Hand surgeons provide update on wild animal bites

The article was prompted by the authors' experience in treating an elderly man who developed a progressive infection of the hand after being bitten on the finger by an opossum. The patient recovered after hospitalization including treatment with intravenous antibiotics.

Dr. Rao and colleagues performed a research review to identify studies of rare animal bites and stings. While many reports have discussed treatment of dog, cat, and snake bites, there has been no recent, comprehensive review focusing on the recommended treatment and potential adverse effects of less-common types of animal bites and injuries.

The review identified 71 articles, including a total of 214 patients, describing less frequently seen bite and sting injuries of the upper limb (hand and arm). Most of the studies were case reports and patient series

Aquatic animals were by far the most commonly reported type of injury, accounting for two-thirds of studies. Stings from jellyfish, lionfish and sea anemones, as well as other venomous aquatic animals, can not only cause severe pain and swelling but may sometimes lead to severe or even life-threatening complications.

Ten percent of studies reported bites by reptiles. Bites by some of these animals, such as beaded lizards, can cause envenomation leading to systemic shock.

Other reports described serious complications resulting from bites caused by small mammals and rodents such as ferrets, skunks, and squirrels. Other categories, including just a few cases each, included serious injuries caused by large mammals, scorpions and centipedes and birds.

The studies suggested that most infections resulting from animal bites are "polymicrobial," caused by several different bacteria or other germs. Infections with multiple, often unfamiliar microbes have the potential to cause tissue destruction and systemic (body-wide) reactions.

Based on the available evidence, Dr. Rao and colleagues outline quick reference principles for the treatment of wild animal bites and stings. These include specific recommendations for preventive antibiotics, providing coverage for unusual bacteria that may be present in infected wounds.

See:

Jacqueline S. Israel, James E. McCarthy, Katherine R. Rose, Venkat K. Rao. Watch Out for Wild Animals. Plastic and Reconstructive Surgery, 2017; 140 (5): 1008 DOI: 10.1097/PRS.0000000000003754

Posted by Dr. Tim Sandle

Bacteria Have a Sense of Touch

Bacteria may not have a central or sensory nervous system as we know it, but they can still physically "feel" the world around them, according to a new study.

By MICHELLE STARR

It turns out the tiny microorganisms don't just respond to chemical signals - they also have a sense of touch, and can recognise surfaces and respond to them. Our sense of touch is a very important tool for living in the world. It helps avoid hazards and dangerous surfaces, and keeps you from crushing delicate objects.

For bacteria, it helps them determine which type of surface they're in contact with - such as a mucous membrane or intestinal wall - and therefore colonise and attack host cells.

It's what happens in the first few seconds after coming into contact with a surface that's crucial for successful infection, the researchers say.

To explore the mechanism by which bacteria sense surfaces, they studied a harmless species called Caulobacter crescentus.

"We have little knowledge of how bacteria read out mechanical stimuli and how they change their behaviour in response to these cues," says senior researcher Urs Jenal of the University of Basel's Biozentrum.

"Using the non-pathogenic Caulobacter as a model, our group was able to show for the first time that bacteria have a 'sense of touch'. This mechanism helps them to recognise surfaces and to induce the production of the cell's own instant adhesive."

Some bacteria have an appendage called the flagellum - whip-like structures that propel them around. Some have just one flagellum, others have many. The juvenile C. crescentus has a single flagellum that it sheds after a set period, or after it finds a suitable surface to adhere to.

By rotating this flagellum, the bacteria can travel through liquids. But these microorganisms don't have muscles - movement is enabled by energy generated by the transfer of protons down the cell membrane.

And it's this mechanism that allows the bacteria to "feel", the researchers have found. When cells come into contact with surfaces, the motor that drives the flagellum is interrupted. This in turn interrupts the proton flow.

Within seconds of this occurring, the bacteria responds, producing the adhesinthat will anchor it in place.

This knowledge could help us understand dangerous bacteria too, says Jenal.

"Even though Caulobacter is a harmless environmental bacterium, our findings are highly relevant for the understanding of infectious diseases," he explains. "What we discovered in Caulobacter also applies to important human pathogens."

The research has been published in the journal Science.

Microbiological Culture Media: A Complete Guide for Pharmaceutical and Healthcare Manufacturers

Did you realize that 90 percent of quality control microbiology remains reliant upon culture-based methods? Taking this fact into account, Microbiological Culture Media: A Complete Guide for Pharmaceutical and Healthcare Manufacturers serves as an excellent reference because it focuses on microbiological culture media as applied to pharmaceutical microbiology.

