RESEARCH LETTER
Characterization of a novel Morganella morganii bacteriophage
FSP1 isolated from river water
Shogo Yamaki, Takuo Omachi, Yuji Kawai & Koji Yamazaki
Laboratory of Marine Food Science and Technology, Faculty of Fisheries Sciences, Hokkaido University, Hakodate, Japan
Correspondence: Koji Yamazaki, Laboratory
of Marine Food Science and Technology,
Faculty of Fisheries Sciences, Hokkaido
University 3-1-1, Minato, Hakodate
041-8611, Japan.
Tel./fax: +81 138 40 5574;
e-mail: [email protected]
Received 11 May 2014; revised 28 July 2014;
accepted 29 July 2014. Final version
published online 28 August 2014.
DOI: 10.1111/1574-6968.12560
Editor: Aharon Oren
MICROBIOLOGY LETTERS
Keywords
Morganella morganii; bacteriophage; phage
therapy; characterization.
Abstract
Morganella morganii has been identified as a causative agent of opportunistic
infections and histamine poisoning. Bacteriophage is a virus and has recently
been considered an alternative agent to antibiotics for the control of bacteria
that have developed antibiotic resistance. In this study, a novel M. morganii
bacteriophage isolated from river water was characterized. The isolated phage,
termed FSP1, was purified by polyethylene glycol precipitation followed by
cesium chloride density-gradient centrifugation. FSP1 has infectivity against
only M. morganii and was identified as a Myoviridae bacteriophage through
morphological analysis with transmission electron microscopy. According to
the one-step growth curve, the FSP1 latent period, eclipse period, and burst
size were 30, 20 min, and 42 PFU infected cell 1, respectively. The genome size
of FSP1 was estimated to be c. 45.6–49.4 kb by restriction endonuclease analyses. Moreover, challenge testing against M. morganii in vitro revealed that FSP1
had high lytic activity and that the viable cell count of M. morganii was
reduced by 6.12 log CFU mL 1 after inoculation with FSP1 at a multiplicity of
infection (MOI) = 10. These results suggested that FSP1 could be used as a
biocontrol agent against M. morganii for treatment of infectious disease treatment or food decontamination.
Introduction
Morganella morganii, a motile gram-negative rod belonging to the family Enterobacteriaceae, has low pathogenicity, but compromised patients can develop diarrhea,
wound infections, urinary tract infections, bacteremia,
and sepsis due to M. morganii (Tucci & Isenberg, 1981;
Chang et al., 2011). In addition, M. morganii is known as
a major histamine producer due to its powerful histidine
decarboxylase. Morganella morganii is responsible for histamine accumulation on food (L
opez-Sabater et al., 1996;
Rodtong et al., 2005), which often causes histamine poisoning (also referred to as scombroid poisoning) in
decomposed fishery products (Becker et al., 2001). Morganella morganii was detected in the gill and skin of
mackerel, sardine, and albacore. Moreover, the surface of
conveyer belts and plastic totes contacted with mackerel
and sardine is contaminated during processing although
no M. morganii was found in the processing plant before
processing (Kim et al., 2003). Although many cases present with mild symptoms, rare cases present with severe
ª 2014 Federation of European Microbiological Societies.
Published by John Wiley & Sons Ltd. All rights reserved
symptoms that can be potentially life-threatening
(Sanchez-Guerrero et al., 1997; D’Aloia et al., 2011).
Therefore, the inhibition of M. morganii growth in food
is important to reduce the risk of histamine poisoning.
Bacteriophages (phages) are bacterial virus and a notable antimicrobial agent. The therapeutic potential of
phages has not been an area of interest since the discovery
of antibiotics. However, phages have recently been suggested as an alternative antibacterial agent to counteract
the emergence of antibiotic-resistant bacteria (Kutateladze
& Adamia, 2010). In fact, many phages, including Staphylococcus aureus phage P-27/HP (Gupta & Prasad, 2011)
and Pseudomonas aeruginosa phage M4 (Fu et al., 2010),
have been investigated for their ability to control human
pathogenic infections. Phages are also available for food
safety. The United States Department of Agriculture and
Food and Drug Administration approved LISTEXTM P100
and SALMONELEXTM (Micreos Food Safety, Netherlands)
to control food pathogens. The major components are
Listeria monocytogenes phage P100 (LISTEXTM P100) and
Salmonella phages (SALMONELEXTM) and are a ‘Generally
FEMS Microbiol Lett 359 (2014) 166–172
167
Characterization of Morganella morganii bacteriophage
Recognized As Safe’ food processing aid. Moreover,
phage-based biocontrol strategies to control Escherichia
coli, Bacillus cereus, and Campylobacter jejuni were also
developed (Atterbury et al., 2003; Viazis et al., 2011;
Bandara et al., 2012). To date, no studies have focused on
the use of phages to control M. morganii, although only a
few studies related to M. morganii phages have been
reported (Schmidt & Jeffries, 1974; Zhu et al., 2010).
