Sunday 24 December 2017

Season's Greetings!

Season's Greetings!

Thank you for supporting Pharmaceutical Microbiology over this past year. The website and the various groups continues to grow and feature a range of microbiology, pharmaceutical and healthcare related news stories.

All the best for the holiday season.

Dr. Tim Sandle

Wednesday 20 December 2017

Draft EU GMP Annex 1 released

The PIC/S Secretariat has notified that the revised EU-PIC/S GMP Annex 1 on the Manufacture of Sterile Medicinal Products has reached Step 2 of the revision process and on 20 December 2017, the PIC/S and EMA published the draft revision of Annex 1 for public consultation.

The consultation period will last 3 months and run from 20 December 2017 to 20 March 2018.

The revised Annex 1 has been prepared in co-operation with the EMA, World Health Organization (WHO), and PIC/S in order to maintain global alignment of standards, and provide assurance of product quality. The document is subject to parallel public consultation by the European Commission (EC), WHO and PIC/S.

Key changes from the earlier PIC/S Annex are:
  • Introduction of new sections: scope, utilities, environmental and process monitoring sections and glossary.
  • Introduction of the principles of Quality Risk Management (QRM) to allow for the inclusion of new technologies and innovative processes.
  • Restructuring to give more logical flow.
  • Addition of detail to provide further clarity.
The revised Annex 1 is downloadable on the PIC/S website (site link) or via this direct link.

The draft has been formatted with prescribed line and page numbers to support a joint international consultation within TGA, PIC/S, WHO and the EC.

Posted by Dr. Tim Sandle

2017 Author of the Year – Journal of GXP Compliance

Tim Sandle has been awarded '2017 Author of the Year – Journal of GXP Compliance' by the Instiute of Validation Technology. This is for Dr. Sandle's contributions to the Journal of GXP Compliance, includig:

Biodecontamination of Cleanrooms and Laboratories Using Gassing Systems V21 #1

Matrix Approach for the Qualification of a Pharmaceutical Facility Autoclave V21 #4

Pharmaceutical Microbiology: Current and Future Challenges V21 #4

Tuesday 19 December 2017

How the immune system identifies invading bacteria

The body's homeland security unit is more thorough than any airport checkpoint. For the first time, scientists have witnessed a mouse immune system protein frisking a snippet of an invading bacterium. The inspection is far more extensive than researchers imagined: the immune system protein, similar to those in humans, scans the bacterial protein in six different ways, ensuring correct identification.


Jeannette L. Tenthorey et al. The structural basis of flagellin detection by NAIP5: A strategy to limit pathogen immune evasionScience, 2017; DOI: 10.1126/science.aao1140

Posted by Dr. Tim Sandle

Monday 18 December 2017

How to gown properly for cleanrooms

As shown below, three videos showing how to gown properly when entering cleanrooms.

Practical workshop on good cleanroom gowning from Simon Fiala – Key Account Manager, COMPREI Reinraum.

Posted by Dr. Tim Sandle

Saturday 16 December 2017

Strain Collection for Improved Expression of Outer Membrane Proteins

An article of interest in Frontiers in Cellular and Infection Microbiology. Here is the abstract:

Almost all integral membrane proteins found in the outer membranes of Gram-negative bacteria belong to the transmembrane β-barrel family. These proteins are not only important for nutrient uptake and homeostasis, but are also involved in such processes as adhesion, protein secretion, biofilm formation, and virulence. As surface exposed molecules, outer membrane β-barrel proteins are also potential drug and vaccine targets. High production levels of heterologously expressed proteins are desirable for biochemical and especially structural studies, but over-expression and subsequent purification of membrane proteins, including outer membrane proteins, can be challenging.

Here, we present a set of deletion mutants derived from E. coli BL21(DE3) designed for the over-expression of recombinant outer membrane proteins. These strains harbor deletions of four genes encoding abundant β-barrel proteins in the outer membrane (OmpA, OmpC, OmpF, and LamB), both single and in all combinations of double, triple, and quadruple knock-outs. The sequences encoding these outer membrane proteins were deleted completely, leaving only a minimal scar sequence, thus preventing the possibility of genetic reversion.

Expression tests in the quadruple mutant strain with four test proteins, including a small outer membrane β-barrel protein and variants thereof as well as two virulence-related autotransporters, showed significantly improved expression and better quality of the produced proteins over the parent strain.

Differences in growth behavior and aggregation in the presence of high salt were observed, but these phenomena did not negatively influence the expression in the quadruple mutant strain when handled as we recommend. The strains produced in this study can be used for outer membrane protein production and purification, but are also uniquely useful for labeling experiments for biophysical measurements in the native membrane environment.

For further details see: Frontiers

Posted by Dr. Tim Sandle

Friday 15 December 2017

How Helicobacter pylori causes gastric cancer

Gastric cancer is one of the five most fatal types of cancer. According to the statistics of the World Health Organization (WHO), about 750,000 patients die each year after developing the disease. The main cause is thought to be the bacterium Helicobacter pylori (H. pylori). At present, there are no effective therapies for gastric cancer and growing spread of antibiotic resistances is further complicating treatment of the infection. Researchers at Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU) have now identified two mechanisms through which this bacterium can cause gastric cancer. Their findings could result in the development of new therapeutic approaches.

The international team of scientists headed by Dr. Nicole Tegtmeyer of the Chair of Microbiology at Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU) investigated how bacteria destroy the stomach's protective layer. This protective layer is composed of densely packed epithelial cells that protect the stomach against the effects of gastric acid. The researchers have now discovered that H. pylori secretes an enzyme, a protease called HtrA, which it uses much like a weapon to penetrate this protective layer. HtrA cleaves the three proteins occludin, claudin-8 and E-cadherin, rupturing the layer of epithelial cells. As a result, the H. pylori bacteria can access deeper, normally pathogen-free tissue layers, and inflict further damage. This is the first step towards gastric cancer starting to develop.