In 23 informative chapters, this book covers how media is used in the modern pharmaceutical microbiology setting and recaps the past, signals the future, and helps interpret the present. The book has been written by Tim Sandle.

Pre-order this book through Dec. 15 and save 15%. Enter campaign code MBCM to apply discount during checkout.

This book also takes into consideration that innovations continue to arise with new media recipes that are formulated for the selection of new strains for the application of media in conjunction with rapid microbiological methods.

If you are a microbiologist working in the pharmaceutical and healthcare sectors, you can’t afford not to own this book!

Contents:

1. Application of Culture Media in Pharmaceutical and Healthcare Microbiology
2. History and Development of Microbiological Culture Media
3. The Science of Culture Media
4. Common Types of Microbiological Culture Media for Pharmaceutical Microbiology
5. The Media Kitchen and the Preparation of Microbiological Culture Media
6. Sterilization of Microbiological Culture Media
7. Quality Control of Culture Media
8. Microbial Cultures
9. The Use of Environmental Isolates in Pharmaceutical Microbiology
10. The Colony Forming Unit
11. Microbial Identification and Visual Assessment of Colonies
12. Qualification of Culture-Based Environmental Monitoring Methods
13. Incubation Strategies for Environmental Monitoring
14. Culture Media for Sterility Testing
15. Culture Media for Media Simulation Trials
16. Culture Media for Microbial Controls During Pharmaceutical Manufacture
17. Assessment of Culture Media for Water Testing
18. Culture Media for Cell Culture Work
19. Diluents and Neutralizers Required for the Pharmaceutical Microbiology Laboratory
20. Data Integrity, Computerized Systems and Microbiological Culture Media
21. Auditing Culture Media Suppliers
22. Industry Practices Relating Culture Media Use
23. Growth and Culture Based Rapid Microbiological Methods

For more details see the PDA Bookstore

Posted by Dr. Tim Sandle

Risk Based Approach to Environmental Monitoring (webinar)

Regulators frequently cite concerns with environmental monitoring and the lack of a well-thought out rationale. This shortfall can be overcome through the application of risk assessment and scientific approaches. The application of risk assessment applies to selecting location for monitoring and frequencies of monitoring; and for data assessment. This presentation outlines the primary tools that can be used to achieve this.

Instructor: Tim Sandle
Product ID: 504486

Date: Tuesday, 19 December 2017 | Time: 10:00 AM PST, 01:00 PM EST | Duration: 60 Minutes

• Understanding what environmental monitoring sets out to show, in relation to environmental control
• Appreciating the limitations of monitoring
• Understanding risk assessment tools like FMEA and HACCP and how they can be applied to environmental monitoring
• Worked examples of how to apply risk based approaches for setting monitoring frequencies
• Worked examples of how to apply risk based approaches for determining monitoring locations
• Understanding how risk assessment can assist with out-of-limits investigationsWhy Should you Attend
• To understand different approaches for environmental monitoring through the adoption of risk based methodologies. These tools can either be applied to the workplace or used to benchmark current practice against. The approaches discussed have been presented to FDA and European medicines inspectors.

Areas Covered

• An overview of environmental monitoring and a biocontamination control strategy
• Discussion of environmental monitoring methods
• Introduction to risk concepts, hazard identification and risk assessment
• Introduction to FMEA
• Introduction to HACCP
• Application of risk tools to elements of the environmental monitoring program, such as assessing contamination risks, setting monitoring frequencies, assessing monitoring locations
• How to determine if too little or too much monitoring is being undertaken

Who will Benefit

• Microbiologists (QA and QC)
• QC managers
• Production managers
• Quality Assurance personnel
• Cleanroom engineer

For details se: Online Compliance

The people factor: investigating the gown

An effective environmental monitoring programme is designed to estimate the microbial content of the room air and surfaces (by incident rate, against alert and action levels, and by assessment of different species) for operations performed within a cleanroom or controlled environment. While individual results are rarely of significance, a well-designed environmental monitoring programme signals conditions contributing to rises in microbial levels. Shifts in microbial trends can be due to ineffective cleaning, disinfection, faulty air handling systems, material and equipment transfer, and the result of personnel related issues. With this latter point the majority of contaminants dispersed into cleanrooms derived from personnel.

Based on this, Tim Sandle has undertaken a review of cleanroom gowns, gown use and gown locations, in relation to microbial contamination. The detection of contamination on the gown either indicates a concern with the practices of an individual operator or a problem with the gown itself. The paper looks at several aspects of gown wearing through a review of data collated over a one year period. The data was studied for four considerations:

Locations most likely to indicate contamination.

