Staphylococcus epidermidis is an opportunistic pathogen that commonly forms biofilms on medical devices. These biofilms make infections very difficult to treat as bacteria in biofilms are up to 1000 times more resistant to antibiotics. The document discusses various strategies to control S. epidermidis biofilms, including using antibiotic combinations to prevent resistance development, targeting mechanisms of biofilm antibiotic resistance, and exploring natural compounds and their synergistic effects with antibiotics.
Blood stream infections- clinical microbiologySijo A
Blood stream infections (BSI) refers to the presence of organisms in blood which are threat to every organ in the body.
It causes shock, multiple organ failure and DIC (Disseminated Intravascular Coagulation).
The presence of bacteria in blood is called Bacteremia.
The bacteria circulate and actively multiply in the blood stream is called Septicemia.
The presence of virus in blood is called Viremia.
The presence of parasite in blood is called Parasitemia.
The presence of fungi in blood is called Fungemia.
Blood stream infections- clinical microbiologySijo A
Blood stream infections (BSI) refers to the presence of organisms in blood which are threat to every organ in the body.
It causes shock, multiple organ failure and DIC (Disseminated Intravascular Coagulation).
The presence of bacteria in blood is called Bacteremia.
The bacteria circulate and actively multiply in the blood stream is called Septicemia.
The presence of virus in blood is called Viremia.
The presence of parasite in blood is called Parasitemia.
The presence of fungi in blood is called Fungemia.
Pneumonia is an infection of one or both lungs caused by bacteria, viruses and fungi. An infection of lung that involves the small air alveoli and the tissue around is called pneumonia.
Los documentos pertenecen a los docentes y alumnos de la facultad de medicina de la Fundación Barceló de Buenos Aires. Los autores de dichos documentos son los responsables de todo su contenido.
Pneumonia is an infection of one or both lungs caused by bacteria, viruses and fungi. An infection of lung that involves the small air alveoli and the tissue around is called pneumonia.
Los documentos pertenecen a los docentes y alumnos de la facultad de medicina de la Fundación Barceló de Buenos Aires. Los autores de dichos documentos son los responsables de todo su contenido.
Estafilococos. Generalidades. Clasificación y Descripción. Estructura y Funciones Celulares. Estafilococos y su importancia en Odontología. Toxinas. Exoenzimas. Patogenia. Datos Clínicos. Identificación.
The effect of silver nanoparticles on Staphylococcus epidermidis biofilm biom...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Bacterial biofilm has been considered responsible for many deaths and high health costs worldwide. Their better protection against antibacterial agents compared to free living cells leads to poor treatment efficiency. Nanotechnology is promising approach to combat biofilm infections. The aim of the present study was to eradicate Staphylococcus epidermidis biofilm with silver nanoparticles (SNPs).
Materials and Methods:
SNPs were used at different concentrations (two fold dilutions) and incubation times (24, 48, 72 h). The crystal violet staining and pour plate assays were used to assess biofilm biomass and bacterial viability, respectively. The ability of SNPs on biofilm matrix eradication was assessed through optical density ratio (ODr). Positive control was defined as an ODr =1.0.
Results:
The crystal violet assay indicated that the biofilm matrixes were intact at different concentrations of SNOs and incubation times. There were no significant differences between these parameters (P >0.05). Bacterial enumeration studies revealed that higher concentrations of SNPs were more effective in killing bacteria than lower ones. Although, longer incubation times led to enhancement of anti-biofilm activity of SNPs.
Conclusion:
The anti-biofilm activity of SNPs was concentration- and time-dependent. The results of this study highlighted that SNPs were effective against cell viability; however they were ineffective against biomass.
International Journal of Pharmaceutical Science Invention (IJPSI) inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Evaluation of resistance profile of pseudomonas aeruginosa with reference to ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
A double-blind study was designed to confirm the antibacterial effect of Pure Bee Venom (PBV) and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris.
