1

1

2

Phage specificity of the freshwater fish pathogen Flavobacterium 3

columnare 4

5

Elina Laanto, Lotta-Riina Sundberg and Jaana K. H. Bamford* 6

7

Department of Biological and Environmental Science and Nanoscience Center, 8

University of Jyväskylä, Jyväskylä, POBox 35, 40014-University of Jyväskylä, Finland 9

10

11

Running title: Bacteriophages of Flavobacteria 12

Key words: F. columnare, fish pathogen, bacteriophages, phage ecology 13

14

Address for correspondence: 15

Jaana Bamford 16

Department of Biological and Environmental Science and Nanoscience Center 17

University of Jyväskylä, 18

P.O.Box 35, 40014 University of Jyväskylä 19

FINLAND 20

Tel: +358 14 260 2272 21

Fax: +358 14 260 2221 22

E-mail: jaana.bamford@jyu.fi 23

Copyright © 2011, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.Appl. Environ. Microbiol. doi:10.1128/AEM.05574-11 AEM Accepts, published online ahead of print on 2 September 2011

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Flavobacteria and their phages were isolated from Finnish freshwaters and fish 24

farms. Emphasis was set in finding phages infecting the fish pathogen 25

Flavobacterium columnare for further purposes as phage therapy agents. Host 26

ranges of the flavobacterial phages varied, phages infecting F. columnare being 27

more host-specific than the other isolated phages. 28

29

Species of the genus Flavobacterium are widely distributed in nature and they have been 30

recorded from diverse habitats (3, 19, 30, 32, 36, 37, 38, 39). In general flavobacteria are 31

non-pathogenic, but some species are opportunistic pathogens (2). Columnaris, a disease 32

caused by a fish pathogen Flavobacterium columnare, can cause up to 100% mortality 33

among the salmonid fingerlings (28). Antibiotic treatment must be applied to prevent 34

mass mortalities at fish farms. Despite effective treatment columnaris disease occurs 35

repeatedly during summer (18). Therefore the number of antibiotic treatments and the 36

amount of antibiotics used can be extremely high. Increased resistance to antibiotics at 37

fish farms and their surroundings has been recorded in environmental bacteria (10, 24, 26, 38

31), and antibiotic resistant fish-pathogenic flavobacteria have also emerged (7). To 39

avoid risks related to antibiotic use, enrichment of bacteriophages could be an ecological 40

method to decrease the number of F. columnare infections. To our knowledge this is the 41

first study on European F. columnare phages. 42

43

In this study, a total of 53 flavobacterial isolates were received during the warm water 44

period (May to August) in years 2008-2009 from water samples that included both open 45

freshwater environments (rivers and lakes not connected to fish farming) and three inland 46

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land based fish farms rearing mainly salmonid fingerlings in Finland (Fig. 1, Tables 1 and 47

2). One bacterium (B67) was isolated from diseased fish during 2007. Water samples 48

were cultured on Shieh agar (25) and 1/5 Luria Bertani agar (22), and yellow and orange 49

pigmented colonies selected for analyses. The flavobacterial isolates were subjected to 50

UP-PCR (for methods see references 5, 6, 13) and the 16S rDNA of different groups 51

were sequenced (Table 1). All F. columnare isolates fell into same UP-PCR group, and 52

thus they were further analysed with Ribosomal Intergenic Spacer Analysis (RISA) (8, 53

29). According to RISA, the isolates were assigned into 5 groups (Table 1). According to 54

our data, occurrence of F. columnare seems to be connected to fish farming environment; 55

it was not isolated in natural waters. However, it is likely, that the initial source of F. 56

columnare at the farms is nature, because there is evidence that it is present also outside 57

fish farms (20, 21, Kunttu et al., unpublished data). 58

59

Phages infecting the genus Flavobacterium and their interaction with the bacterial host 60

have been previously described mostly in marine environments (4, 9, 12, 14) but also the 61

phage-host relationship of a fish pathogen Flavobacterium psychrophilum has been 62

studied (27). A total of 49 bacteriophages were isolated from water samples (Table 2). 63

Phages were enriched using flavobacterial isolates from freshwaters, fish farms and 64

previously described F. columnare strains. Phage stocks were prepared and selected 65

phages were grown by infecting the host bacterium (multiplicity of infection 5-10) at 66

proper cell density and concentrated and purified (22). Many of the isolated phages 67

produced low titer lysates, and they were used only for infection tests (see section for 68

host range studies). Phage genomic DNA was extracted (for methods see references 1 and 69

