PowerPoints 1-5
How are microbes classified in the clinical laboratory?
Phenotypical characteristics and Genotypic characteristics
What are phenotypical characteristics?
characteristics that can be observed and can be seen
What are some examples of phenotypical characteristics?
morphology, staining, environmental requirements, nutritional requirements, resistance profiles, antigenic properties
What are genotypical characteristics?
genetic information
What are some example of genotypical characteristics?
DNA base composition, DNA or RNA sequence
What is a species?
Most basic taxonomic group
What is a genus?
Different species with many common phenotypic and genotypic features
What is a subspecies?
share minor characteristics; called biotypes, subtypes, strains, or genotypes
What happens in the cytoplasm?
metabolic processes
What are plasmids?
small, circular extrachromosomal DNA found in bacterial chromosomes
What are the functions of the bacterial cell wall?
Provides stability and strength to the bacterial cell
Regulates transport of macromolecules in and out of the cell
What are exotoxins secreted by?
living bacterial cell
What are some examples of exotoxins?
Cholera toxin, Toxic Shock Syndrome, Diphtheria Toxin
How do microorganisms adjust to environmental changes?
Recombination
Mutations
Genetic exchange
What is an infection?
bacteria making you sick
What is a pathogen?
organism that has a capability of causing infection
What is normal flora
good bacteria that compete with pathogenic bacteria
What are some advantages of normal flora?
Attach to receptors on host
Compete for nutrients
Stimulate development of immune system
Produce antimicrobial substances
Synthesize vitamins
What is a true pathogen?
always causes infection
What is an opportunistic pathogen?
organism that does not typically cause infections but can in certain situations ie immunocompromised
What are some examples of virulence factors?
Adherence factors
Capsules
Endotoxins
Exotoxins
What is direct mode of transmission?
person to person
What are some examples of direct modes of transmission?
Congenital
Hand-to-Hand
Sexual
Respiratory droplets or secretions
What is indirect modes of transmission?
when bacteria is transferred from something like an object or animal
What are some examples of indirect transmission?
Fomites
Animals, insects, arthropods
Food
Water
Airborne
What is incidence?
Number of those affected in a population
What is prevalence?
Percentage of infected individuals in a population at a given time
What is an Outbreak?
Disproportionately larger number of infected individuals in short period of time
What is endemic?
Disease that consistently present but is limited to a certain area
What is an epidemic?
Sudden increase in disease in certain bacteria
What is a pandemic?
Outbreak of disease across several countries or continents
What is nosocomial?
Healthcare associated infection
Acquired in healthcare setting
What is community-acquired?
Present at time of admission
Acquired within community
What is colonization?
Presence and multiplication of microorganism in host, with no clinical signs of infection
What is morbidity?
Number cases of disease in population
What is mortality?
Number death from disease in population
What is surveillance?
Collection of data pertaining to disease occurrence
What is an index case?
First case
What is an exposure control plan?
describes risk of exposure to infectious agents and explains exposure reduction methods
What are the factors of exposure control
Safety education
Universal/Standard precautions
Engineering controls
Personal protective equipment
Postexposure procedures
Disposal of hazardous
What are some examples of safety education?
Orientation for new employees
Continuing education for current employees
Safety Manual
What is universal precautions?
A set of preventative measures designed to reduce risk of transferring HIV, Hep B, and other bloodborne pathogens
Applies to all blood and body fluids containing visible blood
What is standard precautions?
Applies to ALL patients
Designed to reduce risk of ALL infections in healthcare setting
ALL blood, tissue, body fluids, secretions, excretions (except sweat) are potentially infectious
What is BSL 1
No known pathogenic potential
Adhere to standard lab techniques
What is BSL 2
Follow Level 1 practices, plus lab coat, gloves, decontamination of infectious waste, biohazard warning signs
What is BSL 3?
Follow Level 2 practices, plus special lab clothing, controlled access
Air movement carefully controlled
What is BSL 4
Level 3 practice, plus entrance through separate room which street clothing is replaced with lab clothing
Maximum containment
Decontamination of all personnel and material before leaving area
What are Biological Safety Cabinets (BSC)?
Used for any procedures that create aerosols
What is BSC I?
Open fronted, negative pressure, ventilated cabinet
Unsterilized air
Have HEPA filter
What is BSC II?
Open-fronted, ventilated cabinet
Sterilizes air entering and circulating within cabinet and exhaust air
Have HEPA
What is BSC III?
Highest level of safety
What are examples of Personal Protective Equipment?
Disposable gloves
Laboratory gowns
Eye Protection
Face masks
What are some protocols with a postexposure plan?
All laboratory accidents/incidents must be reported
Appropriate medical treatment given immediately
Potential exposure to Hepatitis B or HIV should be followed clinically and serologically
Documentation required
Review incident and determine preventive measures
What is sterilization?
