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https://us-lti.bbcollab.com/collab/ui/session/playbackBiochemical analysis of unknown bacteria
Aerotolerance Test
Fluid Thioglycollate broth (FTB) is a medium designed to test the aerotolerance of bacteria.
Along with nutrients to support bacterial growth, it contains sodium thioglycollate, thioglycollic acid, L-cystine, methylene blue, and 0.05\% agar.
The sodium thioglycollate, thioglycollic acid, and L-cystine reduce the oxygen to water.
Methylene blue is an indicator that is colorless in an anaerobic environment and greenish-blue in the presence of oxygen.
The agar helps retard oxygen diffusion and helps maintain the stratification of organisms growing in different layers of the broth.
Oxygen is driven out of the broth by autoclaving, but as the broths sit at room temperature, oxygen begins to diffuse back into the tube.
Obligate aerobes will only grow in this oxygen-rich top layer. On another hand, obligate anaerobes will only grow in the lower areas of the tube. Microaerophiles will grow in a thin layer below the richly-oxygenated layer. Facultative or aerotolerant anaerobes can grow throughout the medium but will primarily grow in the middle of the tube, between the oxygen-rich and oxygen-free zones
Reactions typically take up to 1-2 days to develop at 37⁰C
Media is inoculated using an inoculating loop
(A) Escherichia coli and (C) Staphylococcus aureus: both are Facultative Anaerobe, grows both aerobically and anaerobically and growth is seen throughout the tube. Some are capable of growth respiring with oxygen and anaerobically by fermentation.
(B) Clostridium botulinum: Obligate Anaerobe: can not grow in the presence of oxygen, growth is seen approximately 1/4 to 1/2 of the way from the top of the tube.
(D) Neisseria sicca: Microaerophile, requires oxygen but at concentrations below atmosphere, grows just below the surface of the media but not at the top.
(E) Pseudomonas aeruginosa: Obligate Aerobe: oxygen is required for growth and grows at the top of the tube only. The Organism will “settle” and sink into the media if grown longer than 24 hrs.
Aerotolerance Test
Phenol red test
Phenol red broth is a differential test medium prepared as a base to which a carbohydrate such as sucrose, lactose, dextrose or glucose is added.
Included in the base medium are peptone and the pH indicator is phenol red. Phenol red is yellow below pH 6.8, pink to magenta above pH 7.4, and red in between. During preparation, the pH is adjusted to approximately 7.3 so it appears red.
Deamination of peptone amino acids produces ammonia which rises the pH and turns the broth pink.
An inverted Durham tube is added to each tube as an indicator of gas production.
Gas production, also from fermentation, is indicated by a bubble or pocket in the Durham tube where the broth has been displaced.
Acid production from fermentation of the carbohydrate lowers the pH below the neutral range of the indicator and turns the medium yellow. Deamination of peptone amino acids produces ammonia which rises the pH and tSheet1
Gram Positive Chart
B Cereus B megaterium B subtilis E faecalis L lactis M luteus M roseus M phlei S ureae S aureus S epiderm S pyogenes S salivarius
Morphology rod>3um rod rod>3um coccus coccus coccus coccus rod coccus coccus coccus coccus coccus Morphology
Arrangement pairs,chains pairs,chains pairs,chains pairs,chains pairs,chains tetrads/clusters tetrads/clusters mycelium-like pairs,tetrads irreg clusters irreg clusters pairs,chains pairs,chains Arrangement
Pigments - - - - - Yellow Red - - - - - - Pigments
Motility + + + - - - - - + - - - - Motility
Starch [+] + + + - - [-] - [-] [-] d d Starch
Catalase + + + d - + + + + + + - - Catalase
Oxidase [+] - - - - + d + - - - - Oxidase
Indole - - - - - - - - - - - - Indole
VP + - + + + d d - + + + + VP
Citrate [+] + [+] [-] - + - + - d [-] - - Citrate
Urease [-] [+] - - - d - + + + - [+] Urease
Gelatinase + + + d - [+] [-] - + [+] - - Gelatinase
Glucose + + + + + [-] + - + + + + Glucose
Lactose - d [-] + + - - - + + + + Lactose
Sucrose + + + + + [-] + - + + + + Sucrose
Gas - - - - - - - - - - - - Gas
H2S - - - - - - - - - - - - H2S
Thioglycollate(o/f) Thioglycollate(o/f)
gram stain purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue purple/blue gram stain
Morphology rod bacillus rod bacillus rod bacillus spherical coccus rod bacillus spherical coccus spherical coccus spherical coccus spherical coccus spherical coccus spherical coccus spherical coccus spherical coccus Morphology
arrangement CHAINS, STREPTO CHAINS, STREPTO CHAINS, STREPTO clusters, staphylo CHAINS, STREPTO TETRADS TETRADS TETRADS TETRADS, sarcinae clusters, staphylo clusters, staphylo CHAINS, STREPTO CHAINS, STREPTO arrangement
pigment - - - - - Yellow red - - - - - - pigment
colony morphology flat undulate translucent FLAT, IRREGULAR ROUND, SMOOTH CONVEX MUCOID MUCOID, LOBATE ROUND, ENTIRE,SHINY, CONVEX SMOOTH, CONVEX, ENTIRE SMOOTH, CONVEX, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE colony morphology
