Monday, 31 August 2020

Review of the causes of antimicrobial resistance

Antimicrobial agents particularly antibiotics have been critical in the fight against infectious diseases caused by pathogenic microorganisms including bacteria, fungi, viruses and protozoa (1). There usage in clinical medicine for treating infectious diseases has drastically leads to increase in the life expectancy of the human race over the past six decades. This is because the discovery and usage of antibiotics in infectious disease management has helped to reduce the rate of morbidity and mortality caused by infectious disease pathogens in human population. However, in recent years there has been a marked rise in the number and type of antimicrobial resistant organisms (2).

In relation to this subject, Tim Sandle has written an new article.

Here is the abstract:

Antibiotic resistance is one of the biggest challenges to the health sector worldwide, and this medical quagmire threatens our ability to effectively manage and treat some infectious diseases. Microbial resistance to antibiotics and/or antimicrobial agents has been documented not only against antibiotics of natural and semi-synthetic origin such as the penicillins, but also against some purely synthetic compounds (such as the fluoroquinolones) or those which do not even enter the cells (such as vancomycin). And unfortunately, the slow pace in the discovery and development of novel antibiotics have not actually kept pace with the emergence and rate at which bacteria develops and mount resistance to some available antibiotics (3).

Some infectious diseases including but not limited to tuberculosis, bacterial pneumonia, septicaemia, gonorrhoea, wound infections and otitis media are now becoming recalcitrant to treat with some available antibiotics because the causative agents of these diseases are fast becoming resistant to some available antibiotic therapy. These antibiotic resistant organisms have developed several novel ways and mechanisms that allow them to ward-off the antimicrobial onslaught of potent antimicrobial agents and/or antibiotics targeted towards them. This article reviews the primary resistance mechanisms.

The reference is:

Sandle, T.  (2020) Review of the causes of antimicrobial resistance, Microbioz India, 6 (3): 12-20

Access the article for free here:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Sunday, 30 August 2020

Modern myth #6: Low endotoxin recovery affects all products

According to James Cooper, no one disputes that low recovery of lipopolysaccharide (LPS), as used for the LAL test control endotoxin, occurs in certain conditions, such as chelating buffers and detergents. However, this issue does not affect all products. 

Furthermore, the issue at hand is low lipopolysaccharide recovery rather than ‘low endotoxin’ recovery. Endotoxin is more sophisticated, being composed of a hydrophilic polysaccharide covalently linked to a highly conserved, hydrophobic lipid region. LPS and endotoxin do not behave in the same way (LPS activity varies due to the presence of various salts and detergents).

In addition, the use of a uniform screening test can reveal conditions of concern and legitimize the use of alternative naturally occurring endotoxin preparations for endotoxin challenge studies. However, to do required regulatory approval.


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Saturday, 29 August 2020

Modern myth #5: Freeze-dried cultures are safe until they are reconstituted

The number of particles aerosolized upon opening lyophilized cultures depends upon the consistency of the end product. For example, fluffier end products tend to create more concentrated aerosols. Most of the particles aerosolized upon opening lyophilized cultures are larger than 5 um. Dropping a lyophilized culture creates an extremely concentrated aerosol composed predominantly of particles larger than 5 um.

The inclusion of mother liquor in the suspending menstruum reduces the aerosol concentration approximately fourfold.

Two parameters must be established to assess the risk of handling laboratory cultures: the biological decay rate of aerosols under laboratory conditions, and the human infectious dose by the respiratory route. Several common organisms survive at least 1 hour in droplet nuclei at standard laboratory relative humidity.


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Friday, 28 August 2020

Modern myth #4: You need to use a Bunsen burner

Many microbiologists find that the use of pre-sterilized disposable tools to transfer media and bacterial cultures result in lower contamination rates compared with reusable inoculating loops. The latter is a process known to create aerosols that may increase potential air contaminants.

Depending on the organism handled, this can create an element of risk. Several studies have shown that particles of less than 5 um are most effective in establishing airborne infection in laboratory animals; and particles of 1 to 5 um can be deposited in the alveoli, with preferential deposition occurring with 1- to 2-um particles. Particles larger than 3.5 are most probably deposited in the upper respiratory tract. Particles in the 2.0- to 3.5- um range appear to offer equal opportunities for both upper respiratory and alveolar deposition.

