The document discusses various methods for staining bacterial endospores, including Gram staining, modified Ziehl-Neelsen staining, Dorner's method, Schaeffer-Fulton method, Moeller staining, and Abbott's method. Each method uses different primary and counter stains to differentially stain the endospores and vegetative cells. The document provides details on the staining procedures and reagents used for each method as well as the expected color results, with endospores typically staining red or green and vegetative cells staining blue, violet or colorless.
Capsule is an layer around the bacteria cell which gives bacteria the protection and pathogenicity. Staining such an layer is difficult with the normal stains so it is necessary to stain the background and the cell itself which makes the capsule appear colourless.
Capsule is an layer around the bacteria cell which gives bacteria the protection and pathogenicity. Staining such an layer is difficult with the normal stains so it is necessary to stain the background and the cell itself which makes the capsule appear colourless.
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
COLLECTION AND TRANSPORTATION OF CLINICAL SAMPLESNCRIMS, Meerut
Principles of Sample Collection:
Aseptic precautions to minimize chances of
contamination.
Appropriate anatomic sites
Adequate volume
Adequate no. of samples
Appropriate time
Appropriate container with proper labelling
Before initiation of anti-microbials
Adequate information in request form
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
COLLECTION AND TRANSPORTATION OF CLINICAL SAMPLESNCRIMS, Meerut
Principles of Sample Collection:
Aseptic precautions to minimize chances of
contamination.
Appropriate anatomic sites
Adequate volume
Adequate no. of samples
Appropriate time
Appropriate container with proper labelling
Before initiation of anti-microbials
Adequate information in request form
Make drawings of both organisms stained in each staining procedure. L.pdfDhanrajsolanki2091
Make drawings of both organisms stained in each staining procedure. Label each drawing with
the organism name, the total magnification and the name of the staining procedure. For each
drawing, record your observations, including the staining result (acid-fast or non-acid-fast;
spores present or spores absent), cell shape, cell arrangement and cell size. Explain the
mechanism by which the acid-fast stain differentiates between acid-fast and non-acid-fast
organisms. Explain why endospores stain one color and vegetative cells stain a different color in
the endospore stain. Be thoughtful and complete in your explanation. Would you expect to see
endospores in a fresh (24 hour) culture of a Bacillus species? Explain your answer. Suppose you
prepare a Gram stain using a 7-day old culture of a Bacillus species. You suspect endospores
might be present. What would you expect the spores to look like in a Gram stain and why?
Explain when, why and/or how the acid-fast and spore stains are important or useful. In other
words, how do we use that information? (Go farther than just stating that these procedures
identify acid-fast and spore-forming organisms, respectively.)
Solution
1) It is totally dependent on your lab observations.
2) Acid fast staining involves, primary staining with Carbolfuchsin followed by
decolourisation with Acid-alcohol and at last counter staining with Methylene blue. In primary
staining both acid fast and non-acid fast bacteria stained red. But in second decolourisation step
non-acid fast bacteria loses red colour whereas acid fast bacteria remain red. In third counter
staining step non-acid fast bacteria who initially lost red colour in decolourisation step will be
counter stained blue with methylene blue whereas acid fast bacteria retain its red colour
(Carbolfuchsin).
Acid fast bacteria: Mycobacterium and many Nocardia species
3) When vegetative cell of certain bacteria is exposed to harsh conditions, to protect cell forms
an endosperm to survive and as the favourable conditions come again cell behaves like
vegetative cell.
Endospore staining has following major steps:
1. Making bacterial cell smear on slide
2. Heating slide gently for fixing it
3. malachite green staining along with heating gently
4. Rinsing slide with tap water
5. Counterstain with safranin
Spores resist staining. Therefore heating is done to allow stain to penetrate in endospore.
Malachite green is a water soluble dye which is removed easily in washing step from cell wall of
vegetative cells but not from the spores. And that’s why spores appear blue (due to primary dye
malachite green) and vegetative cell retain the secondary stain safranin and appear pin/red.
