This document discusses Gram-negative rods of the family Enterobacteriaceae. It provides characteristics of the family including being facultative anaerobes that are Gram-negative rods and test positive for catalase and nitrate reduction. It also describes identification methods for Enterobacteriaceae including growth on selective media like MacConkey agar and biochemical tests. Key members are discussed and divided into lactose fermenters versus non-fermenters.
Most medically important family of non–spore-forming gram-negative rods.
Most species are normal flora of the GI tract. Salmonella, Shigella, and Yersinia are not normal GI flora.
Major cause of nosocomial infections
Diseases include UTIs, gastroenteritis, septicemia, food poisoning, wound infections, peritonitis, pneumonia, and meningitis
The family exhibits four serological characteristics:
O (somatic) antigen-A cell wall antigen-LPS (heat stable), Used for serological grouping of Salmonella & Shigella.
K (envelope) antigen-Capsular antigen (heat labile)
H (flagellar) antigen-Flagellar antigen-protein (heat labile), Used to serotype Salmonella.
Vi antigen-Capsular antigen of Salmonella Typhi-polysaccharide (heat labile), Role in preventing phagocytosis, may mask O Ag, removed by heating.
Enterobacteriaceae are facultative anaerobes, ferment glucose. Positive nitrate and catalase, non-hemolytic. Except for Plesiomonas, they are oxidase negative.
Most medically important family of non–spore-forming gram-negative rods.
Most species are normal flora of the GI tract. Salmonella, Shigella, and Yersinia are not normal GI flora.
Major cause of nosocomial infections
Diseases include UTIs, gastroenteritis, septicemia, food poisoning, wound infections, peritonitis, pneumonia, and meningitis
The family exhibits four serological characteristics:
O (somatic) antigen-A cell wall antigen-LPS (heat stable), Used for serological grouping of Salmonella & Shigella.
K (envelope) antigen-Capsular antigen (heat labile)
H (flagellar) antigen-Flagellar antigen-protein (heat labile), Used to serotype Salmonella.
Vi antigen-Capsular antigen of Salmonella Typhi-polysaccharide (heat labile), Role in preventing phagocytosis, may mask O Ag, removed by heating.
Enterobacteriaceae are facultative anaerobes, ferment glucose. Positive nitrate and catalase, non-hemolytic. Except for Plesiomonas, they are oxidase negative.
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks..
Qalification
AHLAD T O
MSc MLT (Biochemistry)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
#Proteus microbiology
#Medical
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#Mallu Medicos Lounge
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#PROTEUS - causes, symptoms, diagnosis, treatment, pathology
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.
The lecture was presented to the students of Saudi board of Community Medicine to help them know about the various serological methods applicable in the diagnosis of infectious diseases in general with attention upon the specificity and sensitivity of various diagnostic modalities. The lecture covers the basic principles of each test and the clinical applications with the advantages and disadvantages of each.
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks..
Qalification
AHLAD T O
MSc MLT (Biochemistry)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
#Proteus microbiology
#Medical
#Microbiology
#Biochemistry
#Mallu Medicos Lounge
##MalluMedicosLounge
#MLT
#Channel introduction
#HealthAndVoyage
#New Youtube Channel introduction
#Gram-negative
#Enterobactericea
#Weil Felix Test
#PROTEUS - causes, symptoms, diagnosis, treatment, pathology
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.
The lecture was presented to the students of Saudi board of Community Medicine to help them know about the various serological methods applicable in the diagnosis of infectious diseases in general with attention upon the specificity and sensitivity of various diagnostic modalities. The lecture covers the basic principles of each test and the clinical applications with the advantages and disadvantages of each.
A culture media is a special medium used in microbiological laboratories to grow different kinds of microorganisms. A growth or a culture medium is composed of different nutrients that are essential for microbial growth.
Since there are many types of microorganisms, each having unique properties and requiring specific nutrients for growth, there are many types based on what nutrients they contain and what function they play in the growth of microorganisms.
A culture may be solid or liquid. The solid culture media is composed of a brown jelly like substance known as agar. Different nutrients and chemicals are added to it to allow the growth of different microorganisms.
Classification of enterbacteriaceae and biochemical reactionsDAKhan4
slide 2
Commonly present in large intestine •
Non sporing ,
Non Acid fast,
Gram – bacilli. •
A complex family of organisms, •
Some are non pathogenic •
A few are highly Pathogenic, •
Some commensals turn out to be pathogenic as in UTI after catheterization.
slide 3
Characters of Enterobacteriaceae •
All Enterobacteriaceae
a) – Gram-negative rods
b) – Ferment glucose with acid production
c) – Reduce nitrates into nitrites
d) – Oxidase negative
Facultative anaerobic
Motile except Shigella and Klebsiella
Non-capsulated except Klebsiella
Non-fastidious
Grow on bile containing media (MacConkey agar
slide7
Conserved signature inserts and deletions (CSIs) in protein sequences provide an important category of molecular markers for understanding phylogenetic relationships. ... The CSIs that are restricted to a particular clade or group of species, generally provide good phylogenetic markers of common evolutionary descent
slide 8
The family Enterobacteriaceae harbors many important pathogens, however it has proven difficult to reliably distinguish different members of this family or discern their interrelationships.
