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. 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 carbon 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