Effects of cosmetics containing purified honeybee (Apis mellifera L.) venom on acne vulgaris
Multidrug Resistance Pattern of Staphylococcus Aureus Isolates in Maiduguri ...Scientific Review SR
Multi drug-resistant (MDR) isolates of Staphylococcus aureus are on rise and are becoming a
challenge for timely and appropriate treatment. The present study was carried out with an objective to isolate
Staphylococcus aureus from clinical samples and determine their sensitivity. Out of 110 samples collected, 44
were shown to contained S. aureus. The isolates were subjected to antibiotic sensitivity tests using 10 different
and commonly used antibiotics by modified Kirby- Bauer disc diffusion technique. Out of the total isolates (42)
tested, only 7.1% were susceptible to all the antibiotics. Multiple resistance was eminent in over 92% with
highest occurrence in 4.8% where the entire antibiotics were resisted. Multiple antibiotic resistance indixes
(MAR index) indicated that 0.6 index occurred most (23.8%) followed by 0.5 (19.0%). On the other hand, 0.1
and 0.8 indexes were the lowest with 0.0% and 1.0% occurrence respectively. Ciprofloxacin was resisted by
most of the organisms (64.3%) while amoxicillin (64.3%) and streptomycin (61.9%) were most efficacious. With
over 90% isolate having MAR index ≥ 0.2, the multiple drug resistance by the S. aureus is quite alarming and
might suggest inappropriate antibiotic usage by the sampled population. Therefore, the need to strategize the
nature of antibiotic treatment against S. aureus and massive campaign on indiscriminate antibiotic use is urgent.
Multidrug Resistance Pattern of Staphylococcus Aureus Isolates in Maiduguri M...Scientific Review
Multi drug-resistant (MDR) isolates of Staphylococcus aureus are on rise and are becoming a challenge for timely and appropriate treatment. The present study was carried out with an objective to isolate Staphylococcus aureus from clinical samples and determine their sensitivity. Out of 110 samples collected, 44 were shown to contained S. aureus. The isolates were subjected to antibiotic sensitivity tests using 10 different and commonly used antibiotics by modified Kirby- Bauer disc diffusion technique. Out of the total isolates (42) tested, only 7.1% were susceptible to all the antibiotics. Multiple resistance was eminent in over 92% with highest occurrence in 4.8% where the entire antibiotics were resisted. Multiple antibiotic resistance indixes (MAR index) indicated that 0.6 index occurred most (23.8%) followed by 0.5 (19.0%). On the other hand, 0.1 and 0.8 indexes were the lowest with 0.0% and 1.0% occurrence respectively. Ciprofloxacin was resisted by most of the organisms (64.3%) while amoxicillin (64.3%) and streptomycin (61.9%) were most efficacious. With over 90% isolate having MAR index ≥ 0.2, the multiple drug resistance by the S. aureus is quite alarming and might suggest inappropriate antibiotic usage by the sampled population. Therefore, the need to strategize the nature of antibiotic treatment against S. aureus and massive campaign on indiscriminate antibiotic use is urgent.
Food hygiene is more than cleanliness ......
Protecting food from risk of contamination, including harmful bacteria, poison and other foreign bodies.
Preventing any bacteria present multiplying to an extent which would result in the illness of consumers or the early spoilage of the food.
Destroying any harmful bacteria in the food by thorough cooking
or processing.
Discarding unfit or contaminated food.
T-Cell Activation
• Concept of immune response
• T cell-mediated immune response
• B cell-mediated immune response
I. Concept of immune response
• A collective and coordinated response to the introduction of foreign substances in an individual mediated by the cells and molecules in the immune system.
II. T cell-mediated immune response
• Cell-mediated immunity is the arm of the adaptive immune response whose role is to combat infection of intracellular pathogens, such as intracellular bacteria (mycobacteria, listeria monocytogens), viruses, protozoa, etc.
Major Histocompatibility Complex
MHC:
• Major Histocompatibility Complex
– Cluster of genes found in all mammals
– Its products play role in discriminating self/non-self
– Participant in both humoral and cell-mediated immunity
• MHC Act As Antigen Presenting Structures
• In Human MHC Is Found On Chromosome 6
– Referred to as HLA complex
• In Mice MHC Is Found On Chromosome 17
– Referred to as H-2 complex
• Genes Of MHC Organized In 3 Classes
– Class I MHC genes
• Glycoproteins expressed on all nucleated cells
• Major function to present processed Ags to TC
– Class II MHC genes
• Glycoproteins expressed on macrophages, B-cells, DCs
• Major function to present processed Ags to TH
– Class III MHC genes
• Products that include secreted proteins that have immune functions. Ex. Complement system, inflammatory molecules
Antigen Processing and Presentation MID
Antigens and “foreignness”
• Antigens (or, more properly, immunogens) have a series of features which confer immunogenicity.