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23) and digested with BamHI, EcoRI, HindIII and PstI. For the genomes that were cut, 70

the genome size was calculated from the resulting restriction profile (Table 2). For the 71

phages that were sequenced (FKj-2, FL-1, FCL-2, FCV-1), approximately 3000 base 72

pairs of each genome (except from FCV-1 1600 base pairs) and BLASTed 73

((http://blast.ncbi.nlm.nih.gov/Blast.cgi, May 2011), no significant DNA-sequence 74

sequence similarity was found in the database. Putative protein-coding genes were 75

analyzed using Vector NTI 11.0.0 (Invitrogen). Best matches for FCL-2 were to a 76

hypothetical protein of the Vibrio phage VP16T (score 57.8) and to a hypothetical protein 77

of the Vibrio phage VP16C (score 55.1). For FCV-1 best match was to a hypothetical 78

protein B40-8030 of a Bacteroides phage B40-8 (score 53.9). Transmission electron 79

microscopy (TEM) was used to study phage morphology. All of the Flavobacterial 80

phages that were characterized were tailed phages of the families of Myoviridae, 81

Podoviridae or Siphoviridae (Fig. 1 and Table 2). Most of these phages had an average 82

head size from 50 to 70 nm but some of the Myovirus isolates had capsid sizes around 83

and larger than 100 nm (FJy-3, FKo-2, FKj-2, FKy-1 and FKy-3). 84

85

Phages infecting F. columnare were isolated only from fish farms during disease 86

outbreaks. These phages might survive inside the host cell during cold water period and 87

start a lytic cycle when nutrients come available for the host cell and enough amount of 88

energy is available. 89

90

Studies on aquatic phage-host interplay have been conducted extensively in the marine 91

environments (15) but is less known in freshwaters (15, 34, 35). In our study the host 92

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ranges differed greatly between the phage isolates. In the initial screening for 93

susceptibility of the bacteria for phages, each bacterium was infected with each phage 94

isolate by spotting the phage lysate on top agar containing the host bacterium. Each 95

bacterium was then infected with each phage using plaque assay. Some of the isolated 96

phages infecting Flavobacterium sp. species were detected to have broad host range. 97

These phages infected bacteria isolated from both freshwater and fish farm samples 98

although it has been suggested that single host enrichments select for phages of narrow 99

host range from sewage and marine environments (11, 33). Some of the phages 100

(especially all F. columnare phages) were more host-specific according to our collection 101

of Flavobacterium strains. F. columnare strains isolated from the same location as the 102

phage were the only ones susceptible for that specific phage. 103

104

No infection was observed in F. columnare strains by Flavobacterium sp. phages and 105

vice versa. However, 11 Flavobacterium sp. phage lysates (FTs-1, FKo-2, FL-1, FV-1, 106

FKy-1, FKy-2, FKy-3, FV-3, FV-4, FV-5, FV-6 and FV-8) inhibited the growth or lysed 107

the underlying bacterial culture on all F. columnare strains tested but produced no 108

individual plaques (data not shown). The phages could have been able to bind to the 109

bacteria and cause death but were not able to produce progeny, or the clear spots could be 110

an indication of bacteriocin activity. The causative agent of this strong inhibition or lysis 111

should be studied further for the possibility of developing antimicrobial agents. One of 112

the phages (FCL-1) was isolated from a fish suffering from columnaris disease. The 113

presence of the F. columnare phages in the fish indicates the possibility to control a fish 114

disease by enrichment of these phages. A number of successful reports on phage therapy 115

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against fish diseases have been published when the phages are applied directly to the 116

water (16, 17). Other possible benefits related to phages are that they could be developed 117

to be used as diagnostic tools. In the present study we show that bacteriophages and 118

Flavobacteria are widespread also in the northern fresh waters. We found phages of F. 119

columnare to be host specific making them good candidates for phage therapy. 120

121

All sequences have been submitted to EMBL Nucleotide Sequence Database, 122

http://www.ebi.ac.uk/embl/, under the accession numbers given in Table 1 and 123

#FR714876 (FCL-2), #FR714877 (FL-1), #FR714878 (FKj-2) and #FR865436 (FCV-1). 124

125

Mr Petri Papponen, Ms Katja Ryymin and Ms Irene Helkala are thanked for their 126

excellent technical assistance. Some bacterial strains used in this study were kindly 127

donated by Dr. Päivi Rintamäki and Finnish Food safety authority EVIRA. Station 128

manager Kari Saikkonen and field master Esa Karpoff from The Kevo Research Station 129

