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Diagnostic Microbiol

Diagnostic Microbiology

QuestionAnswer
Diagnostic Microbiology The study of microbes
Diagnostic Microbiology Too small to be seen by the naked eye
Diagnostic Microbiology Bacteria
Diagnostic Microbiology Viruses
Diagnostic Microbiology Fungi
Diagnostic Microbiology Most are nonpathogenic
Bacterial Cell Morphology Obligate aerobes
Bacterial Cell Morphology Obligate anaerobes
Bacterial Cell Morphology Facultative anaerobes
Bacterial Cell Morphology Microaerophilic
Bacterial Cell Morphology Capnophilic
Bacterial Cell Morphology Nutritional requirements vary
Bacterial Cell Morphology Temperature requirements vary
Bacterial Cell Morphology 4 groups according to shape
4 groups according to shape Coccus Bacillus Spiral Pleomorphic
FA- can survive in the absence of O2 but their growth is limited
Microaerophilic-prefer reduced 02 tensions
Capnophilic require high levels of CO2
Coccus—plueral is cocci spherical cells
Bacillus—shaped like rods or cylinders
occur singly and can be subdivided into loose spirals, tight spirals, and comma shaped spirals spiral
shape ranging from cocci to rods Pleomorphic
Variety of Arrangements Single Pairs Clusters/bunches Chains Palisades
Spores Resistant to heat, desiccation, chemicals, and radiation
Spores Classifications
Clusters— Look like a bunch of grapes
Chains short or long
Palisades —”chinese letter pattern”
Spores— intracellular refractive bodies
Classification of cells Central: present in the center of the cell Subterminal: present near the end of the cell Terminal: present at the end or pole of the cell
Bacterial Growth Single DNA strand
Bacterial Growth Binary fission
Bacterial Growth 4 phases of growth on plates
4 phases of growth on plates Lag Phase Exponential Growth Phase Stationary Phase Logarithmic Decline, AKA Death Phase
Classification of cells Central: present in the center of the cell Subterminal: present near the end of the cell Terminal: present at the end or pole of the cell
Bacterial Growth Single DNA strand
Bacterial Growth Binary fission
Classification of cells Central: present in the center of the cell Subterminal: present near the end of the cell Terminal: present at the end or pole of the cell
Bacterial Growth Single DNA strand
Lag Phase: bacteria adapt their metabolism to use the resources on their new media
Bacterial Growth Binary fission
Bacterial Growth 4 phases of growth on plates
4 phases of growth on plates Lag Phase Exponential Growth Phase Stationary Phase Logarithmic Decline, AKA Death Phase
spore formation occurs Death
Lag Phase: bacteria adapt their metabolism to use the resources on their new media
Separate room The Laboratory
EGP: Growth is referred to as doubling time or generation time, variable between species and environmental conditions, continues until essential nutrients are depleted, toxic waste products accumulate, and/or space becomes limiting
Minimal traffic The Laboratory
Total numbers of cells show no net increase or decrease, length of phase is variable amoung species Stationary
Good lighting/ventilation The Laboratory
spore formation occurs Death
Separate room The Laboratory
Easily cleaned surfaces The Laboratory
2 work areas The Laboratory
Access to incubator and fridge The Laboratory
Electrical outlets The Laboratory
Minimal traffic The Laboratory
Storage The Laboratory
Good lighting/ventilation The Laboratory
2 work areas, one for processing the samples and one for culture work
Easily cleaned surfaces The Laboratory
Tx as if zoonotic Laboratory Safety
2 work areas The Laboratory
Aseptic technique Laboratory Safety
Electrical outlets The Laboratory
PPE Laboratory Safety
Storage The Laboratory
Autoclave all tools Laboratory Safety
Access to incubator and fridge The Laboratory
No eating/drinking Laboratory Safety
2 work areas, one for processing the samples and one for culture work
Goggles Laboratory Safety
Most are potentially pathogenic Laboratory Safety
Hair tied back Laboratory Safety
Tx as if zoonotic Laboratory Safety
Any accidents report ASAP to vet Laboratory Safety