• One of these features is “foreignness.”
• So, we can infer that – most often – antigens – ultimately – originate externally.
• (There are exceptions, of course. Some cells become transformed by disease [e. g., cancer] or by aging. In such instances, the antigens have an internal origin.)
Extinction of a particular animal or plant species occurs when there are no more individuals of that species alive anywhere in the world - the species has died out. This is a natural part of evolution. But sometimes extinctions happen at a much faster rate than usual. Natural Causes of Extinction.
Difference between In-Situ and Ex-Situ conservation
Conservation of biodiversity and genetic resources helps protect, maintain and recover endangered animal and plant species. There are mainly two strategies for the conservation of wildlife: In-situ conservation and Ex-situ conservation. Although, both the strategies aim to maintain and recover endangered species, they are different from each other. Let us see how they differ from each other!
Evolution Of Bacteria
Bacteria have existed from very early in the history of life on Earth. Bacteria fossils discovered in rocks date from at least the Devonian Period (419.2 million to 358.9 million years ago), and there are convincing arguments that bacteria have been present since early Precambrian time, about 3.5 billion years ago. Bacteria were widespread on Earth at least since the latter part of the Paleoproterozoic, roughly 1.8 billion years ago, when oxygen appeared in the atmosphere as a result of the action of the cyanobacteria. Bacteria have thus had plenty of time to adapt to their environments and to have given rise to numerous descendant forms.
Impact of Environment on Loss of Genetic Diversity and Speciation
Genetic variation describes naturally occurring genetic differences among individuals of the same species. This variation permits flexibility and survival of a population in the face of changing environmental circumstances. Consequently, genetic variation is often considered an advantage, as it is a form of preparation for the unexpected. But how does genetic variation increase or decrease? And what effect do fluctuations in genetic variation have on populations over time?
GENE ENVIRONMENT INTERACTION
Subtle differences in one person’s genes can cause them to respond differently to the same environmental exposure as another person. As a result, some people may develop a disease after being exposed to something in the environment while others may not.
As scientists learn more about the connection between genes and the environment, they pursue new approaches for preventing and treating disease that consider individual genetic codes.
How to store food in hot
The Good News
To maximize benefit of preservation, keep your food as fresh as possible for as long as possible. You can do this, even in the heat, by creating a “cooler” made from two basic terra cotta pots, one larger than the other. Put the smaller pot in the larger one, fill the gap with sand, and saturate the sand with water. Then cover it with a cloth. To add additional insulation from the heat, bury the pot up to its rim. The evaporation of moisture from the wet sand will cool the air around the food and help keep it fresh.
What is IUPAC naming?
In order to give compounds a name, certain rules must be followed. When naming organic compounds, the IUPAC (International Union of Pure and Applied Chemistry) nomenclature (naming scheme) is used. This is to give consistency to the names. It also enables every compound to have a unique name, which is not possible with the common names used (for example in industry). We will first look at some of the steps that need to be followed when naming a compound, and then try to apply these rules to some specific examples.
IUPAC Nomenclature
IUPAC nomenclature uses the longest continuous chain of carbon atoms to determine the basic root name of the compound. The root name is then modified due to the presence of different functional groups which replace hydrogen or carbon atoms in the parent structure.
Hybridization describes the bonding atoms from an atom's point of view. For a tetrahedral coordinated carbon (e.g. methane CH4), the carbon should have 4 orbitals with the correct symmetry to bond to the 4 hydrogen atoms.
INTRODUCTION:
Hybrid Orbitals
Developed by Linus Pauling, the concept of hybrid orbitals was a theory created to explain the structures of molecules in space. The theory consists of combining atomic orbitals (ex: s,p,d,f) into new hybrid orbitals (ex: sp, sp2, sp3).