(University of Turku) are acknowledged for providing water samples. This work was 130

supported by the Finnish Centre of Excellence Program of the Academy of Finland 131

(2006-2011) grant 1129648 (J.K.H.B), Academy of Finland grant 127500 (L.-R.S.) and a 132

grant from Maj and Tor Nessling Foundation (J.K.H.B.) 133

134

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257

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Tables and Figures 258

Table 1. Bacterial strains isolated and used in the study 259

aThe previously studied F. columnare strains (genomic groups from A to H and colony 260

morphology from 1 to 4) are referred in this study by colony morphology rhizoid, Rz 261

(previously 1), rough, R (previously 2 and 3) and smooth, S (previously 4), following the 262

genomic group A to H, for example Rz-C (previously C1). 263

bUP-PCR group in numbers for Flavobacterium sp. , RISA group in letters for 264

Flavobacterium columnare, ND = not determined 265

cfor the partial 16S rRNA sequence of the bacteria 266

267

Table 2. Bacteriophages isolated and characterized in this study 268

a ND = not determined 269

b First letters in the phage name indicate the isolation host, F for Flavobacterium sp. and 270

FC for F. columnare, following letters refer to the sampling site 271

c L; much larger than the typical tailed phage genome (50 kb) 272

d Letter refers to the F. columnare RISA group that the phage is specific to 273

274

Figure 1. Panel A shows the isolation places of Flavobacteria and their phages from 275

Finland. Sampling sites are marked on the free map obtained from the National Land 276

Survey of Finland © Maanmittauslaitos 2005. The sites, their abbreviations and 277

coordinates are listed on the right. In the panel B are electron micrographs of purified and 278

negatively stained tailed Flavobacterium phages. (a) FCV-1; (b) FCL-2; (c) FJy-3; (d) 279

FKo-2; (e) FKy-1. Scale bar 50 nm. 280

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281

Figure 2. Host ranges of the phages infecting different Flavobacterium sp. strains. Grey 282

squares indicate infection; white indicates no infection and light grey squares mark the 283

strain which was originally used for isolation of the phage. The numbers at right and 284

bottom are the total number of different host ranges of the phage and susceptibility of 285

bacteria to phages, respectively. The approximate titer of each phage on the bacteria is 286

marked with plusses (+ ≤ 105, ++ 106-108 and +++ ≥ 109, all pfu/ml). The titer (PFU/ml) 287

of the phage on the isolation strain is marked on the light grey squares. In parenthesis 288

after the name of the bacterial strain is the abbreviation of the isolation place, which is 289

also included in the phage nomenclature (see Figure 1). F.columnare phages infected 290

only and specifically to one F. columanre RISA group and they are listed in Table 2. 291

292

293

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KeKjTs

Äk

Vj

Jy

Ky

Va

KoLe

Äy

AB

C

KeKjTs

Äk

Vj

Jy

Ky

Va

KoLe

Äy

AB

C

KeKjTs

Äk

Vj

Jy

KyVa

KoLe

ÄyL

V

I

In

Location Abbrevation Longitude Latitude

Lake Konnevesi Ko E 26° 35.644' N 62° 31.446'

Lake Jyväsjärvi Jy E 25° 44.339' N 62° 13.779'

River Vantaanjoki Va E 24° 58.072' N 60° 15.948'

River Kevojoki Ke E 26° 59.429' N 69° 45.337'

River Tsarsjoki Ts E 26° 59.447' N 69° 45.430'

Lake Kevojärvi Kj E 26° 59.608' N 69° 45.430'

Lake Valtimojärvi Vj E 28° 51.000' N 63° 40.688'

River Kymijoki Ky E 26° 49.926' N 60° 41.391'

Lake Leppävesi Le E 25° 54.633' N 62° 16.064'

Lake Äkässaivo Äk E 24° 8.640' N 67° 41.840'

River Äyskoski Äy E 26° 40.756' N 63° 0.665'

Lake Inari In E 27° 53.097' N 68° 47.783'

Fishery L 1) L

Fishery V 1) V

Fishery I 2) I1) Central Finland2) Northern Finland

A

B a b

c d e

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Bacteria

Phages B80

(Jy)

B18

7(L

)

B20

7(I

n)

B28

(Ko)

