Pglo Lab Report

Pglo Lab Report
How does the addition of pGLO plasmid to a solution containing E. coli bacteria affect the growth
and characteristics of the bacteria? Genetic transformation is the incorporation of foreign DNA into
an organism to potentially change the organism's trait. Plasmids are small circular DNA that
replicate separately from the bacterial chromosome. In nature, these plasmids can be transferred
between bacteria allowing for the sharing of beneficial genes. Due to this characteristic, plasmids
allow for genetic manipulation and can be moved between bacteria easily. The pGLO plasmid
utilized in this experiment encodes the gene for Green Fluorescent Protein (GFP), which under the
right conditions can produce a glow. The gene regulation system present in the pGLO plasmid
requires ... Show more content on Helpwriting.net ...
coli bacteria affects bacterial growth and gene expression in several environmental conditions. To
begin, two micro test tubes were labeled; one with +pGLO and one with –pGLO. 250 µl of
transformation solution was added to each microtube, with use of a sterile pipette. Afterwards, the
pipette was placed in a bleach solution and the microtubes were placed into the foam rack and onto
crushed ice. Next, a colony of bacteria was removed from the starter plate with a sterile loop and
thoroughly mixed within the +pGLO microtube. This step was repeated with a second sterile loop
and the –pGLO microtube. After their use, all loops were placed in the bleach solution for
decontamination and the microtubes were returned to the foam rack and ice.
Before the pGLO plasmid was added to the +pGLO microtube, the pGLO plasmid was observed
under a UV light. It was determined that the pGLO plasmid did not glow when placed under the UV
light. Following this observation, a single loopful of +pGLO plasmid was added to only the +pGLO
microtube and mixed thoroughly. After returning the +pGLO microtube to the ice, there was a ten
minute wait
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The Effect of Environmental Factors Such as Temperature,...
The effect of environmental factors such as temperature, osmotic pressure, oxygen concentration
and pH on microbial growth and survival
Abstract:
Microbial growth can be affected by different environmental factors such as temperature, osmotic
pressure, oxygen concentration and pH. Six experiments were carried out in this report testing for
microbial growth against different environmental factors. Good aseptic techniques were used to
prevent contamination, resulting in a uniform set of results that are in line with the literature.
Introduction
Bacteria vary greatly in terms of their characteristics and morphology. Colonies can be classified
according to their colour, form, elevation, margin and size. Pure cultures of microorganisms ... Show
more content on Helpwriting.net ...
Broth cultures of E.coli, Micrococcus luteus and Vibrio natriegens were streaked onto the respective
sections. The plate was then incubated at 37oC for 48 hours. The process was then repeated on
nutrient agar plates with 5, 6.5 and 10% salt.
Experiment 5: Effect of oxygen concentration on bacterial growth
A fresh pipette was used to transfer 0.5ml of broth culture of E. coli, to be inoculated, into a tube of
molten agar previously boiled to drive off oxygen. The tube was then rolled to distribute the bacteria
and allowed time for the agar to harden. The tube was incubated at 37oC for 48 hours. The
procedure was repeated for broth cultures of the bacterium Clostridium sporogenes and B. Subtilis.
Experiment 6: Effect of environmental factors on microbial growth – pH
A sterile pipette was used to add 0.1ml of E. coli culture to the pH 3.0 tube. This was then repeated
for the tubes at pH 7.0 and pH 9.The tubes were then incubated at 37oC for 48 hours. This was then
repeated for saline culture of Saccharomyces cerevisiae but incubated for 72 hours at 25oC.
Results
Six experiments were carried in this report concerning the effect that different environmental factors
have on microbial growth. The results were recorded into tables where (+) symbolises growth and
(–) symbolises no growth.
Experiment 1: Streak plate technique to

Summary Of Lab 2 Isolation Of Microorganisms
LAB 2: Isolation of Microorganisms
11 October, 2016
20 September, 2016
Melvin Espinoza–Rosa
INTRODUCTION
Microorganisms usually live in environments where there are a variety of different types of
microbes. In this lab, Isolation of Microorganisms we inoculated bacteria in different types of
medias which were described as selective media and differential media. Selective media permits
specific organisms to grow on top of having ingredients that do not allow the growth of any
organism that is not desired. In comparison, differential media does not hinder or boost the growth
of specific organisms. In its place it includes ingredients that aid in telling one type from another in
comparing bacteria. Differential media elements ... Show more content on Helpwriting.net ...
Through proper sterile technique and inoculation, I was able to capture maximum success in plate
streaking for selective or differential medias and isolation of bacterium. In order for this lab to be an
overall success the sterile technique, inoculation, and plate streaking were to be done with patience
making it the much harder task in this lab. Observing the final plates for differential or selective, as
well as for isolation were quite difficult to figure out as well. The media that I had more trouble in
differentiating through the variety of characteristics and morphology was Sheep Blood Agar (SBA).
Figuring out specific colonies was difficult due to beta hemolysis which is seen as a zone of clearing
around the colonies. The easiest to distinguish of all three medias was obviously the Tryptic soy
agar (TSA). Being the general growth media, we knew what we were expecting which was to see all
three bacteria to grow thus making the media not selective nor differential. We knew in the case of
Mannitol Salt Agar (MSA) that it could be selective or differential. Through the experiment we were
able to notice which bacteria were salt tolerant and those that weren't. In conclusion it was easily
noticeable that S. aureus was able to ferment mannitol being very salt tolerant while the other
bacteria struggled to prosper in

Assignment 2: Identifying Bacteria
UNKNOWN 65
Identifying bacteria is beneficial for several reasons. In the food industry, the discovery of bacteria
and other microorganisms helped develop processes to prevent food spoilage, such as pasteurization,
and bacteria is used to make certain foods, such as cheese, butter, vinegar, and alcohol (1). The
discovery of bacteria that break down waste products is beneficial to the waste treatment industry,
and the discovery of nitrifying, ammonifying, and nitrogen–fixating bacteria are important for
growing crops in the agricultural industry (1). Bacteria and other microorganisms that can produce
antibiotics have saved countless lives since the first antibiotic, penicillin, was discovered by
Alexander Fleming (1). One of the most important ... Show more content on Helpwriting.net ...
From the oxidase test, unknown 65 appeared to be of the family Enterobacteriaceae. The
Microbiology Laboratory Manual, chart II, showed very close matches for unknown 65 with
Escherichia coli, as did Bergey's Manual. Escherichia had identical tests results as unknown 65,
except for urease and phenylalanine tests (appendix A–16). Also, E. coli is resistant to tetracycline
and unknown 65 was not on the antibiogram (4). Unknown also matched Morganella morganii in all
performed tests, except for the arabinose and lactose fermentation tests. Bergey's manual showed the
same results. The urease test was positive, and since this test can distinguish urease–positive
Proteus, Morganella, Klebsiela, and Providencia from other Enterobacteriaceae, such as E. coli,
which is urease negative, unknown 65 is likely to be Morganella (2). The phenylalanine test is also
used to distinguish between the phenylalanine deaminase–positive genera Proteus, Morganella, and
Providencia from other Enterobacteriaceae, such as E. coli, which is phenylalanine deaminase–
negative (2). Considering these results and the lactose tests, which were contradictory, unknown 65
is most likely in the family Enterobacteriaceae, the genus Morganella, and the species morganii

