Unknown Lab Report Free Essays

Unknown Lab Report #1 Unknown #1 April 25, 2012 Microbiology Spring 2012 MCB2010C Unknown #1 Introduction Identity of a microorganism has proven to be very significant. Doing so can help identify diseases and created treatment and cures for such diseases. As a result, various laboratory tests were performed to an unknown microbe (Unknown #1) found in the water of a nearby pond. We will write a custom essay sample on Unknown Lab Report or any similar topic only for you Order Now By identify the microbe, the safety of the water will be known to those around it. Materials and Methods Unknown #1 was found in a nearby pond that was created by an earthquake. Some of the various methods introduced and practiced in class were applied in identifying the microbe. Procedures were followed as stated in the index of the Virtual Unknown Software. First the shape and color of the microbe was noted. This is done to know the morphology of the microbe. After concluding that the microbe was Gram negative rods, it was inoculated into a previously prepared test tube. Then various tests were performed to isolate and identify the unknown microbe. Table 1 shows the tests, purpose, reagents and results of the various tests performed. Each test was performed according to the index of the Virtual Unknown software and the following tests were performed: 1) Adonitol Fermentation 2) Cellobiose Fermentation 3) Maltose Fermentation 4) Lactose Fermentation 5) Raffinose Fermentation 6) Sorbitol Fermentation 7) Sucrose Fermentation 8) Melibiose Fermentation 9) Indole Production Results Table 1: Biochemical Tests Results Test| Purpose| Reagents| Observations| Results| Adonitol Fermentation| To determine if the microbe can ferment the carbohydrate (sugar) adonitol as a carbon source. | None| Turned pink| Negative adonitol test| Cellobiose Fermentation| To determine if the microbe can ferment the carbohydrate (sugar) cellobiose as a carbon source. | None| Turned Yellow| Positive cellobiose fermenter| Maltose Fermentation| To determine if the microbe can ferment the carbohydrate (sugar) maltose as a carbon source. None| Turned Yellow| Positive maltose fermenter| Lactose Fermentation| To determine if the microbe can ferment the carbohydrate (sugar) lactose as a carbon source. | None| Turned Pink| Negative lactose fermenter| Raffinose Fermentation| To determine whether the microbe can use sugar raffinose for carbon and energy. | None| Turned pink| Negative raffinose fermenter| Sorbitol Fermentation| To determine if the microbe can ferment the carbohydrate sorbitol as a c arbon source. None| Turned yellow| Positive sorbitol fermentation| Sucrose Fermentation| To determine if the microbe can ferment the carbohydrate sucrose as a carbon source| None| Turned Pink| Negative sucrose fermenter| Melibiose Fermentation| To determine if the microbe can ferment the carbohydrate (sugar) melibiose as a carbon. | None| Turned Pink| Negative melibiose fermenter| Indole Production| To determine whether the microbe can produce indole from the amino acid tryptophan. | Add five to ten drops of Kovac’s reagent to the test tube. The reagent does not mix with water and forms a thin layer above the broth. The reagent reacts with idole to produce a cherry red ring. | Red ring at the top of broth| Positive indole test| Flowchart Unknown#1 Gram Stain Gram Negative Rod Adonitol Test (-) Positive Negative Klebsiella ornithinolyticaCitrobacter koseri Klebsiella oxytocaEnterobacteraergenes Klebsiella oxytocaEscherichia fergusonii Klebieslla pneumoniae sspKlebsiella ornithinolycia pneumoniaKlebsiella pneumonia Klebsiella oxytocassp ozaenae Klebsiella terrigenaKlebsiella pneumonia ssp Leclercia adecarboxlatarhinosclermatis Klebsiella terrigena Leclercia adecarboxylata Mollerlla wisconsinisis Providencia alcalifaciens Providencia rettgeri Serratia fonticola Serraria rubidea Cellobiose fermentation (+) Positive Negative Buttiauxella agrestis Edwardsiella hoshinae Enterbacter cloacae Edwardsiella ictaluri Enterbacter intermedius Edwardsiella tarda Enterbackter sakazakii Edwarsiella tarda Kluyvera ascorbata biogroup 1 Rahenlla aquatilis Escherichia blattae Escherichia coli Ewingella Americana Morganella morganii ssp morganii Morganella morganii ssp sibonii Proteus mirabilis Proteus myxofaciens Proteus penneri Proteus vulgaris Providencia rustigaianii Providencia stuartii Salmonella bongori Salmonella cholerasuis ssp arixonae Salmonella paratyphi A Salmonella typhi Serratia marcescens Serratia proteamaculans Shingella prtoeamaculans Shingella dysnteriae Shingella flexneri Shingella sonnei Tatumella ptyseos Yersinia pestis Yersina pseudotubercilosis Maltose Fermentation (+) PositiveNegative NoneNone Lactose Fermentation (-) Positive Negative Buttiauxella agresitsCitrobacter amalonaticus bigroup 1 Enterobacter cloacae Enterobacter amnigenus bigroup 1 Enterbacter intermedius Enterbacter vulneris Enternbacter sakazakii Serratia odonifera bigroup 