B17

6(L

e)B

223

(Ky)

B22

2(K

y)

B18

3(L

)

B16

7(J

y)

B22

4(K

y)

B24

1(V

)

B25

7(V

)

B26

0(V

)

B26

2(V

)

B16

9(J

y)

B17

8(Ä

k)

B11

4(K

e)

B21

8(V

)B

110

(Va)

B17

1(V

j)

B22

5(V

)

B24

3(V

)B

105

(Va)

B13

0(K

j)

B17

4(K

y)

B18

0(Ä

y)

B20

9(I

n)

B21

4(V

)

B12

7K

j)

B12

1(T

s)

B22

6(V

)

FTs-1 ++ ++ ++ +++ ++ ++ ++ ++ ++ ++ ++ +++ ++ ++ + ++ +++ +++ +++ ++ ++ ++ + 10824

FJy-3 + + ++ ++ ++ ++ + ++ 1010+ + ++ + ++ + + + + + + ++ ++ + + 24

FV-10 ++ ++ ++ + ++ + + ++ ++ ++ ++ ++ ++ 103

++ ++ + ++ 18

FV-5 ++ ++ ++ + ++ + + ++ ++ ++ 109

++ ++ ++ ++ ++ +++ 17

FV-6 ++ ++ ++ + ++ + ++ ++ ++ ++ ++ ++ ++ ++ ++ ++ 108

17

FV-8 ++ ++ ++ + ++ + ++ ++ ++ ++ ++ 108

++ ++ ++ ++ +++ 17

FJy-2 ++ ++ +++ 104++ ++ + ++ ++ ++ + ++ ++ + 14

FKj-2 ++ ++ ++ ++ ++ + + ++ ++ + ++ 1010++ 13

FKy-2 ++ ++ +++ ++ 1011

+++ ++ ++ + ++ ++ + 12

FKo-1 ++ ++ ++ 106++ + ++ ++ + ++ 10

FKy-1 + ++ 1010

++ ++ ++ ++ 7

FLe-1 1010+++ +++ +++ + + ++ 7

FV-9 ++ + + + 109

+++ + 7

FKo-2 ++ 106++ + ++ ++ 6

FKj-1 ++ ++ ++ + + 1036

FV-11 +++ ++ + + +++ ++ 6

FV-1 + ++ + ++ +++ 1010

6

FKy-3 ++ + 108

++ ++ 5

FL-1 ++ + 109+ + 5

FV-12 +++ + + +++ 4

FV-2 + + 106

3

FV-3 + 109

+++ 3

FKe-1 ++ 1082

FV-4 + 109

2

FJy-1 1091

18 12 12 12 11 11 10 9 9 9 9 9 9 8 8 7 7 7 6 6 6 6 6 6 4 4 4 4 3 3 1

on April 11, 2020 by guest

http://aem.asm

.org/D

ownloaded from

Bact

eria

l st

rain

aU

P-PC

R/

RIS

A g

roup

bEM

BL A

cces

sion

nu

mbe

rcSa

mpl

ing

site

Sour

ce/

Ref

eren

ceBa

cter

ial

stra

ina

UP-

PCR

/

R

ISA

gro

upb

EMBL

Acc

essi

on

num

berc

Sam

plin

g si

teSo

urce

/ R

efer

ence

B67

AFi

sher

y L

This

stud

yB2

4327

FR69

6354

Fish

ery

VTh

is st

udy

B28

1FR

6963

28La

ke K

onne

vesi

This

stud

yB2

44N

DFi

sher

y V

This

stud

yB8

02

FR69

6329

Lake

Jyvä

sjär

viTh

is st

udy

B245

CFi

sher

y V

This

stud

yB1

053

FR69

6330

Riv

er V

anta

anjo

kiTh

is st

udy

B247

CFi

sher

y V

This

stud

yB1

104

FR69

6331

Riv

er V

anta

anjo

kiTh

is st

udy

B257

26FR

6963

55Fi

sher

y V

This

stud

yB1

145

FR69

6332

Riv

er K

evoj

oki

This

stud

yB2

59C

Fish

ery

VTh

is st

udy

B121

6FR

6963

33R

iver

Tsa

rsjo

kiTh

is st

udy

B260

ND

FR69

6356

Fish

ery

VTh

is st

udy

B127

7FR

6963

34La

ke K

evoj

ärvi

This

stud

yB2

61C

Fish

ery

VTh

is st

udy

B130

8FR

6963

35La

ke K

evoj

ärvi

This

stud

yB2

62N

DFR

6963