1. Why Firefly give light during night?
2. Why atomic mass and Atomic numbers are given to elements ?
3. Why elements have been characterized and classified into different groups?
4. What is the transition of elements and what they play their role in elements stability?
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
The Gram stain is a fundamental technique in microbiology used to classify bacteria based on their cell wall structure. It provides a quick and simple method to distinguish between Gram-positive and Gram-negative bacteria, which have different susceptibilities to antibiotics
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
CDSCO and Phamacovigilance {Regulatory body in India}NEHA GUPTA
The Central Drugs Standard Control Organization (CDSCO) is India's national regulatory body for pharmaceuticals and medical devices. Operating under the Directorate General of Health Services, Ministry of Health & Family Welfare, Government of India, the CDSCO is responsible for approving new drugs, conducting clinical trials, setting standards for drugs, controlling the quality of imported drugs, and coordinating the activities of State Drug Control Organizations by providing expert advice.
Pharmacovigilance, on the other hand, is the science and activities related to the detection, assessment, understanding, and prevention of adverse effects or any other drug-related problems. The primary aim of pharmacovigilance is to ensure the safety and efficacy of medicines, thereby protecting public health.
In India, pharmacovigilance activities are monitored by the Pharmacovigilance Programme of India (PvPI), which works closely with CDSCO to collect, analyze, and act upon data regarding adverse drug reactions (ADRs). Together, they play a critical role in ensuring that the benefits of drugs outweigh their risks, maintaining high standards of patient safety, and promoting the rational use of medicines.
Basavarajeeyam is an important text for ayurvedic physician belonging to andhra pradehs. It is a popular compendium in various parts of our country as well as in andhra pradesh. The content of the text was presented in sanskrit and telugu language (Bilingual). One of the most famous book in ayurvedic pharmaceutics and therapeutics. This book contains 25 chapters called as prakaranas. Many rasaoushadis were explained, pioneer of dhatu druti, nadi pareeksha, mutra pareeksha etc. Belongs to the period of 15-16 century. New diseases like upadamsha, phiranga rogas are explained.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
1. Prepared By Amjad Khan Submitted to
1
StaphylococcusEpidermidis
Introduction
Staphylococcus epidermidis is a Gram-positive bacterium that normally colonizes the human
skin and mucous membranes. Previously regarded as an innocuous commensal microorganism
it is now seen as an important opportunistic pathogen, becoming,in the past few decades, the
most frequent causative agent of nosocomial infections. This is mainly due to the increasing use
of medical devices, allowing for biofilm formation in such surfaces 1. S. epidermidis biofilm-
related infections normally begin with the introduction of bacteria from the skin of the patient
or health care personnel during device insertion.
While these infections rarely lead to mortality, they are associated with increased patient
morbidity 1.An important aspect of the biofilm-related infections is their economic burden on
the public health system at an annual cost of over 2 billion Dollar in the US alone 2. Microbial
biofilms are communities of bacteria that live adhered to a surface and are surrounded by an
extracellular polymeric matrix, in an increased antibiotic resistance and tolerance to the
immune system3. Multiple mechanisms have been proposed for the high resistance of bacterial
biofilms to antibiotics, including 1) the low diffusivity of antibiotics through the matrix, 2) the
inactivation of the antibiotics by matrix components, 3) the presence of a sub-population of
bacteria, known as persisters, that are unaffected by antibiotics, or 4) the heterogeneous
nature of the biofilm composition and tridimensional structure 4.These mechanisms only
partially explain the increased resistance and probably this phenotype is the result of more than
one specific mechanism. Increased bacterial resistance toward antibiotics has led to the search
for new antimicrobial therapeutic agents such as essential oils fromplants. Farnesol is a
naturally-occurring sesquiterpene that was originally isolated from essential oils found in many
plants 5. Farnesol has also been found to be produced by Candida spp, being involved in
quorum sensing 6. Of high importance is the fact that farnesol has been shown to have
antimicrobial potential against several bacteria, including S. epidermidis7.However, the
mechanism of action of farnesol is not yet fully understood, but it seems to
be related to cell membrane integrity 8.
We recently described a bacterial strain where farnesol had no detectable antibiotic effect but
strongly reduced biofilm biomass. We hypothesized that farnesol could be inducing biofilm
detachment9. In this manuscript we tested this hypothesis and assayed the effect of farnesol in
biofilm-forming clinical isolates of S. epidermidis, representing a wide diversity in terms of
genetic background, geographic and clinical origins.
General introduction
The coagulase-negative staphylococci Staphylococcus epidermidis is a human skin commensal
microorganism.
