The document discusses bacteriology of water and air. It provides details on:
1. Indicator organisms used to detect fecal contamination in water, including E. coli, enterococci, and Clostridium perfringens.
2. Methods for analyzing water samples, including multiple tube fermentation and membrane filtration.
3. Parameters for evaluating air quality in operating theaters, including particle counts, air changes per hour, and temperature/humidity.
4. Surface surveillance involves sampling high-touch sites to identify potential pathogens and inform outbreak investigations.
Antimicrobial sensitivity testing (AST) or Antibiotic Sensitivity Testing.
Contents:
1. Need of AST
2. Bacterial Resistance
3. Preperation of test: selection of antibiotic and bacteria
4. Types of tests
5. Process of tests
Microbiology of E coli giving basic of Escherichia coli, its morphology, cultural and biochemical characteristics, Antigenic character, pathogenesis, laboratory diagnosis, prevention and control
Antimicrobial sensitivity testing (AST) or Antibiotic Sensitivity Testing.
Contents:
1. Need of AST
2. Bacterial Resistance
3. Preperation of test: selection of antibiotic and bacteria
4. Types of tests
5. Process of tests
Microbiology of E coli giving basic of Escherichia coli, its morphology, cultural and biochemical characteristics, Antigenic character, pathogenesis, laboratory diagnosis, prevention and control
Methods to detect potability of water samplevimala rodhe
Water is precious and it is the base for living, Several disease causing pathogens are transmitted through water. There are various methods to detect the presence of pathogens in drinking water samples.Some of the methods to detect microbiological quality of water are discussed.
Water quality describes the condition of the water, including chemical, physical, and biological characteristics, usually concerning its suitability for a particular purpose such as drinking.
Most probable number or multiple tube fermentation techniqueSamsuDeen12
multiple tube fermentation or most probable number is a microbiological technique used to check the portability of water. microbial analysis of water is determined, and distinguished between faecal and non faecal contaminated water.
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
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
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This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
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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
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- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
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4. WATER SURVEILLANCE
Need for water surveillance
• To know water is fit for consumption
• To rule out water borne infections
5. Water borne pathogens
Bacterial Vibrio cholerae, Salmonella Typhi, S. Paratyphi A, B and C,
Shigella species, Yersinia enterocolitica, Campylobacter
jejuni ,Diarrheagenic Escherichia coli
Viral Hepatitis A virus ,Hepatitis E virus ,Polio virus, ,Rota virus
Protozoal Entamoeba histolytica, Giardia lamblia, Balantidium coli,
Cryptosporidium, Isospora
Helminthic Ascaris lumbricoides, Enterobius vermicularis
Trichuris trichiura
Helminths
transmitted through
aquatic hosts
Dracunculus medinensis Diphyllobothrium latum
Schistosomes
6. When to do??
• Outbreaks
• Dialysis units
• Routinely: optional
7. Criteria for indicator organisms:
• Present in feces in abundant number
• Should be a member of the intestinal micro-
flora of humans
• Longer survival time
• Should have some direct relationship to the
degree of faecal pollution
8. Indicator organisms
• Fecal (thermotolerant) Escherichia coli
– Confirms recent fecal contamination of water
– Most sensitive indicator
• Other indicators:
– Coliform other than fecal E.coli
– Fecal Streptococci
– Clostridium perfringens
– Pseudomonas aeruginosa
– Bacteriophages
9. Indicator organisms of fecal pollution
of water
Indicator organisms Interpretation-
(Presence in water indicates-)
Coliform (other than
E.coli)
Remote contamination- either fecal
(presumptive) or soil and vegetation
Fecal (thermotolerant)
Escherichia coli
Confirms recent fecal contamination of
water
Most sensitive indicator
Fecal streptococci Confirms remote fecal pollution
Clostridium perfringens Remote contamination
Pseudomonas aeruginosa Least reliable indicator
Useful in hospitals and food
establishments
Bacteriophages Phage specific for E.coli- indicate fecal
pollution of water Indirectly indicates
viral pollution
10. Methods of analysis
• Standard tests usually employed for
bacteriological analysis of water are-
Presumptive coliform count ( Multiple tube
method)
Differential coliform count (Eijkman test)
Membrane Filtration method
11. Multiple tube method
• Used for the estimation of presumptive coliform count
expressed as the most probable number (MPN) of
coliform organisms in 100 ml water.
