3.1. Screening for Bi NPs producing microorganism
The obtained results of the screening step revealed that one sample [taken from salty marsh land around Shahr-e Babak, Kerman, Iran] contained a bacterial strain (Fig. 1a) capable to reduce Bi3+ ions into Bi0 which was evident from color change from yellowish into dark brown (Fig. 1b). Similar results were observed when the selected isolate was cultivated in the culture flask including Bi3+ ions compared to that of culture media without Bi3+ ions (Fig. 1c and Fig. 1d). Lack of brown color after culturing the selected isolate in the Bi3+ ions free culture media (Fig. 1a and Fig. 1c) confirmed that the produced color was not related to the pigment production by the selected isolate but
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Among the bacteria, Yersinia enterocolitica deposited elemental bismuth during the exponential phase of growth [28]. The anaerobic bacterium C. acetobutylicum was found to be able to produce hexagonally shaped Bi2S3 nanostructures with the average size of 440–500 nm [16]. Nazari et al. [5] isolated a marine Gram-negative bacterial species (S. marcescens) capable to form Bi NPs with the average particle size of 150 nm. The ability of bacterial strains of Delftia family like D. acidovorans for reduction of Au3+ ions to Au0 has been previously reported [29]. It was showed that D. acidovorans was able to increase its survival in the presence of toxic Au3+ ions by secreting delftibactin which facilitates the reduction of Au3+ ions to less toxic elemental Au0 nanoparticles [29].
In order to determine the related possible mechanism(s) involve in Bi NPs biosynthesis in the present study, Delftia sp. SFG was cultivated in the presence of nitrate reductase inhibitor (piperitone) and Bi3+ ions and the color change of both the culture broth and produced biomass was evaluated. The culture broth of Delftia sp. SFG which isolated after 72 h cultivation from the cells was not able to reduce the added bismuth subnitrate to Bi NPs. Thus, the presence of microbial cells is critical and Bi NPs was biosynthesized by Delftia sp. SFG biomass. Different intracellular mechanism for biologically synthesis of NPs by microorganisms has been
There are many differents ways to identify a bacterial unknown and many different situations where identification would be beneficial. One way to identify bacterial unknowns is to perform biochemical tests. In this experiment multiple biochemical tests were done, by performing these tests on the bacterial unknown received the two different bacteria were then identified. The citrate test is done to test the ability of organisms to use citrate as a carbon source. This test uses Simmons citrate agar, the agar contains sodium citrate as the only carbon source and has bromothymol blue as the pH indicator. The organisms that use citrate as a carbon source use the enzyme to transport the citrate into the cell. The cells converts ammonium dihydrogen
The first result of importance was the result of the Gram stain. The observations of the unknown bacteria from the slant culture after Gram staining showed that the unknown bacteria were Gram negative bacilli (Image 1). After determining the unknown bacteria was Gram negative, an oxidase test was conducted on a sample from the slant culture. The cotton swap with the sample of bacteria did not change color when the oxidase reagent was applied, thus providing a negative result. With a negative oxidase test, further tests were conducted to determine various characteristics of the unknown bacteria. A MR-VP broth was inoculated with a sample from a slant culture of unknown bacteria. After incubation, the methyl red reagent was added to the broth, and the broth turned red, providing a positive result (Image 2). An EMB agar streak plate was inoculated with a sample from a slant culture of the unknown bacteria, and after incubation, growth was found on the plate, providing a positive result (Image 3). A Citrate agar slant was inoculated, and after incubation, growth was found on the media, providing a positive result (Image 4). A Urea agar slant was inoculated, and after incubation, the agar had changed from a peach color to a bright pink color, providing a positive result (Image 5). Using the flowchart (Figure 1) developed from the Table of Expected Results, the lab partners started at the oxidase test. Given the negative result of the oxidase test, the flowchart is
Nitrate reduction was tested for by inoculating a nitrate broth with the unknown gram (-) culture, and allowing growth to take place. Adding 2 drops of both sulfanilic acid and α-napththylamine to the medium if the first test to see if nitrite is present. If nitrite is present, the medium turns red, indicating a positive test. However, if the medium does not change, a second test is performed to see if nitrite was further reduced. In this second test, zinc powder is added to the broth to catalyze the reduction of any nitrate present to nitrite. If nitrate is present when the zinc is added the reduction of this compound will cause the medium to turn red, from the previously added reagents. Red medium on the second addition indicates nitrate was not reduced and a negative test result. However, if the medium does not change after the addition of the zinc, the unknown is positive for nitrate reduction, as the nitrite has just been further reduced, preventing its detection. The result that yielded was positive on the first step.