Where Bunsen burners are used, it is optimal to flame a loop in the airspace underneath the burner flame.

Moreover, the idea that the Bunsen burner creates an upward draft of air to prevent contaminants in the air from settling on the work surface below does not always result in more robust practices, as sometimes airflow disruption leads to particles settling out.


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Thursday, 27 August 2020

Modern myths #3: Alcohol hand sanitisers cause bacterial mutation and help create resistant strains

The alcohol-based antibacterial rubs are effective enough that they do not create resistant strains, although antibacterial soaps may present a hazard.

But some antimicrobial soaps present challenges...

While the alcohol rub stays on the hands and is not meant to be rinsed off, the antibacterial triclosan is rinsed off before it can do all its work and then enters the water supply. In addition, products like triclosan can cause problems once they are in the water supply, and resistant strains of bacteria have been created in labs using triclosan, although it remains to be seen if it will happen in the natural environment.

Generally, antibacterial soap does not do enough to justify its use. The objective of hand washing, by rinsing in soap and water for at least 20 seconds, is not to kill bacteria, but simply to get germs and viruses off our hands. Using a sink and washing hands thoroughly 15 to 20 seconds with regular soap and then rinsing that is the most effective method of 'de-germing', or removing bacteria and viruses from your hands.

Hand washing with soap and water does not remove all the microbes from our hands, because some are an important part of our skin, and even if we did kill them, they would return.
Given that regular soap and water removes the organisms, there is often no need for an antibacterial agent, and it probably will not work anyway. Hand sanitisers are best reserved where hand washing facilities are not readily accessible.


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Wednesday, 26 August 2020

Modern myths #2: We understand microbes best in pure culture and as single colonies.

A common image associated with microbiology is that of the plastic Petri dish.  The ability to grow microorganisms like this is in fact the foundation of microbiology and much of our knowledge about microbial biochemistry, genetics, physiology, etc. comes from culture-based work. 

However, there are several limitations to this approach, some of which can now be addressed with alternative methods. 

Culturing organisms in a Petri dish meaning growing them in non-natural conditions, not only are the organisms in an artificial environment with a pre-defined collection of nutrients, they are usually grown in isolation… cut off from the normal community of microbes surrounding them in nature. 

In addition, we now know that only around 1% of the microorganisms in a typical environment can even be cultured at all. 

New sequence-based techniques such as metagenomics and rRNA sequencing can help address both of these concerns to varying degrees, and are an important complement to culture-based techniques.

 Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Tuesday, 25 August 2020

Modern myths #1 E. coli bacteria are all the same

Bacterial strains showcase a diversity of shapes, sizes, and functions. The bacterium Escherichia coli has caused outbreaks of lethal foodborne disease. However, only certain varieties of E. coli can make people sick. For example, strain “O157:H7” is particularly dangerous to people. Other strains of E. coli help out with the normal human digestive processes. Most people carry safe strains in their guts all the time without getting sick.

Therefore testing simply for the presence of E. coli is not sufficient; it is necessary to differentiate between pathogenic and non-pathogenic strains. 

For further details see:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Monday, 24 August 2020

Assessing Gram-stain error rates within the pharmaceutical microbiology laboratory

The implications of errors with the Gram-stain can influence the selection of the test method (and test kit) for the next stage of identification, whether this is a manual biochemical identification method (such as API) or a semi-automated method (such as Vitek or Omnilog). Getting an identification wrong could lead to an incorrect root cause analysis (which impacts on all types of pharmaceutical processing, including sterile products) and potential errors relating to batch release (especially with non-sterile pharmaceuticals where understanding the pathogenic nature of the organism is a key requirement).

In relation to this fundamental aspect of pharmaceutical microbiology, Tim Sandle has written an article.

The abstract reads:

Gram-staining remains the fundamental method for determinative bacteriology, dividing bacteria into Gram-positive and Gram-negative organisms. This test provides information as to the origin of any contamination and is a pre-requisite for many microbial identification methods. Despite the longevity of the test, the test is highly reliant upon analyst technique and therefore errors occur. While there are a few studies looking at errors in the clinical context, research has not been extended to the pharmaceutical microbiology laboratory context. In this study, we present a review of over 6,000 Gram-stains and establish an error rate of around 3%, with the most common reason for error being an over-decolourisation step resulting in organisms that should be Gram-positive appearing as Gram-negative. The analysis enables others to benchmark their facilities against.