The Effect Of Standard Curve Developed Using A...
period. The results were compared against standard curve developed using a concentration gradient
of Na2So4 with BaCl2 (De Zoysa et al., 2007).
3.15. FTIR of purified polysaccharide
FTIR (Fourier Transform Infrared Spectroscopy) spectra of purified PS fractions were determined
using FT–IR spectrophotometer model 5700 (M/S Thermo electron Corporation, USA).
Polysaccharide powder (2–3 mg) was mixed with KBr and pressed into a disk. The whole IR
spectrum was analyzed with a scan range of 4000–400 cm–1. Thirty scans were taken with 4 cm–1
resolution. CO2 and H2O corrections were incorporated. Reproducibility of the normalized spectra
was ±2%. (Shanthi et al., 2014).
3.16. Testing isolates probiotic properties
3.16.1. Blood hemolysis
Hemolysis test was performed according to the method described by Guttmann and Ellar (2000).
Overnight cultures of isolated Enterococcus durans and Enterococcus hirae were streaked on blood
agar and incubated at 37°C for 24 h. Blood agar plates were examined for signs of hemolysis. Blood
hemolysis test was performed in duplicates.
3.16.2. Resistance to low pH
Isolates Enterococcus durans and Enterococcus hirae were tested for their ability to resist low pH
values as follow, 25 ml of sterile MRS broth adjusted to pH 6.4, 4, 3 and 2 was inoculated using 1%
(v/v) of an overnight culture, then incubated at 37ºC for 6 h. The absorbance at 620 nm was
monitored using spectrophotometer (Unico, USA) at hourly intervals (Nawaz et al., 2011).
3.16.3. Bile

The Theory Of Microorganisms Exist
Introduction
Microbiology is the study of microorganisms; whether it is bacteria, fungi, algae, protozoan, viruses
or archaea (Leboffe & Pierce, 2006). Microbes come in all different structures and classifications.
Before the 17th century the idea or microbes existed, but was never proven until a scientist provided
something that allowed us to see what the naked eye alone could not see (Pelczar, 2015).
It all began when Anton van Leeuwenhoek invented a microscope and began observing objects
under it (Boundless, 2015). The theory of microorganisms existed prior to Leeuwenhoek's discovery
of them but they were never documented, just imagined. Leeuwenhoek was the first person to see
and explain a single celled organism, bacteria and even how blood flows in the capillaries (ICO,
2015). Robert Hooke was actually the first person reported to view a microbe under a microscope
and even discovered the cells of plants. Multiple experiments were done including and experiment
that proved that there are bacteria in the air contaminating things all around us. Broth was boiled and
one pot was exposed to the air while the other pot was not (History, 2012). The pot that was exposed
to the air showed that there were bacteria and life forms present; giving reason to believe that there
were some sort of microbes in the air, some of which can cause disease and some that are harmless.
The harmless microbes are actually extremely beneficial to the environment for many reasons
(Pelczar, 2015).

Alderson Smear Procedure
The microbe that I received was on plate B section 5. I began the project by creating pure cultures of
my microbe on a sterile nutrient agar plate, a nutrient agar slant and an agar broth by using
Alderson's procedure for aseptic techniques (Alderson, 2015, p. 53). After inoculating the media
with my microbe, I incubated them at 37°C for 48 hours to allow for growth. After 48 hours,
utilizing Alderson's technique for making smears from solid cultures I created 3 smears (Alderson,
2015, pp. 93–94). Using these smears, I created a gram stain (Alderson, 2015, pp. 100–101), a spore
stain (Alderson, 2015, pp. 114–115) and an acid–fast stain (Alderson, 2015, pp. 120–121). Despite
using Alderson's techniques for creating the smears and stains, my techniques differed slightly.
When creating a smear, Alderson's technique says to spread the inoculated drop of water to allow the
slide to dry however, for the purpose of speeding up the drying time I placed my slides in the ____.
In addition, when heat fixing my slides I used a clothespin to hold the slides over the flame when
heat fixing. When creating my stains, contrary to Alderson's techniques, I did not blot any of my
slides ... Show more content on Helpwriting.net ...
I inoculated an Eosin–Methylene Blue (EMB) agar plate and a MacConkey agar plate using
Alderson's streak plate technique (Alderson, 2015, pp. 64–65). Next, using Alderson's procedure for
aseptic techniques for inoculation (Alderson, 2015, p. 53) along with the procedures for each
individual test I conducted the biochemical tests on 1 lactose Durham broth (Alderson, 2015, p.
267), 1 glucose Durham broth (Alderson, 2015, p. 267), 1 TSIA slant (Alderson, 2015, p. 267), 1
SIM agar (Alderson, 2015, p. 268), 2 MRVP broths (Alderson, 2015, p. 268) and 1 citrate agar
(Alderson, 2015, p. 268). I also tested my microbe's ability to produce oxidase (Alderson, 2015, p.
267). Finally, I tested for the presence of catalase (Aryal, 2015, para.

Bacteria Lab
Lab Report: Unknown #1
Paola Rosales
302672960
Professor Youssef
INTRODUCTION:
Bacteria are the smallest living organisms, they are prokaryotic and have a simple cell structure.
They do not contain a nucleus, and are unicellular. Bacteria are the most abundant microorganism.
They are among the earliest forms of life that appeared on earth billions of years ago. They can grow
in different temperatures and many exist naturally in our human flora, such as staphylococci.
Growth within different temperatures can range from 0 degrees Celsius such as psychrotrophs, to
hyperthermophiles who grown best above 70 degrees Celsius (Ritchey, Exercise 16). Bacteria have
many shapes, size, and multicell arrangements. The main bacteria shapes are cocci (spherical or
ovoid shaped), bacilli (straight rods), coccobacillus (short rods), fusiform bacillus (rod shaped
bacilli with tapered ends), vibrio (curved rods), spirillum (single spiral bacteria), and spirochete (if it
is flexible and undulating). Bacteria can form multicell arrangements, such as diplococcus (two–cell
arrangements) (. Bacteria are usually named for their shape or multicell arrangement, but cannot be
truly identified by just those characteristics (Strelkauskas, Edwards, Fanhert, Pryor, 2016). Bacteria
can be classified by the way they stain. They are clear under a microscope and require staining to be
seen. To stain a bacteria, one takes a dye to color the cells. There are basic dyes that contain

Culture And Sensitivity Of Bacteria
Culture and Sensitivity If your patient has an infection of any kind it 's critical to know which
antibiotics will be effective against the particular pathogen or disease–causing agent. This means
that the species and strain of bacteria, fungus, or other pathogen must be identified and he drugs that
will be the most effective at clearing up the infection must be determined. The only way this can be
done is by running a culture and sensitivity test. Just keep in mind that there are many different
types of culture medias and hundreds of different sensitivity discs that correlate to different drugs. A
negative test on one of these media plates does not meant that there is nothing there. Which is why a
lot of clinics, like our own, run them in house and also send them to a lab such as Idexx to have
them run a more complete test there. A culture is a specific test to find and identify organisms such
as bacteria or fungus, that can cause an infection. A sensitivity test is a specific test that checks the
results of a culture to see what kind of medicine, commonly antibiotics, that will work best to treat
those organisms. A culture is made when a sample is added to a substance, or "Media" that promotes
the growth of fungus, bacteria, or whatever it is you are looking for. There are different types of
media for different types of tests, but the most common type of plate is called an AGAR plate,
which is made up of a nutrient substance gel derived from algae. Bacteria usually