• Medium- MacConkey purple broth (double strength and
single strength) in bottles or tubes is the standard
medium of choice.
• Durham's tube is used to detect gas production
• Bromocresol purple is used as indicator
12. Multiple tube method (cont..)
• Procedure - Measured amounts of water samples are added
to tubes containing MacConkey purple broth by sterile
graduated pipettes as under:
o 50 ml of water to one bottle of 50 ml double strength medium
o 10 ml of water each to five tubes of 10 ml double strength
medium
o 1 ml of water each to five tubes of 5 ml single strength medium
• Result:
o Inoculated tubes are incubated at 37° C for 48 hours
o Positive test is indicated by a color change in the medium to
yellow from purple (due to lactose fermentation) and gas
collected in the Durham’s tube.
13. • Interpretation- The interpretation of presumptive coliform
count is as follows:
o Presumptive Coliform count (Most probable number)- An
estimate of coliform count per 100 ml is calculated from
the tubes showing acid and gas production using the
McCradey’s probability table.
Multiple tube method (cont..)
14. • Quality of water supply is determined by the
presumptive coliform count. The of most probable
numbers of 0, 1-3, 4-9 and ≥10 per 100 ml are
interpreted as:
o Excellent
o Satisfactory
o Intermediate
o Unsatisfactory respectively.
• Detection of coliform bacteria does not always indicate
fecal contamination as some of them may be found in
environment.
• Hence, it is further tested by differential coliform count
to detect the fecal E.coli.
Multiple tube method (cont..)
15. Presumptive coliform count
Most probable number (MPN)/100mL
McCrady’s table
1 tube of
50 ml
5 tube of
10 ml each
5 tube of
1 ml each
MPN/
100ml
0 0 0 <1
0 0 1 1
0 0 2 2
0 1 0 1
0 1 1 2
0 1 2 3
0 2 0 2
0 2 1 3
0 2 2 4
0 3 0 3
0 3 1 5
0 4 0 5
16. Presumptive coliform count
Most probable number (MPN)/100mL
1 tube of
50 ml
5 tube of
10 ml each
5 tube of
1 ml each
MPN/
100ml
1 0 0 1
1 0 1 3
1 2 2 10
1 0 3 6
1 1 0 3
1 1 1 5
1 1 2 7
1 1 3 9
1 2 0 5
1 2 1 7
1 2 2 10
1 2 3 12
1 3 0 8
1 3 1 11
1 3 2 14
1 3 3 18
1 3 4 21
17. Presumptive coliform count
Most probable number (MPN)/100mL
1 tube of
50 ml
5 tube of
10 ml each
5 tube of
1 ml each
MPN/
100ml
1 4 0 13
1 4 1 17
1 4 2 22
1 4 3 28
1 4 4 35
1 4 5 43
1 5 0 24
1 5 1 35
1 5 2 54
1 5 3 92
1 5 4 161
1 5 5 >180
18. Classification of quality of drinking water supply according
to bacteriological tests
Quality of
drinking water
supply
Most probable number
(MPN)/100 ml of water
Coliform
counts / 100
ml
E.coli count /
100 ml
1. Excellent 0 0
2. Satisfactory 1 – 3 0
3. Intermediate 4 – 9 0
4.
Unsatisfactory
≥10 ≥1
19. Differential coliform count
(Eijkman test)
• Done to confirm that the coliform bacilli detected
in the presumptive test are fecal E.coli.
• This is done by:
o Sub culturing the positive tubes on lactose containing
medium such as brilliant green bile broth for detection
of lactose fermentation with production of acid and
gas at 440C and
o Demonstrating positive indole test at 440C
20. Fecal streptococci detection
• When presumptive coliforms are present but
fecal E.coli are absent, detection of fecal
streptococci would confirm the fecal origin of
coliform bacilli.
• Subcultures are made from positive tubes - tubes
containing 5 ml of glucose azide broth and
incubated at 45°C for 48 hours.
• Presence of acid - indicates fecal streptococci
• Further confirmation can be done by plating onto
bile esculin azide agar (black colonies formed).