The reference is:

Sandle, T. (2020) Assessing Gram-stain error rates within the pharmaceutical microbiology laboratory, European Journal of Parenteral and Pharmaceutical Sciences, 25 (1):

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Saturday, 22 August 2020

New warning over ineffective hand sanitizers

Not all hand sanitisers are effective: How to spot a substandard or dangerous product. Many regulators have dropped some of the requirements for hand sanitisers in order to help to address supply problems. At the same time poor quality products have entered the market, including many containing toxic methanol. In this video, Tim Sandle explains what purchasers and members of the public need to watch out for:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Friday, 21 August 2020

Effect of COVID-19 pandemic on TB response

A new report released today finds that the global response to the COVID-19 pandemic is having unintended yet drastic consequences on tuberculosis (TB) services, with lockdowns and limitations on diagnosis, treatment and prevention services expected to increase the annual number of TB cases and deaths over the next five years; at least five years of progress on TB response will be lost. The modeling analysis released by the Stop TB Partnership shows that under a three-month lockdown and a protracted 10-month restoration of services, the world could see an additional 6.3 million cases of TB between 2020 and 2025 and an additional 1.4 million TB deaths during that same period.

“We never learn from mistakes. For the past five years, TB, a respiratory disease, has remained the biggest infectious disease killer because the ‘TB agenda’ consistently became less visible in front of other priorities,” said Dr Lucica Ditiu, Executive Director of the Stop TB Partnership. “Today, governments face a torturous path, navigating between the imminent disaster of COVID-19 and the long-running plague of TB. But choosing to ignore TB again would erase at least half a decade of hard-earned progress against the world’s most deadly infection and make millions more people sick.”

The new study was commissioned by the Stop TB Partnership in collaboration with the Imperial College, Avenir Health and Johns Hopkins University, and was supported by USAID. The modeling was constructed on assumptions drawn from a rapid assessment done by The Stop TB Partnership on the impact of the COVID-19 pandemic and related measures on the TB response in 20 high-burden TB countries—representing 54% of the global TB burden.

The modeling focused on three high burden countries—India, Kenya, and Ukraine—and extrapolated estimates from those countries to create global estimates of the impact of COVID-19 on TB. The authors note that the model can be replicated in any other country and that the findings can be used by countries for data-driven decisions and financial requests.

TB is a forgotten respiratory disease that still kills 1.5 million people each year, more than any other infectious disease. Incidence and deaths due to TB have been declining steadily over the last several years as a result of intensified activities by high burden countries to find people with TB early and provide appropriate treatment.

In 2018, during the UN General Assembly (UNGA) High-Level Meeting on TB, Heads of States and governments committed to significantly scale up the TB response. In 2018, this resulted in identifying an additional 600,000 people who could access TB care. In 2019, we also saw very promising progress. The COVID-19 pandemic, especially considering the mitigation measures put in place, has proven to be a major setback in achieving the UNGA targets, as TB case detection has dramatically fallen, treatments have often been delayed and the risk of interruption of treatment and potential increase of people with drug-resistant TB has increased.

According to the new study, with a three-month lockdown and a protracted 10-month restoration of services, global TB incidence and deaths in 2021 would increase to levels last seen in between 2013 and 2016 respectively, implying a setback of at least five to eight years in the fight against TB.

To minimize the impact of the COVID-19 pandemic on TB, save millions of lives and get the world back on track in achieving the UNGA targets, national governments need to take immediate measures that ensure the continuity of TB diagnostic, treatment and prevention services during the lockdown period and undertake a massive catch-up effort to actively diagnose, trace, treat and prevent TB.

Stop TB Partnership and partners call upon the leadership of all countries—particularly those with high TB burdens—to ensure the continuity of the TB response in the time of COVID-19, to take proactive measures that include those who are most vulnerable and to provide protection against economic hardship, isolation, stigma and discrimination. We urge governments to secure the human and financial resources needed for seamless continuation of TB services amid the COVID-19 response.