Destroys ALL pathogens (spores, vegetative cells, fungi, viruses
What is disinfectant?
Destroys or inactivates bacteria, viruses, and fungi (not spores)
What is antiseptic?
Chemical germicide used on skin; not to be substituted for disinfectant
List the organisms most resistant to least resistant to killing agents.
Prions
Bacterial spores
Mycobacteria
Nonlipid viruses
Fungi
Bacteria
Lipid viruses
What are the criteria for specimen rejection?
Received in Formalin
24 hour specimen collection
Sputum with >10 SEC/lpf
Smear from cervix, vagina, or anus for Neisseria gonorrhoeae
Improper, nonsterile container
Specimens for anaerobes that are contaminated with normal flora
What are requirements for GI Tract specimens?
Fresh stool
Specimen collected from endoscopy
What are the requirements for blood specimens?
Collect when patient is febrile
2-4 specimens collected from separate venipuncture sites at least one hour apart
Aerobic & anaerobic bottle
20 mL
What are requirements for Urine specimens?
Clean-catch midstream
Catheterized
Always quantitate (perform colony count)
Inoculate plate with 1 micrometer loop
# colonies x 1000 = CFU /mL
Requirements for Cerebrospinal Fluid
3rd tube for micro
No delay
What are the requirements for the genital tract specimens?
Wet mount for trichomonas vaginalis
Selective plates for N. gonorrhoeae
What are the requirements for wound and abscess specimens
Ideal —> collect with needle and syringe
What is a broth?
Media in liquid form
What is an agar?
Gel or semisolid media
What is a colony?
Bacterial population derive from single bacteria cell
What are isolated colonies?
not clustered in one big clump
What is general isolation media?
Supportive media
Supports growth on non-fastidious bacteria
What are examples of general isolation media?
Nutrient agar
Trypticase soy agar
Nutrient broth
What is non-selective media?
Also called enriched media
Contain nutrient supplement
Contain 5% defibrinated sheep red blood cells
What are some examples of non-selective media?
Sheep blood agar
Chocolate agar
What is sheep blood agar?
See presence of hemolysis by bacteria
What is chocolate agar?
Contain X and V factors for Haemophilus
What is differential media?
Allows grouping of microbes based on colonial appearance
What are some examples of differential media?
Sheep blood agar
MacConkey Agar
Eosin-methylene blue (EMB) agar
What is selective media?
Inhibits growth of all bacteria except those that are sought
What are examples of Selective media
Hektoen enteric (HE) agar
Xylose-lysine-deoxycholate (XLD) agar
Colistin-nalidixic acid (CNA) agar
Modified Thayer-Martin (MTM) agar
Salmonella-Shigella (SS) agar
What is enrichment broth?
Liquid media used to inhibit growth of one organism while enhancing that of another organism by providing nutrients
What are examples of enrichment broth
Gram negative broth
Thioglycolate broth
Lim Broth
What is prophylactic therapy?
treating patients who are at high risk for infection
What is empiric therapy?
Patient who has all the symptoms and signs of infection and antibiotic is given to most likely cause or most common cause of infection
What is definitive therapy?
the pathogen has been IDed and the physician knows that the patient is on the correct antimicrobial treatment
What is bacteriostatic?
Inhibits growth
Antimicrobial removed —> multiplication
Successful if patient has effective immune system
What is bactericidal?
Kills bacteria
Preferred in serious infections (endocarditis, meningitis, osteomyelitis)
Neutropenic patients
Are penicillins bacteriostatic or bactericidal?
bactericidal
Are Cephalosporins bacteriostatic or bactericidal?
bactericidal
Are carbapenems bacteriostatic or bactericidal?
bactericidal
Are monobactams bacteriostatic or bactericidal?
bactericidal
Is vancomycin bacteriostatic or bactericidal?
bactericidal (slowly)
Are Fluoroquinolones bacteriostatic or bactericidal?
Bactericidal
Are aminoglycosides bactericidal or bacteriostatic?
bactericidal
Is metronidazole bacteriostatic or bactericidal?
bactericidal
Is daptomycin bacteriostatic or bactericidal?
bactericidal
Are macrolides bacteriostatic or bactericidal?
bacteriostatic
Are tetracyclines bacteriostatic or bactericidal?
bacteriostatic
What is the mode of activity for B-lactam antibiotics?
Inhibits cell wall synthesis by inhibition of enzymes needed for peptidoglycan formation
What is nitrocefin?
Agent used to test for beta lactamase production
What are examples of B-lactam antibiotics?
Penicillins
Monobactams
Carbapenems
Cephalosporins
What are example(s) of Penicillins?
Penicillin
Ampicillin
Methicillin
Piperacillin