitoring program. With the publication of FDA Warning Letters and 483 That form shows environmental monitoring procedure and practice of the staff belong to top 10 of the gaps found During inspections.

Benedict RAMOND has written an interesting article for the magazine La Vague. The article can be found here.

Here is an extract:

“This article, for the Environmental Monitoring program (EM) in aseptic processing as well in Conventional Clean Rooms (CCR) or using Isolator / RABS technology: Describes the main expectations of the règlements in place, presents the hot topics in microbiology and Gives solutions for implementation of best practices. From the gaps highlight highlighted by FDA During inspections, Microbiology The Following topics will be Described: samples rent justification, missing samples and data interpretation, trending system and CAPA system.”

Posted by Dr. Tim Sandle

Bromate and water

The determination of the bromate content of water is a challenge that can be met either by using an elaborate ion chromatography system or a photometric method. Both approaches demand careful handling and a critical appraisal of the result that is yielded. If the necessary working steps are carefully followed, photometry constitutes a true alternative to chromatography for those laboratories that do not possess ion chromatography systems.

Laboratory Manager is offering a free white paper on this subject, which can be requested here.



Posted by Dr. Tim Sandle

Best Practices in Environmental Monitoring:

A new Pharmig one day course - 'Best Practices in Environmental Monitoring'.

For details see:

Best Practices in Environmental Monitoring

Posted by Dr. Tim Sandle

Has A New Form Of Microbial Life Been Discovered?

The human intestines are home to trillions of microorganisms (many bacteria, some fungi, and by-products – collectively known as the microbiome). Several strands of research are showing how the numbers and types of certain bacteria are important for the regulation of immune function and how disruption can lead to autoimmune diseases.

New research suggests we may need to re-consider how we track and categorize the microorganisms within this ecosystem. Scientists at the Pierre and Marie Curie University in Paris, France are looking at how to redefine the classification of bacteria found within the colon. This is through examining 86 gene families using next-generation sequencing methods.

The findings suggest the current three categories for microbial life – bacteria, archaea, and eukaryotes (which includes fungi and single-celled parasitic animals) – could be insufficient. In short, a case could be made for a fourth type of microbial life. These microbes are lurking in what is termed “microbial dark matter.”

According to New Scientist magazine, the research group found 230,000 DNA sequences that are related to known sequences in those 86 gene families. This analysis found “an additional 80,000 stretches of microbial DNA that belonged in the 86 gene families.”

There is a lot of work to do, and evidence gathering to complete, before a firm case is made for a fourth domain and before this becomes universally accepted in the microbiology world. This includes isolating and culturing the organisms. Finding a means to do this, given most microbes are unculturable, will be difficult.

The researchers have put forward the notion in the journal Biology Direct. The research paper is headed “Highly divergent ancient gene families in metagenomic samples are compatible with additional divisions of life.”



Posted by Dr. Tim Sandle

Microbiomes and stress

If you are feeling stressed, your microbiome – the communities of microbes living in and on you – may be suffering too. A team at the University of Guelph, Canada, studied the link between stress and microbiomes in wild red squirrels, and found that individuals with low stress hormone levels had a more diverse microbiome, while increased stress caused a spike in potentially harmful bacteria, but less bacterial diversity.

See:

• University of Guelph
• Biology Letters

 Posted by Dr. Tim Sandle

ISO 14644 Part 13 - public comment

Part 13 of the revised international cleanroom standard is available for public comment. The details are:

‘Surface’ refers to the interface between two phases. For the purpose of this document, the surface is a solid. “Clean surface” is where one or more of the contamination categories (particles, chemical) are under control due to cleaning / decontamination. The degree of cleanliness is specified in the corresponding surface cleanliness classifications (ISO 14644-9:2012 Surface cleanliness by particle concentration SCP; ISO 14644- 10:2013 Surface cleanliness by chemical concentration SCC). Different cleaning methods are necessary depending on the degree of cleanliness (cleanliness class) required. This standard gives guidance on the selection of cleaning methods to achieve specified cleanliness levels. For the selection procedure, the aspects of surface description, cleanliness specifications, types of contamination, cleaning techniques, material compatibility, and assessment methodology are taken into consideration. Most of the methods are suitable for removal of more than one contamination category at the same time therefore a common standard for the selection of a cleaning method for both particles as well as chemical contamination is needed.