Recognizing that this is an unprecedented situation, the Stop TB Partnership is continuing support for national TB Programmes and partners through its multiple technical, innovative and people-centered platforms. To ensure access to TB and COVID-19 resources, the Stop TB Partnership is sharing actions, experiences and recommendations from countries and partners through a dedicated TB and COVID-19 webpage and has recently published interactive maps with TB and COVID-19 situations in countries.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Thursday, 20 August 2020

Endotoxin Limits for Investigational Oncology Drugs and Biological Products

The U.S. FDA has issued an new guidance document titled “Setting Endotoxin Limits During Development of Investigational Oncology Drugs and Biological Products.” The document is currently in draft form (dated July 2020).

This guidance describes FDA’s recommendations to investigational new drug sponsors for setting endotoxin limits during the development of investigational drugs intended for use in combination with other approved drugs or for the codevelopment of two or more investigational drugs.

In keeping with the principles of facilitating drug development for serious and life-threatening diseases, this guidance outlines FDA’s current thinking on a risk-based approach to setting acceptance criteria for endotoxins during the clinical development of drugs intended to treat serious and life-threatening cancers.

For details see:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Wednesday, 19 August 2020

A Practical example of applying Quality Risk Management in GDP – Transportation Risks

This is about identifying and defining the risks. The MHRA write:

“Many companies present such summaries as a spreadsheet which assists communication, an essential part of QRM. The summary should be regularly re-evaluated and potential changes assessed during quality management review meetings. It is rare to find sound scientific justification for acceptance of a load subject to an excursion, and in the uncommon instances where a supplier or customer contacts the marketing authorization holder for stability information, it is often not directly comparable to the excursion experienced. Unfortunately, the most common reason for accepting a consignment with a temperature excursion is purely commercial, which may put patients at risk and undermines any risk management carried out by the company.

In some cases of quality risk management, attempts were made to inappropriately apply mean kinetic temperature to underestimate impact rather than develop good control and preventive measures.”

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Tuesday, 18 August 2020

ICH guideline M7 - mutagens

ICH guideline M7 assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk.

Q&A Step 2b Since the ICH M7 Guideline was finalized, worldwide experience with implementation of the recommendations for DNA reactive (mutagenic) impurities has given rise to requests for clarification relating to the assessment and control of DNA reactive (mutagenic) impurities.

The Q&A document is intended to provide additional clarification and to promote convergence and improve harmonization of the considerations for assessment and control of DNA reactive (mutagenic) impurities and of the information that should be provided during drug development, marketing authorization applications and/or Master Files.

The scope of the Q&A document follows that of ICH M7. “Applicant” is used throughout the Q&A document and should be interpreted broadly to refer to the marketing authorization holder, the filing applicant, the drug product manufacturer, and/or the drug substance manufacturer.


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Monday, 17 August 2020

Minimizing microbial contamination on cleanroom surfaces

One design aspect is with the incorporation of antimicrobial materials into surfaces as a means to reduce microbial numbers (or at least to prevent microorganisms from growing). This addition has become commonplace in many hospitals1 and within food factories;2 however, the adoption has been slower within pharmaceuticals (where the use of antibacterial materials used to coat cleanroom surfaces is sometimes referred to as “biotrunking”). A second design aspect lies with the selection of surface properties of materials, so that surfaces can reduce the possibility of microbial attachment, making disassociated organisms easier to kill by disinfection. Combined, antimicrobial surfaces with specific topography has the potential to reduce microbial survival in cleanrooms.

In relation to this, Tim Sandle has written an article. The reference is:

Sandle, T. (2020) Minimizing microbial contamination on cleanroom surfaces, American Pharmaceutical Review, 23 (2): 30-35

In this article, different antimicrobial technologies together with physical properties are considered together with a review of available literature to examine the efficacy of such surface materials.

For details, see:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Sunday, 16 August 2020

Microbiome's role in attacking cancerous tumors

Researchers with the Snyder Institute for Chronic Diseases at the Cumming School of Medicine (CSM) have discovered which gut bacteria help our immune system battle cancerous tumours and how they do it. The discovery may provide a new understanding of why immunotherapy, a treatment for cancer that helps amplify the body's immune response, works in some cases, but not others. The findings, published in Science, show combining immunotherapy with specific microbial therapy boosts the ability of the immune system to recognize and attack cancer cells in some melanoma, bladder and colorectal cancers.

