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Chapter 13 : Phages and Bacterial Vaginosis

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Abstract:

Bacterial vaginosis (BV) was previously called nonspecific vaginitis or -associated vaginitis. Symptomatic women report an offensive, fishy-smelling discharge, which is most marked after unprotected intercourse or at the time of menstruation. It is unknown how BPDE promotes the induction of prophages in lysogenic lactobacilli. The study of the lambda phage, however, may shed some light on this question. The number of vaginal lactobacilli can be reduced in a smoking woman: by inducing the lysogenic strains to lyse and release phages or when the released phages may in turn lyse other strains by infection. The relationship between smoking and BV may be much more complicated, since smoking can weaken the human immune system and also cause many other health problems. The author and coworkers have documented that phages infect vaginal under in vitro conditions. This suggests that phage infection may occur in vivo as well. DNA evidence of phages were sought in different vaginal that would suggest an infection and/or transmission of phages in women and evidence that phages are released from the male urinary tract. The major difference is that lysogenic bacteria are capable of releasing phages, either spontaneously or upon induction, such as by cigarette chemicals. Collectively, this study provides initial evidence that vaginal phages can be transmissible infective agents. In summary, some vaginal and yogurt lactobacilli are lysogens that spontaneously release phages. Future studies should include establishing phage taxonomy, characterizing phage diversity, determining phage genomic sequences, and studying interactions between phages and lactobacilli.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13

Key Concept Ranking

Bacterial DNA Replication
0.4436546
Linear Double-Stranded DNA
0.4288957
Pelvic Inflammatory Disease
0.42570385
Agarose Gel Electrophoresis
0.42497495
Urinary Tract Infections
0.4198723
0.4436546
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Figures

Image of FIGURE 1
FIGURE 1

Protein profiles of some representative strains by SDS–10% PAGE. M, protein molecular weight markers. Lanes 1 to 17, vaginal strains: 1, KC23T; 2,TL152; 3,TL145a; 4,TL143b; 5,TL114; 6,TL127a; 7,TL109b; 8,TL60a; 9,TL27; 10,TL23b; 11,TL23a; 12,TL33a; 13,TL13; 14,TL102; 15,TL76; 16,TL74c; 17,TL34c.At the bottom of the gel, the species identification of each strain is indicated by a letter, as follows: C, ; G, ; J, .

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 2
FIGURE 2

Electron micrographs of vaginal phages.A1, ϕkc21T;A2, ϕkc12a; B1, ϕkc39; B2, ϕkc7a. Bar, 50 nm.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 3
FIGURE 3

DNA fingerprinting of vaginal phages. The phage DNAs were digested with RI. Lanes: M, molecular weight DNA markers; 1, ϕkc5a; 2, ϕkc7a; 3, ϕkc12a; 4, ϕkc21T; 5, ϕkc23T; 6, ϕkc31; 7, ϕkc39; 8, ϕTL32b; 9, ϕTL33a; 10, ϕTL34; 11, ϕTL72a; 12, ϕTL74c; 13, ϕTL75a; 14, ϕTL76; 15, ϕTL122b; 16, ϕTL125; 17, ϕTL138; 18, ϕTL141. Note that lanes 10, 12, and 14 show identical DNA patterns.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 4
FIGURE 4

Electron micrograph of negatively stained phage ϕy8 from a name-brand yogurt in the U.S. food market. Bar, 100 nm.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 5
FIGURE 5

DNA fingerprinting of phages isolated from sewage. Lanes: 1, ϕskc2; 2, ϕskc3; 3, ϕskc5; 4, ϕskc6.The estimated genomic sizes of the four phages are between 31 and 44 kb.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 6
FIGURE 6

phage induction by BPDE. The phage induction frequencies were plotted as a function of BPDE dosage in twofold increments. The zero dose indicates the spontaneous induction frequency.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 7
FIGURE 7

Comparison of maximal phage release frequencies among spontaneous, BPDE, and mitomycin C inductions. The concentration of mitomycin C was constant, at 200 g/ml.The concentrations of BPDE for maximal phage release were 1 to 2 µg/ml for ϕkc5a, 16 ng/ml for ϕkc7a, and 6 µg/ml for ϕy8.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 8
FIGURE 8

phage analysis by agar-drop assay. The phage ϕkc5a (released from the vaginal strain KC5a) was dropped onto another vaginal strain, KC1b, which served as an indicator. Upper left region, plaques caused by mitomycin C as a positive control; upper right region, phages released spontaneously; bottom region, phages induced by BPDE.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 9
FIGURE 9

In vitro BV model. (A) Without phage, KC13 remained dominant throughout the 12-h growth period in the coculture system. (B) After the addition of phage ϕkc39, the bacterial dominance shifted from to .

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 10
FIGURE 10

Evidence of possible in vivo phage infection of vaginal lactobacilli in one woman. (A) A protein profile analysis of the two host lactobacilli (1, KC12a; 2, KC12b) showed different patterns, indicating that they are different strains. M, protein molecular weight standards. (B) Restriction enzyme analysis of DNAs isolated from two phages (1, ϕkc12a; 2, ϕkc12b) of the two vaginal lactobacilli of this woman, showing identical electrophoretic patterns.M, DNA molecular weight standards. (C) Electron microscopic image of phage ϕkc12a. Bar, 50 nm.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Image of FIGURE 11
FIGURE 11

Evidence of possible phage transmission among different women.(A) Protein profile analysis of three host lactobacilli (1, TL34; 2, TL74c; 3, TL76) showing two different patterns. M, protein molecular weight standards. (B) Restriction enzyme analysis of DNAs isolated from three phages (1, ϕTL34; 2, ϕTL74c; 3, ϕTL76) of vaginal lactobacilli of the three women, showing identical electrophoretic patterns. M, DNA molecular weight standards. (C) Electron microscopic image of phage ϕTL76. Bar, 50 nm.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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Tables

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TABLE 1

Lysogens among different species in vaginal isolates from women in the United States and Turkey

Among Turkish lactobacilli, the obligate anaerobes were and ; among U.S. lactobacilli, the obligate anaerobes were and .

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 2

Infection of vaginal lactobacilli by 67 phages from the United States and Turkey

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 3

Morphotype-specific primers for vaginal phage classification

For determinations of phage sequence data for PCR analysis, genomic DNAs of four phages, representing each morphotype, were digested with Sau3A1.The DNA fragments were cloned into pUC18.A plasmid that carried an insert of about 1 kb was selected for each phage.The sequence was determined by the automated sequencing facility at the University of Missouri, Kansas City.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 4

Commercial products analyzed for phages

Among the 20 yogurt products tested, 11 were found to release phages, of which 7 infected vaginal lactobacilli under in vitro conditions. For the protection of the reputations of these commercial products, however, the sources of these phages are not specifically indicated.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 5

Comparison of three prolate-headed phages ( )

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 6

Origin and isolation of sewage phages

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13
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TABLE 7

Two groups of women and their vaginal floras, species, and phage types

The three Turkish cities Giresun,Trabzon, and Gumushane are separated by about 100 km.The three Turkish women are not related.

Citation: Tao L, Pavlova S, Kiliç A. 2005. Phages and Bacterial Vaginosis, p 256-279. In Waldor M, Friedman D, Adhya S (ed), Phages. ASM Press, Washington, DC. doi: 10.1128/9781555816506.ch13

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