Differences between re-laundered and single-use gowns.

Variations of gowns when re-laundered.

Variations in efficiency of gowns when worn over time.

The reference of the paper is:

Sandle, T. (2017) The people factor: investigating the gown, European Pharmaceutical Review, 22 (4): 23-26

For further details, please contact TimSandle

Posted by Dr. Tim Sandle

Technical Guide for Design, Control and Monitoring of Single-Use Systems

The Bio-Process Systems Alliance (BPSA) announced today that its new technical guide to Design, Control and Monitoring of Single-Use Systems for Integrity Assurance is now available for download at http://bpsalliance.org/technical-guides.

The uptake of single-use technologies (SUT) in more critical good manufacturing practices (cGMP) processes and applications has made assurance of integrity a critical quality attribute for both suppliers and end-users.

"I expect this to be one of the most significant documents written in the last five years to support our mission of driving the adoption of single-use technologies worldwide," said Kevin Ott, BPSA Executive Director.

The document provides recommendations to both suppliers and end-users in the single-use technology industry regarding strategies, tools and procedures that can assist in providing enhanced assurance of integrity of single-use systems. It can help end-users convey their specific requirements to the supplier. In turn, suppliers can use the document to demonstrate what they can provide to the end-user.

The technical guide divides best practices into two separate but complementary sections – a risk-based approach and practical tests for SUTs.

The document was written by a working group of end-users and suppliers who recognized the need to provide guidance in the absence of industry standards.

Cleanroom Management in Pharmaceuticals and Healthcare - special offer

Dear Reader,

Euromed Communications have recently brought out a new 2017 edition of the Cleanroom Management in Pharmaceuticals and Healthcare. Since the first edition of this book in 2013 there have been many changes to the approach and methods for cleaning and certifying cleanrooms, most notably the revisions to Parts 1 and 2 of the ISO 14644 series of global cleanroom standards. In addition to setting out the principal changes in these revised standards, many of the other chapters in the book have been updated to reflect their requirements, bringing current practices and Good Manufacturing Practice regulations up-to-date. The book is edited by Tim Sandle and Madhu Raju Saghee.

This book was reviewed in the May issue of Pharmig News and full details of the book can be found on the Euromed Communications website.

The publishers are offering a special discount to readers of this site and of the Pharmig, Sterility Assurance & Pharmaceutical Microbiology LinkedIn Groups at 20% off the cover price.

Thus the special offer costs for the new manual are as follows:

Hard back: £196 (~$260)

Paperback: £152 (~$200)

If you wish to take up this offer simply send an email to jill.monk@euromedcommunications.com mentioning the code ‘Pharmig20’ and you or your company will be invoiced accordingly.

Please note this offer ends on 10 December 2017.

Tim Sandle, on behalf of Euromed Communications

Examining the lifestyles of microbes

Scientists are identifying and characterizing more microbes each year using DNA sequencing technologies. As each new species is sequenced, scientists add it to the microbial "tree of life," creating a virtual census of what's there.

Turns out it's not an easy job. To put things in perspective, scientists aren't sure how many microbes even exist. Estimates vary widely from millions to trillions.

University of Delaware professor Jennifer Biddle and Rosa Leon-Zayas, who completed post-doctoral work at UD earlier this year, recently described new details about microbes known as Parcubacteria in a paper published in Environmental Microbiology.

The Parcubacteria were found in sediment samples collected by James Cameron within the Challenger Deep region of the Mariana Trench during the Deepsea Challenge Expedition. Leon-Zayas' doctoral advisor, Doug Bartlett at Scripps Institution of Oceanography, was a chief scientist on the expedition.

"From a scientific perspective, Challenger Deep was an invaluable opportunity to collect samples from the deepest part of the ocean," said Leon-Zayas, the paper's lead author, now an assistant professor at Willamette University.

Scientists traditionally have learned how microbes work by growing and studying them in petri dishes and beakers. It wasn't until DNA sequencing advanced to include the ability to separate and test microbes present in environmental samples (such as soils or sediments) that scientists realized they had missed a huge portion of bacteria now called the Candidate Phyla Radiation (CPR).

One group of CPR microbes called the Parcubacteria had been seen in the groundwater and shallow sediments of a few places on land, but it had only been intensively studied in sediment samples from an aquifer near Rifle, Col.

When Cameron collected sediment samples at the bottom of the trench, the scientists discovered that many different species of Parcubacteria live there, too.

"We were interested in seeing if the microbes living at the bottom of the ocean had the same lifestyle as the microbes living in soils in Rifle, Colorado," said Biddle, a marine microbiologist and associate professor in the College of Earth, Ocean, and Environment's School of Marine Science and Policy.