Sterile Chips Lab Report
Methodology: Food items to be tested (ham, cheese, chip). You will need four of each. Sterile swabs
Sterile gloves Timer Agar plates (you will need 12 of them.) Notebook to record results
Experimental Procedure: 1. Put on sterile gloves ( do not touch any non–sterile surfaces when
wearing gloves) 2. Drop the first item of food (ham) on the first surface (tiles) 3. Start the timer 4.
Remove the item from the surface after 5 seconds 5. Swab the item with a sterile swab (do not touch
anything else with the swab) 6. Gently run the swab back and forth in a zigzag motion over the agar.
Do not touch any part of the agar twice 7. Put the lid of the petri dish back on and label as 'Ham
Tiles' 8. Change your gloves 9. Repeat the test with ... Show more content on Helpwriting.net ...
The ham agar plate grows the most bacterial colonies because this food has the most moisture.
Germs like living in moisture and this is why the test could not be done on a normal tiled floor but
instead had to be done with artificially covered surfaces. It also makes sense that germs would stick
to a food that was wet instead of a food that was dry. Cheese had the second most bacterial colonies,
and we could say this could be because of it being the second moistest food. Lastly, was the chip.
The chip got less bacterial colonies because it had no moisture what so ever. The difference in the
lowest and the highest on the ham varied widely from the lowest and the highest on the chip. The
lowest in the ham is 12 (ham on a bricked, or tiled surface) and the highest is 25 (ham on a bricked
surface) whereas the lowest for a chip is 9 (chip on a tiled surface) and the highest is 15 which is a
carpeted surface. The average amount of bacterial colonies on the ham throughout the entire
experiment is 16.83 whereas it is 12.08 on the chip showing that ham generally picks up more
bacteria than a

Handling Procedure
Procedure:
1. Find a clean counter or table to work on. Make sure you have everything you will need and that
all of the items are clean.
2. Put on a pair of disposable gloves.
3. Make a paper towel damp with some 70% isopropyl rubbing alcohol. Wipe down the work
surface with the rubbing alcohol–dampened paper towel.
4. Using a permanent marker, label the five baby food jars #1 to #5.
5. Perform a 1:10 serial dilution of the colloidal silver. Notice that the bottle you purchased is
labeled 500 PPM. This means that the concentration of silver nanoparticles in the bottle is
approximately 500 parts per million (PPM), which is the equivalent of 500,000 micrograms per liter
(µg/L). Although you are using colloidal silver, you are interested in the silver nanoparticle
concentration, which is 500,000 micrograms in every liter. ... Show more content on Helpwriting.net
...
Use the cotton swab to streak the five labeled nutrient agar plates with the E. Coli K12 bacteria from
the tube.
36. Remove a tiny bit of E. coli from the tube using the cotton swab.
37. Hold the tweezers with the cotton swab (or just the cotton swab alone if you are using extra–
long cotton swabs) in one hand.
38. Hold the swab in your right hand
39. Hold the E. coli tube in your other hand.
40. Using the hand that is holding the cotton swab, unscrew the lid of the tube against your pinky,
wrapping your pinky around the lid.
41. Hold the lid in this pinky. Do not set the lid down.
42. Using your index finger and thumb, dip the cotton swab into the bacteria tube. Rub the tip of the
cotton swab against the agar slant.
43. Get a small piece of the agar (smaller than a grain of rice) on the end of the cotton swab.
44. Visible representation of the bacteria is not needed.
45. Screw the lid back on the tube.
46. While holding the cotton swab in one hand, remove, and hold the lid of an agar plate using your
other hand.
47. Spread the E. coli over the surface of the agar plate
48. Streak a vertical line across the surface of the agar

Mixed Culture Report
Introduction The purpose of this experiment was to allow each student to perform certain procedures
and utilize the skills accumulated over the semester in order to interpret and identify two unknown
organisms within a mixed culture. Each student was given a mixed culture containing one Gram
negative organism and one Gram positive organism. I received unknown mixed culture #15. The
possible Gram negative organisms within the mixed culture include: Enterobacter aerogenes,
Pseudomonas aeruginosa, Neisseria perflava, Proteus vulgaris, and Moraxella catarrhalis. The
possible Gram positive organisms within the mixed culture include: Corynebacterium xerosis,
Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, and Clostridium
perfringens.
Methods
Using the mixed culture given, each student performed two Gram stains, which is described in the
lab manual Microscopy And A Survey Of Microorganisms in Exercise 6 (McPherson pg. 53).
During each procedure, every student followed aseptic technique in order to minimize
contamination. This process is explained in Exercise 4 (pg. 37). Following the two Gram stains,
each slide was viewed using a bright light microscope in order to find out the initial morphologies
and different Gram ... Show more content on Helpwriting.net ...
66). The two TSA plates were incubated both aerobically and anaerobically because it displayed
how oxygen affected the growth of each organism. Both aerobically and anaerobically incubated
plates displayed identical white, convex elevated isolated colonies with entire margins and glossy,
convex elevated isolated colonies with entire margins. Therefore, this procedure did not eliminate
any of the possible organisms due to the fact that S. aureus, S. epidermis, E. faecalis, E. aerogenes,
P. vulgaris, and P. aeruginosa all grow under aerobic and anaerobic

Manipulation of Bacteria Essay
MANIPULATION OF BACTERIA
INTRODUCTION:
In this experiment that we performed, there were many methods that were used to help us
manipulate and identify the bacteria E.coli on a MacConkey agar plate. The first part of the
experiment involved the methods of manipulating, identifying and counting the bacteria and the
second part was to find out whether the bacteria E.coli was the only type found in the given area by
gram staining.
E.coli was the chosen bacteria for this type of experiment. It is a gram negative bacterium that will
grow rapidly given 'any culture medium with the necessary energy source, nutrients, pH, and
temperature'. Therefore, MacConkey Agar being the medium for its growth will enable us to achieve
the experiment ... Show more content on Helpwriting.net ...
http://www.suite101.com/content/api20e–bacterial–id–test–strip–a55408
METHOD FOR MANIPULATION AND IDENTIFICATION OF BACTERIA
 Lab coat and gloves have to be worn for safety
 Start of by ensuring the work area and equipments are clean so that there is no contamination to
your work.
 Taking the sterile loop without holding the loop side dip inside the given bacteria culture provided
and streak the MacConkey agar plate shown below.
 The first loop L1 is to be streaked as shown. The loop is then to be put into the sharps bin
immediately to prevent contamination of the work area and other equipments.
 Taking a fresh loop streak L2a then using the other side of the same loop, streak L2b also.
 A third loop L3a is then used to streak the bottom right, then flip the other side and streak the
bottom left corner L3b like the image shows.
 Lastly, take the last loop L4 and create a squiggle in the middle of the plate. This is the streaking
procedure so that the bacteria will be spread out as much as possible for better results during the
count.
 This is then covered and labelled ready for inoculation to make it easy to identify.
 For the viable count, a set of dilutions have to be made. Place the 9 tubes in their stand and add
900ul of PBS into each bottle adding it to the neat sample of bacteria culture first.
 Sequentially, add 100ul from the previous sample to the next e.g. take 100ul from the neat sample
dilution into the following 900ul forming 10