In this study, we describe the novel M. morganii phage
FSP1 isolated from river water, and we characterize FSP1
for use as an antimicrobial agent against M. morganii.
Materials and methods
Bacterial strains and growth conditions
The strains used in this study are listed in Table 1. All
bacteria listed in Table 1 were incubated at 30 °C for
Table 1. List of bacterial strains in this study and host range of FSP1
Bacterial speacies
Strain
Gram-negative bacteria
Morganella morganii ssp. morganii
Morganella morganii ssp. morganii
Morganella morganii ssp. morganii
Aeromonas hydrophila
Citrobacter freundii
Cronobactor sakazakii
Enterobacter gergoviae
Escherichia coli
Escherichia coli
Escherichia coli
Hafnia alvei
Klebsiella pneumoniae
Proteus hauseri
Proteus mirabilis
Proteus mirabilis
Proteus vulgaris
Providencia alcalifaciens
Providencia rettgeri
Providencia rustigianii
Providencia stuartii
Pseudomonas fluorescens
Raoultella ornithinolytica
Salmonella Enteritidis
Salmonella Infantis
Salmonella Typhimurium
Yersinia enterocolitica
Gram-positive bacteria
Bacillus cereus
Listeria monocytogenes
Paenibacillus chibensis
Staphylococcus aureus
+, clear plaque;
, no plaque.
FEMS Microbiol Lett 359 (2014) 166–172
NBRC3848T
NBRC3168
ATCC25829
NBRC3820
JCM1657T
ATCC51329
JCM1234
JCM1649T
ATCC51446
ATCC51813
JCM1666T
ATCC13883
ATCC13315
JCM1669T
ATCC33583
ATCC33420
ATCC9886
ATCC9250
JCM3953T
ATCC33672
JCM5963T
ATCC31898
NBRC3313
ATCC51741
IID1000
ATCC9610
JCM2152T
ATCC7644
NBRC15958T
NBRC14462
Plaque
formation
+
+
+
18 h in tryptic soy broth (TSB, BD, Franklin Lakes, NJ)
or TSB supplemented with 0.6% yeast extract (BD).
Phage isolation
Phages were isolated by the method of Carvalho et al.
(2010a) with modifications. Three hundred milliliters of a
river water sample was added to 300 mL of double strength
TSB supplemented with 400 lg mL 1 CaCl2 and
400 lg mL 1 MgSO4, and the host strains M. morganii
NBRC3848T, M. morganii NBRC3168, and M. morganii
ATCC25829, at the exponential phase, were inoculated.
After incubation at 30 °C for 12 h, the cultures were
centrifuged (3500 g, 30 min, 4 °C), and the supernatants
were filtrated using 0.45-lm polyvinylidene fluoride filters
(Merck Millipore, Billerica, MA). For detection of phage,
filtrates were spotted on a lawn of M. morganii NBRC 3848T
made using the double agar overlay method and incubated
at 37 °C for 24 h. Clear zones were picked and suspended in
1 mL of SM buffer (100 mM NaCl, 8 mM MgSO47H2O,
50 mM Tris-HCl, and 0.01% gelatin, pH 7.5), and single
plaques were formed using the double agar overlay method.
This procedure was repeated at least three times.