To understand the interrelationships among the Enterobacteriaceae species, we have constructed two comprehensive phylogenetic trees for 78 genome-sequenced Enterobacteriaceae species based on 2487 core genome proteins, and another set of 118 conserved proteins.
The genome sequences of Enterobacteriaceae species were also analyzed for genetic relatedness based on average amino acid identity and 16S rRNA sequence similarity.
In parallel, comparative genomic studies on protein sequences from the Enterobacteriaceae have identified 88 molecular markers in the form of conserved signature indels (CSIs) that are uniquely shared by specific members of the family.
slide 10 identification of enterobacteriaceae
biochemical reaction
Oxidase test
All members of Enterobacteriaceae are oxidase negative
Pseudomonas is oxidase positive
O/F test
All members of Enterobacteriaceae are O+/F+
Pseudomonas is O+/F-
Nitrate reductase
All members of Enterobacteriaceae are nitrate reductase positive
Pseudomonas is nitrate reductase negative
slide 11
identification of enterobacteriaceae
Differentiation between LF & NLF by Growth
Method:
on MacConkey agar
MacConkey agar is inoculated with tested organism using streak plate technique
Incubate the plate in incubator at 37 C/24 hrs
Results:
LF organism appears as pink colonies (e.g. E. coli)
NLF organism appears as colorless colonies (e.g. Shigella)
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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
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
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.
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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.
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.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
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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
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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
4. Characters ofCharacters of EnterobacteriaceaeEnterobacteriaceae
AllAll EnterobacteriaciaeEnterobacteriaciae
– Gram-negative rodsGram-negative rods
– Ferment glucose with acid productionFerment glucose with acid production
– Reduce nitrates into nitritesReduce nitrates into nitrites
– Oxidase negativeOxidase negative
– Catalase positiveCatalase positive
Facultative anaerobicFacultative anaerobic
All motileAll motile exceptexcept Shigella, KlebsielShigella, Klebsiel andand YersiniaYersinia
Non-capsulated exceptNon-capsulated except KlebsiellaKlebsiella
Non-fastidiousNon-fastidious
Grow on bile containing media (MacConkey agar(Grow on bile containing media (MacConkey agar(
5. Microscopic and ColonyMicroscopic and Colony
MorphologyMorphology
Gram negative bacilli or coccobacilliGram negative bacilli or coccobacilli
Non-spore formingNon-spore forming
Colony morphology on BA or MH of littleColony morphology on BA or MH of little
value, as they look the same, except forvalue, as they look the same, except for
KlebsiellaKlebsiella
Selective and differential media are usedSelective and differential media are used
for initial colony evaluation (ex.for initial colony evaluation (ex.
MacConkey, Hektoen Enteric (HE),MacConkey, Hektoen Enteric (HE),
Xylose lysine deoxycholate (Xylose lysine deoxycholate (XLDXLD(( agarsagars((
6. EnterobacteriaceaeEnterobacteriaceae
SomeSome EnterobacteriaceaeEnterobacteriaceae are true pathogensare true pathogens
– SalmonellaSalmonella spp.spp.
– ShigellaShigella spp.spp.
– YersiniaYersinia spp.spp.
– Certain strains ofCertain strains of E. coliE. coli (ETEC, EPEC, EIEC, EHEC((ETEC, EPEC, EIEC, EHEC(
Most members of theMost members of the EnterobacteriaceaeEnterobacteriaceae areare
opportunistic or cause secondary infections ofopportunistic or cause secondary infections of
wounds, the urinary and respiratory tracts, andwounds, the urinary and respiratory tracts, and
the circulatory system e.g.the circulatory system e.g. E. coliE. coli..
Enterobacteriaceae divided into TWO mainEnterobacteriaceae divided into TWO main
groups according to action on LACTOSEgroups according to action on LACTOSE
– Lactose Fermenters (LF(Lactose Fermenters (LF(
E. coli, Citrobacter, Klbesiella, EnterobacterE. coli, Citrobacter, Klbesiella, Enterobacter
– Lactose Non-Fermenters (LNF(Lactose Non-Fermenters (LNF(
Salmonella, Shigella, Proteus, YersiniaSalmonella, Shigella, Proteus, Yersinia
7. Identification ofIdentification of EnterobacteriaceaeEnterobacteriaceae
Gram stainGram stain
– AllAll EnterobacteriaceaeEnterobacteriaceae are Gram-negative rodsare Gram-negative rods
– Arranged in singleArranged in single
8. Identification ofIdentification of EnterobacteriaceaeEnterobacteriaceae
Based on Biochemical reactionsBased on Biochemical reactions
Can be grouped into threeCan be grouped into three:-:-
A) Ability to ferment carbohydrates to acid orA) Ability to ferment carbohydrates to acid or
Acid and gasAcid and gas..