Common Microorganisms Essay
INTRODUCTION
Microorganisms such as bacteria, fungus, mold, and yeast are present and common in almost every
environment on earth. The normally microscopic organisms can easily be seen using differing types
of agar, which creates an ideal environment for the organisms to form colonies, which are groups of
hundreds of organisms that can be seen with the naked eye. In order to see individual
microorganisms, it is necessary to use the magnification of a high–powered microscope.
These techniques of microbiology are used in the following five experiments. The first experiment
used Trypticase Soy Nutrient Agar (TSA), which can grow a wide variety of organisms and contains
casein and soybean meal and a minute NaCl, to study the effectiveness of ... Show more content on
Helpwriting.net ...
Label them A, B, C, and D.
3) Press the pad of a dirty thumb onto quadrant A.
4) Swab the thumb with sterile water, allow to dry, and imprint it onto quadrant B. The sterile water
acts as a control.
5) Press the other dirty thumb onto quadrant C.
6) Swab this thumb with the alcohol swab, allow to dry, and imprint on quadrant D.
Experiment 2. Effectiveness of mouthwash as an antiseptic
1) Divide a petri plate with Trypticase soy agar into 2 quadrants.
2) Swab the inside of your mouth with a sterile cotton swab, inoculate one half of the agar.
3) Rinse your mouth with Listerine brand mouthwash. Wait 5 minutes after rinsing.
4) Swab the inside of your mouth with a clean sterile swab, inoculate the second half of the agar.
Experiment 3. Normal human flora
1) Obtain two petri dishes containing one half Trypticase Soy Agar and one half Mannitol Salt Agar.
2) Moisten two sterile swabs in sterile water, remove excess water, and then use one of them to swab
the inside of your nose, and the other to swab a patch of skin, like your neck.
3) Inoculate each dish of agar with either the nose or shin swab on both sections.
Note: For experiments one through three, each petri plate should be taped shut and stored upside–
down for one week at room temperature to incubate the

The Organisms And Its Effects On Our Daily Environmental...
Introduction
A microbe is described in the dictionary as "a microorganism, especially a bacterium causing
disease or fermentation." The thought of these bacteria make some people shudder; the thoughts of
unseen enemies crawling on surfaces and primary areas of life. To prevent the spread an overgrowth
of the organisms, we as a race have invented and developed many innovations. Although these fears
are justified, could it be that these microbes we fear so much, exist in greater numbers within us than
on our daily environmental surfaces. If samples were taken from different areas on a college campus
that are assumed to be dense with bacteria, would the intensity of growth be greater than samples
taken from the human body?
Material and Methods
Students were provided with the following equipment in a sterilized Microbiology Lab. Cotton
Swabs Distilled Water Test Tubes with caps Petri Dishes Nutrient Agar Blood Agar Inoculation
Loop Bunsen Burner Slides Pipette Compound Light Microscope Various Reagents.
Students were asked to gather samples from four sites on a college campus where it was believed
bacteria would be most concentrated. Students were also asked to collect samples from their inner
nose and throat. The four environments that were supposed to have a dense congregation of
microbes on campus were: the "Science Building Main Entrance door handle", a "Vending Machine
Cash receiver", an "Elevator Button" and a "Rubbish Bin Lid." Two prepared

An Unknown Disease Caused By Microorganisms
Introduction Whenever there is an unknown disease caused by microorganisms, tests are usually
made in order to identify the organism causing the disease. There are several tests that need to be
made and they include tests such as performing a gram stain, streaking a plate to isolate colonies,
inoculating a broth culture, inoculating API strip, and performing oxidase and catalase tests. Having
knowledge on how to identify these tests are of high importance in the medical field so it would be
to the advantage of those individuals who know how to examine microorganisms and be able to
identify it by correctly performing tests on organisms. To begin with, a good way to get exposed to
tests that will be needed by people entering the medical field would be to perform practice test like
being assigned an Unknown that students could identify by performing tests. The Unknown that was
assigned to me for example was Unknown #22. In order to identify Unknown #22 I had to perform
several test and make sure that all the results matched with each other to be sure that I had
performed my tests right. The goal of my experiment is to learn how to properly perform tests on an
Unknown organism and be able to identify it. By learning to identify a specific microorganism and
performing various testing techniques I will be able to identify a microorganism if required when I
am working in the medical field. When a microorganism is identified, the doctors' job is made easier
because they are able to

Pglo Transformation Essay
Connor Lauffenburger
3/17/13
pGlo Transformation Lab Report
I Introduction
The purpose of this experiment was to show the genetic transformation of E. coli bacteria with a
plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that
confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate
independently of chromosomes. In this lab, the Green Fluorescent Protein, which is typically found
in the bioluminescent jellyfish Aequorea Victoria, was cloned, purified, and moved from one
organism to another with the use of pGlo plasmids. It was hypothesized that if bacteria that were
transformed with +pGlo plasmids are given the gene for GFP, then transformed cell colonies ...
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Next, the transformation efficiency was calculated by dividing the total amount of DNA on the agar
plate in μl by the number of cells growing on the +pGLO LB/amp/ara plate.
During the purification section of this lab, the LB/amp/ara agar plate was examined for well–
isolated green colonies and the LB/amp plate was observed for white colonies with space between
each other. These colonies were circled on the outside of the plates using a marker. Next, two 15
milliliter culture tubes containing 2 milliliters of nutrient growth media were obtained and labeled
"+" and "–". Using a new inoculation tube, the circled colonies from each plate were scooped out
and immersed in their respective culture tubes. Once the bacteria was mixed into the solution, the
tubes were sealed and placed horizontally into the 32⁰ incubator for 24 hours.
After labeling a new microtube with a marker, the culture tubes were removed from the incubator
and observed under the ultraviolet lamp. Using a sterile pipette, the contents of the (+) culture tube
were transferred to the 2 milliliter mictrotube and the (–) culture tube was disposed of. The (+) tube
was then spun in the centrifuge at maximum speed for 5 minutes. Once it was removed from the
centrifuge, a noticeable bacterial pellet had formed at the bottom of the tube and the remaining
liquid was disposed of. After the

A Study On Blood Hemolysis Essay
A portion of (2–3 mg) from the partially purified polysaccharide powder was mixed with KBr then
pressed into a disk. The whole infrared spectrum was analyzed at a scan range of 4000–400 cm–1.
Thirty scans were taken with 4 cm–1 resolution. CO2 and H2O corrections were incorporated.
Reproducibility of the normalized spectra was ±2%. (Shanthi et al., 2014).
3.16. Testing isolates probiotic properties
3.16.1. Blood hemolysis
Hemolysis test was performed according to the method described by Guttmann and Ellar (2000).
Overnight cultures of isolated Enterococcus durans and Enterococcus hirae were streaked on blood
agar and incubated at 37°C for 24 h. Blood agar plates were examined for signs of hemolysis. Blood
hemolysis test was performed in duplicates.
3.16.2. Resistance to low pH
Isolates Enterococcus durans and Enterococcus hirae were tested for their ability to resist low pH
values as follow, 25 ml of sterile MRS broth adjusted to pH 6.4, 4, 3 and 2 was inoculated using 1%
(v/v) of an overnight culture, then incubated at 37ºC for 6 h. The absorbance at 620 nm was
monitored using spectrophotometer (Unico, USA) at hourly intervals (Nawaz et al., 2011).
3.16.3. Bile salts tolerance
The selected isolates were examined for their ability to grow in presence of different concentrations
of bile salts. The two concentrations 0.1% and 0.3% (w/v) were used for this purpose. An aliquots of
20 ml sterile MRS broth supplemented with 0, 0.1% and 0.3% bile salts were inoculated with