The researchers identified bacterial species that were associated with colorectal cancer tumours when treated with immunotherapy. Working with germ-free mice, they then introduced these specific bacteria along with immune checkpoint blockade, a type of cancer immunotherapy. Research revealed that specific bacteria were essential to the immunotherapy working. The tumours shrank, drastically. For those subjects that did not receive the beneficial bacteria, the immunotherapy had no effect.


Lukas F. Mager, Regula Burkhard, Nicola Pett, Noah C. A. Cooke, Kirsty Brown, Hena Ramay, Seungil Paik, John Stagg, Ryan A. Groves, Marco Gallo, Ian A. Lewis, Markus B. Geuking, Kathy D. McCoy. Microbiome-derived inosine modulates response to checkpoint inhibitor immunotherapy. Science, 2020; eabc3421 DOI: 10.1126/science.abc3421

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Saturday, 15 August 2020

Sustainable nylon production - from bacteria

Nylon manufacture could be revolutionised by the discovery that bacteria can make a key chemical involved in the process, without emitting harmful greenhouse gases.

Scientists have developed a sustainable method of making one of the most valuable industrial chemicals in the world -- known as adipic acid -- which is a key component of the material.

More than two million tonnes of the versatile fabric -- used to make clothing, furniture and parachutes -- is produced globally each year, with a market value of around £5 billion.

Industrial production of adipic acid relies on fossil fuels and produces large amounts of nitrous oxide -- a greenhouse gas three hundred times more potent than carbon dioxide. A sustainable production method is urgently required to reduce the damage caused to the environment, the team says.

Scientists from the University of Edinburgh altered the genetic code of the common bacteria E.coli in the lab. The modified cells were grown in liquid solutions containing a naturally occurring chemical, called guaiacol, which is the main component of a compound that gives plants their shape.

Following a 24-hour incubation period, the modified bacteria transformed the guaiacol into adipic acid, without producing nitrous oxide.The environmentally friendly approach could be scaled up to make adipic acid on an industrial scale.


Jack T. Suitor, Simon Varzandeh, Stephen Wallace. One-Pot Synthesis of Adipic Acid from Guaiacol in Escherichia coli. ACS Synthetic Biology, 2020; DOI: 10.1021/acssynbio.0c00254

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Thursday, 13 August 2020

For bacteria, a small genome means some serious decluttering — even in the ribosome

Researchers from Skoltech, Lomonosov Moscow State University, and the Kharkevich Institute for Information Transmission Problems have studied the genomes of some 200 strains of bacteria to determine which proteins in the ribosome, part of the key cell machinery, can be safely lost and why. The paper was published in the journal Molecular Biology and Evolution.

The ribosome is a universal cellular machine, present in all eukaryotes and prokaryotes, that builds proteins in a process called translation. The two major components of the ribosome, the so-called small and large ribosomal subunits, consist of ribosomal RNA (rRNA) molecules and ribosomal proteins. 

The composition of these fundamental ‘protein factories’ is fairly consistent across cells, but there is evidence that some bacteria function without a complete set of ribosomal proteins, so researchers have been looking to determine which of the proteins are truly essential for a working ribosome.

Skoltech professor and vice president for biomedical research Mikhail Gelfand and his colleagues analyzed ribosomal protein composition in 214 relatively small bacterial genomes. They identified a set of frequently lost proteins and showed that only nine ribosomal proteins were completely conserved, while each of the remaining 48 was lost in at least one strain from the dataset.

“Tiny genomes are characteristic of endosymbionts, bacteria that live within other bacteria or eukaryotic cells. In this non-changing environment and under weak selection they tend to lose non-essential (even if necessary for free-living bacteria) genes — similar to multicellular parasites that often miss entire organs. The ribosome has been assumed to be the most conserved organelle with a standard set of proteins; but if you have only 121 genes — the present bacterial record for simplicity — you cannot encode all fifty-something ribosomal proteins, so some of them have to be lost. We have demonstrated that the patterns of this loss are not random,” Professor Gelfand says.