Leon-Zayas used a sorting technique to separate the microbial cells from the sediment particles so that scientists could amplify and sequence the microbe DNA. The researchers then characterized the individual microbial genomes. Based on the genes that are present in the genome -- sections of DNA that define what metabolisms a cell is capable of -- scientists can infer what the bacteria is doing.

This genomic sequencing revealed that Parcubacteria from the deep sea have a fairly simple metabolism; but the genomes were larger than that of their terrestrial cousins and even had a few extra features. In particular, these features indicated the bacteria may be able to perform anaerobic respiration, using things like nitrate to breathe instead of oxygen.

Parcubacteria also seemed to have more proteins and enzymes associated with cold environments, not surprising since the bottom of the Mariana Trench is cold and dark.

"It makes sense that organisms at the bottom of the ocean might have to be more self-sufficient. The environment is extreme and there isn't as much food," Biddle said.

See:

Rosa León-Zayas, Logan Peoples, Jennifer F. Biddle, Sheila Podell, Mark Novotny, James Cameron, Roger S. Lasken, Douglas H. Bartlett. The metabolic potential of the single cell genomes obtained from the Challenger Deep, Mariana Trench within the candidate superphylum Parcubacteria (OD1). Environmental Microbiology, 2017; 19 (7): 2769 DOI: 10.1111/1462-2920.13789



Posted by Dr. Tim Sandle

ISO/IEC 17025 moves to final stage of revision

Calibration as well as testing and analysing a sample is the daily practice of more than 60 000 laboratories worldwide, but how can they reassure customers about the reliability of their results?

Over the years, ISO/IEC 17025, General requirements for the competence of testing and calibration laboratories, has become the international reference for testing and calibration laboratories wanting to demonstrate their capacity to deliver trusted results. The International Standard, published jointly by ISO and IEC (International Electrotechnical Commission), contains a set of requirements enabling laboratories to improve their ability to produce consistently valid results.

News from ISO:

However, the laboratory environment has changed dramatically since the standard was last published, leading to the decision to revise the standard and integrate significant changes. Steve Sidney, one of the Convenors of the working group revising the standard, explains: “The last version of ISO/IEC 17025 was published in 2005. Since then, market conditions have changed and we felt we could bring some improvements to the standard.”

Heribert Schorn, working group Convenor who also participates in IECEE (System of Conformity Assessment Schemes for Electrotechnical Equipment and Components), adds: “The revision was needed to cover all the technical changes, technical developments and developments in IT techniques that the industry has seen since the last version. Additionally, the standard takes into consideration the new version of ISO 9001.”

This standard is of high significance for the IEC Conformity Assessment Community as it outlines the basic requirements for testing within all Conformity Assessment Schemes and Programmes operating within the IECEE, IECEx, IECQ and IECRE Conformity Assessment Systems.

The review was started in February 2015 as a result of a joint proposal by the International Laboratory Accreditation Cooperation (ILAC) and the South African Bureau of Standards (SABS), who is a member of ISO and hosts the IEC National Committee. The standard’s revision process has now reached the Final Draft International Standard (FDIS) stage, the last leg of development before publication.

The main changes:

The revision of ISO/IEC 17025 takes into account the activities and new ways of working of laboratories today. The main changes are as follow:

The process approach now matches that of newer standards such as ISO 9001 (quality management), ISO 15189 (quality of medical laboratories) and ISO/IEC 17021-1 (requirements for audit and certification bodies). The revised standard puts the emphasis on the results of a process instead of the detailed description of its tasks and steps.

With a stronger focus on information technologies, the standard now recognizes and incorporates the use of computer systems, electronic records and the production of electronic results and reports. Modern-day laboratories work increasingly with information and communication technologies and the working group felt it was necessary to develop a chapter on this topic.

The new version of the standard includes a chapter on risk-based thinking and describes the commonalities with the new version of ISO 9001:2015, Quality management systems – Requirements.

The terminology has been updated to be more in step with today’s world and the fact that hard-copy manuals, records and reports are slowly being phased out in favour of electronic versions. Examples include changes to the International Vocabulary of Metrology (VIM)and alignment with ISO/IEC terminology, which has a set of common terms and definitions for all standards dedicated to conformity assessment.

A new structure has been adopted to align the standard with the other existing ISO/IEC conformity assessment standards such as the ISO/IEC 17000 series on conformity assessment.

The scope has been revised to cover all laboratory activities including testing, calibration and the sampling associated with subsequent calibration and testing.