However, this bacterium can become an opportunistic pathogen and as such is often associated
with bacteremia and hospital acquired infections, particularly in patients with catheters or
2. Prepared By Amjad Khan Submitted to
2
others medical devices [1]. The main cause of their pathogenicity is their ability to adhere and
form biofilms on the surfaces of the medical devices formerly mentioned. A biofilm is an
aggregate of cells adhered to living or non-living surfaces and embedded in a self-produced
matrix of extracellular polymeric substances (EPS). In biofilm form, bacteria are protected from
antimicrobial agents and the host immune system contributing to the persistence of biofilm
infections, and can be up to 1 000 fold more resistant to antibiotics and other antimicrobial
agents, than their planktonic counterparts [2-4].
The high resistance presented by
Staphylococcus epidermidis biofilm cells to antimicrobial agents boosted the persistant demand
for new control strategies against this pathogenic bacterium in this mode of growth.
In fact, new strategies to control S. epidermidis biofilm-mediated infections have attracted the
attention of many research groups and several studies have and are being done in order to find
antimicrobial agents effective against this bacterium, specifically in this mode of growth.
Natural subtances with possible antimicrobial properties have raised
researchers’ interest and are pointed as potential alternatives to antibiotics. Some examples
are farnesol and other terpene alcohols, N-acetylcysteine (NAC), berberine, casbane diterpene,
salvipisone, etc. that have been tested as novel agents against several pathogenic
microorganisms such as bacteria, fungi and also virus, and namely against S.
epidermidis. The development of adaptative resistance to antibiotics used in common clinical
practice has also increased the interest on the new generation of antibiotics such as tigecycline,
daptomycin, linezolid, arylomycins, etc.,
which are seen as therapeutic alternatives. Furthermore, the rapid emergence of resistance has
highlighted the importance of antibiotics combination or with other antimicrobial agents,
aiming at the reduction of the likelihood of
resistance development and the enhancement of the effects of individual antimicrobial agents
by synergistic action. Some of these strategies showed encouraging in vitro results in controlling
S. epidermidis biofilms and appear to be promising alternatives to standard antibiotics usually
used in the treatment of S. epidermidis related infections.
Due to the increased involvement of S. epidermidis in foreignbody-related infections, the rapid
development and exhibition of multiple antibiotic resistance as well as its great propensity to
lead to persistent, chronic and recurrent infections, this pathogen remains a challenge and a
subject of study by several research groups and continues to receive significant attention.
Materials and Methods
Bacterial strains
Two well known biofilm-forming strains were selected to be used as control strains, based on
our previous work:S. epidermidis 9142 biofilm biomass is reduced by farnesol while S.
3. Prepared By Amjad Khan Submitted to
3
epidermidis 1457 biofilms shows no reduction7.Furthermore, we also used 25 clinical strains
previously characterized by several different molecular typing techniques, namely
staphylococcal chromosome cassette mec (SCCmec) and multilocus sequence typing (MLST),
were selected from a total of 217 nosocomial isolates collected between 1996 and 2001 in 17
different countries, from disease (107 isolates) and from carriage (87 isolates). Isolates were
selected in order to include the highest diversity as possible in terms of genetic backgrounds,
geographic and clinical origins. Finally a subset of 9 clinical strains isolated from indwelling
devices in Boston, MA, USA, were also included. All strains are listed in the results section.
Biofilm quantification
Biofilms were quantified using 3 different approaches. TSB was inoculated with one single
colony of each S. epidermidis control strain, in a 10 mL sterile tube, and incubated at 37°C in a
shaker at 120 rpm for 24 ± 2 h. Then a 1:200 dilution was performed in fresh TSB supplemented
with 1% (w/v) of glucose (TSBG), and incubated at 37°C in a shaker at 120 rpm in 96 well culture
plates (Orange Scientific, Braine-l’Alleud, Belgium) for 24 h. Biofilms were then washed twice
with 0.9% NaCl and fresh TSBG was added with or without 300 μM of farnesol and allowed to
incubate in the same conditions for further 24 h. For biofilm biomass determination, the
standard crystal violet staining method was used as described elsewhere 10. To determine cell
viability, biofilms were washed twice with 0.9% NaCl and resuspended in 0.9% NaCl followed by
gentle sonication as described above. CFUs were determined by the standard plating method,
using TSA plates. To determine the percentage of growth of bacteria inside the biofilm after 48
h, we quantified the relative population density of biofilms and the respective suspension
formed during the last 24 h of growth, on each 96 well, as described before 11. These
experiments were repeated three to five times with 8 replicates.