Apparently, ribosomal proteins of the small subunit were more likely to be retained than the large subunit proteins, and most frequently lost proteins were located on the ribosome surface, where they formed fewer contacts with other ribosome components. They were also incorporated in the ribosome late in evolution, so it seems that bacteria tend to practice the ‘last in, first out’ approach when it comes to dropping ribosomal proteins.

The researchers also found that the three bacteria with the shortest genomes in the group lost the largest number of proteins; there was a correlation between genome size and a number of retained ribosomal proteins. Yet since ribosomal proteins are in the cell’s essential toolkit, they are generally among the last to leave a ‘downsizing’ bacterial genome.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Wednesday, 12 August 2020

Mouthwashes could reduce the risk of coronavirus transmission

Sars-Cov-2 viruses can be inactivated using certain commercially available mouthwashes. This was demonstrated in cell culture experiments by virologists from Ruhr-Universität Bochum together with colleagues from Jena, Ulm, Duisburg-Essen, Nuremberg and Bremen. High viral loads can be detected in the oral cavity and throat of some Covid-19 patients. The use of mouthwashes that are effective against Sars-Cov-2 could thus help to reduce the viral load and possibly the risk of coronavirus transmission over the short term. This could be useful, for example, prior to dental treatments. However, mouth rinses are not suitable for treating Covid-19 infections or protecting yourself against catching the virus.

The results of the study are described by the team headed by Toni Meister, Professor Stephanie Pfänder and Professor Eike Steinmann from the Bochum-based Molecular and Medical Virology research group in the Journal of Infectious Diseases, published online on 29 July 2020. A review of laboratory results in clinical trials is pending.

Eight mouthwashes in a cell culture test

The researchers tested eight mouthwashes with different ingredients that are available in pharmacies or drugstores in Germany. They mixed each mouthwash with virus particles and an interfering substance, which was intended to recreate the effect of saliva in the mouth. The mixture was then shaken for 30 seconds to simulate the effect of gargling. They then used Vero E6 cells, which are particularly receptive to Sars-Cov-2, to determine the virus titer. In order to assess the efficacy of the mouthwashes, the researchers also treated the virus suspensions with cell culture medium instead of the mouthwash before adding them to the cell culture.

All of the tested preparations reduced the initial virus titer. Three mouthwashes reduced it to such an extent that no virus could be detected after an exposure time of 30 seconds. Whether this effect is confirmed in clinical practice and how long it lasts must be investigated in further studies.

The authors point out that mouthwashes are not suitable for treating Covid-19. "Gargling with a mouthwash cannot inhibit the production of viruses in the cells," explains Toni Meister, "but could reduce the viral load in the short term where the greatest potential for infection comes from, namely in the oral cavity and throat -- and this could be useful in certain situations, such as at the dentist or during the medical care of Covid-19 patients."

Clinical studies in progress

The Bochum group is examining the possibilities of a clinical study on the efficacy of mouthwashes on Sars-Cov-2 viruses, during which the scientists want to test whether the effect can also be detected in patients and how long it lasts. Similar studies are already underway in San Francisco; the Bochum team is in contact with the American researchers.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Monday, 10 August 2020

Simultaneous detection of Bacterial, Fungal and Pseudomonas aeruginosa contamination in Pharmaceutical products

Rapid detection of microbial contaminants in pharmaceutical products is play a vital role in contamination control. The purpose of this study is to standardize multiplex polymerase chain reaction (mPCR) assay to detect the pharmaceutical contaminants (either bacterial or fungal) and Pseudomonas aeruginosa specifically in single PCR reaction.

A new article of interest has been published.

A total of 13 pharmaceutical samples of ophthalmic products, chemotherapeutic, psychiatric, cardiac and gastrointestinal drugs were selected and artificially contaminated with <10 CFU of microorganisms. After enrichment, DNA was extracted, targeting the conserved region in eubacterial genome (16SrRNA), panfungal genome (28S rRNA) and P. aeruginosa (oprL). Specificity and sensitivity of the technique were verified. The mPCR assay was standardized by varying concentration of PCR reagents and conditions. In conclusion, there was no PCR inhibition and false positive results were observed. This assay can easily be incorporated for pharmaceutical products to quickly screen for microbial contamination.

