Vibrio parahaemolyticus is an extremely dangerous pathogen that lives in the ocean. Vibrio is known to be an extremely motile and are used to various environments with fluctuating calcium concentrations (Gode-Potratz, Chodur, McCarter). Vibrio’s ability to move extremely efficiently and be able to survive in multiple environments allows for this pathogen to be extremely flexible and infect individuals multiple ways. Thus that is why Vibrio is the leading cause of seafood-borne gastroenteritis worldwide (Gode-Potratz, Chodur, McCarter). Vibrio’s ability to be diverse and unique with its outer membrane proteins is what allows it to be a good test subject in understanding how genes are turned on and off in various environments. Therefore, if we …show more content…
To study the variation gene expression, we used Vibrio parahaemolyticus, calcium, centrifuge, spectrophotometer, mini-protean unit, SDS-PAGE, and UV transilluminator. During the first week of the procedure, you must come in around 24 hours before actually starting your procedure so you have adequate time for bacterial growth. The process consists of multiple steps of adding different chemicals to your bacteria and centrifuge it after add a chemical each time for different lengths of time. After all of the steps, I added 209 mL of LSB (Laemelli Sample Buffer) so that all test runs have equal amount of cells. Final step of this week is freezing our samples. While week two consisted of using our sample from previous week to make a gel plate that will go into the mini-protean unit. The mini-protean unit will distribute the vibrio in a way we can read its gene expression. After the gel is created in the mini-protean unit you must stain and de-stain the gel so it can be taken a picture of. To take the picture we use a UV transilluminator. For the standard procedure, reference (Department of Biology.
I identified the genus and species of an unknown bacterial culture, #16, and I applied the following knowledge of morphologic, cultural and metabolic characteristics of the unknown microorganism according to the laboratory manual as well as my class notes and power point print outs. I was given an incubated agar slant labeled #16 and a rack of different tests to either examine or perform myself; the tests are as follows: Gram Stain; Nutrient Gelatin Test; Carbohydrate Fermentation; Dextrose, Lactose and Sucrose; IMVIC tests; Citrate, Indole, Mythel-Red and Vogues Proskauer test; as well as a Urease and TSI Test. Materials and Methods/Results Upon receiving the Microorganism (M.O.) #16, I prepared a slide by cleaning and drying it. Then, using a bottle of water I placed a sterile drop of water on the slide and used an inoculating loop, flame sterilized, I took a small sample of the unknown growth in my agar slant and smeared it onto the slide in a dime sized circle and then heat fixed it for ten minutes.
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
Making H. pylori a vital microorganism to research in order to expand the study of microbiology and its interaction with humans. According to Blaser, the H. pylori “is a group of extremely varied strains cooperating and competing with one another. They compete for nutrients, niches in the stomach and protection from stresses.” There can be a variety of strains found in a single stomach, and even though they appear identical, their genes are very different.
The first day an unknown sample was assigned to each group of students. The first test applied was a gram stain to test for gram positive or gram-negative bacteria. The morphology of the two types of bacteria was viewed under the microscope and recorded. Then the sample was put on agar plates using the quadrant streak method for isolation. There were three agar plates; one was incubated at room temperature, the second at 30 degrees Celsius, and the third at 37 degrees Celsius. By placing each plate at a different temperature optimal growth temperature can be predicted for both species of bacteria.
Streak plate technique was used to isolate pure culture for each bacteria (2). The Gram stain was used to determine Gram reaction and morphology of each bacteria (2) Selective and differential media such as, salt agar, MacConkey agar and blood agar were used for bacterial identification (2). Gelatin deeps were inoculated to detect production of gelatinase (2). Starch Agar plate were inoculated to detect amylase (2). Ocular reticle used to determine bacteria size (2). Motility deeps were inoculated to detect motility on bacteria (2). Thioglycollate broth used to determine oxygen requirements (2). Carbohydrate fermentation
...et light. If the LAA plate glows green under exposure to ultraviolet light, then we can conclude that our unknown insert piece of DNA would be the kan gene. If it does not glow green under exposure to ultraviolet light, then then we streak the colony from our LAA plate onto the LAC plate using a sterile glass spreader. When the LAC plate is dray, we place it upside down in the microfuge rack so that it can be incubated at 37 ºC. Incubation at 37 ºC will allow the transformed bacterial cells to grow. If we see bacterial growth on the LA plate containing chloramphenicol, we can conclude that our unknown insert piece of DNA would be the cat gene, since the cat gene is resistant to chloramphenicol. Afterwards, we then grab the microfuge tube labeled NP and repeat the aforementioned steps shown above pertaining to the LA plates. This would be considered our control.