Molecular characterization of the clinical
strains
All strains were characterized by multilocus sequence typing (MLST)following the scheme
proposed by Thomas et al.12. The MLST data wereanalysed using the goeBURST algorithm
(http://goeBURST.phylowiz.net).This analysis was performed on March 21st, 2012. Isolates
were considered as belonging to the same clonal complex (CC) if sharing six out of seven
loci.The SCCmec type was determined by the combination of the class of mec complex and the
type of ccr complex. SCCmec was considered non-typable when either mec complex or ccr
complex, or both, were nontypable by the methods used or when the isolate carried more than
one ccr type. SCCmec was considered to be new if a new combination of mec complex and ccr
complex was found. The presence of the genes associated to biofilm formation, namely icaA,
aap and bhp was detected by PCR, using DyNAzyme II PCR mix (Finnzymes, Vantaa, Finland), in
the following thermal conditions: initial denaturation at 94ºC for 5 min followed by 35 cycles of
94ºC at 30 s, 54ºC at 30 s and 72ºC at 45 s. A final extension step was performed for 10 min at
72 ºC. Negative results were re-checked with a second set of primers. Oligonucleotide primers
4. Prepared By Amjad Khan Submitted to
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were designed using the Primer3 software, having S. epidermidis RP62A genome as template:
icaA (Fw - tgcactcaatgagggaatca, Rv- tcaggcactaacatccagca; amplicon size of 417), aap (Fw -
gctctcataacgccacttgc, Rv - ggacagccacctggtacaac; amplicon size of 617), bhp (Fw -
tggactcgtagcttcgtcct, Rv - tctgcagatacccagacaacc; amplicon size of 213).For biofilm biomass
determination, the standard crystal violet staining method was used 10.
GRAPH
Effect of farnesolon S. epidermidisbiofilms. Bacteria were allowed to form
biofilms for 24 h, after which fresh medium was added with or without 300 μM of farnesol and
allowed to grow for further 24 h. Top: crystal violet staining; middle: CFUs determination;
5. Prepared By Amjad Khan Submitted to
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bottom: percentage of planktonic cells living outside the biofilms.* significant difference, paired
samples t-student, p<0.05.
Staphylococcus epidermidis biofilm control strategies
Traditional antibiotic combination
As it was already referred, a worrying current problem is the increasing bacterial resistance to
traditional antibiotics commonly used in hospitals to treat coagulase-negative (CoNS)
infections, which is aggravated when the cells are in biofilms. Generally, antimicrobial agents
alone are ineffective against S. epidermidis biofilms. Rifampicin, a RNA
synthesis inhibitor, is among the most effective molecules for treating biofilm-related infections
[5,6]. However, this antibiotic has a high propensity for rapid development of resistance [7,8,9].
This fact makes rifampicin unfeasible as monotherapy. A solution to this problem is the
combination of antimicrobial agents, avoiding the use of monotherapy.
Antibiotic combination represents a therapeutic option in the treatment of S. epidermidis
infections [10]. In fact, some authors demonstrated that, for example, the combination of
rifampicin with quinolones or fusidic acid could prevent the emergence of rifampicin resistance
during therapy [8,11].
Furthermore to avoid resistance, this strategy can also potentiate the effect of individual
antimicrobial agents by synergic action. Gomes et al. tested the susceptibility of S. epidermidis
biofilms in vitro to traditional antibiotics alone and in double combinations at breakpoint
concentration and demonstrated that there are some combinations that can be
potentially considered for therapeutic use for an efficient control of S. epidermidis biofilm
related-infection (data not shown). Rifampicin is always present in these combinations, being
rifampicin+gentamicin and rifampicin+clindamycin the most active combinations in terms of
CFU reduction and with the broadest range of action
Stationary phase Log phase
Inoculum cell density control farnesol control farnesol
109 / mL 0.27 ± 0.13 0.14 ± 0.13 N/A N / A
108 / mL 0.24 ± 0.26 0.02 ± 0.30 N/A N / A
107 / m L 0.13 ± 0.26* -1.12 ± 0.41* 0.85 ± 0.10** 0.03 ±
0.07**
106/ mL 0.75 ± 0.56* -1.42 ± 0.76* 1.56 ± 0.27** 0.29 ±
0.27**
105 / mL 0.93 ± 0.13* -1.01 ± 0.47* 1.50 ± 0.06* -1.07 ±
0.07*
104 / mL 0.63 ± 0.38* -1.50 ± 0.39* 1.16 ± 0.08* -0.61 ±
0.09*
Antibiotic resistance of bacteria in biofilms
6. Prepared By Amjad Khan Submitted to
6
Bacteria that adhere to implanted medical devices or damaged tissue can encase themselves in
a hydrated matrix of polysaccharide and protein, and form a slimy layer known as a biofilm.