Serial Dilutions: Salmonella Typhimurium Culture
Serial dilutions, also known as limiting dilution series, help to determine an estimation of how many
phages/viruses are in a given culture (Zelterman et al., 2010). In this experiment we determined how
many phages were in our given Salmonella typhimurium culture by doing three serial dilutions of an
original phage suspension. This is important because it will allow us to get a rough estimation of
how many phages where in our original sample and also provide us with knowledge on how to
perform serial dilutions with phages which can be used to determine an unknown sample in the
future. We expect that there will be less plaques on the last dilution than in the first plated dilution
because the more diluted each test tube is the less phages are ... Show more content on
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Each test tube containing 9–ml of a dilution blank (distilled water) was aseptically inoculated with
1ml of Salmonella typhimurium phage. One test tube was a control that contained no phage and was
diluted into a tube and poured on the plate. The first inoculation was the original phage suspension
and from there we diluted it three more times in 9–ml dilution blank test tubes. Once the test tubes
were inoculated .5–ml of each dilution was placed in soft agar tubes quickly, so that it would not
solidify, and then were poured on the trypticase–soy agar plates. The TS agar plates were left to
incubate for 6–10 hours. Once done incubating each plate was then counted to see how many
plaques had formed on the bacteria lawn.
Results
The results of the plaque assay showed that the 10^–2 plated dilution contained more than 300
plaques and was too many to count. For the 10^–3 plated dilution there were 37 plaques which
indicated there were 3.7*10^4 pfu/ml. The last plate which was a 10^–4 dilution had four plaques on
it and was too small of an amount to use to find the original phages in the suspension.

Lab Report On Microbial Unknowns Essay
Microbial Unknowns Objective: In the past couple of weeks, our class received a single broth that
contained two species of bacterium. One bacterium was gram positive and one bacterium was gram
negative. We were required to use our techniques and knowledge we learned in class to separate and
identify the two species we were given in our broths. Methodology: Day One: On the first day of
our Biochemical Unknown Lab we were given a tube that contained broth media which held two
different types of bacterium. Once we were given our broths, we had to streak two Nutrient Agar
plates with the mixed culture tube by using a loop. After I streaked my Nutrient Agar plates, I
decided to perform a gram stain. First I flamed the loop and set it aside to cool. After I flamed the
loop I prepared two slides with a drop of water in the middle. Once the loop was cool I dipped the
loop into the broth and smeared the loop onto both slides mixing the unknown bacterium with the
water. I flamed my loop, set it aside, heat fixed both slides, and let the slides air dry. Once the slide
were dry I set them on two rods above the drainage area and covered the smears with Crystal Violet.
I let the Crystal Violet sit on the smear for one minute and rinsed off the Crystal Violet. Once the
Crystal Violet was rinsed I applied Grams Iodine and let that sit for one minute. After one minute
passed I rinsed the slide again and moved on to applying the alcohol. I dripped the alcohol onto the
slide until the

Microbiology Unknown Lab Report Essay
Lab12–Medical Microbiology– Part1– Differential Media/Biochemical Tests, Sp2012 (Set all of
your margins to ½")
Purpose: The purpose of this lab is to help you become a little familiar with some of the tests that
can be typically performed in a clinical or research lab facility. These tests may help in determining
a particular pathogen's growth needs.
There are several sections to this lab. Find each section and complete the "Preparing for Class"
sections.
Preparing for class – Day 1
Read in your lab manual about the following agar mediums: Blood Agar (pg 168), EMB Agar (pg
170), Mannitol Salt Agar (MSA)(pg 172) ), MacConkey Agar (pg 174), and PEAAgar (pg 176) to
answer the following:
1. What does the blood agar select for? ... Show more content on Helpwriting.net ...
|bacteria, or |enterococci |
| | | |greenish/gray hue | |
| | | |around | |
|EMB Agar | |Distinguishes bacteria that ferment |Dark blue colonies with|E. coli and P. |
| |Gram–negative bacteria |lactose and or sucrose and those that|green metallic sheen or|aeruginosa |
| | |do not. |pink. | |
|MSAAgar |For organisms that are |Isolates for mannitol fermentation |Yellow color change in
|Staphylococcus aureus |
| |halotolerant. | |surrounding media. |and Staphylococcus |
| | | | |epidermidis |
| | | |

Streak Plate Essay
Student: Yi–Ren Wang
Course: BIO–205 BD2 Microbiology
Instructor: Dirk VandePol
Date: 6/21/2013
Streak Plate Isolation for Obtaining Pure Culture
1. When an agar plate is inoculated, why is the loop sterilized after the initial inoculation in put on?
Ans: We use agar plate to inoculate microbes by zipping the loop on the agar several times. We
streak on the agar plate four time, propose is to isolate the unknown bacteria. Therefore, the first
time to streak on the plate, there are million of bacteria on the loop. For that reason, we need to
sterilize the loop before next streaking. Then we can get small group of colonies out from the large
group of colonies to observe and distinguish the unknown bacteria.
2. Define ... Show more content on Helpwriting.net ...
6. Why is necessary to isolate individual colonies from a mixed growth?
Ans: Every individual colony in a mixed growth might present different bacteria. We isolate them to
help us effectively to determine on their differences.
7. Why was blood agar, rather than a nutrient agar plate used for the culture from your mouth?
Ans: Because in a blood agar contains more nutrient than a nutrient agar. In our mouth it could have
more different species bacteria that blood agar will be a great place for them to grow.
8. Are a large number of microorganisms found in the mouth a cause for concern? Explain.
Ans: In my opinion, it might not necessary to worry about microorganisms in our mouth. There are
some of them are harmless bacteria, and keep balance in our mouth. Therefore, depending on the
species we may take more concern.
9. How do microorganisms find their way into the mouth?
Ans: there are a lot of ways microbe can go inside our mouth, such as eating food, drinking water,
putting fingers on mouth. All of them could covered with microorganisms to pass through to our
mouth.
Date Collection

Nutrient Agar Blood Agar
Describe
Nutrient Agar: We use a mixed culture on this nutrient agar. As the simple we use, there are two
different colonies. First of all, small circular size and light white color. Secondly, large and flat ring
size

High School Bacteria Experiment
In this experiment, we wanted to investigate which surfaces in the school had the most bacteria, and
we thought that the surfaces used more would have the most bacteria. For this experiment we used
agar plates, cotton swabs and distilled water in order to see how much bacteria was on each surface.
Cotton swabs were wet with distilled water and then the swabbed ws wied on a 2 inch space of the
surface and twisted to cover the entire swab.once we swabbed the surface, we wiped the swab on the
plate in a zigzag motion holding it upside down. The girls bathroom sink handle had 389 bacteria,
and the boys bathroom by the football field had 0. We saw that places commonly used such as the
bathroom sinks, door handles and railings contracted the most bacteria. ... Show more content on
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Things that are used commonly by students and staff should be routinely disinfected to ensure the
health and safety of the people using the provided materials. Bacteria is related to disease (American
Association for the Advancement of Science, 1885) Previous studies on the surface of a high school
telephone showed that the phones had an uncountable number of bacteria colonies (Yalowitz, 2003).
We know that surfaces used by the public often become contaminated because of the bacteria on
peoples hands. A study on the bacteria on peoples hands showed that 28% of people have fecal
matter on their hands (Judah, 2010). A study done on the contamination on public doorknobs
showed that over 86.7% were contaminated (Nworie, 2012). This experiment was to show us how
much bacteria is on the surface of the things we use