1 Kluyvera ascorbata Serratia plymuthica Rahnella aquatillis Raffinose Fermination (-) PositiveNegative Citovacter amalonaticus biogroup 1Cedecea davisae Enterbacter amnigenus biogroup 1Cedecea lapagei Enterobacter gergoviaeEnterbacter cancerogenus Escherichia vulnerisEscherichia hemannii Serratia odorifera biogroup 1Hafrina alvei Serratia plymuthicaYersinia kristenernii Sorbitol fermentation (+) PositiveNegative Cedecea neteriCedecea davisae Serratia ficaria Cedeca lapagei Yersinia enterocolitca Enterobacter cancerogenus Yersinia frederiksenii Escherichia hermannii Hafnia alvei Yersinia kristensenii Sucrose fermentation (-) PositiveNegative Cedcea neteriEnterbacter amnigenus biogroup 2 Serratia ficariaSalmonella choerasuis ssp hotenae Yersinia enteroclitica Yersinia frederiksenii Melibiose Fermentation (-) PositiveNegative Enterbacter amnigenus biogroup 2Citrobacter freudii Salmonella cholerasuis ssp houtenaeCitrobacter amalonaticus Indole production (+) PositiveNegative Citrobacter amalonaticusCitrobacter freundii Discussion/Conclusion After conducting a variety of various tests, it was concluded that Unknown #1 was Citrobacter amalonaticus. Once the morphology of the microbe was identified, it was isolated and inoculated into a test tube. The nine tests performed on the microbe were done according to the index of the virtual unknown software and the results of each test correlated to the results anticipated. As a result, it was concluded that the unknown #1 microbe was Citrobacter amalonaticus. Citrobacter amalonaticus has a gram negative rod-shape. They can be found in the digestive system and are linked with digestive disorders such as diarrhea. They can also be present in the urinary tract and most commonly causes Urinary Tract Infections. Citrobacter amalonaticus is also known as an opportunist pathogen. Opportunist pathogens are pathogens that do not cause much damage to healthy individuals; however, it can cause very severe damage to those with a weakened immune system. Besides being found in the intestines of humans, Citrobacter amalonaticus is usually found in the ground (soil), in the air and in the intestines of animals such as bats. (1) Citrobacter amalonaticus also has some positive effects. It has the ability to recycle hydrogen. In addition it is capable of breaking down nitrates into nitrites in the nitrogen cycle. This process is commonly used in the biodegration industry. There tannic acid degrades into tanneries. (2) Citrobacter amalonaticus is also capable of combing metals and phosphates. This allows for the harmful substances such as uranium to be removed from water, soil and uranyl phosphate crystals. (2) References: Citrobacter amalonaticus A cousin to Citrobacter Freundii. http://www. citrobacterfreundii. org/citrobacter-amalonaticus/citrobacter-amalo How to cite Unknown Lab Report, Essay examples Unknown Lab Report Free Essays Background: In Jane Horack’s article â€Å"Staphylococcus epidermidis†, S. epidermidis is described as â€Å"gram-positive cocci bacteria that are part of the normal flora on the skin and nasal passages. † The article goes on to say that the species was originally named Staphylococcus Albus by microbiologist Rosenback in 1884. We will write a custom essay sample on Unknown Lab Report or any similar topic only for you Order Now When viewed under a microscope S. epidermidis will appear in chains, pairs, or grape-like clusters (Horak 1). Taxonomically, the species S. epidermidis falls in the genus Staphylococcus, which is in the bacterial family Staphylococcaceae. S. pidermidis is in the phylum Firmicutes, under the Bacillales order. Like many members of the genus Staphylococcus, S. epidermidis is non-motile, as well as non-spore forming (Horak 1). The species is also facultative anaerobes, but not all strains of S. epidermidis will ferment. S. epidermidis is catalase positive, and this sets them apart from other gram-positive cocci, such as Streptococcus. They are also urease positive, cannot utilize Mannitol, and are resistant to several antibiotics (Horak). Staphylococcus epidermidis is considered â€Å"an opportunistic pathogen. It usually has a symbiotic relationship with its host, and for this reason it rarely causes diseases and is usually considered nonpathogenic (Avdic, Habes, and Avdic 3885). Rece ntly though, the microorganism is becoming the common cause of nosocomial infections. In â€Å"Microbiology: with diseases by taxonomy†, Richard Bauman defines a nosocomial infection as â€Å"a disease acquired in a healthcare setting. † These infections are often found with implants and plastic items that have inserted into the body, such as catheters, pacemakers, and urinary catheters (Avdic, et. l 3885). â€Å"The ability of this microorganism to causes infections is primarily due to its ability to form biofilms on synthetic surfaces of implanted medical devices† (Avdic, et. al 3885). Biofilms are considered the primary residence of microorganisms in nature, and