Guidance is provided on the assessment of cleaning methods for achieving the required surface cleanliness levels specified in ISO 14644-9: Classification of surface cleanliness by particle concentration and ISO 14644-10: Classification of surface cleanliness by chemical concentration. Different cleaning methods may be necessary depending on the degree of cleanliness required. The selection procedure considers aspects such as surface description, cleanliness specifications, types of contamination, cleaning techniques, and material compatibility. Guidance is provided on assessing cleaning efficacy.

The reference is: 

Draft for Public Comment 16/30280732 DC Draft ISO 14644-13 "Cleanrooms and associated controlled environments Part 13 Cleaning of surfaces to achieve defined levels of cleanliness in terms of particle and chemical classifications."

Interested parties should contact their local standards committee to obtain a copy.

Posted by Dr. Tim Sandle

Striving for the highest Grade

As a prototype for the future, is it possible to consider an energy efficient filter system that cost-effectively maintains airflow towards a reduced rate of change? Michael Rodd, Chief Sales Officer at M+W Products (www.products.mwgroup.net) looks at elements of life sciences in the clean room and envisages how future practice can radically change the concept of clean room procedures.

It is common to expect the output of a clean room operation to end as a microprocessor in your phone, laptop or car, but in fact, the first measured effort to control an environment was used for medicinal purposes on a table in a hospital. British surgeon Joseph Lister in 1867, at a time when multiple surgeries were performed using the same contaminated equipment, sterilised his hand using carbolic acid and experimented implementing this technique on wounds and needles to discover the elimination of bacteria.

Although much has changed since, and both our efforts and realisations of scientific procedures have adapted, the dedication of modern life sciences was reinvigorated as a fairly new discipline of the clean room subdivision over the last century. This process encapsulates everything utilised by the way of clean room technologies in the production of medical substances, pharmaceuticals and biological compounds.  

As the history of the pharma world displays, our requirement for a clean room stems from the need for contamination control. Whether that’s using gloves during primitive operations or alcohol as an antiseptic to sterilise needles or creating paint and lacquer work out in uncontaminated air near the Pacific Ocean, our need to benefit from such processes has been fuelled by the requirement to control and clean our imminent environment.

This demonstrates something else too – that the clean room has developed from a permutation of science and engineering – assessing scientific processes that have worked and engineering them to deliver better results. This multidisciplinary approach has been encased by technological practices available at certain periods of time.

Fundamental to the field of life sciences and pharmaceutical micro biotechnology is contamination control. First and foremost, the apparatus used for pharmaceuticals in clean room laboratories should be dedicated and separated from other areas. Nailed down, this involves the classification between protecting the working environment and protecting the operator from any potential contamination. This is because humans remain the biggest cause of contamination; that may never change. To put this into perspective, the outer layer of human skin can host up to 1 million microorganisms per square cm, and equally, human saliva up to 1 billion per square ml.

The frenzy of microbes is difficult to keep on top of at every stage of the process. Basic negligence can cause the microorganisms from our clothes, mobile phones or bare skin to contaminate working stations. For instance, hands, whether gloved or un-gloved, are one of the main sources of spreading infection or transferring microbial contamination. Thus, an important part of good contamination control within a cleanroom requires the use of cleaning and disinfection agents. It goes without saying that just as personnel may be the biggest contaminants, they are also critical to the maintenance of asepsis in a controlled environment. Therefore diligence and training in clean room technology is essential throughout the entire process.

The improper analysis of microbiological inspections may cause inadvertent contamination. Therefore, thorough aseptic processing like product or microbial bioburden to calculate viable organisms is needed to prevent any sort of contamination during the process stream. In some cases, even measuring total particulate count within a vessel does not cater for the continuous generation of organisms by individuals; therefore it does not always quantitate all contaminants and provide the whole picture of microbiological content. Microorganisms will associate with physical particulates and therefore it is necessary to include monitoring techniques that satisfy both the classification and regulatory requirements by differentiating the microbiological components of an operation. Stringent optimisation of tests will give assurance that bioburden of the environment is apt for clean laboratory practices.

In its simplest form, a product being operated within a clean room during a process line includes an enclosed vessel or large container that is sealed from external air temperature. Throughout this process, it is the aim of every operative to maintain conditions to a level where they can be declared sterile or aseptic.

A clean room is classified based on the cleanliness of its air so as far as clean rooms go, anyone wants to keep the environment as clean as possible. One of the trends we’ve seen across this spectrum (and it is a large spectrum) is on the lower end quality scale concerned with areas that are Clean Not Classified (CNC). These are areas that can be regarded clean within definitive purposes of the word in regards to the production area, but they are not actually classified because they work on a process we call Good Manufacturing Practice (GMP) or Current Good Manufacturing Practice (CGMP).