4) A 24 hour culture will not show any endospore. Endospores forms when cell exposed to
harsh conditions like harsh temperature, lack of nutrients etc. But as it faces favourable
conditions of temperature and nutrition, it again gains its vegetative characteristics. As bacteria
are in culture from 24 ho.
Hereby you can get all about bacterial staining.
MICROBIAL STAINING
introduction
Microbial Staining - giving color to microbes. Because microbes are colorless and highly transparent structures.
Staining process in which microbes are getting color.
STAINES / DYES
Staines / dyes - organic compounds which carries either positive charges or negative charges or both .
Based on the charges
Basic stain / dyes :- stain with + ve charge .
Acidic stain / dyes - stain with -ve charge .
2. Based on function of stain :
Neutral stain / dyes - stain with both charges .
Simple staining only one dye is used differentiation among bacteria is impossible Eg . Simple Staining.
Differential staining- more than one dye is used- Differentiation among bacteria is possible- Eg. Gram's staining, Acid - fast staining.
Special staining - more than one dye used - Special structures are seen. Eg. Capsule staining , Spore staining .
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
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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.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
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
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
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
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.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
2. HISTORY
Endospores were first described by Cohn (1872) in Bacillus
subtilis and later by Koch(1876) in the pathogen, Bacillus
anthracis.
Cohn demonstrated the heat resistance of endospores in B.
subtilis, and Koch described the developmental cycle of
spore formation in B. anthracis.
3. HISTORY CONTD….
In 1922, Dorner published a method for staining endospores. It
employed a lengthy heating step but resulted in differential
staining of endospores and vegetative cells in the same sample.
Shaeffer and Fulton modified Dorner’s method in 1933 to
make the process faster.
4. ENDOSPORES…..
Endospores are so named because they are formed intracellularly,
although they are eventually released from this mother cell or
sporangium as free spores.
An endospore is a spherical/oval, thick-walled, highly resistant
structure formed in certain bacterial species that represent a
dormant or resting stage in the growth cycle of an organism. e.g.
Bacillus sp., Clostridium sp.
5. ENDOSPORES…..
Exhaustion of nutrients (Carbon and Nitrogen)
Sub-optimal temperatures
NOT a method of reproduction
The endospore consists of the bacterium's DNA and part of
its cytoplasm, surrounded by a very tough outer coating.
Triggering
factors
6. COVERINGS OF SPORE
Spore wall :
• delicate membrane from
which cell wall of future
vegetative bacterium develop
Spore cortex
Spore coat
• tough, multi-layered
Exosporium
7. RESISTANT TO….
Ultraviolet radiation
Desiccation
High temperature
Extreme freezing
Chemical disinfectants
Starvation
DESTROYED BY….
Autoclaving
Tyndallisation
ENDOSPORES…..
8. Germination of spores-
Increase in metabolic
activity of spore
Endospore swells
Germ cell appears by
rupture or absorption of
spore coat
Elongates to vegetative
bacterium
9. Distend bacillary body –
Clostridium spp.
Do not distend bacillary body –
B. anthracis
Central – B. anthracis,
C. bifermentans
Terminal - C. tetani, C. tertium
Sub-terminal – C. perfringens
(club-shaped)
Oval - B. anthracis, C. tertium
(tennis racket)
Spherical - C. tetani. (drum-stick)
SHAPE, POSITION AND SIZE OF
SPORE RELATIVE TO PARENT CELL
13. Endospores are best observed in unstained wet films
under the phase contrast microscope.
They appear as large, refractile, oval or spherical within
the bacterial mother cells or elsewhere free from the
bacteria.
17. MODIFIED ZIEHL-NEELSEN STAINING
METHOD
Spores – red , Bacteria – blue
Counterstain - Loeffler’s methylene
blue for 1-2 min
Decolourise using 0.25% to 0.5%
sulphuric acid for 7-10 min
Wash with water
Heat the slide intermittently until steam
rises for 5 min
Cover the slide with carbol fuchsin
Heat fix the smear
18. DORNER METHOD
Vegetative cells – colourless Endospores – red
Background – black.