Antibiotic resistance of bacteria in the biofilm mode of growth contributes to the chronicity of
infections such as those associated with implanted medical devices.
The mechanisms of resistance in biofilms are different from the now familiar plasmids,
transposons, and mutations that confer innate resistance to individual bacterial cells. In
biofilms, resistance seems to depend on multicellular strategies. We summarise the features of
biofilm infections, review emerging mechanisms of resistance, and discuss potential therapies.
Resistance mechanisms
The familiar mechanisms of antibiotic resistance, such as efflux pumps, modifying enzymes, and
target mutations,12 do not seem to be responsible for the protection of bacteria in a biofilm. Even
sensitive bacteria that do not have a known genetic basis for resistance can have profoundly
reduced susceptibility when they form a biofilm. For example, a -lactamase-negative strain of
Klebsiella pneumoniae had a minimum inhibitory concentration of 2 _g/mL ampicillin in
aqueous suspension.13 The same strain, when grown as a biofilm, was scarcely affected (66%
survival) by 4 h treatment with 5000 _g/mL ampicillin, a dose that eradicated freefloating
bacteria.13 When bacteria are dispersed from a bacteria.13 When bacteria are dispersed from a
biofilm they usually rapidly become susceptible to biofilm they usually rapidly become
susceptible to antibiotics,14,15 which suggests that resistance of bacteria in biofilms is not
acquired via mutations or mobile genetic
Strategies to control Staphylococcus epidermidis biofilms
F. Gomes, B. Leite, P. Teixeira and R. Oliveira
IBB- Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University
of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Staphylococcus epidermidis is the
staphylococci species most commonly associated with bacteremia and hospital-acquired
infections and has recently arisen as the leading cause of infections related to indwelling
medical devices such as vascular catheters, prosthetic joints and artificial heart valves. The
prevalence of S. epidermidis in hospital-acquired infections is due to its ability to adhere and
form biofilms on biomaterial surfaces. This feature is one of the most important virulence
factors found in S. epidermidis. In biofilm form, bacteria are protected from antimicrobial
agents and the host immune system contributing to the persistence of biofilm infections. In
addition, the emergence of S. epidermidis resistance to conventional therapies, based in the
use of traditional antibiotics, leads to the failure of the current treatments used in the
combat of S. epidermidis infections and is becoming a major concern. These facts are
stimulating the continuous search for novel agents able to eradicate S. epidermidis biofilm
infections or that can work in synergy with the currently available antimicrobial agents. New
strategies have been showing encouraging in vitro results in controlling S. epidermidis biofilms
and seem to be promising alternatives to standard antibiotics usually used in the treatment of
S. epidermidis related infections.
7. Prepared By Amjad Khan Submitted to
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Conclusions
Although new antibiotics have been developed in order to overcome the growing problem of
bacteria resistance, namely of S. epidermidis, this does not seem to be sufficient due to the rapid
emergence of resistance caused by the overuse of antibiotics, the high diversity of these bacteria,
their short replication time, the horizontal transfer of resistance genes, etc. Another problem to
take into consideration is the strain diversity and response to the different antimicrobial agents.
The effect of antimicrobial agents is highly strain-dependent and the rate of success will be
strongly dependent on the infectious S. epidermidis strain. The only way to avoid or slow the
rapid emergence of resistance remains the prudent use of antibiotics and/or the development of
alternatives to control S. epidermidis-related infections. Therefore it is still needed a continuous
search for new strategies against S. epidermidis biofilms. Overall, there are some promising
therapeutic strategies, such as some natural compounds and antimicrobial agents combinations,
that can be possibly used for medical purposes especially in the combat of infections caused by
S. epidermidis biofilms.
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