The ability to manufacture medical devices with consistent high quality relies on well implemented and well documented GMP. The GMP code sets out a guideline to achieve sterility assurance. Any business that delivers clean room technology requires constant monitoring and up-to-date certification.

The measurement most universally applied is the Grade A-D standard, whereby a cubic foot sample is taken of the environment and the number of particles greater than 0.5mm measured within it.  Areas are classified on a graded system from A onwards, with A being the cleanest. So the purpose of a low quality CNC is to effectively get as close as possible to classification. D certified environments accept the inclusion of certain particles even in dynamic conditions (i.e. when people are operating within contained vessels) with turbulent airflow, whereas A certified environments deal with linear laminated airflow that is non-turbulent. Cleanliness is commonly achieved through the development of effective HEPA (High Efficiency Particulate Air) filtration and this is considered as clean as necessary for a GMP Grade A condition.

Let’s complicate things a step further. Introduced here is the onion concept. Its name is derived from the idea that as you peel the onion, a processor must past through successive cleaner areas to reach the centre, non-turbulent, Grade A area. In essence, the onion concept caters for the disposal of multiple grade conditions within the same vessel or container.

Furthermore, the application of disinfectants on specimens reduces the microbial bioburden as we discussed earlier. Therefore, when collecting environmental monitoring samples, personnel should begin with Grade A locations and then move on to areas that are away from technical apparatus but still within the controlled zone.

This grade system is applicable in cases where processors and products are exposed during transportation, or operations that are required to be performed outside of sealed compartments. Grade B is commonly of high purity but does not necessarily involve laminate air flow. So the trick here lies in maintaining airflow to a certain speed. Essentially, imagine the onion concept: each time a layer of the onion is peeled, the amount of particulate or contaminated risk to the product is reduced.

For any sales or financial operative, however, this all costs money and clean rooms are an expensive market already. The integration of new technology and efficient solutions has ignited consideration of future controlled environmental procedures to limit the number of air changes per hour. Can future systems be both more efficient and challenge the tradition of multiple air changes and still ensure constant clean air? This is a radical thought and not on the agenda yet, but with a global drive for efficiency and cost-saving mechanics already found within homes and our cars, is there a need for such ideas to be infused as part of the life science debate? A future model to strive for would be to find an energy efficient procedure of controlled environments that maintains the right quality and temperature at the lowest price possible. The future of pharmaceutical trends in the clean room is exciting.

New developments with antimicrobial viruses

Scientists from the National Physical Laboratory (NPL) and University College London (UCL) have converted a breast milk protein into an artificial virus that kills bacteria on contact.

The antimicrobial activities of this protein are mainly due to a tiny fragment, less than a nanometer across, made up of six amino acids. Based on the metrology of antimicrobial mechanisms, the team predicted that copies of this fragment gather at the same time, and at the same point, to attack bacterial cells by targeting and disrupting microbial membranes.

For further details, see the news report from the U.S. National Physical Laboratory.


 Posted by Dr. Tim Sandle

Tuberculosis Is Coming Back – Here’s Why

Tuberculosis is a disease caused by bacteria that infects the lungs. It is highly contagious and can potentially be fatal, but until recently it was on the way out.

A special report by Megan Ray Nichols.

During the last 20 years, overuse of antibiotics has helped to create drug-resistant strains of the bacterium that have turned tuberculosis from a Victorian-era disease that was often romanticized into something that needs to be feared and respected once again.

Concentration in Cases

While the overall number of tuberculosis cases in the U.S. is dropping, a number of population subsets are increasingly affected by the disease.

A number of factors put individuals at risk for contracting TB, including weakened immune systems or medical professionals pass on the infection after treating infected patients. A 2014 report by the CDC revealed some worrisome trends, including the fact that more than 50 percent of TB cases are concentrated in four states: Florida, New York, Texas and Florida.

What do those four states have in common? They are also the main entry points for immigrants, both legal and illegal, who are entering the United States. As of 2013, more than 64 percent of TB cases were diagnosed in foreign-born individuals who had come to the United States.

Since a standard case of TB often costs around $17,000 to treat, many people who don’t have access to health care or insurance often go untreated, spreading the disease to anyone that they may come into contact with. Drug-resistant TB can cost even more, with a full treatment reaching $134,000. Extreme drug-resistant TB, which doesn’t respond to any of the traditional treatments, can cost nearly half a million dollars and even this cost doesn’t guarantee that the patient will be cured of the disease.