Propagation and purification of phage
FSP1 was propagated and purified by polyethylene glycol
precipitation followed by cesium chloride density-gradient
centrifugation. FSP1 was inoculated into log phase culture
of M. morganii NBRC3848T. The culture was incubated
at 30 °C for growth of FSP1 followed by centrifugation
(10 000 g, 30 min, 4 °C) and filtration with 0.45-lm
polyvinylidene fluoride filters. After DNase I and RNase
A (Sigma-Aldrich, St. Louis, MO) were added at
1 lg mL 1 and incubated at room temperature for 1 h,
the FSP1 lysate was treated with 1 M sodium chloride for
1 h on ice. Furthermore, polyethylene glycol 6000 (Wako
pure chemical industries, Osaka, Japan) was added to
final concentration of 10%. The mixture was incubated at
4 °C for 12 h and was then centrifuged (10 000 g,
20 min, 4 °C) to precipitate phage particles. The pellet
was suspended in SM buffer, and the polyethylene glycol
was removed by chloroform extraction and centrifugation
(10 000 g, 10 min, 4 °C). Purification of FSP1 was carried out by cesium chloride density-gradient centrifugation (100 000 g, 2 h, 4 °C), and the sample was dialyzed
against SM buffer. The purified phage was stored at 4 °C
for further investigations.
Host range determination
The host range of FSP1 was determined using a spot test
method. One hundred microliters of each bacterial
ª 2014 Federation of European Microbiological Societies.
Published by John Wiley & Sons Ltd. All rights reserved
168
culture listed in Table 1 was added to 3.5 mL of 0.5%
molten agar, and this mixture was overlaid onto a tryptic
soy agar (BD) plate. Ten microliters of FSP1 solution
(109 PFU mL 1) was spotted onto each agar plate. Plates
were incubated at 37 °C for 6 h and were then examined
for clear zones on the bacterial lawn.
Transmission electron microscopy
The morphology of FSP1 was analyzed by negative staining with 2.5% samarium triacetate, which was reported as
a good negative-staining reagent (Nakakoshi et al., 2011).
The FSP1 solution was dropped on a 200-mesh Cu grid
(Nisshin EM, Tokyo, Japan) for 3 min. The excess solution was absorbed with filter paper, and 2.5% samarium
triacetate was dropped on the grid. The excess solution
was absorbed with filter paper, and FSP1 was examined
using a transmission electron microscope (JEM-1011,
JEOL, Tokyo, Japan).
Isolation and restriction analysis of phage DNA
FSP1 was suspended to TENS buffer (50 mM Tris-HCl,
100 mM EDTA, 100 mM NaCl, 0.3% sodium dodecyl
sulfate, pH 8.0, Stenholm et al., 2008). Proteinase K was
added to a final concentration of 200 lg mL 1, and the
mixture was incubated at 65 °C for 10 min for extraction
of phage DNA. Purification of the phage DNA was carried out using the Phage DNA Isolation Kit (Norgen Biotek, Thorold, Ontario, Canada) according to the
manufacturer’s instruction. For genome size estimation,
isolated phage DNA was digested with the restriction
endonucleases EcoRI, HindIII, and XbaI (Nippon gene,
Tokyo, Japan) according to the manufacturer’s instructions. The digested DNA fragments were separated with
agarose gel electrophoresis using 0.5% SeaKem gold agarose (Takara Bio, Shiga, Japan). The DNA bands were
visualized by SYBR green nucleic acid stain (SigmaAldrich), and the sum of each band size was calculated as
the phage genome size. Gene ladder wide I (Nippon gene)
and 2.5 kb DNA ladder (Takara Bio) were used as the
molecular size markers.
Temperatures and pH stability test
To determine the thermal stability of FSP1, FSP1 was suspended to give a final concentration of 107 PFU mL 1 in
TSB. The mixture was incubated at 55–70 °C for 1 h,
samples were collected every 10 min, and the PFU of
FSP1 was determined using serial dilution and the double
agar overlay method. To determine the pH stability of
FSP1, FSP1 was suspended to give a final concentration
of 107 PFU mL 1 in TSB (pH 1.0–12.0) and incubated at
ª 2014 Federation of European Microbiological Societies.
Published by John Wiley & Sons Ltd. All rights reserved
S. Yamaki et al.
30 °C for 24 h. The PFU of FSP1 was determined using
serial dilution and the double agar overlay method.
Adsorption assay
Morganella morganii NBRC3848T was incubated at 30 °C
until reaching an OD600 nm = 0.1 in TSB. The FSP1 suspension was added at a multiplicity of infection (MOI) of
0.01, and the mixed culture was incubated at 30 °C.