What Is 5. 5 Antimicrobial Agar Diffusion Assay
5.5 Antimicrobial Activity
The antimicrobial agar diffusion assay was performed according to disc diffusion method against
four bacterial strains; Bacillus subtilis , Shigella flexneri, Escherichia Coli ,Enterobacter cloacae and
four species of fungi Saccharomyces cerevisiae, Aspergillus candidus, Aspergillus niger and
Pencillium Species.Potato Dextrose Agar for fungus and NAM(Nutrient agar media) for bacteria
was prepared according to the accurate composition and immediately after autoclaving, it was
cooled in a 45 – 50°C.The freshly prepared and cooled medium was poured into petri plates. The
agar medium was cooled to room temperature unless the plate is used the same day; and stored in a
refrigerator (4°c).
5.5.1 SPREADING OF BACTERIAAND FUNGUS ON THE PLATES
100 µL of bacteria and fungus from freshly prepared culture was taken in the pipette and poured in
the middle of the respective petri plate. Remove excess inoculum by lightly pressing the swab
against the tube wall at a level above that of the liquid.Using a cotton swab that has already put in
UV light, the bacteria and fungus was spread evenly on the surface of the plate so that bacteria and
fungus were spread in each corner of the plate and dried for 4–5 minutes Inoculate the agar by
streaking with the swab containing the inoculum. Rotate the plate by 60° and repeat the rubbing
procedure. Repeat two times. This will ensure an even distribution of the inoculum. Allow the
surface of the medium to dry for

Seven Agar Plates
For this experiment we utilized a variety of materials. We first utilized a sharpie pen to write down
the locations of our experiments and our names among other information on seven different Agar
plates. We then used seven different containers of sterilized water to make sure there was no
contamination between different samples. We also utilized seven different disposable swabs for this
exact same reason. We also used seven packages of Para–film in order to seal the plates. Lastly we
made use of a freezer in order to keep our plates at the proper temperature for bacteria incubation
and growth. These are all the physical materials we made use of while conducting our own
experiment. For our methods, first collected all of our materials. We

Dna And Human Dna Has Transformed Genetics
Done in the past decades, progress of new and great techniques for learning and manipulating DNA
has transformed genetics. The cloning plasmid shown in figure 16.1 is one example of the tools that
are now available. These techniques have permitted biologist to occur directly in the genetic fate of
organisms for the first time, the ability to directly isolate and manipulate genetic material was one of
the most profound change in the twenty first century, plasmid vectors (small, circular chromosomes)
are typically used to clone relatively small pieces od DNA, up to a maximum of about 10 kilo bases
(kb).
A plasmid vector must have (1) an origin of replication to allow it to be replicated in E. coli
independently of the chromosome, and (2) a selectable maker , usually antibiotic resistance. The
selectable maker allows the presence of the plasmid to be easily identified through selection, cells
that contain the maker will live when plated on antibiotic –containing growth media, and cells that
lack the plasmid will not live (they are killed by the antibiotic). A fragment of DNA is inserted by
the techniques described in figure 16.2 in a region of the plasmid called the multiple cloning site
(MCS).
This region contains a number of unique restriction sites such that when the plasmid is cut with any
of these enzyme, it will produce a linear plasmid. The DNA interest can then be ligated into this site,
after which plasmid DNA is introduced into cells. Next, the presence of

A Study On Nutrient Agar
Nutrient Agar To observe the colony morphology, the original unknown culture was streaked for
isolation on nutrient agar using the quadrant streaking technique, inverted, and incubated at 37° C
for 48 hours. This allowed for observation of the colony morphology. Separation of Gram–positive
and Gram–negative The original unknown culture was streaked for isolation on Columbia CNA agar
and MacConkey agar using a quadrant streak, inverted, and placed in a 37° C incubator for 48 hours.
CNA agar contains a mix of colistin and nalidixic acid, as well as sheep blood. If there is poor or no
growth on CNA agar, then the organism was inhibited by colistin and nalidixic acid and is Gram–
negative. However if there is good growth, the organism was not inhibited by colistin and nalidixic
acid, and is Gram–positive. MacConkey agar contains bile salts and crystal violet. If there is poor
growth or no growth, the organism was inhibited by crystal violet and/or bile, and is Gram–positive.
If there is good growth, the organism was not inhibited by crystal violet or bile, and is Gram–
negative. Gram Stain This protocol was performed once using the culture on the Columbia CNA
agar, and once using the culture on the MacConkey agar. The Gram stain uses crystal violet as a
primary stain, ethanol–acetone as a negative stain, and safranin as a counterstain. A small amount of
the culture was heat–fixed to a glass microscope slide. The culture was flooded with methylene blue,
and allowed to sit for one

Microbial Diversity Of Soil And Determine The Population...
Soil samples were collected from a construction site in North Norman and the duck pond at the
University of Oklahoma. To demonstrate the microbial diversity of soil and determine the
population density of bacteria, actinobacteria, and fungi, the samples were cultured in the laboratory.
Serial dilutions of the samples were prepared and grown on different growth media for selective
culturing of the three groups. Following incubation of the plates at 37C for two days then at 4C for
five days, it was observed that the colonies on the agar plates had different morphologies. Colony
counts were used to determine the original cell density of the three microbial groups. Although the
calculated population densities were inconsistent because of multiple countable plates, the consistent
trend was that the population density of simple bacteria was the highest in soil sample 1 and that
fungi outnumbered simple bacteria and Actinobacteria in soil sample 2.
Introduction
The soil matrix is a microbial storehouse in which a variety of microorganisms co–exist. Within the
soil habitat, microorganisms are taking in nutrients, respiring, and participating in biogeochemical
cycles. There are prokaryotic microbes, eukaryotic microbes, and viruses. The most abundant
microbial populations in soil are simple bacteria, Actinobacteria, and fungi (1). The eukaryotic
populations in soil include algae, fungi, and protists. Fungi participate in recycling biomass and
stimulating plant growth (2). Soil fungi

Exercise 1-3 Aseptic Report
Exercise 1–3: Aseptic Technique, Transfers, and Inoculations The Goal: The goal of this exercise is
to learn and apply Aseptic techniques, which will be used throughout this course, by aseptically
transferring cultures to three types of media: broth, plate, and agar slant. Materials: Lab coat 10%
bottle of bleach Sharpie Masking tape Vortex mixer Sterile Tryptic soy broth test tubes Sterile
Tryptic soy agar slants Tryptic soy broth culture of Serratia marcescens Tryptic soy agar slant
culture of Serratia marcescens Inoculating loop Bunsen Burner Striker Methods: Before you start
any kind of Transfer you must: 1) Wash hands, place on lab coat, and disinfect work area with the
ten percent bleach. 2) Set up your Bunsen burner and light flame using a striker ( make sure the
flame has two cones) 3) ... Show more content on Helpwriting.net ...
4) Make sure that the inoculating loop is a complete loop, if broken obtain a new inoculating loop.
Collecting microbes from a Broth: 1) Using your writing hand, pick up the inoculating loop and heat
it, from the base to the loop end, until it is glowing orange. 2) Allow the inoculating loop to cool for
a few seconds. 3) With your non–writing hand grab the desired broth culture tube. 4) While the
inoculating loop is cooling, mix the broth culture by placing the tube in the vortex mixer (hold the
tube and not the lid). 5) After the broth is mixed and the inoculating loop is cooled, take the cap off
of the broth culture tube by using the two fingers, on your writing hand, that are not holding the
inoculating loop. 6) Briefly pass the top of the tube through the flame a couple of times. 7) Hold the
inoculating loop still, move the broth culture tube up until the loop is within the