This first phase, however, is followed by one that is even more dangerous, as the team discovered. Needle-like protrusions, termed type IV secretion systems, are activated and function as 'molecular syringes'. Using a receptor-dependent mechanism, these inject a bacterial toxin, the CagA protein, through the basolateral membrane of the host cells. The injected CagA subsequently reprograms host cells, making them potentially cancerous. Another effect of this protein is that it prevents the human immune system from recognising and eliminating the bacteria -- a crucial mechanism for the long-term survival of H. pylori in the human stomach.


Anne E. Robinson, Nathan E. Thomas, Emma A. Morrison, Bryan M. Balthazor, Katherine A. Henzler-Wildman. New free-exchange model of EmrE transport. Proceedings of the National Academy of Sciences, 2017; 201708671 DOI: 10.1073/pnas.1708671114

Posted by Dr. Tim Sandle

Tuesday 12 December 2017

Novel antibiotic resistance gene in milk

A new antibiotic resistance gene has been found in bacteria from dairy cows. This gene confers resistance to all beta-lactam antibiotics including the last generation of cephalosporins used against methicillin-resistant Staphylococcus aureus. A transfer to S. aureus which is likely according to the researchers would jeopardize the use of reserve antibiotics to treat human infections caused by multidrug-resistant bacteria in hospitals.

Macrococcus caseolyticus is a harmless bacterium naturally found on the skin of dairy cows which can spread to milk during the milking process. It can also be present in dairy products made from raw milk like e.g. cheese. Researchers of the Institute of Veterinary Bacteriology of the University of Bern have identified a new methicillin resistance gene in strains of M. caseolyticus isolated from milk. Transfer of the gene to Staphylococcus aureus, a bacterium found on the skin and mucosa of animals and humans, would have dramatic consequences for public health. This methicillin resistance gene would turn this bacteria into a hazardous methicillin-resistant S. aureus (MRSA), which is known to cause difficult-to-treat infections in hospitals. Acquired methicillin resistance in bacteria is associated with genes mecA, mecB, or mecC. However, none of these genes were present in the M. caseolyticus strains -- they carried the novel resistance gene mecD.


Sybille Schwendener, Kerstin Cotting, Vincent Perreten. Novel methicillin resistance gene mecD in clinical Macrococcus caseolyticus strains from bovine and canine sources. Scientific Reports, 2017; 7: 43797 DOI: 10.1038/srep43797

Posted by Dr. Tim Sandle

Friday 8 December 2017

Pharmig News #69

A new edition of Pharmig News has been issued, edition 69. In this issue:
  • Industry reflections by David Keen.
  • Part 2 of Pharmig’s industry review of culture media practices.
  • Review of MHRA deficiencies – trends and topics.
  • Regulatory round-up
  • New Pharmig courses
  • And more…
For a copy or further details, contact Pharmig (

The articles by Tim Sandle in the new edition are:

Sandle, T. (2017) Industry practices relating to culture media use: The Pharmig survey (part 2), Pharmig News, Issue 69, pp2-5

Sandle, T. and Leavy, C. (2017) A focus on regulatory trends: MHRA annual deficiency review, Pharmig News, Issue 69, pp2-5

Posted by Dr. Tim Sandle

Thursday 7 December 2017

5 Keys To Aseptic Processing Improvement & Efficiency

Today, life science products should also be affordable to patients and a reasonable business proposition for manufactures. These objectives can present challenges to manufacturers as they strive to gain production and process efficiencies.

Hal Baseman has written an interesting article for Pharmaceutical Online. Here is an extract:
  • Science- and risk-based approaches should be used to obtain information needed to make decisions related to the evaluation, design, qualification, operation, and monitoring of sterile product manufacturing processes.
  • Technology should be considered to mitigate or reduce the risk to product quality identified in sterile product manufacturing processes and operations.
  • Traditional testing and monitoring methods as control strategies should be challenged to ensure they are the best means for aseptic processes.
  • New products, therapies, and technologies will present challenges to traditional and existing methods for development, manufacture, validation, and testing of sterile products.
  • Global health authority technical and regulatory guidance and requirements should be harmonized with regard to technical language and definitions.
The article can be accessed here: Pharmaceutical Online

Posted by Dr. Tim Sandle

Wednesday 6 December 2017

Data-Driven Risk Management For Quality By Design

The main hope of ICH Q8-Q10 pertaining to operational excellence is to enable (bio)pharmaceutical companies to achieve product realization. The goal is to do so by establishing and maintaining a state of control and facilitating continual improvement while responding to pressures for efficiency and profitability improvements.

Peiyi Ko, Ph.D. and Peter Calcott, Ph.D. have written an interesting article on the subject for Pharmaceutical Online. Here is an extract:

“risk management is open to individual interpretations and at times has varied and been time-consuming and not-informative for prioritization, re-inventing risk assessments and leading to wasted resources and unsatisfactory outcomes. Therefore, it is proposed to use a quantitative approach with probabilistic calculations and monetized harm to account for occurrence and severity, respectively. Specifically, failure modes and effects analysis (FMEA) is a classic tool for summarizing the modes of failure, factors causing these failures, and the likely effects of theses failures to reduce process complexity for management. It generates a risk priority score for a failure mode by multiplying the ratings for severity, occurrence, and detection.”

To access the article, see Pharmaceutical Online.

Posted by Dr. Tim Sandle

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