The reference is:

Vijaykumar, R., Alfaiz, F. and Sandle, T. (2020) Simultaneous detection of Bacterial, Fungal and Pseudomonas aeruginosa contamination in Pharmaceutical products using Multiplex PCR Simultaneous detection of microbial contamination in pharmaceutical products using Multiplex PCR, Chimica Oggi - Chemistry Today, 38(2): 20-22

For details, please see:

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (

Sunday, 9 August 2020

Asia-Pacific (APAC) — Future Hub for Sterilization Technology?

One of the most common words that we usually associate with hospitals and other medical facilities is ‘sterilization’. This word has become synonymous with healthcare solutions, so much so, that the first question usually asked by people during a medical procedure or test is “Are the instruments sterilized?” After all, sterilization is specifically adopted for low fatality during surgeries and other treatment procedures. 

A guest post by Pramod Kumar.

It has been more than 150 years since sterilization was first adopted in medical procedures, and it continues to remain an integral part of healthcare solutions and services all over the world. Furthermore, with the growing incidence of hospital-acquired infections (HAI) in several countries, the adoption of sterilization methods will increase even more in the coming years. According to the Centers for Disease Control and Prevention (CDC), “Every year, Americans contract 1.7 million infections while being treated in hospitals. These infections are associated with approximately 99,000 deaths annually. In addition to the significant toll on patients’ lives, HAIs represent an estimated $30 billion in added healthcare costs.” 

Recommended book: Sterility, Sterilisation and Sterility Assurance for Pharmaceuticals: Technology, Validation and Current Regulations

Hence, with the growing prevalence of HAIs in many countries and mushrooming requirement for sterilization, the global sterilization technology market will exhibit rapid progress in the future years.

As noted above, the increasing prevalence of HAIs is a major factor fueling the requirement for sterilization technology across the world. The growing incidence of these infections has massively pushed up the mortality rate and caused heavy financial losses for the healthcare industry. According to the World Health Organization (WHO), “Of every 100 hospitalized patients at any given time, seven in developed and 10 in developing countries will acquire at least one healthcare-associated infection”. Furthermore, the prevalence of HAIs is considerably higher in middle- and low-income countries than the high-income ones.

Growing Surgery Volume also Playing Important Role

As per a report published by the WHO in 2016, almost 266.2 to 359.5 million surgical procedures were performed all over the globe in 2012. Considering the boom in the population and increase in the access to healthcare, it is assumable that the surgery volume would have burgeoned further in the last 7­–8 years. Furthermore, the increasing incidence of obesity is causing a sharp rise in the requirement for bariatric surgeries, on account of these surgeries being highly effective in treating obesity. The rapid rise in the number of surgeries being performed all over the world is a major factor propelling the demand for sterilization products and solutions.

Sales of Sterilization Devices To Rise Steeply in Future

In addition to the growing prevalence of HAIs and the increasing number of hospitals around the world, the rapid advancements in the biotechnology, medical device, and pharmaceutical industries are creating a huge requirement for various sterilization devices across the world. There are multiple types of such devices used in medical settings, such as those based on the low-temperature, steam, radiation, heat, liquid, ultrasound, and filtration technologies.

Out of all the devices, the usage of heat sterilizers is predicted to increase at the highest rate in the coming years. This is ascribed to the growing incorporation of heat sterilizers in instrument reprocessing and biotechnology and pharmaceutical manufacturing plants, for the sterilization of bulk products, moisture-sensitive materials, and products having medicinal properties. Moreover, heat has been the most effective way to kill microbes, which makes heat sterilizers vastly popular.

What Are Major Application Areas of Sterilization Technology?

Pharmaceutical sterilization and medical devices are the most important application areas of the sterilization technology. The adoption of sterilization methods is predicted to be quite high for medical devices in the forthcoming years. This is mainly credited to the surging requirement for advanced surgical devices, on account of the soaring geriatric population and ballooning number of surgical procedures around the world. Additionally, the increasing awareness on sterilization amongst both healthcare practitioners and patients is boosting the need for the proper sterilization of medical devices.

