Lab Practical Part 1 Flashcards
Bacterial species have characteristic patterns of colony growth, but can this be used as a diagnostic tool to identify a colony on a plate or broth?
- Morphology gives clues, but can’t identify the species . Different species night have similarities
- Medium, temp, environment also affect growth patterns
Pinpoint, small, moderate or large
Size
Color of colony
Pigmentation
Shape of colony
Form
Unbroken, peripheral edge
Circular
Indented peripheral edge
Irregular
Cultural characteristics
differences in macroscopic appearance of growth
What is used to separate microorganisms into taxonomic groups?
Cultural characteristics
Rootlike, spreading growth
Rhizoid
The appearance of the outer edge of the colony is called
Margin
Sharply defined, even
Entire
Marked indentations
Lobate
Wavy indentations
Undulate
Toothlike appearance
Serrate
Threadlike, spreading edge
Filamentous
Degree to which the colony growth is raised on the agar surface
Elevation
Elevation not discernable
Flat
Slightly elevated
Raised
Dome shaped elevation
Convex
Raised, with elevated convex central region
Umbonate
A culture containing a single unadulterated species of cells is called a
Pure culture
Cluster of cells that originates from the multiplication of a single cell and represents the growth of a single species of microorganism
Colony
A solution containing these nutrients is a
Culture medium
All culture media are
- Broth culture
- Semisolid medium
Broth medium
Lacks a solidifying agent
Semisolid medium
A concentration of less than 1% agar
A solid can exist as 3 forms as agar
- Agar slant
- Agar deep
- Agar plate
Sterility is the hallmark of successful work in the microbiology laboratory and _______ is the process of rendering a medium or material free of all forms of life
Sterilization
Using correct aseptic technique minimizes the likelihood that
Bacterial cultures will be contaminated
Using correct aseptic technique minimizes the likelihood that bacterial cultures will be contaminated and reduces the opportunity that
you will be exposed to potential pathogens.
Heat = __________or __________such as the autoclave used in class which is adding steam with pressure over time.
Dry (hot air)
Moist (wet heat)
Types of sterilization techniques
- Heat
- Filtration
- Chemicals
- Radiation
normal flora that does not cause negative effects in healthy individuals but instead symbiotic relationships with your skin.
Resident flora
can be removed with good handwashing techniques, even adding surgical scrubbing.
Transient flora
Why are resident flora more difficult to remove?
They are more difficult to remove because they are accustomed to living inside or on the human body. They have evolved and adapted to withstand the conditions of the human body. The immune system might not react to them because it recognizes them as part of the body. They are also covered by hair follicles, oil, and dead skin cells which obstruct their removal.
Clinical Application: What are nosocomial infections?
Nosocomial infections are infections that someone contracts while in a medical facility. It is important to clean surfaces thoroughly and use appropriate protective gear while working in a medical environment so nosocomial infections can be prevented.
How is handwashing affected by water only, soap and water, soap and scrubbing?
- water removes polar
- soap is surfactant, amphipathic
S. scrubbing, physically removes the substances
How does resident flora influence infections?
- Using up nutrients
- outcompete for space and resources.
- Can cause infection when it goes somewhere else
How does handwashing affect residential versus transient flora?
Both types of flora can be removed through handwashing, however, transient flora is more easily removed from the hands than resident flora. Some resident flora resides on the skin and has adapted to be hard to remove from the skin. Resident flora are covered by hair follicles, oil, and dead skin cells which obstruct their removal
If medical and surgical personnel wear gloves, then why do they still have to wash their hands?
There may be an accidental transfer of microbes when removing PPE. Or PPE may not be donned appropriately, exposing the medical personnel and patients to microbes.
subculturing
The transfer of microorganisms from one medium to another.
What are you preparing and maintaining?
We are practicing subculturing to learn to prepare and maintain stock cultures in various media.
What is an inoculum?
An inoculum is a microbial growth that is used to start another culture of that same microbe. This allows microbiologists to examine the growth of the microbe in a specific medium under controlled conditions.
What is inoculation?
inoculation is the process of introducing a microbe to a medium.
Identify three rules that you would tell another student to always remember to do when you are completing an aseptic technique for handling and transferring microbes.
Flame the loop or needle before and after inoculation.
Flame test tubes before and after using inoculating loop.
Never place the caps on the laboratory bench. The caps must be kept in the hand that holds the sterile inoculating loop or needle.
