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Chapter 1 : Isolation and Identification of O1 from Fecal Specimens

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Isolation and Identification of O1 from Fecal Specimens, Page 1 of 2

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Abstract:

The introduction of cholera into Latin America during 1991 and 1992 has underscored the need for public health and clinical microbiologists worldwide to be familiar with methods for the isolation and characterization of . This chapter summarizes basic laboratory techniques for cholera and is intended to provide a reference for the microbiologist who has not specialized in these procedures. Isolates of serogroups other than O1 or O139 are occasionally associated with sporadic diarrheal illness, limited outbreaks of diarrhea, and extraintestinal infection. Fecal specimens (stool or rectal swabs) should be collected as early as possible in the illness, as the number of O1 in the stool begins to decrease soon after the onset of symptoms. Alkaline peptone water (APW) is a simple and inexpensive medium that has been used for transport as well as enrichment of specimens for . Although V cholerae O1 grows on a variety of commonly used agar media, isolation from fecal specimens is more easily accomplished by use of a selective plating medium. Typically, strains of ferment glucose and sucrose without producing gas. Fermentation tests should be read at 24 h but if negative may be incubated for up to 7 days. Blood agar plates containing 5 to 10% sheep blood may be used to test O1 for hemolysis.

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1

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Clinical and Public Health
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Antimicrobial Susceptibility Testing
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Figure 1

Overnight colonies of on TCBS agar are yellow because of sucrose fermentation. The colonies are typically large (2 to 4 mm in diameter), circular, smooth, glistening, and slightly flattened.

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
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Image of Figure 2
Figure 2

On TTGA, colonies of are gray, flattened, and surrounded by a cloudy zone caused by the production of gelatinase.

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
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Figure 3

Procedure for recovery of from fecal specimens. , If the APW cannot be streaked after 6 to 8 h of incubation, subculture at 18 h or sooner to a fresh tube of APW, incubate for 6 to 8 h, and then streak to TCBS.

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
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Figure 4

The string test is a simple method for differentiating species, which are almost always positive, from species, which are negative.

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
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Figure 5

will grow in nutrient broth without added NaCl (0% NaCl; tube B), but growth of this species is stimulated by the addition of 1 % NaCl (tube A). V. parahaemolyticus and other halophilic vibrios will not grow in nutrient broth with 0% NaCl (tube C).

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
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Tables

Generic image for table
Table 1

Characteristics of

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
Generic image for table
Table 2

Serotypes of serogroup O1

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
Generic image for table
Table 3

Differential characteristics of selected members of the and

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
Generic image for table
Table 4

Differentiation of classical and El Tor biotypes of serogroup Ol

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
Generic image for table
Table 5

Hemolytic activity of Ol classical and El Tor biotypes

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1
Generic image for table
Table 6

Zone size interpretative standards for the for selected antimicrobial disks (not validated for O1)

Citation: Kay B, Bopp C, Wells J. 1994. Isolation and Identification of O1 from Fecal Specimens, p 3-25. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch1

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