Sales of Sterilization Products Booming among Pharmaceutical and Biotechnology Firms

Out of all the major end users of sterilization products, namely ambulatory surgical centers and clinics, hospitals, academic and research organizations, medical device companies, and pharmaceutical and biotechnology companies, the highest procurement of these products is currently being observed among pharmaceutical and biotechnology companies. The main reasons behind the adoption of these products by these organizations are the mushrooming production of biotechnology and pharmaceutical products and increasing number of such companies across the world. Additionally, such companies are subject to some of the most stringent safety regulations of any industry, and failing to meet the set standards can not only lead to a loss of patient life, but also strong regulatory action.

North America Is the Most Sterile Continent

Globally, North America observed the highest adoption of sterilization technology in the past, with the U.S. recording huge sales of enabling products. This is ascribed to the existence of numerous medical device and pharmaceutical companies and hospitals and growing prevalence of HAIs in the country. In addition to these factors, the surging investments being made in the healthcare industry by private and public organizations and increasing number of surgical procedures being performed are also pushing up the requirement for these products in the region.

But Asia-Pacific (APAC) Is Quickly Catching Up!

Amongst all regions, the adoption of the sterilization technology is increasing at the highest rate in the Asia-Pacific (APAC) region. This is attributed to the surging healthcare expenditure, growing geriatric population, rising pollution levels, advancing pharmaceutical industry, and the presence of a flourishing medical tourism sector in the region. As per the observations of the World Bank, the total healthcare spending in South Asia was 3.9% of its gross domestic product (GDP) in 2000, and this share grew to 4.4% in 2014. The increasing healthcare spending in South Asian countries is creating huge growth opportunities for medical device manufacturing companies, which will, in turn, cause a swift expansion of the sterilization technology industry in the region, in the years to come.

Hence, it can be concluded that the demand for sterilization methods and products will rise tremendously all over the world in the coming years, mainly on account of the growing incidence of HAIs, increasing number of surgical procedures, and rising public awareness on hygiene.

Source: P&S Intelligence


Friday, 7 August 2020

Covid-19: Infectious coronaviruses 'circulating in bats for decades'

Greater horse shoe batImage copyrightGETTY IMAGES
Image captionBats may harbour viruses, but should not be persecuted, say experts

Coronaviruses capable of infecting humans may have been circulating undetected in bats for decades.

By Helen Briggs (BBC)

Research suggests one of the closest known ancestors of the virus that causes Covid-19 emerged in bats more than 40 years ago.

It has been poised for human crossover for some time, the scientists said.

And this casts further doubt on conspiracy theories that the virus causing Covid-19 was bioengineered or escaped from a laboratory, they added.

Prof David Robertson, of the University of Glasgow, worked on the study, published in the journal Nature Microbiology.

He said that while Sars-CoV-2 (the pandemic coronavirus) is genetically very close to the nearest known bat viruses, they are separated in time by several decades.

"That suggests that these viruses with potential to emerge in humans have been around for some time," he told BBC News.

"We really do need to understand where or how the virus has crossed into the human population. If we now believe there's this generalist virus circulating in bats we need to get better at monitoring that."

The work points to the need for further surveillance of emerging diseases in humans and to carry out more sampling within wild bat populations, if we are to prevent future pandemics, he said.

"If these viruses have been around for decades that means that they've had lots of opportunity to find new host species, including humans," said Prof Robertson.

The researchers compared the genetic make-up of Sars-CoV-2 with that of a close relative in bats, a virus known as RaTG13, and other related bat viruses.

They dated the time the two shared a common ancestor, and found they went along their own evolutionary pathways several decades ago.

Prof Mark Pagel of the University of Reading, who was not part of the study, said the work suggests that coronaviruses capable of infecting humans have been present in bats for perhaps 40 to 70 years but have gone undetected.

"This is significant in pointing to the scale and nature of the problems that zoonotic transmission presents to humans - there may be numerous and as yet undetected viruses capable of infecting humans that reside in animal hosts."

Bats migrating in ZambiaImage copyrightGETTY IMAGES
Image captionBats are found across the world and can migrate long distances

The viruses may have gone on to infect other wildlife, particularly those coming into close contact with each other through illegal wildlife trade. But to date, all the evidence points to bats being the important reservoir.

Previous research has suggested that pangolins might have played a role in the evolution of Sars-CoV-2, but the latest study suggests this is not the case.

Instead, pangolins may have picked up the virus more recently through contact with other wild animals through wildlife trafficking into China.


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