Using ISO/IEC 17025 facilitates cooperation between laboratories and other bodies. It assists in the exchange of information and experience and helps harmonize standards and procedures, as Warren Merkel, another Convenor of the working group, explains. “ISO/IEC 17025 impacts the results delivered by laboratories in a number of ways. The standard requires them to meet criteria for competence of their personnel, the calibration and maintenance of their equipment and the overall processes they use to generate the data. This requires laboratories to think and operate in a way that ensures their processes are under control and their data are reliable.” Results also gain wider acceptance between countries when laboratories conform to the standard.

Developed jointly by ISO and IEC in the Committee on conformity assessment (CASCO), the new version of ISO/IEC 17025 will replace the 2005 version and is scheduled for publication at the end of this year.

Posted by Dr. Tim Sandle

FDA approves first ever ‘digital pill’

An innovative medical device, in the form of a pill containing a built-in sensor has been developed and been given approval, based on safety and efficacy data, by U.S. authorities.

by Tim Sandle

What is being heralded as the world’s first ‘digital’ pill has gained approved by the U.S. Food and Drug Administration (FDA). The pill is called Abilify MyCite. The pill contains a medication called aripiprazole, which treats conditions like schizophrenia, bipolar disorder and depression. Also combined with the pill is an ingestible sensor. The pill comes after several years of research and is a venture between the Japanese pharmaceutical company Otsuka and digital medicine service Proteus Digital Health.

It is the sensor that creates the ‘digital’ or ‘smart’ pill. The purpose of the sensor, according to The Verge, is to record when the pill has been taken. This happens by a signal being sent to a wearable patch (fixed to the left rib cage); and then from the patch to a mobile device such as a smartphone via Bluetooth. The patch has additional functionality. It records activity levels, sleeping patterns, steps taken, and heart rate. The patch needs to be replaced every seven days.

The purpose of this is to determine when a pill has been taken. This either acts as a reminder  to patients that they have (or to verify that they have not) taken their required dose of medication; or, for more serious cases where a patient has been sectioned as the result of a mental disorder, to enable medics to record the fact that a medication has been taken.

Failure to take medications has societal and economic consequences, such as putting a strain on the hospital system. According to Dr. William Shrank, chief medical officer of the health plan division at the University of Pittsburgh Medical Center, who spoke with the New York Times on this subject: “When patients don’t adhere to lifestyle or medications that are prescribed for them, there are really substantive consequences that are bad for the patient and very costly.”

The sensor is only the size of a grain of sand. It is manufactured from silicon, copper, and magnesium. In terms of how the sensor works, an electrical signal is activated when the sensor comes into contact with stomach acid.

It is a common problem with medications, either through forgetfulness or as a conscious act, when patients do not taken the medicines prescribed to them. The digital pill aims to redress this.

Commenting on the go-ahead for the digital pill to be marketed, Mitchell Mathis, who is the director of the Division of Psychiatry Products in the FDA, told PharmaPhorum: “Being able to track ingestion of medications prescribed for mental illness may be useful for some patients.”

The regulator added: “The FDA supports the development and use of new technology in prescription drugs and is committed to working with companies to understand how technology might benefit patients and prescribers.”

The agreement relates to both parts of the smart medication: Abilify and Proteus Health’s sensor and patch, for the U.S. market. A label warning will accompany the product. This will state that the combined smart system has not been shown to improve patient compliance and that there are concerns about the effectiveness of the tracker in real-time since detection may be delayed. Nevertheless, the digital pill is likely to become popular with the medical establishment. The price of the pill has yet to be announced.

The Wall Street Journal opines that there could now be a raft of approval requests for other digital pills. The paper also notes that the FDA are preparing to hire more staff with understanding of software development in relation to medical devices.

Imagining a world without species

Categorizing species can get especially hazy at small, microbial scales. After all, the classical definition of species as interbreeding individuals with sexually viable offspring doesn't apply to asexual organisms. Examining shared DNA doesn't help either: collectively, E. coli bacteria have only 20 percent of genes in common. The classification process gets even trickier as many microbes work so closely that it is unclear what to call separate organisms, let alone separate species.

The woes of classification generate contentious debates in the biology community. But, for postdoctoral fellow Mikhail Tikhonov, one field's contentious debate is another's theoretical playground. In new research, he asks: Could organism interactions be described without mentioning species at all?

Key question: “how does evolution act on the structure within a community, rather than on a species?"

This question is not only interesting on a theoretical level, but could have real-world implications in understanding and treating human disease. While some diseases (like pneumonia or meningitis) have specific culprits, many others (like obesity or type II diabetes) seem to be associated to a community-level dysfunction of our microbiome -- the highly diverse bacterial communities that live on and inside our bodies. To understand these diseases, researchers must understand how the system works as a whole.

