Antimicrobial susceptibility testing (AST) determines the resistance of microorganisms to antimicrobial agents. There are several methods for AST including disk diffusion, dilution methods, and E tests. Guidelines for AST are provided by CLSI and EUCAST. Mueller Hinton agar is the recommended medium for testing and inoculum preparation follows the 0.5 McFarland standard. Quality control using standard reference strains is important to ensure accurate and reproducible AST results.
The PPT is mainly all about Mycobacterium Tuberculosis. Agents causing the disease Tuberculosis, pathogenesis, laboratory diagnosis, treatment and prophylaxis. It was made for both BSc and MSc students.
Susceptibility testing is used to determine which antimicrobials will inhibit the growth of the bacteria or fungi causing a specific infection. The results from this test will help a healthcare practitioner determine which drugs are likely to be most effective in treating a person's infection.
The PPT is mainly all about Mycobacterium Tuberculosis. Agents causing the disease Tuberculosis, pathogenesis, laboratory diagnosis, treatment and prophylaxis. It was made for both BSc and MSc students.
Susceptibility testing is used to determine which antimicrobials will inhibit the growth of the bacteria or fungi causing a specific infection. The results from this test will help a healthcare practitioner determine which drugs are likely to be most effective in treating a person's infection.
University Institute of Pharmaceutical Sciences is a flag bearer of excellence in Pharmaceutical education and research in the country. Here is another initiative to make study material available to everyone worldwide. Based on the new PCI guidelines and syllabus here we have a presentation dealing with the quality control tests of parenteral as referred in the pharmacopoeia.
Thank you for reading. Hope it was of help to you.
UIPS,PU team
Principles and methods of different microbiological assay, methods for standa...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IV Part-2 Principles and methods of different microbiological assay, methods for standardization of antibiotics.
Introduction: Principles Advantages of Microbial Assay: Disadvantages of Microbial Assay: MICROBIOLOGICAL ASSAY OF ANIBIOTICS PRINCIPLE Media used for antibiotics assay Standard Preparation. Buffer Solutions Preparation of the Sample Solution: Test Organisms Preparation of inoculum: Methods of preparation of test organism suspension: Assay Methods: Method A: Cup-plate or Cylinder Plate Method.
Method B: Turbidimetric or Tube assay Method
Assessment of microbial contamination and spoilage. PHARMACEUTICAL MICROBIOLO...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-2
Assessment of microbial contamination and spoilage.
Assessment of microbial contamination and spoilage
1. Physical and chemical changes:
2. Assessment of viable microorganisms in non-sterile products:
3. Sterility test:
4. Estimation of pyrogens:
Microbial Limit Tests:
Total Aerobic Microbial Count:
Membrane Filtration.
Plate Count Methods.
Pour Plate Method.
Surface spread Method.
Most Probable Number(MPN)
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T) Unit-IV Part-3
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic: Introduction:
Principle
Microbiological Assay of Cynocobalamin (Vitamin B12):
Tritrimetric Method.
Turbidimetric Method.
Preparation of Standard Cynocobalmine stock solution:
Preparation of Basal Medium Stock Solution:
Test Solution of the material to be assayed Preparation of inoculum: Procedure of Titrimetric method: Turbidimetric Method: Microbiological assay of Amino acids. Assessment of a New Antibiotic.
Introduction:
MIC of an antibiotic is tested either by one of the following ways,
Liquid Dilution Method.
Solid Dilution Method
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.
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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
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
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
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
- 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
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
2. Introduction
• Once an organism is isolated , characterization
frequently includes tests to detect antimicrobial
resistance, which is the prime important key
component for the physician.
• The procedures used to produce the
antimicrobial susceptibility profiles and detect
resistance to therapeutic agents are referred to as
antimicrobial susceptibility testing (AST)
methods.
3. TESTING METHODS:
• Methods that directly measure the activity of
one or more antimicrobial agents against a
bacterial isolate
• Methods that directly detect the presence of a
specific resistance mechanism in a bacterial
isolate
• Special methods that measure complex
antimicrobial organism interactions
6. MEDIA IN AST
BEST MEDIUM – MHA (Mueller Hinton Agar)
1. Shows acceptable batch to batch
reproducibility for susceptibility testing
2. Low in sulphonamide, trimethoprim and
tetracycline inhibitors
3. Gives satisfactory growth of most non
fastidious pathogens
Media
7. • Store the plates at 2 to 8 deg C
• Use within 7 days of preparation
• Each batch of MHA plates should be checked for
sterility control
• MHA added with 2% NaCl
• MHA added with defibrinated sheep blood 5%s
9. Antibiotic Stock solutions
• Buy commercial pure source of antibiotics
• Don’t use injectable solutions
• Accurate weighing of powders is must
• Standard strains of stock cultures should be
used to evaluate the stock solution
• After preparing the stock solution, make 5 ml
aliquots and frozen it
Disc
11. DRIED FILTER PAPER DISCS:
• Whatmann no.1 filter paper is made to form a
disc size of 6mm
• Keep in petridish and sterilize in a hot air oven
• With the help of antibiotic delivery loop which
has a 20G wire with diameter of 2mm the
antibiotic is delivered (0.005ml)
12. Storage of discs
• Refrigerate at 2 to 8 deg C
• Beta lactam class drugs should be frozen
• The drugs should be kept outside at RT 1 to 2
hours before work
• The dispensing apparatus which is used to
deliver the drugs should also be refrigerated
• Check the expiry of drugs
13. Inoculum – Standard:
• O.5 Mcfarland standard
• Prepared by 0.5ml of 0.048mol/L BaCl2 and
99.5ml of 0.18mol/L of H2SO4
• Added with constant stirring
• Turbidity standard is checked by
spectrophotometer – 625nm – absorbance
should be 0.008 to 0.10
Inoculum
14. • Seal the tubes containing the Mcfarland
standards and store in dark at RT
• The standard turbidity should be mixed
throroughly every time before use
• Check the density monthly and replace it
monthly
15. AST METHODS
• Disk diffusion method
• MIC method
• E Test
• Automated systems
16.