What Has Been Done?

TB causes more deaths globally than any other infectious diseases. It is airborne and released when an infected patient coughs or sneezes, and can easily be caught by anyone who is in close proximity to the infected individual.

The current immigration polices both in the U.S. and in areas like London do not allow for the screening and/or treatment of potentially infected individuals. Instead, large numbers of these individuals are living together, often in small or substandard housing which creates the perfect transmission conditions for TB and other similar diseases.

The Centers for Disease Control have guidelines in place requiring medical examinations for any refugees or immigrants coming into the United States, whether they are here on a temporary visa or permanently. Unfortunately, they can only perform these exams if the individuals are entering the country legally. Individuals who carry TB and other similar diseases but are not able to be examined are carrying a disease that was nearly eradicated in many countries into fresh breeding grounds.

What Precautions Need to be Taken?

While there is a great call right now for countries to open their borders in the wake of the events in Syria that left many as refugees, the borders should be opened intelligently.

• Housing needs to be improved for refugees or immigrants so that large numbers of people are not crammed into small spaces.
• Individuals need to submit to a medical exam before being allowed to enter a country. A basic TB test usually takes less than 24 hours if the patient isn’t symptomatic. 
• Medical establishments need to be more approachable — if an individual is presenting symptoms of TB, they should be able to approach a medical establishment for treatment, if for no other reason than to prevent the disease from spreading farther. 
• The public as a whole, including refugees and immigrants, needs to be educated on the symptoms and effects of the disease. Someone might go about their day just thinking they have a bad cough while transmitting TB to everyone they meet.

These precautions might seem a bit harsh, but with many people crossing borders into countries where TB was nearly a thing of the past, heavy-handed precautionary measures may be just the thing to help us wipe out TB once and for all.

Bingeing on health foods won’t boost your immune system

An interesting statement: "There are only two ways the human body can deal with the invading pathogens and infections that can cause colds and other illnesses – and neither involves vitamins or ‘superfoods’ that claim to offer protection."

This comes from an article published in The Guardian, by Dara Mohammadi.

Here is an extract: "The immune system can be broadly split into two parts, the innate and the acquired response. On detection of infection, it’s the innate response that acts first. Though fast, it lacks in finesse, and deals with an invading pathogen in much the same way that the Ghostbusters might try to remove a ghost from a haunted hotel. It gunges the halls and doorways to try to flush it out (that’s why you fill up with phlegm and snot), it yanks up the thermostat to try to boil it (why you run a fever), and it shuts down the building until the problem is solved (it makes you depressed and lethargic so you don’t go out and pick up another infection while your immune system is at work)."

To view the full article, see The Guardian.

Posted by Dr. Tim Sandle

On pathogenic bacteria

Some of the most pathogenic bacteria in the world possess the ability to quickly overwhelm and kill their prey cells, by injecting toxic proteins into them – often using a process called type VI secretion (T6S). When first discovered in 2006, scientists believed this system was unstoppable, but a new study from Princeton University, USA, and the University of Basel, Switzerland, revealed that if the target cells are a large enough group, the predatory bacteria cannot overpower them all. Although the outer organisms are vulnerable to attack, if the group is big enough the inner cells can reproduce at a rate that means the pathogen is unable to take over.

See:

• Princeton University
• PLOS Computational Biology

 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.

In an article for Controlled Environments magazine, Tim Sandle examines eight key steps to be followed, in relation to cleaning and disinfection, in helping to keep cleanrooms “clean.”

To view the article, see CE.



Posted by Dr. Tim Sandle

Bacterial Jiggle: Curious Motions of Syphilis and Lyme Disease (video)

The bacteria that cause syphilis and Lyme Disease have something extraordinary in common: they manage to propel themselves through their environment in spite of the fact their tails are located inside their bodies.

Jennifer Frazer has written an interesting article on the curious motions of specific bacteria for Scientific American.

The bacteria that cause syphilis and Lyme Disease have something extraordinary in common: they manage to propel themselves through their environment in spite of the fact their tails are located inside their bodies.

For bacteria, they're also unusually shaped and active. In this movie, you can see the bacteria that cause Lyme Disease moving like living, squirming cavatappi.

To read the article in full, see Scientific American.



Posted by Dr. Tim Sandle