B) Ability to utilize a substrate as the soleB) Ability to utilize a substrate as the sole
source of carbonsource of carbon
C) Ability to enzymaticaly degrade aC) Ability to enzymaticaly degrade a
compoundcompound
9. Identification ofIdentification of EnterobacteriaceaeEnterobacteriaceae
Biochemical reactionsBiochemical reactions
Oxidase testOxidase test
– All members ofAll members of EnterobacteriaceaeEnterobacteriaceae are oxidase negativeare oxidase negative
– PseudomonasPseudomonas is oxidase positiveis oxidase positive
Oxidative/Fermentation testOxidative/Fermentation test
– All members ofAll members of EnterobacteriaceaeEnterobacteriaceae are O+/F+are O+/F+
– PseudomonasPseudomonas is Oxidative+ / Fermentation -is Oxidative+ / Fermentation -
Nitrate reductaseNitrate reductase
– All members of Enterobacteriaceae are nitrate reductase positiveAll members of Enterobacteriaceae are nitrate reductase positive
– PseudomonasPseudomonas is nitrate reductase negativeis nitrate reductase negative
See & compare these tests forSee & compare these tests for Pseudomonas sppPseudomonas spp in the Labin the Lab
10. Classification ofClassification of EnterobacteriaceaeEnterobacteriaceae
Enterobacteriaceae
Lactose fermenters
E. coli, Citrobacter,
Klebsiella, Enterobacter
Non-lactose fermenter
Salmonell, Shigella
Proteus, Yersinia
There are several selective and differential media used to
isolate distinguishes between LF & LNF
The most important media are:
MacConkey agar
Eosin Methylene Blue (EMB) agar
Salmonella Shigella (SS) agar
In addition to Triple Sugar Iron (TSI) agar
11. Differentiation between LF & NLF byDifferentiation between LF & NLF by Growth on MacConkey agarGrowth on MacConkey agar
MacConkey Agar
Contains
Bile salts Crystal violet Lactose Neutral red
MacConkey agar is selective & differential medium forMacConkey agar is selective & differential medium for EnterobacteriaceaeEnterobacteriaceae
Inhibit growth of G+ve bacteria
Cause of selectivity
Cause of differential
pH indicator
Acidic: Pink
Lactose feremnters
Pink colonies
Lactose non feremnters
colorless colonies
12. Classification ofClassification of EnterobacteriaceaeEnterobacteriaceae according toaccording to
lactose fermentation (growth on MacConkey Agar)lactose fermentation (growth on MacConkey Agar)
Enterobacteriaceae
Lactose Fermenters Lactose Non-Fermenters
Escherichia coli
Klebsiella spp
Enterobacter spp
Citrobacter spp
Salmonella spp
Schigella spp
Proteus spp
Yersinina spp
Pink colonies
Colorless colonies
Acid
Neutral red
No acid
13. Identification ofIdentification of EnterobacteriaceaeEnterobacteriaceae
Differentiation between LF & NLF byDifferentiation between LF & NLF by Growth on MacConkey agarGrowth on MacConkey agar
Method:Method:
– MacConkey agar is inoculated with tested organismMacConkey agar is inoculated with tested organism
using streak plate techniqueusing streak plate technique
– Incubate the plate in incubator at 37 C/24 hrsIncubate the plate in incubator at 37 C/24 hrs
Results:Results:
– LF organism appears as pink colonies (e.g.LF organism appears as pink colonies (e.g. E. coliE. coli))
– NLF organism appears as colorless colonies (e.g.NLF organism appears as colorless colonies (e.g.