Environmental Lab Report Sample
Exposed to Sunlight Does Not Reduce the Microbial Communities in an Aquatic Ecosystem
Sekora Martinez
Wendy Guzman, Jasmine Jones, Lou Lindley
BIOL 1442_002; Danielle Rivera; July 27, 2016
Clean water is the key component to surviving life. Yet, there is some water that is contaminated that
can cause disease or death if not treated right away. To determine rather the sun has a positive or
negative effect on rather or not microbial grow more in an aquatic environment, three samples of
water were tested from a local creek. Water that was primarily in a shaded area, where no sunlight
could reach and water that was completely exposed to sunlight, with a control variable of sterilized
water from the class room. Each sample was left in a stable ... Show more content on
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There were different sizes of each microbial but the average size for the water exposed to sunlight
was 0.7mm in section one, 1.5mm in section two, and 0.5 in section three (table 1). The agar plate
with a tap, sterilized water, had the largest microbial in size at about 1.7 mm. The light exposed
water had an increase in size from section one at 0.5 mm to 1.1 in section 3.
Analyzing each plate, there was a physical feature that was easily identify for example the surface,
texture color and elevation. In the lighted plate, each section had a smooth surface appearance,
section 1 had a rigid texture but sections 2 and 3 had a buttery texture. Each section had a yellow
color to it and its elevation was raised just enough to be noticed. The shaded plate had sections 1 and
3 with a smooth surface, buttery texture, yellow–whitish color and both raised in elevation. While as
section 2 with a rough surface and rigid texture, a tan color and was completely flat in elevation.
The plate with sterilizing water had no growing microbial in sections 1 or 2 but in section 3 there
was one large microbial that was rough, fuzzy, yellow and convex. There is a clear difference
between each agar plate with size, diversity, and

How The Clean Water Is A Very Important Aspect Of Health
This report represents my individual effort. I did not receive or offer aid to anyone when performing
this assignment, nor did I plagiarize any material.
Signed: __________________________________________________________
Introduction Having clean water is a very crucial aspect of health. Many people don't have access to
clean water, this can be life threatening. There are lethal bacterial infections that are present in
water. There are many different diseases that can acquire by drinking contaminated water like:
cholera, typhoid fever, and dysentery. The biggest concern and issue is the ingestion of water that is
contaminated with fecal matter (1). It has been discovered that the main source of bacteria is this
fecal contamination. Fecal matter can enter water sources by runoff from farms or any area that
contains many animals. This contamination can also come from insects, rats, or other small animals
entering wells or water sources (2). Testing the quality of water, it is often common to test if the
water contains coliforms. It is easier to test for coliforms, instead of testing for bacteria that can
cause each waterborne disease. Coliforms are gram negative, facultative anaerobic bacteria that do
not form endospores. They are also rod shaped and will ferment lactose (slides). When they ferment
lactose, this will produce acid and gas. This acid and gas production will allow the total coliform
count to be determined.

Phage From The Environment
In the experiment "Isolate a Novel Phage from the Environment: Enrichment of Environmental
Samples" it first started out by adopting a phage from a classmate. Then proceeded to put buffer into
two samples about 5mL each then used the centrifuge for 10 seconds each. The samples were then
left to settle for 10 minutes. After the 10 minutes were over, a 5mL pipette was used to drain the
"clear" liquid that was in the sample to put into a sterilization filter unit to filter the phages into a
micro centrifuge tube. When that was completed. Proceed to add .5mL of m.smegmatis and 4.5 mL
of top auger into a culture tube with the liquid from the soil sample. Pour the mixture into an agar
plate, wait for the TA to harden and then invert. The next experiment ... Show more content on
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Then add 90 micro liters of phage buffer to each tube and then take 10 micro liters of the mixture in
10^0 and put that in 10^–1 then put 10uL of 10^–1 into 10^–2 and vortex well. Take 10uL of 10^–2
and put that into 10^–3 and vortex well. Repeat for 10^–3 into 10^–4 and vortex well. To infect M.
smegmatis with the diluted phage solutions. Label Culture tubes 10^0, 10^–1, 10^–2, 10^–3, and
10^–4. Inset 0.5mL of M. smegmatis into each culture tube and insert 10uL of the dilutions 10^0,
10^–1 through 10^–4 into the corresponding culture tubes. Allow 15 to 30 minutes for the phage to
infect the bacteria. Be sure to record the time allowed to infect. After the time of 15 minutes had
passed, label five agar plates to correspond with the culture tubes. After the plates are labeled,
remove top agar from a 55 degree Celsius water bath. For each sample a 5mL pipette attached to a
pipette is used to aseptically transfer 4.5mL of TA to the culture tube containing the bacteria. As
soon as the TA is transferred into the tube, immediately pull it back up into the pipette and transfer
the entire mixture to the corresponding agar plate. Do this for all the agar plates and their
corresponding numbers. Make sure to use a sterile pipette each time something is transferred the
mixture to avoid contamination. The TA starts to harden after a few moments, after it harden, invert
the plate and store the plates in a 37 degrees Celsius incubator for a

Sample Characterization, Identification And Performing...
Phase I–Phenotypic characterization 1. Collection of microbial isolates from various sample,
characterization, identification and performing antibiotic susceptibility testing. 2. Performing
Metallo Beta Lactamase (MBL) screening and confirmatory tests. 3. Testing carbapenemase
production phenotypically. 4. Testing microbial isolates against higher and multidrug antibiotics.
Phase II–Molecular Characterization 1. To identify the type of carbapenemase and MBL genes by
PCR 2. To identify the expression of different types of carbapenemase encoding gene in the MDR–
Acinetobacter baumannii. 3. To determine the genetic relatedness of the different strains of
Acinetobacter baumannii typed by pulse field gel electrophoresis (PFGE). 4. To identify the new
variants and sub types of MBL and carbapenemase genes analysed by bionumerics software. Phase
III– Data analysis and Publication of results 1. Collection of all the test results, analysis using SPSS
and deriving study conclusion. 2. Publication of the research in high impact factor ISI research
journals. RESEARCH METHODOLOGY: 1. Place of study: The present study will be conducted in
Government General Hospital located which is a tertiary care multi–specialty hospital receiving
patients from the surrounding regions of Wadi Al Dawaser, Saudi Arabia. The genotypic study will
be carried out in collaborative laboratory. The data processing and result analysis will be carried out
in the Department of Medical Laboratory Sciences,

Finding The Minimal Inhibitory Concentration
Finding the Minimal Inhibitory Concentration:
The Process and the Importance
Melissa Bañaga
San José State University
Abstract The purpose of this project was to determine the minimal inhibitory concentration of our
given antibiotic called tetracycline. Tetracycline is a bacteriostatic agent and instead of directly
killing the bacteria, it prevents the growth mainly by inhibiting the synthesis of proteins. We
hypothesized that the minimal inhibitory concentration for our test would range from 4 g/mL to 8
g/mL. This process spanned across four lab meetings located in a biology lab at Duncan Hall of San
José State University. The first lab introduced us to sterile and plating techniques as we prepared and
plated a serial ... Show more content on Helpwriting.net ...
There are many antibiotics out there but it is not enough just to administer as much of an antibiotic
that you want. There are particular numbers needed to achieve the desired and more longer lasting
result.
Introduction
Disease and plague had once completely and disastrously overwhelmed humanity for countless
years. During the era when medicine was not as advanced and sickness and disease had run rampant,
people had resorted to dangerous, unethical and unsanitary methods of healing. Desperate to
alleviate the pain, doctors and healers of the time did what they could to make the pain and
discomfort of the ill disappear. Though the intentions of many doctors was to heal the sickly, they
did not realize there was a more scientific approach of solving many of their problems. On a
microscopic level, there are bacteria and microbes that are attacking the body, causing diseases to
ensue. In order to deal with the issue at hand, scientists had to figure out what could be used to fight
these particular pathogens and not just what, but how much of these antimicrobials should be used.
To better understand this idea of knowing the proper dosage needed for an antibiotic, we exercised
this concept of minimal inhibitory concentration. The minimal inhibitory concentration is the lowest
amount of the antimicrobial concentration needed to prevent the noticeable growth of the
microorganism after incubating the sample overnight. Each group was given a specific antibiotic of