57Fi

sher

y V

This

stud

yB1

679

FR69

6336

Lake

Jyvä

sjär

viTh

is st

udy

B263

ND

FR69

6358

Fish

ery

VTh

is st

udy

B169

10FR

6963

37La

ke Jy

väsj

ärvi

This

stud

yB2

67C

Fish

ery

VTh

is st

udy

B171

11FR

6963

38La

ke V

altim

ojär

viTh

is st

udy

B268

CFi

sher

y V

This

stud

yB1

7412

FR69

6339

Riv

er K

ymijo

kiTh

is st

udy

B269

ND

Fish

ery

VTh

is st

udy

B176

13FR

6963

40La

ke L

eppä

vesi

This

stud

yB2

70C

Fish

ery

VTh

is st

udy

B178

14FR

6963

41La

ke Ä

käss

aivo

This

stud

yB2

71C

Fish

ery

ITh

is st

udy

B180

15FR

6963

42R

iver

Äys

kosk

iTh

is st

udy

B272

JFi

sher

y I

This

stud

yB1

8316

FR69

6343

Fish

ery

LTh

is st

udy

B273

CFi

sher

y I

This

stud

yB1

85G

FR69

6344

Fish

ery

LTh

is st

udy

B274

CFi

sher

y I

This

stud

yB1

8717

FR69

6345

Fish

ery

LTh

is st

udy

B275

CFi

sher

y I

This

stud

yB2

0718

FR69

6346

Lake

Inar

iTh

is st

udy

Rz-

AA

49B2

0919

FR69

6347

Lake

Inar

iTh

is st

udy

R-B

B49

B214

20FR

6963

48Fi

sher

y V

This

stud

yR

z-C

C49

B218

21Fi

sher

y V

This

stud

yS-

CC

49B2

2222

FR69

6349

Riv

er K

ymijo

kiTh

is st

udy

R-D

D49

B223

23FR

6963

50R

iver

Kym

ijoki

This

stud

yS-

DD

49B2

2424

Riv

er K

ymijo

kiTh

is st

udy

Rz-

EE

49B2

2524

FR69

6351

Fish

ery

VTh

is st

udy

R-E

E49

B226

25FR

6963

52Fi

sher

y V

This

stud

yS-

EE

49B2

30I

Fish

ery

VTh

is st

udy

S-F

FF.

col

umna

re ty

pe st

rain

NC

IMB

2248

B234

CFi

sher

y V

This

stud

yR

z-G

G49

B235

CFi

sher

y V

This

stud

yR

-GG

49B2

36N

DFi

sher

y V

This

stud

yS-

GG

49B2

37C

Fish

ery

VTh

is st

udy

R-H

H49

B241

26FR

6963

53Fi

sher

y V

This

stud

yS-

HH

49

bU

P-PC

R g

roup

in n

umbe

rs fo

r Fla

voba

cter

ium

sp.

, R

ISA

gro

up in

lette

rs fo

r Fla

voba

cter

ium

col

umna

re, N

D =

not

det

erm

ined

cfo

r the

par

tial 1

6S rR

NA

sequ

ence

of t

he b

acte

ria

aTh

e pr

evio

usly

stud

ied

F. c

olum

nare

stra

ins (

geno

mic

gro

ups f

rom

A to

H a

nd c

olon

y m

orph

olog

y fro

m 1

to 4

) are

refe

rred

in th

is st

udy

by c

olon

y m

orph

olog

y rh

izoi

d,

Rz

(pre

viou

sly

1), r

ough

, R (p

revi

ousl

y 2

and

3) a

nd sm

ooth

, S (p

revi

ousl

y 4)

, fol

low

ing

the

geno

mic

gro

up A

to H

, for

exa

mpl

e R

z-C

(pre

viou

sly

C1)

.

TABL

E 1.

