Alexandra Barbera, Lauren Schoeppner

Diagnosis of an Enteric Infection

Providencia alcalifaciens

Table 1: Bacterial Assays on Patient Sample

Assay Observations Interpretation (Positive/Negative)

Lactose Fermentation Cloudy pink broth, no gas bubbles, alkaline Negative

Glucose Fermentation Cloudy yellow broth, no gas bubbles. Acidic. Positive

Sucrose Fermentation Cloudy yellow broth, no gas bubbles, acidic. Positive

Indole Production Cherry red when Kovacs reagent was added Positive

Methyl Red Presented with a redish-orange color. Uniformly transparent Positive

Citrate Use The agar tuned blue, indicative of alkaline Positive

H2S production The medium turned a light brown color, but no black precipitate was present Negative

Motility The medium was heavily concentrated at the stab, but the medium was slightly cloudy Positive

Urease Presented with a tan-orange color indicative of a neutral pH Negative

Lysine Decarboxylase The medium was equivalent to that of the control tube. No purple was present. Brown with a neutral Negative
pH.

The lactose tests determined that the organism was not capable of fermenting lactose, but both the

glucose and sucrose fermentation tests revealed that the organism was capable of fermenting glucose and

sucrose (2). The indole test is part of the SIM assay and looks for the organisms ability to break down

tryptophan into indole, pyruvate and ammonia. The organism was found to break down tryptophan by the

Kovacs reagent reacting with indole to produce a cherry red appearance (2). The SIM assay also tests for

the production of hydrogen sulfide, H2S, by Fe2+ ions in the medium reacting with any sulfide in the

organism to form a black precipitate that is visible to the human eye (2). This particular organism was

found to be negative for the production of sulfide, but the medium did turn a brownish tint, which may

have resulted from incubating the tube longer than the desired time. The last test that is used by the SIM

agar looks at the organisms ability to be motile, or possession of a flagellum to move about in the

semi-solid agar (2). This organism tested positive for motility because there was a slight cloudiness to the

agar and that the stab, where it was inoculated, was not clearly defined in the agar. The next test, the
methyl red assay, differentiated between species producing small and large amounts of organic acids as a

result of glucose fermentation using methyl red, a pH color indicator that utilizes a ratio of red to yellow

molecules to determine the concentration of organic acids (2). This organism was a orange-red color

making the test a slight positive with a lower pH than those that did not produce as much acids. The

citrate assay detects the organisms ability to use citrate as its sole carbon energy source by the pH

indicator bromthymol blue displaying a blue color if positive for ammonium salt and citrate metabolism

and a green color if negative (2). The organism being tested displayed a positive result meaning that it

was able to utilize both the citrate and ammonium salts. The urease test determined the organisms ability

to produce urease, an enzyme that breaks down urea, by using phenol red as the indicator (2). The

organism in question resulted in a negative urease test, meaning that there was a slow or absent

production of urease. The last test utilized was the lysine decarboxylase assay, which looks for the

production of cadaverine and CO2 from the removal of a carboxyl from lysine. The test has a negative

result as it was identical to the control with no purple present.

These tests helped to diagnose the patient, P, with the bacteria Providencia alcalifaciens as all test

results correctly matched with what is characteristic of P. alcalifaciens. The genus Providencia is a

member of the family Enterobacteriaceae. This family contains species that are found in the large

intestine as a part of the normal microbiota in healthy people but also contains species that are pathogens

to the body (2).

Where the pathogen may be found outside the body: (1)

Providencia species are particularly found in water sources and soil that animals and humans

have came into contact with.

Which biochemical assays might be relevant to the activity of the organism within the body and

explanation:
 The phenol red tests are useful in determining the organisms ability to ferment sugars such as

sucrose, glucose, and lactose and if they produce a gas while doing so (2). Since the organism

tested negative for lactose but positive for glucose fermentation, this could show that the

organism does not produce the enzyme beta-galactosidase (2). The phenol red test also shows that

the organism is an aerotolerant anaerobe by the production of acids through the fermentation

process in an open test tube (2).

The motility test is useful in determining the organisms ability to move from place to place

within the body (2).

The urease test and MacConkey agar plating may be useful in differentiating between P.

alcalifaciens and other providencia species. This is because most providencia bacteria appear

urease positive. They also show lactose fermentation on MacConkey agar. However, P.

alcalifaciens is urease negative and does not show lactose fermentation on MacConkey agar (1).

Determine if your organism is normally found in humans or if it is only associated with disease:

P. alcalifaciens is an invasive pathogen and is typically not seen as part of the normal microbiota

of the intestines (2).

P. alcalifaciens strains have been shown to be a cause of traveler's diarrhea (5).

The invasive strains caused actin condensation in the infected tissues (3).

In a study involving 14 strains, 17% were highly invasive, 22% were moderately

invasive, and 61% were weakly or noninvasive (4).

Infects the intestinal mucosal epithelium (5).

Describe the signs and symptoms of the disease: (1)

Diarrhea, most common sign and symptom

Gastroenteritis (4)
 Can either be mild or moderately severe (1) and typically resolves on its own with no

further treatment necessary

Abdominal pain

Low fever (present in about 25% of infected patients)

Tenesmus is not a common sign but it is associated with P. alcalifaciens infection

Describe the mode(s) of transmission of the disease: (1)

P. alcalifaciens is transmitted through the fecal-oral route

Typically a food borne illness caused by unsanitary practices of food preparers

Has an incubation period of about 69 hours

Describe the treatment of the disease: (1)

Typically resistant to some penicillins, and tetracyclines

Can be treated with aminoglycosides, and most cephalosporins

Susceptibility testing should be completed to ensure that the particular strain of P. alcalifaciens is

not resistant to certain drugs of choice

Very little is known about treatment of P. alcalifaciens

Based on in vitro testing though, it has been found that P. alcalifaciens is most likely be

susceptible to a second generation cephalosporin

Also, all isolates of P. alcalifaciens were sensitive to chloramphenicol, gentamicin, neomycin and

norfloxacin

Diarrhea due to P. alcalifaciens can also be treated with ampicillin taken four times a day

at a dosage of 500mg per dose

Replacement of electrolytes and fluids are also incredibly important to the treatment of P.

alcalifaciens because of the possibility of dehydration due to excessive diarrhea

 Works Cited

1. Armbruster, C. E., & Mobley, H. (2017). Providencia species. Retrieved November 3, 2017, from

E-Sun Technologies Inc.

2. Keating, S. (2016). Microbiology: the laboratory experience. New York, NY: W.W Norton and

Company Inc.

3. Guth, B. E., & Perrella, E. (1996). Prevalence of invasive ability and other virulence-associated

characteristics in Providencia alcalifaciens strains isolated in Sao Paulo, Brazil. Journal of

Medical Microbiology, 45(6), 459-462. doi:10.1099/00222615-45-6-459

4. Janda, J. M., Abbott, S. L., Woodward, D., & Khashe, S. (1998). Invasion of HEp-2 and Other

Eukaryotic Cell Lines by Providenciae: Further Evidence Supporting the Role of Providencia

alcalifaciens in Bacterial Gastroenteritis. Current Microbiology, 37(3), 159-165.

doi:10.1007/s002849900357

5. Albert, M., Alam, K., Ansaruzzaman, M., Islam, M., Rahman, A., Haider, K., . . . Montanaro, J.

(1992). Pathogenesis of Providencia alcalifaciens-induced diarrhea. Infection and Immunity,

60(12), 5017-5024. Retrieved November 3, 2017.