Pink Bacilli Research
Results
Observing under 1000X magnification, the gram stain depicts pink bacilli, as seen in Figure 1.
Figure 2 depicts the growth of a brown bacteria on a MacConkey Agar plate. Figure 3 depicts the
growth of a similarly brown bacteria surrounded by a very thin clear area on a blood agar plate. As
seen in Figure 4, the slant of gram–negative isolate appears mainly clear, with a slight green tint.
Figure 5 shows the methyl red test, which produced no color change. Figure 6 shows the Voges–
proskauer test, which also produced no color change. Figure 7 depicts the lactose test, which
produced no color change and no gas. The Indole test resulted in a red ring and the Hydrogen
Sulfide, H2S, test resulted in a black precipitate, both of which can ... Show more content on
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It is also shown to not ferment lactose. It does not use the mixed acid fermentation or butylene
glycol pathways during breakdown of glucose. According to the gram–negative table, this indicates
that the unknown gram–negative microbe is Proteus vulgaris (5). Comparing these results to known
biochemical characteristics, all results shown are consistent except for the methyl red test, which is
supposed to be positive, indicating that the bacterium does use the mixed acid pathway during
breakdown of glucose. This error could be due to contamination, incorrect amount of methyl red
indicator added, or not being incubated for a long enough

Examples Of Selective Media In Microbiology
Microbiology Lab Report
"Use of Differential, Selective, and Enriched Media in Microbiology"
BIO 3150–700
Dr. Marilu Santos
Introduction:
The purpose of this experiment is to display the numerous special media used to isolate bacterial
types, the differences between closely related groups of bacteria, the practice of sanitation and
aseptic technique in microbiology, the assessment of naturally occurring antibiotics and other
substances, and to correctly pinpoint the characteristics of bacteria and the changes that occur when
placed in different media.
Three different types of media were used in this experiment. Selective media allows for the growth
of one type of bacteria while preventing the growth of another type. An example of selective media
that was used in this experiment is the Eosin Methylene–Blue agar used for the growth of
Escherichia coli. These bacteria are commonly found in places of fecal contamination (CDC). A
control group of Escherichia coli was used as well as swabs from the student's phone and hand.
Another type of media used in this experiment is Differential/Selective media. This media makes
detection easier after the incubation period has occurred. An example of differential/selective media
used in this experiment was Mannitol Salt agar used to isolated the bacterium Staphylococcus
aureus and Staphylococcus epidermidis. The Mannitol Salt agar was streaked with these two control
groups as well as a swab obtained from the student's forehead using a wet cotton swab.
Another type of differential/selective media is the Mueller–Hinton agar plate. In this experiment
three Mueller–Hinton plates were used to compare the antibiotic resistance of different types of
bacteria. The first plate was streaked with Escherichia coli, the second plate was streaked with
Staphylococcus aureus, and the third plate was streaked with Pseudomonas aeruginosa.
The final type of media used in this experiment was an enriched media agar plate. Enriched media is
used to provide an environment in which bacteria is able to grow. These mediums contain nutrients
that other agar plates lack, making growth difficult for certain types of bacteria. A blood agar plate
was used to obtain a swab of the Streptococcus bacteria

Streaking Lab Report
Lab Three: Streaking and Spreading Plates
Introduction
In this laboratory experiment, we was introduce to an introduction to streaking and spreading of
bacteria in agar plates such that single cells can be isolated from one another, each cell can
reproduces to form a visible colony composed of genetically identical clones. Streaking and
spreading bacteria is to obtain individual colonies is usually the first step in genetic manipulation of
microorganisms.
Materials
Petri dishes
Metal transfer loop
Bunsen burner
Test tube rack
Glass spreader
Alcohol
E. coli
Methods
Streaking Procedure
1. The petri dish was labeled.
2. The metal transfer loop was sterilized before obtaining the specimen.
3. The culture was opened and the specimen ... Show more content on Helpwriting.net ...
The spreader was placed in contact with the inoculum on the surface of the plate and was positioned
to allow the inoculum to run evenly along the length of the spreader. The even pressure was applied
to the spreader and the plate was spun by hand.
3. After the spread plate was permitted to absorb the inoculum for 10 minutes, they were inverted
and incubated.
Results
Streaking and spreading bacteria over the surface of the plate diluted the amount of bacteria diluted
until the individual cells were streak and spread of the surface of the plate. From theses, individual
cell a single colony arises. All cells in the colony genetically identical. However, the streaking and
spreading was not quite properly performed, but there were some visible colonies that arise.
Discussion
In order to obtain well–isolated discrete colonies, the quadrant streak and spread technique was
used. This allowed dilution of the original microbial material over the entire surface of the plate. As

the original sample was diluted by streaking and spreading it, over successive quadrants the number
of organism decreases. Usually by the third or fourth quadrant only a few organism were transferred
on the by the inoculating loop and theses produce a few isolated

Antimicrobial Effectiveness Of Spices And Herbs
From the ancient times spices and herbs has been mainly used as natural food preservatives, as food
additives and as flavouring agents. Various spices indicates antimicrobial action against distinctive
sorts of microorganisms. This project proposal gives a literature review considering antimicrobial
action of vital oils generally utilized as spices and herbs. For e.g.: garlic, mustard, bay, cumin,
thyme, basil, oregano, pepper, ginger, sage, rosemary etc. Against most regular bacteria and fungi
that contaminated food (Staphylococcus spp, Salmonella spp, Escherichia spp and many others).
Antimicrobial activity is depending on the content of extracts and essential oils, types of food and
microorganisms, type of spice and herb and chemical composition.
Spices and Herbs Inhibitory Effect
Cinnamon, cloves, mustard Strong
Allspice, bay leaf, caraway, coriander, cumin, oregano, rosemary, sage, thyme Medium
Black pepper, red pepper, ginger Weak Table 1.
Table 1 shows the Antimicrobial Effectiveness of Spices and Herbs from the results of different
investigations.
Introduction:
In the later years, buyers have more knowledge about the food they eat and how it processed.
Manufactured foods are chemical substances which used in food industry to reduce the factors of
discoloration, spoilage and contamination by bacteria, organisms or pathogens. There are more
disadvantages of having manufactured foods therefore there is an interest of replace the
manufactured foods

Potato Dextrose Agar Lab Report
each to be plated on 0.4 g potato dextrose agar (PDA). The PDA (10.26% potato extract, 51.28%
glucose, 38.46% agar, 35.5366 Nm cycloheximide) uses potatoes as a source of nitrogen, potassium,
and glucose as the carbon source, while cycloheximide is used to inhibit fungal growth. Of each
dilution 0.1 ml was plated on PDA and spread with the hockey stick method. Each plate was then
labelled with the dilution factor and placed in the incubator at 37°C for 24–48 hours dependent upon
growth. A seventh plate was created with soil from wet sample, plated in the same manner on PDA.
Four master plates were then created from the serial dilution plates. Potato dextrose agar was
selected and master plates were divided into a grid for isolation of

Pglo Lab Report

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