21. Clostridium perfringens detection
• By first heating the water sample (which kills all vegetative
bacteria retaining the spores)
• Then performing multiple tube method by sub culturing it
in differential reinforced clostridial medium (DRCM).
22. Enzyme methods
• Detection of specific enzymes for the detection and
confirmation of coliform bacilli and fecal E.coli is rapid and
novel method described recently.
o β galactosidase – is a coliform bacilli specific enzyme
o β glucuronidase- is a fecal E.coli specific enzyme
23. Membrane filtration method
• Measured volume of the water sample is filtered through a
membrane of pore size small enough to retain the indicator
bacteria to be counted on its surface.
• Membrane is then placed on a suitable selective indicator
medium and incubated
• Indicator bacteria grow into colonies on its upper surface
24. CDC recommends targeted air
surveillance
• Investigation of an outbreak
• For research purpose
• After reconstruction or newly constructed buildings
• After fumigation (fumigation itself is not routinely
recommended)
• For short term evaluation of a change in infection-control
practice.
25. Evaluation of the quality of air in OT
Microbiological parameters
1. Microbiological sampling method
2. Particle count method
26. Particle count method
• Airborne particle concentrations in OT is measured by means of
a laser light scattering instrument (particle counter)
• The particle count of an OT is considered acceptable only when
it falls in the acceptable range according to the international
standards system ISO 14644-1.
27. Settle plate Method
• Petri dishes containing an agar medium of
known surface area are left open for 30
minutes to 1 hour.
• Plates are incubated at 37°C for 24 hours
• Colony count: Large bacteria carrying
dust particles settle onto the medium.
28. Slit Sampler Method
• Most efficient and convenient method
• Number of bacteria carrying particles
suspended in a unit volume of air.
• Air is sucked through the equipment at a
rate of one cubic foot (28.3 liter) per
minute for 10 minutes
• Directed onto a plate containing culture
medium through the slit
• The plate is rotated mechanically so as
to allow the organisms to spread out
evenly on the medium
29. Non-microbiological parameters
Properties Recommendations
Air changes per
hour
Minimum 20 numbers of air changes per hour;
out of which four should be fresh air.
Air velocity 25-35 FPM (feet per minute)
It is checked by anemometer.
Positive pressure
(PP)
Difference in positive pressure between OT
and adjoining areas is required to prevent
outside air entry into OT.
PP should be maintained in OT at all times
(operational & non-operational hours)
Minimum PP of 2.5 Pascal is recommended.
Temperature 21 +/- 30C
(ortho 180C +/- 20C)
Relative humidity 20 to 60% (ideal 55%)
30. Non-microbiological parameters
Properties Recommendations
Air handling in the OT There should be dedicated AHU (air handling unit) for each
OT and should not be linked to air conditioning of any other
area/OT.
Window & split AC should never be used in OT as they are
pure re circulating units and have pockets for microbial
growth.
Air is supplied through Terminal HEPA filters in the ceiling
The minimum size of the filtration area should extend one
feet on each side of the OT table to cover the entire OT
table and surgical team.
Validation of HEPA Filters is done biannually by DOP
(Dispersed Oil Particulate) test.
Wall, floor and ceiling Should be anti-static and made up of non-porous, smooth,
seamless materials
Paints used should have antibacterial, anti-fungal
31. SURFACE SURVEILLANCE
• Environmental surface sampling has been used to determine
a) potential environmental reservoirs of pathogens
b) survival of microorganisms on surfaces
c) the sources of the environmental contamination.
32. SURFACE SURVEILLANCE
• Locations: It is required for high risk locations such as
operation theatres and ICU settings.
• Sites for sampling (high touch areas)
• Indications (CDC recommendation)- Indicated for research,
as a part of an epidemiologic investigation, or during an
outbreak investigation.
o Routine periodic surface surveillance is not recommended.
• Method: Moistened sterile swabs (soaked in sterile saline)
are used to take the samples from high risk areas as
mentioned above and then inoculated on to non-selective,
nutrient-rich agar (e.g. blood agar) for the recovery of
aerobic bacteria.
33. SURFACE SURVEILLANCE
• Reporting: Only pathogenic organisms isolated are reported.
• Newer technique such as luminometer (expresses bacterial
contamination as CFU/ml) and glow gel techniques are
available which are easy to perform though expensive.