Citrobacter Freundii is a species of bacteria that can be potentially harmful to humans. It is known to cause meningitis by protruding into the brain and replicating itself (1). The Citrobacter species has also been found as a cause of some urinary tract infections, diarrhea, and even gastrointestinal diseases and symptoms (3). C. Freundii can be located in a wide variety of soils and water (3). Lastly, it is also the cause of many nosocomial infections due to its presence in water (1).
There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that have been learned so far in the microbiology laboratory class for the identification of unknown bacteria. The identification process can be completed with a series of deferential stains and biochemical tests. Creating a dichotomous key helps to limit the amount of biochemical tests done on an unknown organism and by observation
In this lab, the organism that we have been working with is the bacterium, Serratia marcescens. S. marcescens is a member of the Enterobacteriaceae family, and tends to grow in damp environments. S. marcescens is an ideal bacterium to work with in the lab because it reproduces quicker than other bacterium. This bacterium produces a special pigment called prodigiosin, which is red in color. The prodigiosin pigment is intensified when S. marcescens is grown at higher densities. During our experiment, temperature, pH, salinity concentration and oxygen requirements were tested on S. marcescens to measure their optimal growth and prodigiosin production.
Bacteriophages were first discovered about 100 years ago by Frederick Twort and Felix D'Herelle. Bacteriophages or commonly known as “phages” are viruses that infect bacterial hosts. Bacteriophages come in multiple shapes and sizes. But a good amount of them are tailed viruses that contain double stranded DNA. The head of the bacteriophages has a protein shell which is attached to the tail. Some phages differ as some don’t have double stranded DNA they may be single stranded. Phages are incapable of reproducing on their own. In fact, they need a bacterial host to reproduce. Like any virus, bacteriophages are very fixed to discrete hosts. The host bacteria that we are using is called arthrobacter. Arthrobacter is a genus of bacteria found in soil (Pope et al 2016). Arthrobacter can help reduce hexavalent chromium which can cause some symptoms such as irritations to humans. Hexavalent chromium may also cause lung cancer if inhaled. The SEA PHAGE project was broken down into two parts. The first part which we are currently doing is isolating, purifying and amplifying the bacteria from our soil samples that we collected in the beginning of the year.
I inoculated a T-Soy agar with bacteria number 118, for this I used a streak isolation method. Next, in order to distinguish between Gram positive and Gram negative I used a streak isolation technique on a CNA plate, then performed the same exact procedure on a MacConkey plate. The results from the CNA plate showed the Gram Positive bacteria was an Alpha hemolyzer. Next, I used a P Disc on a T-Soy agar inoculated with bacteria 118 and determined the Gram Positive bacteria was not sensitive to P Disc antibiotics. This revealed the Gram Positive bacteria to be Streptococcus Mitis. The results from the MacConkey plate proved the Gram Negative bacteria to be a lactose fermenter. With the Gram Negative bacteria I performed a lysine test with positive results. Next, I performed an ornithine test on the Gram Negative bacteria, with negative results, therefore I concluded the Gram Negative bacteria was Klebsiella pneumoniae.
In the world of microbiology it is vitally important to be able to discern the identities of microorganisms. Not only is it important in a lab setting but as well as in healthcare in general. Properly identify what strain of bacteria a person has will aid in the proper medicine and dose given. Throughout the semester we have learned about different types of bacteria and certain test that can clearly identify them. The purpose of this lab report is to identify a Gram-positive or Gram-negative bacterium. Using all the knowledge of procedures and lab techniques identify the unknown and discuss all the tests you performed.