See:

Mikhail Tikhonov. Theoretical microbial ecology without species. Physical Review E, 2017; 96 (3) DOI: 10.1103/PhysRevE.96.032410

 Posted by Dr. Tim Sandle

Aggressive UTI bacteria hijack copper

Escherichia coli bacteria -- those at the root of hard-to-treat urinary tract infections (UTIs) -- hijack trace amounts of copper in the body and use it as a nutrient to fuel growth. This finding suggests blocking this system may starve E. coli infections, opening the door to treating UTIs using drugs that work differently from traditional antibiotics.

Copper is an essential mineral -- found in shellfish, whole grains, nuts, beans and other foods. It can kill pathogens in high concentrations. But it was unclear how E. coli handles copper ions present in urine, an extremely complex medium containing many trace metals and other compounds.

In past work studying strains of E. coli known to cause difficult-to-treat UTIs, the researchers showed that a molecule called yersiniabactin that is secreted by the bacteria sequesters copper, preventing it from accumulating to antibacterial levels. But what it does with this bound copper has been unknown.

While bacteria are known to bring iron -- another essential mineral -- into the cell, the researchers noted that E. coli have long been thought to lack a method to import copper. Indeed, scientists have assumed that yersiniabactin only imports iron.

In the new study, the researchers showed that yersiniabactin imports copper ions into the cell, where these charged particles help trigger the many biochemical reactions that bacteria require to grow and reproduce. The scientists further showed that once relieved of its mineral cargo, yersiniabactin goes back outside the cell to mop up more copper. The researchers dubbed this strategy "nutritional passivation." In metallurgy, passivation refers to treating or coating metal to make it less reactive.

The researchers also have shown that yersiniabactin can bind to a variety of metals beyond copper and iron, including nickel, cobalt and chromium.

See:

Eun-Ik Koh, Anne E Robinson, Nilantha Bandara, Buck E Rogers, Jeffrey P Henderson. Copper import in Escherichia coli by the yersiniabactin metallophore system. Nature Chemical Biology, 2017; DOI: 10.1038/nchembio.2441

Posted by Dr. Tim Sandle

Antibiotic Alternative Micreos wins Most Impactful Innovation

In the presence of former UN-Secretary-General Kofi Annan and Dutch minister Melanie Schultz of Infrastructure & the Environment, Micreos' alternative to antibiotics was chosen as the most impactful innovation of The Netherlands. Micreos will represent The Netherlands at the semi-finals of Ideas from Europe in Talinn, Estonia on November 22 2017.

Micreos' endolysin technology, enabling targeted killing of only unwanted bacteria - regardless of antibiotic resistance - was selected by a professional jury. Jury-chair Kasja Ollongren, Deputy mayor of Amsterdam, emphasized the impact Micreos' products can have for millions of people.


The Making Waves-conference was attended by hundreds of policy makers, entrepreneurs, investors and developers. Guest of honour, Kofi Annan, winner of the Nobel Peace Prize, said: "I am really excited about Micreos' alternative to antibiotics".


Kasja Ollongren: "We chose Micreos because the company's technology is already helping tens of thousands of people and we're convinced Micreos' alternative to antibiotics can be life-changing for millions."

Micreos CEO Mark Offerhaus: "Antibiotic-resistance is an enormous global problem. We are honoured with the recognition. Gladskin has already been life-changing for people suffering from inflammatory skin diseases such as eczema, acne, rosacea and wound infections caused or aggravated by the Staphylococcus aureus bacteria, including the resistant MRSA. But as far as we're concerned, this marks only the beginning. Our endolysin technology offers a real and sustainable alternative to antibiotics, the broad potential of this technology is immeasurable. There's no need to wait and no time to lose."

Posted by Dr. Tim Sandle

Thank you to Rapid Micro Biosystems

Rapid Micro Biosystems has been involved with the Pharmaceutical Microbiology Resources website and the Pharmaceutical Microbiology LinkedIn Group since the early days. These sites have helped to promote the concept of rapid microbiological methods and some of Rapid Micro Biosystems innovations.

Although the partnership has reached an end-point, I would like to thank Rapid Micro Biosystems for their support and sponsorship over the past six-years.



Posted by Dr. Tim Sandle

The future possibilities of graphene for microbiology

Graphene is the most widely researched new material. It is an allotrope of carbon and due to special properties the material is being tested out within the fields of consumer and medicinal electronics. While these applications have received considerable attention developments relating to microbiology are taking place. This short review article considers bacterial staining and anti-bacterial activity, as two of the most promising future developments.