Observe under oil immersion lens
Dry a thin even film of nigrosin(10%) on the slide
Rinse with water and blot dry
Decolorize with acid-alcohol for 1 minute
Wash with water
Cover the smear with carbol fuchsin and
intermittently heat the slide for 5 min. Do not
allow the stain on the smear to dry
Heat fix the slide
19. VARIATION IN DORNER METHOD
Vegetative cells – colourless Endospores -
red Background – black.
Observe under oil immersion lens.
Mix a loopful of nigrosin on a glass slide
with one loopful of the boiled carbol
fuchsin-organism suspension and air dry to
a thin film.
Immerse the tube in a boiling water bath
for 10 minutes.
Mix an aqueous suspension of bacteria
with an equal volume of carbol fuchsin in a
test tube.
20. SCHAEFFER-FULTON METHOD / MODIFIED
ASHBY METHOD
Vegetative cell – pink or red and Spore – green.
Counterstained with safranine 1 min, vegetative cells stained.
After cooling, outer layer makes the spore resistant to the action of
decolorizing agent (water), but water can easily decolorize the vegetative
cells.
Smear taken from the steam bath and allowed to cool.
Both the spore and vegetative cells appear green.
Smear is heated over a steam bath for 5 min.
The primary stain, Malachite Green, is added over the heat fixed bacterial
smear
23. MOELLER STAIN
Spores red; Bacteria-blue.
The slide is rinsed with acidified ethanol,
and counter-stained with methylene blue
for 30 seconds.
The slide is then heated over a bunsen
burner, or suspended over a hot water
bath, covered with a paper towel, and
steamed for 3 minutes.
Carbol fuchsin is applied to a heat-fixed
slide.
24. MODIFIED MOELLER STAINING
METHOD
Rinse thoroughly in running water
Differentiate with 2 % sulphuric acid for 5-10 sec.
Rinse thoroughly in running water.
Flood with un-steamed Kinyoun’s carbol-fuchsin solution containing Tergitol
7, and stain for 3 min.
Rinse thoroughly in running water.
Immerse in 5% chromic acid solution for 3 min.
Fix in absolute methyl alcohol for 1-3 min.
Spread a small drop of the specimen on a slide, and allow it to air-dry at room
temperature.
25. MODIFIED MOELLER STAINING
METHOD CONTD…
Microscopic examination using oil
objective lens
Rinse with running water and allow
it to air-dry.
Counterstain with 0.1 % Loeffler’s
methylene blue for 1-2 min.
Rinse for 10 sec in running water.
Decolorize with 80 % ethanol until
removal of excess stain dye from the
slides.
26. ABBOTT'S METHOD
Spores blue, bodies of the bacteria red.
Wash in water, dry and mount.
Stain for 8-10 seconds in aniline-fuchsin solution.
Wash in water.
Wash in 95% alcohol containing 0.2 to 0.3% HCl.
Wash in water.
Stain the slide deeply with methylene-blue, heating until the solution boils.
27. HOLBROOK AND ANDERSON
LIPID/SPORE STAINING METHOD(1980)
Wash in water, dry and examine. Spores-green, vegetative bacilli-red,
lipid granules-unstained.
Counterstain with 0.5% safranine for 20 sec.
Wash film with xylene for 5 sec and blot dry.
Stain with 0.3% sudan black B in 70% ethanol for 15 min
Stain the film with 5% malachite green for 2 min while the slide is
held little above the surface of boiling water in a beaker. Then wash
with water and blot dry.
Air dry the film and fix with minimal flaming.
28. Many treatments are known which destroy the permeability barrier,
such as
Severe heat fixation (Bartholomew and 'Mittwer, 1950),
Acid hydrolysis (Robinow, 1951),
Ultraviolet light (Bartholomew and Mittwer, 1952),
Mechanical rupture (Fitz-James, 1953; Rode and Foster, 1960).
OTHER MODIFICATIONS OF
DORNER’S AND SCHAEFFER-
FULTON METHODS….