ShigellaShigella))
Flame & Cool
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4
5
14. Growth of Enterobacteriaceae on
MacConkey agar
Uninoculated plate Lactose non feremters
Salmonella, Shigella,
Proteus
Lactose feremters
E. coli, Citrobacter
Klebsiella, Enterobacter
Colorless colonies Pink colonies
15. Reaction on Salmonella Shigella (SS) agarReaction on Salmonella Shigella (SS) agar
SS agar is a selective & differential medium used for isolation ofSS agar is a selective & differential medium used for isolation of
SalmonellaSalmonella andand ShigellaShigella
The selective agents areThe selective agents are bile saltsbile salts, and, and brilliant green dyebrilliant green dye, which, which
inhibit gram-positive organismsinhibit gram-positive organisms
The medium contains onlyThe medium contains only lactoselactose as a differential agent and thusas a differential agent and thus
differentiates on the basis of lactose fermentationdifferentiates on the basis of lactose fermentation
The formation of acid on fermentation of lactose causes the neutralThe formation of acid on fermentation of lactose causes the neutral
red indicator to make pink coloniesred indicator to make pink colonies
Non lactose fermenting organisms are colorless on the mediumNon lactose fermenting organisms are colorless on the medium
SS agar containsSS agar contains sodium thiosulfatesodium thiosulfate andand ferric ammoniumferric ammonium
citratecitrate allows the differentiation of organisms that produce H2Sallows the differentiation of organisms that produce H2S
– Lactose fermenters, such asLactose fermenters, such as E. coliE. coli, have colonies which are, have colonies which are
pinkpink
– ShigellaShigella appears transparent or amberappears transparent or amber
– SalmonellaSalmonella appears transparent or amber with black centersappears transparent or amber with black centers
due to H2S productiondue to H2S production
Lactose
Lactose fermenter
Acid
Neutral red
Pink colonies
Ferrous sulfide
Black precipitate
H2S + Ferric ammonium citrate
16. Identification ofIdentification of EnterobacteriaceaeEnterobacteriaceae
Differentiation between LF & NLF byDifferentiation between LF & NLF by Growth on SS agarGrowth on SS agar
Method:Method:
– SS agar is inoculated with tested organismSS agar is inoculated with tested organism usingusing
streak plate techniquestreak plate technique
– Incubate the plate in incubator at 37 C/24 hrsIncubate the plate in incubator at 37 C/24 hrs
Flame & Cool
Flame & Cool
Flame & Cool
1 2
3
4
5
17. AA.. Klebsiella pneumoniaeKlebsiella pneumoniae
BB.. Escherichia coliEscherichia coli
CC:: Salmonella spSalmonella sp..
DD:: Proteus mirabilisProteus mirabilis
EE:: Ps. aeruginosaPs. aeruginosa
..
BothBoth are lactose fermentersare lactose fermenters
BothBoth Salmonella sp. & ProteusSalmonella sp. & Proteus product H2Sproduct H2S
PseudomonasPseudomonas colonies are nearly colorlesscolonies are nearly colorless
Growth of Enterobacteriaceae on SS agar
18. Growth ofGrowth of EnterobacteriaceaeEnterobacteriaceae onon
EMB agarEMB agar
Coli-type colonies are very dark,
almost black e.g. E. coli
19. Reaction on Triple Sugar Iron (TSI) AgarReaction on Triple Sugar Iron (TSI) Agar
TSI containsTSI contains
– Three different types of sugarsThree different types of sugars
Glucose (0.1%)Glucose (0.1%)
Lactose (1%)Lactose (1%)
Sucrose (1%)Sucrose (1%)
– Phenol red (acidic: Yellow)Phenol red (acidic: Yellow)
TSI dispensed in tubes with equal butt & slantTSI dispensed in tubes with equal butt & slant
PrinciplePrinciple
– To determine the ability of an organism to attack a specificTo determine the ability of an organism to attack a specific
carbohydrate incorporated into a basal growth medium, with orcarbohydrate incorporated into a basal growth medium, with or
without the production of gas, along with the determination ofwithout the production of gas, along with the determination of
possible hydrogen sulphide production.possible hydrogen sulphide production.
20. Reaction on TSIReaction on TSI
Method:Method:
– Inoculate TSI medium with an organism byInoculate TSI medium with an organism by
inoculating needle by stabbing the butt andinoculating needle by stabbing the butt and
streaking the slantstreaking the slant
– Incubate at 37°C for 24 hoursIncubate at 37°C for 24 hours
21. Reaction on TSI
Result
Example
Butt color Slant color H2
S
Yellow Red
Negative A/Alk/-
(Glucose fermented)
LNF
e.g. Shigella
Yellow Red
Positive
black in butt
A/Alk/+
(Glucose fermented with H2
S)
LNF e.g. Salmonella &
Proteus
Yellow Yellow
Negative
A/A/-
(three sugars are fermented)
LF e.g. E. coli,
Klebsiella,Enterobacter
Red Red
Negative
Alk/Alk/-
(No action on sugars)
Non fermenter e.g.
Pseudomonas
Result
s
22. Practicle WorkPracticle Work
Gram stainGram stain
Oxidase testOxidase test
Indole testIndole test
O/F test *O/F test *
Nitrate reductase test*Nitrate reductase test*
Growth on MacConkey’s agarGrowth on MacConkey’s agar
Growth on EMB agar*Growth on EMB agar*
Growth on SS agarGrowth on SS agar
Reaction on TSIReaction on TSI