growth in slant flat undulate translucent FLAT, IRREGULAR ROUND, SMOOTH CONVEX MUCOID MUCOID, LOBATE ROUND, ENTIRE,SHINY, CONVEX SMOOTH, CONVEX, ENTIRE SMOOTH, CONVEX, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE RAISED, CIRCULAR, ENTIRE growth in slant
growth in broth SEDIMENT SEDIMENT SEDIMENT UFT UFT SEDIMENT PELLICLE PELLICLE UFT UFT SEDIMENT SEDIMENT UFT growth in broth
sulfur (SIM) - - - - - - - - - - - - - sulfur (SIM)
indole (SIM) - - - - - - - - - - - - - indole (SIM)
motility (SIM) + + + - - + - - + - - - - motility (SIM)
catalase + + + ± - + + + + + + - - catalase
oxidase + - - - - + ± ± + - - - - oxidase
Methyl Red (MR) + - - + ± - ± - - - ± - - Methyl Red (MR)
vogues proBiochemical and microscopic
analysis of unknown bacteria
EXPERIMENTAL RESULTS
FOR IDENFICIATION OF THE UNKNOWN
BACTERIUM
Microscopic images:
Gram stain, morphology, arrangement
400X magnification
400X magnification
1000X magnification
Cultural Characteristics:
pigment, colony morphology
Growth in slant
Growth in broth
SIM test results (sulfur, indole, motility)
Catalase test result
On microscope slide
In tube
Oxidase test result
Methyl Red (MR) test result
https://www.bing.com/images/search?view=detailV2&ccid=FUELMhD8&id=4D3B7EBA56F59A72CFAD4A4DCD587018032AB5F7&thid=OIP.FUELMhD8PhSH6gIsQfvqZAEsEs&q=methyl+red+and+voges+proskauer+application&simid=608054868267696835&selectedIndex=1
https://www.bing.com/images/search?view=detailV2&ccid=FUELMhD8&id=4D3B7EBA56F59A72CFAD4A4DCD587018032AB5F7&thid=OIP.FUELMhD8PhSH6gIsQfvqZAEsEs&q=methyl+red+and+voges+proskauer+application&simid=608054868267696835&selectedIndex=1
Voges-Proskauer (VP) test result
https://www.bing.com/images/search?view=detailV2&ccid=3HwOpJNZ&id=AC46E32E3825E673DF38503A6B150A2D69E73AC5&thid=OIP.3HwOpJNZ-vAIogJMYTPDXwEsEs&q=methyl+red+and+voges+proskauer+application&simid=608025585180738723&selectedIndex=0
https://www.bing.com/images/search?view=detailV2&ccid=3HwOpJNZ&id=AC46E32E3825E673DF38503A6B150A2D69E73AC5&thid=OIP.3HwOpJNZ-vAIogJMYTPDXwEsEs&q=methyl+red+and+voges+proskauer+application&simid=608025585180738723&selectedIndex=0
Citrate test result
Urease test result
Gelatinase test result
Phenol red dextrose test result
Phenol red lactose test result
Phenol red sucrose test result
Phenol red Gas formation test result
Aerotolerance test result:
fluid thiolglycollate (FTG)
Obligate Aerobe (OA) or Facultative Anaerobe (FA)??????
Phenylalanine Deaminase test result
Starch Hydrolysis test result
Casein Hydrolysis test result
Lipid Hydrolysis test result
Biochemical and microscopic analysis of unknown bacteria
Microscopic images:�Gram stain, morphology, arrangement
Cultural Characteristics:�pigment, colony morphology
Growth in slant
Growth in broth
SIM test results (sulfur, indole, motility)�
Catalase test result
Oxidase test result
Methyl Red (MR) test result
Voges-Proskauer (VP) test result
Citrate test result�
Urease test result
Gelatinase test result
Phenol red dextrose test result
Phenol red lactose test result
Phenol red sucrose test result
Phenol red Gas formation test result
Aerotolerance test result:�fluid thiolglycollate (FTG)
Phenylalanine Deaminase test result
Starch Hydrolysis test result
Casein Hydrolysis test result
Lipid Hydrolysis test result�
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w or quality improvement; it was just all part of good nursing care. The goal for quality improvement is to monitor patient outcomes using statistics for comparison to standards of care for different diseases
e a 1 to 2 slide Microsoft PowerPoint presentation on the different models of case management. Include speaker notes... .....Describe three different models of case management.
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https://youtu.be/fRym_jyuBc0
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With covid coming into place
In my opinion
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4 In order to get the entire family to come back for another session I would suggest coming in on a day the restaurant is not open
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The word assimilate is negative to me. I believe everyone should learn about a country that they are going to live in. It doesnt mean that they have to believe that everything in America is better than where they came from. It means that they care enough
Data collection
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3 The first thing I would do in the family’s first session is develop a genogram of the family to get an idea of all the individuals who play a major role in Linda’s life. After establishing where each member is in relation to the family
A Health in All Policies approach
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Losinski forwarded the article on a priority basis to Mary Scott
Losinksi wanted details on use of the ED at CGH. He asked the administrative resident