They can be found anywhere and identifying them becomes crucial to understanding their characteristics and their effects on other living things, especially humans. Biochemical testing helps us identify the microorganism present with great accuracy. The tests used in this experiment are rudimentary but are fundamental starting points for tests used in medical labs and helps students attain a better understanding of how tests are conducted in a real lab setting. The first step in this process is to use gram-staining technique to narrow down the unknown bacteria into one of the two big domains; gram-negative and gram-positive. Once the gram type is identified, biochemical tests are conducted to narrow down the specific bacterial species.
Cueto may have a potential claim that the OER’s interpretation of the enabling act and ensuing actions exceeded the authority delegated to the OER by Congress. The OER’s notice of liability refers to Section (3) of the statute, and requires the OER to provide to Cueto “notice of the factual basis for the finding.” However, Cueto may argue that the OER failed to comply with this provision of their own enabling statute. The OER’s notice simply related to Cueto that their liability was assumed due to Cueto’s ownership of the pipeline. If Cueto was to challenge the notice per Sections (3) and (4) of the statute, it is highly unlikely the OER’s only evidence at the hearing would be Cueto’s ownership of the pipeline. It is reasonable to interpret that the “factual basis” described in the statute for Cueto’s liability that would be required for the OER to prevail at the hearing is the same or similar factual basis the OER must provide to a responsible party upon notice. If simply noting Cueto’s ownership of the
The first step to the unknown is selecting an actual organism. The best way to select a culture is based on a high-quality distribution. Equally important, shaking up the broth tube facilitates in the distribution. Upon selection, a gram check for purity is performed. Step by step instructions for this procedure can be found in Benson’s, Microbiological Applications p. 99. Furthermore, an aseptic technique must be performed for this test and the entire tests following the unknown. The purpose of this test is to differentiate between gram positive and gram-negative bacteria. The key indicator of gram-positive bacteria is a purple stain and a pink stain for gram-negative bacteria. A slide is viewed with a microscope under oil immersion. Equally
With Egyptian woman being of one aspect of a multi-dimensional look at woman of the ancient times, woman who wrote classical literature in India also played an important part in providing future generations with stories and poems that were skillfully explained with suggestions, compared to the graphic descriptions that were seen in Ancient Egyptian love poems. One of the ancient Indian authors who perfected this technique is Vikatanitamba. Even though Vikatanitamba is similar in evoking passion within her poems, like the ancient Egyptians, she shows her able show her multi-dimensionality, but discretely. One poem where this was skill was shown was “As he came to bed”. Within the first line, she is able to subtly explain her dress falling off
Feminist theories are a way to review works and analyze the ways in which dominant patriarchal themes and oppression of women are prevalent. By reviewing certain stories with feminist theories in mind, the deeper meaning and hidden agendas of some authors can be found. In certain stories, authors give their female characters a traditional passive voice and glorify the patriarchal ideal. Feminist theories also help to recognize certain gender stigmas that are presented in stories and how the authors or narrators tackle those traditional gender roles.
Vitruvius wrote Document 31 Constructing a Harbor around the first millennium B.C.E. He provides a list of instructions on how to build a good harbor. The passage relates to Humphrey’s transportation section on chapter five because harbors played a huge role in early civilizations. Harbors are extremely important because they protect ships from severe weather conditions. It would be a problem for sailors if they encounter bad storms and rainfall because it can cause a lot of damage on their ships. Ever since early people started using transportation by sea, they always wanted a strong harbor to provide protection for their ships and other properties. One thing the passage shows about the ancient technological process for harbors is “retaining
Stout, M.A, et al. "Microbiology Lab Notebook". Lab handbook. University of Texas. Arlington. 2014. Print.
Every second, approximately 1023 viral infections occur in the ocean(8). These infections affect the mortality of marine organisms and are subsequently a major force behind global geochemical cycles and the structure of microbial populations and communities(4, 5, 8). Microorganisms constitute 90% of the living biomass in the sea, and it is estimated that 20% of this biomass is eliminated by viruses every day(8).Viruses influence the mortality of bacteria in the ocean and therefore regulate both bacterial populations and those in subseq...
In the world, there are approximately 4,740 species of frogs that can most likely be found in warm, tropical regions near the equator (). Frogs mostly remain near water or moist areas because they are amphibians. Frogs breathe and absorb water through their skin to collect vital nutrients that they need to survive. A frog is important to an ecosystem because they serve as a predator and prey, but what will happen to the ecosystem if all the frogs are extinct? Over 100 different species have gone extinct, and approximately 287 species of amphibians around the world have been detected with the newly found fungus known as Chytrid ().