An essay by Tim Sandle

Graphene is a material derived from carbon and it has unique physicochemical properties. Graphene is formed where graphite is taken and atom thick layers are sliced away. The resultant structure is a single-layer of carbon atoms linked in a hexagonal chicken-wire pattern. Within the structure each of the atoms share a cloud of electrons moving freely about the surface. The material is light, transparent, strong and very conductive (Allen et al, 2010). Compared to other carbon allotrope, such as fullerenes, carbon nanotubes and graphite, graphene exhibits many exceptional physical and chemical properties. Graphene related materials are of great interest in the field of biomedicines and applications are underway in biosensing and drug delivery.

Graphene as a Gram-stain alternative

New research, from the University of Illinois at Chicago, suggests that graphene can be used for performing Gram-stains, as part of microbial identification. A review of the properties of graphene shows it can detect variances to cell vibration when a cell comes into contact with the material. So far the tests undertaken with graphene have related to cancer. Here atomic vibration differs depending upon whether the cell is a cancer cell or a normal cell. This happens because the cancer cell’s hyperactivity leads to a higher negative charge, and this causes a higher level of protons to be released. This difference can be detected, helping medical technologists to identify cancerous growth.

Assessing the variances in vibration is possible using an established laboratory method called Raman spectroscopy (a spectroscopic technique used to observe vibrational, rotational, and other low-frequency modes in a system).

According to lead researcher Vikas Berry, who is the associate professor and head of chemical engineering, who led the research along with Ankit Mehta, assistant professor of clinical neurosurgery in the UIC College of Medicine: “We may be able to use it with bacteria to quickly see if the strain is Gram-positive or Gram-negative…We may be able to use it to detect sickle cells.” Berry made this remark to Controlled Environments magazine (Anon, 2017).

In a parallel development, one research group have created a graphene sensor for Escherichia coli. This involved fabricating a flexible substrate onto which a sensor device with O-ring is fitted. Once contact takes place with the suspected organism, Raman spectra is used to indicate the presence (Basu et al, 2014).

As all microbiologists know, the Gram-stain is the key test for distinguishing between two groups of bacteria based on cell wall morphologies (Sandle, 2014). The Gram stain procedure distinguishes between Gram positive and Gram negative groups by coloring these cells red or violet. Gram staining is a common technique used to differentiate two large groups of bacteria based on their different cell wall constituents. The Gram stain procedure distinguishes between Gram positive and Gram negative groups by coloring these cells red or violet. Gram positive bacteria stain violet due to the presence of a thick layer of peptidoglycan in their cell walls, which retains the crystal violet these cells are stained with. Alternatively, Gram negative bacteria stain red, which is attributed to a thinner peptidoglycan wall, which does not retain the crystal violet during the decolouring process (Sandle, 2004).

While the Gram-stain technique is well described it is sometimes prone to error. This can relate to the types of organisms, the age of the cultures, or due to errors made by the person performing the test (such as over decoloursation). A method based on graphene would error proof. Whether such a method becomes commercially available will depend on development costs.

Graphene as an antibacterial agent

As well as using graphene as a potential diagnostic tool, graphene can also be used as an anti-bacterial measure (where liposome-embedded graphene reduces the growth capability of bacteria). Research by Zappacosta and colleagues showed that graphene aqueous dispersion is stable for several days and demonstrates significant antibacterial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) strains, with a reduction in the growth of S. aureus and E. coli as high as 60 and 78%, respectively.

In a similar application, researchers have looked at graphene-iodine nano-composites, formed via electrostatic interactions between positively charged graphene derivatives and triiodide anions, as anti-bacterial agents (Some et al, 2015). With this, the antibacterial potential of these graphene-iodine composites against Klebsiella pneumonia, Pseudomonas aeruginosa, Proteus mirobilis, Staphylococcus aureus, and Escherichia coli has been demonstrated. The success against the organisms relates to the inherent cytotoxicity of the nanocomposite, specifically through electron transfer interaction from microbial membrane to graphene.

Furthermore, scientists are studying graphene oxide with the aim of creating bacteria-killing catheters and medical devices. Here coating surgical tools with this carbon-based compound could kill bacteria, reducing the need for antibiotics, decreasing the rates of post-operative infections and speeding recovery times. This is with graphene oxide, which is a form of graphene with molecular oxygen incorporated into it. This compound protects against infection by destroying bacteria before it gets inside the body. In terms of the process the graphene oxide wraps around the bacteria, puncturing its membrane. A broken membrane prevents the bacteria from growing and often kills it.