Bac

teria

l stra

ins i

sola

ted

and

used

in th

e st

udy

on April 11, 2020 by guest

http://aem.asm

.org/D

ownloaded from

Phag

ebSa

mpl

ing

site

Isol

atio

n str

ain

Phag

e fa

mily

App

roxi

mat

e ge

nom

e siz

e (k

bp)c

RIS

A

grou

pd

FJy-

1La

ke J

yväs

järv

iB

80M

yovi

rida

e30

FJy-

2La

ke J

yväs

järv

iB

167

ND

>48

FJy-

3La

ke J

yväs

järv

iB

169

Myo

viri

dae

LFK

o-1

Lake

Kon

neve

siB

28M

yovi

rida

e25

-48

FKo-

2La

ke K

onne

vesi

B28

Myo

viri

dae

20-3

0FK

e-1

Riv

er K

evoj

oki

B11

4M

yovi

rida

eN

DFT

s-1

Riv

er T

sars

joki

B12

1N

D25

-48

FKj-1

Lake

Kev

ojär

viB

127

ND

>48

FKj-2

Lake

Kev

ojär

viB

130

Myo

viri

dae

25-4

8FK

y-1

Riv

er K

ymijo

kiB

222

Myo

viri

dae

20-4

8FK

y-2

Riv

er K

ymijo

kiB

223

Podo

viri

dae?

25-4

8FK

y-3

Riv

er K

ymijo

kiB

224

Myo

viri

dae?

LFL

e-1

Lake

Lep

päve

siB

176

Siph

ovir

idae

?20

-30

FL-1

Fish

ery

LB

183

Myo

viri

dae

55FV

-1Fi

sher

y V

B21

4M

yovi

rida

e28

FV-2

Fish

ery

VB

218

ND

25-4

8FV

-3Fi

sher

y V

B22

5M

yovi

rida

eN

DFV

-4Fi

sher

y V

B22

6Po

dovi

rida

e25

-48

FV-5

Fish

ery

VB

241

ND

LFV

-6Fi

sher

y V

B24

3N

D30

-40

FV-8

Fish

ery

VB

257

ND

LFV

-9Fi

sher

y V

B26

0Po

dovi

rida

e?L

FV-1

0Fi

sher

y V

B26

2N

DL

FV-1

1Fi

sher

y V

B26

3N

D>4

8FV

-12

Fish

ery

VB

278

Podo

viri

dae?

60FC

L-1

Fish

ery

LB

67M

yovi

rida

e/Po

dovi

rida

e50

AFC

L-2

Fish

ery

LB

185

Myo

viri

dae

30G

FCL-

3Fi

sher

y L

R-G

ND

30G

FCL-

4Fi

sher

y L

R-G

ND

ND

GFC

V-1

Fish

ery

VR

z-C

Myo

viri

dae

50C

FCV

-2Fi

sher

y V

B23

5N

DN

DC

FCV

-3Fi

sher

y V

B23

6N

DN

DC

FCV

-4Fi

sher

y V

Rz-

CN

DN

DC

FCV

-5Fi

sher

y V

Rz-

CN

DN

DC

FCV

-6Fi

sher

y V

Rz-

CN

DN

DC

FCV

-7Fi

sher

y V

Rz-

CN

DN

DC

FCV

-8Fi

sher

y V

Rz-

CN

DN

DC

FCV

-9Fi

sher

y V

B24

5N

DN

DC

FCV

-10

Fish

ery

VB

247

ND

ND

CFC

V-1

1Fi

sher

y V

Rz-

CN

DN

DC

FCV

-12

Fish

ery

VR

z-C

ND

ND

CFC

V-1

3Fi

sher

y V

B26

1N

DN

DC

FCV

-14

Fish

ery

VR

z-C

ND

ND

CFC

V-1

5Fi

sher

y V

Rz-

CN

DN

DC

FCV

-16

Fish

ery

VR

z-C

ND

ND

CFC

V-1

7Fi

sher

y V

Rz-

CN

DN

DC

FCV

-18

Fish

ery

VR

z-C

ND

ND

CFC

V-1

9Fi

sher

y V

Rz-

CN

DN

DC

FCV

-20

Fish

ery

VR

z-C

ND

ND

Ca N

D =

not

det

erm

ined

c L;

muc

h la

rger

than

the

typi

cal t

aile

d ph

age

geno

me

(50

kb)

d L

ette

r ref

ers t

o th

e F.

col

umna

re R

ISA

gro

up th

at th

e ph

age

is sp

ecifi

c to

TAB

LE 2

. Bac

terio

phag

es is

olat

ed a

nd c

hara

cter

ized

in th

is stu

dya

b F

irst l

ette

rs in

the

phag

e na

me

indi

cate

the

isola

tion

host,

F fo

r Fl

avob

acte

rium

sp. a

nd F

C fo

r F. c

olum

nare

, fol

low

ing

lette

rs re

fer t

o th

e

on April 11, 2020 by guest

http://aem.asm

.org/D

ownloaded from