Flaming the inoculating instrument prior to and after each inoculation
These precautions help prevent contamination of the inoculum and help prevent accidental transfer of microbes on other surfaces. They also help protect the microbiologist against contamination.
Holding the test tube caps in the hand as illustrated in Figure 2.1 on page 14
This prevents the spread on microbes onto the lab bench which can lead to accidental infections.
c. Cooling the inoculating instrument prior to obtaining the inoculum
Heat can kill microbes. It is important to cool the instrument to avoid damaging the cultures in the inoculum.
Flaming the neck of the tubes immediately after uncapping and before recapping
These precautions help prevent contamination of the inoculum and help prevent accidental transfer of microbes on other surfaces. They also help protect the microbiologist against contamination.
Identify a pure culture
Pure cultures contain only one type of organism and allow us to study their cultural, morphological, and biochemical properties.
Identify a streak plate
The streak plate technique is a technique used to isolate pure cultures. This dilution technique spreads a loopful of culture over the surface of an agar plate as a means to separate and dilute the microbes and ensure individual colony growth.
In the “Tips for Success” section on page 22, what is the point of number 1?
An isolation plate has isolated distinct, individual colonies. The point of this is to get
individual, isolated colonies somewhere on the plate.
In the “Tips for Success” section on page 22, what is the point of number 2?
2.Pay attention to how well you sterilize your loop and maintain your aseptic technique
If the loop is not sterilized between streaks, or if aseptic technique is not maintained, there will not be a decrease in bacteria leading to individual colonies.
How does this technique apply to Koch’s Postulates? Chapter 1 and Chapter 14 will be helpful to answer this question.
- Isolate, grow and compare the microorganism in people with the same disease.
- Reintroduce in healthy person and see if they get disease
- Singles our microorganism as cause of a disease
The streak plate technique helps make the discrete, isolated, identical bacterial colonies needed to identify bacterial pathogens. which lets observers
It allows observers to collect samples of one type of culture and grow it to observe its characteristics.
____ ______ staining is the use of a single stain, for visualization of morphological shape (cocci, bacilli, and spirilli) and arrangement (chains, clusters, pairs, and tetrads).
Simple staining
Simple staining staining is the use of a ________ stain, for visualization of morphological shape
single
Simple staining staining is the use of a single stain, for visualization of morphological _______ (cocci, bacilli, and spirilli)
shape
Simple staining staining is the use of a single stain, for visualization of morphological shape (cocci, bacilli, and spirilli) and _____________ (chains, clusters, pairs, and tetrads).
arrangement
__________ ___________staining is the use of two contrasting stains, for the separation into groups acid-fast stain and Gram stain, and the visualization of structures (flagella, capsule, spore, and nuclear).
Differential staining
Differential staining staining is the use of _____ contrasting stains, for the separation into groups acid-fast stain and Gram stain, and the visualization of structures (flagella, capsule, spore, and nuclear).
two
Differential staining staining is the use of two contrasting stains, for the separation into groups acid-fast stain and ______ stain, and the visualization of structures (flagella, capsule, spore, and nuclear).
Gram
Differential staining staining is the use of two contrasting stains, for the separation into groups acid-fast stain and Gram stain, and the visualization of ________ (flagella, capsule, spore, and nuclear).
structures
T or F Bacterial smears must be prepared prior to executing of any of the staining techniques.
True
Whether using a broth culture or a culture from a solid medium, both should be air dried before continuing. T o F
True
T o F A broth culture requires water to be added to dilute the broth so it can be spread out to about the size of a nickel.
False
There is no way to avoid the bacterial smear from washing away on the slide. T o F
False
Simple staining methods are a quick procedure for determining
whether a clinical sample has the presence of a foreign bacterial pathogen.
Identify the bacteria that is a chain of spheres
streptococcus
Identify the bacteria that is a curved rod
vibrio
Identify the bacteria that is a chain of rods
streptobacillus
Identify the bacteria that is a cluster of spheres
staphylococcus
Slides should be handled with care during negative staining because
Heat fixation is not applied, leaving the bacteria alive
the first diagnostic test
gram staining
Shape, orientation, gram type, and can be performed on multiple cultures.
gram staining
to determine if an organism possesses a capsule, or demonstrate spore formation
negative staining
frequently used to ID a specific fungi that is found in bird droppings and linked to meningeal and lung infections in humans
negative staining
Gram stains are
differential stains
Imparts color to all cells
crystal violet