17. CONVENTIONAL AST
DISK DIFFUSION METHOD:
• Simplest and most convenient
• Widely used everywhere
• Developed by kirby, sherrris, bauer and turk in
1966
• Types:
1. Kirby Bauer method
2. Stokes method
19. Application of Discs
• 150 mm plate 12 discs
• 100 mm plate 6 discs
• Drug should not be relocated
• Distance from the lid edge 15mm
• Distance between two drug from center to
center 24mm
• Inoculum disc placement incubation (
only 15 minutes delay is acceptable each)
21. INTERPRETATION:
• After 16 to 18 hours
• Confluent lawn of growth
• Zones of inhibition uniformly circular
• Read with reflected light
• For MRSA read in transmitted light
• When proteus is tested thin veil of
swarming growth after the zone of inhibition
should be ignored
22. STOKES METHOD
• Built in controls against many variables and
provide dependable results
• A standard sensitive strain of the bacterium is
inoculated in the middle third of the culture plate
• Standard strains:
• S.aureus ATCC 25923
• E.coli ATCC 25922
• P.aeruginosa ATCC 27853
24. • The test bacterium is inoculated in the upper
and lower third of the plate
• Antibiotic discs are placed between the
standard and test inocula so that zones of
inhibition formed around each disc are
composed of standard and test bacteria.
• The results are reported as Susceptible,
Intermediate susceptible, Resistant
25. DILUTION METHODS
• The minimum concentration of antimicrobial
to inhibit or to kill the microorganism is
determined
• MIC
• Broth dilution method
• Agar dilution method
26. • MIC – lowest concentration of antimicrobial
that will inhibit the visible growth of an
organism in an ideal growth condition
• MBC – least concentration of the test antiiotic
which will completely kill the bacteria tested
27. BROTH DILUTION METHOD
• Media: cation adjusted MH broth with a pH of
7.2 to 7.4
• For Haem.influenzae – HTM
• For TMP-SMX – Thymidine free medium
• For oxacillin Resistance – MH broth with 2%
NaCl
28. Working antibiotic solution
• Take original stock solution
• Prepare stock dilutions of the antibiotic of
concentrations 1000 and 100ug/ml
• Arrange two rows of 12 sterile 7.5*1.3cm
capped tubes in the rack
• Take a 30ml universal screw capped bottle
• Add 8ml broth and required antibiotic
concentration and mix the contents
29. • Take 2ml + 2ml and put it in first tube in both
rows
• Remaining 4ml broth add 4ml fresh broth
• Transfer 2ml+2ml and put in second tube
• Continue this dilution upto 11 tubes
• 12th tube control
30. INOCULATION
• Inoculation of 1st row with one drop of overnight
broth culture 1 in 1000 dilutions
• 2nd row with control with known sensitivity
organisms
• Final inoculum = 5 * 105 cfu/ml
• Incubate at 37degC for 16 to 18 hours
• Inoculate another tube with 2ml broth and keep
at 4degC in a refrigerator overnight to be used as
standard for determinattion of complete
inhibition
32. INTERPRETATION
• Positive control tube – turbidity
• Negative control tube – clear
• MIC end point is read
• Special comments:
1. For TMP-SMX
2. Trailing
33. MICROBROTH DILUTION METHOD
• Use double strength mueller hinton broth
• 4 X strength antibiotic solutions prepared as
serial 2 fold dilutions
• Test organism is 2X106 /ml
• Done in 96 well plate
34. AGAR DILUTION METHOD
• 1ml concentration of the drug + 24 ml of MHA
• Inoculum – make 1:10 dilution of
0.5Mcfarland standard inoculum which
delivers 107 cfu/ml
• Using pipette or calibrated loop , deliver 0.001
ml on the surface of the agar giving the final
inoculum of 104 cfu /spot
• Inoculate a control plate
36. INTERPRETATION
• Examine the drug free control growth of the
test organism for viability and purity
• Place the plate on a dark background and
examine them for the lowest concentration
that inhibits visible growth
• A single colony of a faint haze is not recorded
as growth
37. E test
• Epsilometer test
• An exponential gradient testing methodology
• A predefined stable antimicrobial gradient is
present on a thin inert carrier strip
• Following incubation, the E strip releases drug
and a symmetrical inhibition ellipse is
produced
• MIC = intersection of the inhibitory zone edge
and the calibrated carrier strip
39. QUALITY CONTROL IN AST
• QC: a process in the laboratory designed to
monitor the analytical phase of testing
procedures to ensure that tests are working
properly
• CLSI recommends use of ATCC strains for QC in
AST
40. • QC strains should be included daily with the
test ideally
• Not more than 1 in 20 results should be
outside the accuracy limits
• No zone should be more than 4 SD away from
midpoint between the stated limits
41. • Due to expense, it can be switched as once
weekly testing
• Perform QC for 30 days and with less than
10% inaccuracy , continue for weekly
• Test repeated for each new drug included
• All documentation maintained indefinitely