30. SUMMARY
Method Primary stain Decoloriser Counterstain Interpretation
Grams stain Crystal violet Acetone Saffranin Spore-colourless
Bacteria-violet
Modified Ziehl-
Neelsen Stain
Carbol fuchsin 0.25%-0.5%
H2SO4
Loeffler’s
methylene blue
Spore-red
Bacteria-blue
Dorner stain Carbol fuchsin Acid alcohol Nigrosin Spore-red
Bacteria-colorless
Variation in
dorner stain
Carbol fuchsin Nigrosin Spore-red
Bacteria-colorless
Schaeffer-Fulton
stain
Malachite green Water Safranine Spore-green
Bacteria-red
Bartholomew and
mittwer method
Malachite green Water Safranine Spore-green
Bacteria-red
Abbott's Method Methylene blue Acid alcohol Aniline-fuchsin Spore-blue
Bacteria-red
Moeller stain Carbol fuchsin Acidified
ethanol
Methylene blue Spore-red
Bacteria-blue
Modified moeller
stain
Kinyoun’s
carbol-fuchsin
2%H2SO4
80% ethanol
Loeffler’s
methylene blue
Spore-red
Bacteria-blue
31. REFERENCES
Colour Atlas & Textbook of Diagnostic Microbiology by Koneman
Practical Medical Microbiology by Mackie & McCartney
Monica Cheesbrough
Textbook of Microbiology by Ananthanarayan
http://www.microbelibrary.org/component/resource/laboratory-test/3112-
endospore-stain-protocol
http://chestofbooks.com/health/disease/Pathology/Methods-For-Staining-
Spores.html#
http://www.generalmicroscience.com/microbial-laboratory-
techniques/endospore-staining-by-bartholomew-and-mittwers-method/
M. Hayama, K. Oana, T. Kozakai, S. Umeda, J. Fujimoto, H. Ota, Y.
Kawakami Proposal of a simplified technique for staining bacterial spores
without applying heat-successful modification of Moeller’s method. Eur J
Med Res (2007) 12: 356-359.
Editor's Notes
Bacteria produce a single endospore internally. The spore is sometimes surrounded by a thin covering known as the exosporium, which overlies the spore coat. The spore coat, which acts like a sieve that excludes large toxic molecules like lysozyme, is resistant to many toxic molecules and may also contain enzymes that are involved in germination. The cortex lies beneath the spore coat and consists of peptidoglycan. The core wall lies beneath the cortex and surrounds the protoplast or core of the endospore. The core contains the spore chromosomal DNA which is encased in chromatin-like proteins known as SASPs (small acid-soluble spore proteins), that protect the spore DNA from UV radiation and heat. The core also contains normal cell structures, such as ribosomes and other enzymes, but is not metabolically active.
Up to 20% of the dry weight of the endospore consists of calcium dipicolinate within the core, which is thought to stabilize the DNA. Dipicolinic acid could be responsible for the heat resistance of the spore, and calcium may aid in resistance to heat and oxidizing agents. However, mutants resistant to heat but lacking dipicolinic acid have been isolated, suggesting other mechanisms contributing to heat resistance are also at work.[5]
Visualising endospores under the light microscope can be difficult due to the impermeability of the endospore wall to dyes and stains. While the rest of a bacterial cell may stain, the endospore is left colourless. To combat this, a special stain technique called a Moeller stain is used. That allows the endospore to show up as red, while the rest of the cell stains blue. Another staining technique for endospores is the Schaeffer-Fulton stain, which stains endospores green and bacterial bodies red. The arrangement of spore layers is as follows:
Exosporium
Spore coat
Spore cortex
Core wall
Because of their tough protein coats made of keratin, spores are highly resistant to normal staining procedures.
C. Perfringens also called as C.welchii. Other subterminals- C.sporogenes, c. botulinum, c.novyi type C. other terminals- C. innocuum, C. putrifiucm, C. melanominatum. C.bifermentans and C. sordelli don’t typically bulge. Central – B.cereus, B.megaterium
It is equivalent to combining Burdon’s stain for lipid with Ashby’s stain for spores.