are composed of numerous microorganisms (Bauman 173). Numerous studies clearly show that a large number of hospital strains have the ability to form multilayered biofilms on inert surfaces (Avdic, et. al 3886). The formation of biofilms has serious clinical consequences and is the cause of many persistent and chronic infections particularly in patients who have long been hospitalized or are in critical condition. (Avdic, et. al 3886). These infections are more severe in patients with compromised immune systems, such as the elderly, young children, or cancer patients (Horak 2). One of the biggest concerns with S. epidermidis is the frequency of these nosocomial infections. As these infections become more prevalent, antibiotic resistance is quickly increasing (Horak). In Michael Otto’s article â€Å"Staphylococcus epidermidis – the â€Å"accidental† pathogen†, he suggests the reason for this resistance is due to antibiotic overuse. Vaccination and decolonization has been discussed as preventative measures, but there is currently no vaccine for S. epidermidis, and the effect of decolonization may prove counterproductive. If S. epidermidis was removed from the normal human flora, it might allow other microorganisms to colonize (Otto). For those reasons, it is commonly agreed that best way to deal with S. epidermidis infections is by a series of preventative measures. Staphylococcus Epidermidis biofilm Staphylococcus Epidermidis biofilm Materials and Methods: There were series of tests performed before the unknown could be properly identified. The unknown was first plated using the antiseptic technique. The aim of the antiseptic technique is to separate colonies of each microorganism so it may become easily distinguished and pure colonies will isolated. The process began by flaming the inoculating loop over the Bunsen burner to kill any pre-existing bacteria. The neck of the vial containing cells of the unknown was also flamed to kill any existing microorganism. The inoculation loop was then dipped in the nutrient broth, and neck of the vial was flamed once again. After a brief cool-down, the inoculating loop was streaked across a nutrient agar plate using the Quadrant Streak method. The Quadrant Steak method involves smearing the bacteria in quadrant one. The quadrant begins at the top of the agar plate, the loop is smeared into a downwards motion. Following this process, the inoculating loop was flamed once again. After the loop cooled down, a single colony from quadrant one was targeted and smeared into quadrant two, and was followed by flaming the inoculating loop. The process was repeated to smear bacteria into the remaining quadrants three and four. The agar plate was then turned upside to prevent condensation from dipping onto the culture and causing contamination. The process was repeated four additional times on to a phenylethyl alcohol agar (PEA), a Mannitol slat agar (MSA), a MacConkey agar (MAC), and an eosin methylene blue agar (EMB). Following the incubation of the several agar plates, the growth of colonies allowed for the next step in diagnosis – staining. The simple stain was performed first and can be used to determine cell shape, size, and arrangement. The process required the inoculating loop to be flamed once more, and a loopful of the culture was smeared onto a clean slide. A drop was water was applied onto the slide, and mixed to create a thin film. After the water had dried, the slide was heat-fixed. The heat-fixture ensured the bacteria would not be removed during the staining process. The slide was then covered with the agent Methylene blue for thirty seconds, and the excess dye was removed with water the gentle blotting. S. epidermidis – simple stain S. epidermidis – simple stain Following the simple stain, the Gram Stain was performed. The Gram Stain is a procedure used to determine the presence or lack of peptidoglycan in the cell walls of a bacterium. The process began with flaming the inoculation loop, and smearing a colony of bacteria onto a clean slide. A drop of water was applied and smeared to create a thin film. After the water had completely dried, the slide was then passed through the Bunsen burner. This adhered the bacteria to slide. The slide was then stained with Crystal Violet for sixty seconds, and completely rinsed with distilled water. The slide was rinsed with distilled water after every step. After the crystal violet, the slide was covered in Iodine for sixty seconds, and rinsed. Following the Iodine, the slide was decolorized with 95% ethyl alcohol-acetone for three to five seconds, and rinsed. The last step was the counterstain with Safrain for thirty seconds, rinse, and gently blot off the excess water. If the bacteria retained a purple stain, it was considered Gram-positive. However, if the bacteria possessed a pink or red stain, it was considered Gram-negative. S. epidermidis Gram-stain S. epidermidis Gram-stain How to cite Unknown Lab Report, Papers