Aseptic technique Laboratory Safety
PPE clean, long sleeved, knee-length, white lab coats to prevent contamination of scrubs, disposable gloves, +/- face masks wash coats weekly with hot water and strong bleach, no not mix with other laundry
PPE Laboratory Safety
Gram Stain Categorizes bacteria as Gram+ or Gram-
Autoclave all tools Laboratory Safety
Gram Stain Purple=Gram+
No eating/drinking Laboratory Safety
Gram Stain Red=Gram-
Goggles Laboratory Safety
Potassium Hydroxide Test Gram-variable reaction
Hair tied back Laboratory Safety
Potassium Hydroxide Test Gram- become sticky, gram+ do not
Any accidents report ASAP to vet Laboratory Safety
Acid fast stain Mycobacterium and Nocardia spp
PPE clean, long sleeved, knee-length, white lab coats to prevent contamination of scrubs, disposable gloves, +/- face masks wash coats weekly with hot water and strong bleach, no not mix with other laundry
Gram Stain Categorizes bacteria as Gram+ or Gram-
Gram Stain Purple=Gram+
Gram Stain Red=Gram-
Potassium Hydroxide Test Gram-variable reaction
Potassium Hydroxide Test Gram- become sticky, gram+ do not
Acid fast stain Mycobacterium and Nocardia spp
Giemsa Stain Detects spirochetes and rickettsiae
Culture Media Any material solid or liquid that can support the growth of microorganisms
Types Transport media General purpose media Enriched media Selective media Differential media Enrichment media
Transport media: keep microbes alive while not encouraging growth and reproduction
General purpose: : AKA Nutrient media, not commonly used in vet med
Enriched meet the requirements of the most fastidious pathogens—added blood, serum, or egg
Selective: contain antibacterial substances such as bile salts or antimicrobials which inhibit or kill all but a few types of bacteria—MacConkey
allow bacteria to be differentiated into groups by biochemical rxns on the medium Differential
liquid media that favor growth of a particular group of organisms Enrichment
Blood Agar (Trypitcase Soy Agar) Supports growth
Blood Agar (Trypitcase Soy Agar) Sheep blood most common
Blood Agar (Trypitcase Soy Agar) Detects hemolysis
MacConkey Agar and Eosin-Methylene Blue Agar Selective and differential
MacConkey Agar and Eosin-Methylene Blue Agar Suppresses growth of gram+
MacConkey Agar and Eosin-Methylene Blue Agar Bile salts for lactose fermenting
MacConkey Agar and Eosin-Methylene Blue Agar Turn pink=lactose producing, clear is negative
Thioglycollate Broth Liquid medium
Thioglycollate Broth Determine oxygen tolerance
Urea Tubes Slant tube
Urea Tubes Pink is a positive=urea hydrolysis
Sulfide-Indole Motility Tubes Slant tube
Sulfide-Indole Motility Tubes Stab innoculation
grow in very bottom Anaerobic
facultative are throughout and aerobic are on the top
Brain-Heart Infusion Broth Used to increase number of organisms
Brain-Heart Infusion Broth Add blood to it
Brain-Heart Infusion Broth then plate it
Triple Sugar Iron Agar ID of Salmonella and differentiation of enteric bacteria
Triple Sugar Iron Agar Anaerobic bacteria turns it red to yellow
Salmonella-Shigella agar Pathogenic enteric gram neg bacteria
Mannitol Salt Agar Selective medium for Staph
Mannitol Salt Agar Positive turns medium yellow
Mueller-Hinton Gives standardized results during antibiotic sensitivity testing
Inoculation of Culture Media Sterile technique
Inoculation of Culture Media Streaking culture plates
Sterile technique flame everything, culture plates left closed until being use
Colony Characteristics Size Pigment Density Elevation Form Texture Odor Any hemolysis Figure 4-22 pg 136
Motility Microscopic exam
Catalase Testing Gram + cocci and small gram + bacilli
Catalase Testing Hydrogen peroxide
Catalase Testing + will make gas bubbles
Coagulase Testing Clotting factors
Coagulase Testing + will have a clot
Mycology Most are multicellular
Mycology Hyphae
Mycology Yeast is unicellular
Mycology Most make spores
Mycology Yeast=budding
Mycology Testing: DTM
Mycology Cultures: Sabouraud dextrose agar
-webs of slender tubing which grow towards food surfaces Hyphae
Virology Specialized laboratories
Created by: JODY84