The objective of this experiment was to identify two unknown bacteria from a mixed culture. Which was done by using the aseptic technique which was very important to avoid any contamination and keeping the workspace clean while culturing bacteria for different tests. To start, I chose a tube which had a solution of mixed culture. I used the flame to sterilize the inoculating loop and dipped it into the tube and streaked for isolation on 2 TSA plates and placed them in an incubator at 37 for 24 hours. Next day I observed the growth of 2 different types of colonies one for each unknown on the two plates. So I picked the best one and labeled it as master plate and discarded the other plate. From the master plate, I subcultured each type of colony
In conclusion, after conducting previously mentioned biochemical tests in order to identify the unknown bacteria, became obvious that unknown gram negative was Proteus Vulgaris. Having eliminated all the bacteria that didn’t show expected results and confirming with such tests as catalase, glucose, indole, citrate, urea, and SIM, it was accurate to name the unknown gram negative. Furthermore, gram positive became obvious it was Streptococcus Pneumoniae. Having eliminated all the bacteria that didn’t show expected results and confirming with such tests as blood agar, catalase, and optochin and bacitracin tests, it was accurate to name the unknown gram positive. I have learned that it is extremely important to be to able to identify what kind
The bases of this experiment was to discover the identify of the unknown from three possible specimens: Klebsiella pneumonia, Escherichia coli, and Enterobacter aerogenes. Utilizaing the T streak technique, the bacteria was isolated into pure colonies for further study. The Gram Stain method was used to identity the morhphology of the bacteria such as the shape and whether the bacteria was Gram positive or Gram negative. Biochemical test were also used to help identify the unknown bacteria. The biochemical test used was the Triple Sugar Iron Agar, Sulfur Indole Motility test, Methyl Red test, Voges-Proskauer test, Citrate test, Urease test, and the Gelatin test. After observing the morphology of the bacteria using the Gram Stain method and utilizing all the possible biochemical test, the bacteria was identified to be Enterobacter aerogenes.
On June 25th, 2015 I chose the test tube labeled #19. This test tube contained an unknown bacterium, and the purpose was to determine the unknown bacterium by the end of the semester. Throughout the course, I ran a series of differential tests that would lead me to discovering the characteristics of my unknown. These tests that I will discuss in this paper are vital to understanding the biochemical mechanisms that different bacteria can perform, therefore helping me identify my bacterium based on molecular differences. During the course of this paper, I will refer to my unknown as unk#19. Also, I would note that aseptic technique was performed throughout the entire experiment and subcultures were regularly made.
Being able to control bacterial growth is something that is important in our everyday lives. As shown in the previous labs, bacteria can grow and create colonies extremely quickly especially in the right environments. By acknowledging this, it is then important to get an understanding of how bacterial growth can be controlled by humans. To control microorganisms it means to inhibit their growth (static) and or kill them (cidal) (Kenneth Todar, 2015); therefore since focusing on bacteria the terms bactericidal and bacteriostatic are both extremely important for this lab. One broad method we will use to control bacterial growth is heat. The amount of heat needed to control bacterial growth is different for different species of bacteria (Kenneth Todar, 2015). Bacteria can also respond differently depending if moist heating method such as an autoclave with steam is used, or a dry heating method such as inoculating a loop over a fire is used (Kenneth Todar, 2015). UV works by damaging the cells DNA, without proper DNA, the cells will die and the object
Life on this planet began with microorganisms. Through millions of years microorganisms have found ways to successfully adapt and survive. These adaptations have created a wide biodiversity, allowing them to basically populate in all places. Why are these microbes so important? Because they shape the history of our world. Some microbes can be deathly to humans while some others are favorable, for example, bacteria that lives in the gut of both humans and animals and helps during the process of digestion (Alfred Brown & Heidi Smith, 2006). Understanding these interactions help scientists to find ways to protect humans from potential deathly pathogens. In order to observe microbes, microscope proficiency and microorganisms’ identification are crucial skills in a microbiology lab. During this laboratory session, samples of environmental and human organisms were inoculated into two different rich media and incubated to their according temperature. After this, appropriate use and calibration of the microscope was performed. Lastly, morphology and size of different species of bacteria, algae, fungi and protozoan were recorded.