Studies conducted at the Università Cattolica del Sacro Cuore in Rome indicate that the compound is most effective when paired with salt. Getting the salt balance correct is important. With too little salt and then the graphene oxide is unable to wrap around the bacteria; and with  too much salt and the graphene aggregates, failing to puncture the bacteria's membrane. In order to destroy both Gram positive and Gram negative bacteria a 300 nanometer sheet of graphene oxide solution must be mixed with low molarity (<10 mM) calcium chloride is required (Anon, 2015).

Summary

These two related research strands (for differential microbiology and as an antibacterial agent) signal that graphene, the so-called ‘wonder material’ of our age, is set to make a significant impact upon microbiology. As with the development of any novel method, progress will be slow. However, the research results reported to date suggest that graphene is set to make a major contribution to microbiology.

References

Allen, M. J., Tung, V.C. and R. B. Kaner, R.B. Honeycomb carbon: a review of graphene, Chem. Rev., 2010, 110, 132

Anon. Biophysical Society report “Towards a “green” antimicrobial therapy: Study of graphene nanosheets interaction with human pathogens”, 2015 (http://tinyurl.com/zzgsofu)

Anon. First Use of Graphene to Find Cancer Cells, Controlled Environments, 2017 (http://www.cemag.us/news/2017/02/first-use-graphene-find-cancer-cells)

Basu, P.K., Indukuri, D., Keshavan, S., Bhat, N. (2014) Graphene based E. coli sensor on flexible acetate sheet, Sensors and Actuators B Chemical 190:342-347 (https://www.researchgate.net/publication/256926101_Graphene_based_E_coli_sensor_on_flexible_acetate_sheet)

Sandle, T. (2004) Gram’s Stain: History and Explanation of the Fundamental Technique of Determinative Bacteriology’, IST Science and Technology, No. 54, pp3-4

Sandle, T. (2014). ‘Microbial Identification: Laboratory Techniques and Methods. In Chesca, A. (Ed.) Methods for Diseases: Diagnostic with Applicability in Practice, Lambert Academic Publishing, Germany, pp15-26

Some, S., Sohm, J., Kim, J. et al Graphene-Iodine Nanocomposites: Highly Potent Bacterial Inhibitors that are Bio-compatible with Human Cells, Scientific Reports 6, Article number: 20015 (2016). doi:10.1038/srep20015

Zappacosta, R., Di Giulio, M., Ettorre, V. et al Liposome-induced exfoliation of graphite to few-layer graphene dispersion with antibacterial activity, J. Mater. Chem. B, 2015, 3, 6520-6527 (http://pubs.rsc.org/en/content/articlehtml/2015/tb/c5tb00798d)

by Dr. Tim Sandle

EDQM: Microbiological control symposium

A symposium on microbiology in the pharmaceutical sector was the opportunity for the EDQM to gather feedback from users of the European Pharmacopoeia on alternative testing methods for microbiological control and sterilisation processes. The event, which took place in Strasbourg on 10-11 October 2017, was attended by a wide range of experts in the pharmaceutical and microbiological fields who reviewed the latest trends and innovations in the field of microbiology, in addition to Pharmacopoeial approaches and related regulatory requirements.

Over recent years, the European Pharmacopoeia has given pharmaceutical manufacturers access to new chapters for microbiological control, kick-starting a new era for the world of microbiology and rendering the uptake of modern microbiological methods an evolution already in process, rather than a revolution.

Among the topics covered was the use of modern methods for microbiological control, with specific sessions focusing on sterilisation and biological indicators, rapid microbiological methods and control methods for cell therapy products and pharmaceutical water. Reports on successful new methods gave an overview of the potential benefits in terms of costs and time efficiency and, most importantly, in terms of quality. Authorities, manufacturers and suppliers of new technologies also discussed the current acceptance of these new methods at a regulatory level across the world. Over recent years, the European Pharmacopoeia has given pharmaceutical manufacturers access to new chapters for microbiological control, kick-starting a new era for the world of microbiology and rendering the uptake of modern microbiological methods an evolution already in process, rather than a revolution.

Various chapters of the European Pharmacopoeia have laid the foundations for the use of modern methods for microbiological control and have been available for more than ten years – notably 5.1.6. Alternative methods for control of microbiological quality, 2.6.7. Mycoplasmas, 2.6.12. Microbiological examination of non-sterile products: microbial enumeration tests and 2.6.27. Microbiological examination of cell-based preparations. Chapter 5.1.6 in particular was recently further revised to take into full account the latest technological developments and to provide clear guidance for validating alternative microbiological methods.