Every 5 min, samples were collected and centrifuged at
10 000 g for 2 min, followed by filtration using a 0.45lm polyvinylidene fluoride filter. The PFU of nonadsorbed phage in the supernatant was determined using
serial dilution and the double agar overlay method.
One-step growth experiments
Morganella morganii NBRC3848T was incubated at 30 °C
until reaching an OD600 nm = 0.1 in TSB. FSP1 was
added at an MOI = 0.1 and adsorbed at 30 °C for 5 min.
The free phages in the mixture were removed by centrifugation (10 000 g, 2 min, 4 °C), and the pellet was suspended in the same volume of fresh TSB. The suspension
was incubated at 30 °C, and the samples were collected
after 5 min. The phage titer of the sample was measured
in two ways: A sample was measured either immediately
after sampling or after chloroform treatment (for lysis of
the bacterial cells). The burst size, latent period, and
eclipse period of FSP1 were determined by changes in the
phage titer.
Challenge test
Morganella morganii NBRC3848T was inoculated into
fresh TSB for 6 log CFU mL 1, the culture was incubated
at 30 °C, and the samples were collected at 1-h intervals.
The samples were serially diluted and spread onto tryptic
soy agar plates. When the OD600 nm reached 0.1, which
corresponds to a viable cell count of c. 107 CFU mL 1,
the culture was divided into two sets of samples, and
FSP1 was added to only one sample at a MOI = 1.0 or
10. Both samples were incubated and spread onto tryptic
soy agar plates at 1-h intervals. The tryptic soy agar plates
were incubated at 37 °C for 24 h, and the CFU were
enumerated.
Results
Isolation and host range of an isolated phage
In this study, we isolated a M. morganii phage from a
river water (Odajima river in Hakodate, Japan) sample,
and the isolated phage was designated as FSP1. The host
FEMS Microbiol Lett 359 (2014) 166–172
169
Characterization of Morganella morganii bacteriophage
9
8
Log PFU mL–1
range of FSP1 was determined for 26 strains of gram-negative bacteria and 4 strains of gram-positive bacteria.
FSP1 formed plaques for 3 strains: M. morganii
NBRC3848T, M. morganii NBRC3168, and M. morganii
ATCC25829, but no other strains were infected by FSP1
(Table 1). Moreover, FSP1 formed clear single plaques on
the M. morganii strains used in this study.
7
6
5
4
Morphology of FSP1
The morphology of FSP1 virions was observed by negative staining. Transmission electron microscopy showed
that FSP1 virions had a head measurement of
110 7.2 nm (mean SD) and a contractile tail measurement of 114 5.5 nm (n = 8) (Fig. 1). Therefore, it
is suggested that FSP1 belongs to the Myoviridae family, a
member of tailed phage, which has contractile tail consisting of a sheath and a central tube.
Adsorption and one-step growth of FSP1
For elucidation of the FSP1 growth property, we conducted an adsorption assay and one-step growth curve
analyses. Forty percent of FSP1 was adsorbed to M. morganii NBRC3848T at 5 min. In addition, almost all of the
FSP1 was adsorbed at 40 min (data not shown). According to the one-step growth experiment, the latent period
of FSP1 was 30 min, the rise period was 15 min, the
eclipse period was 20 min, and the burst size was 42 PFU
infected cell 1 (Fig. 2).
3
10
20
30
40
50
60
Incubation time (min)
Fig. 2. One-step growth curve of FSP1 in TSB at 30C. Open
diamond: non-treated, closed diamond: chloroform treated. The
results are shown as the mean standard deviation from three
independent experiments
HindIII, and XbaI. Agarose gel electrophoresis of the
digested DNA showed that FSP1 DNA had many restriction sites for each of the restriction endonucleases
(Fig. 3). The genome size of FSP1 DNA was calculated as
the sum of each band and was c. 45.6–49.4 kb.
Thermal and pH stability of FSP1
To analyze the stability of FSP1, thermal and pH stability
were evaluated. FSP1 was stable at 50 °C but heating to
1
(kb)
2
3
4
5
6
20
30
20
15
10
10
7.0
7.5
5.0
5.0
Genomic size of FSP1
To determine the FSP1 genome size, FSP1 DNA was
digested with the restriction endonucleases EcoRI,
0
4.0
3.0
2.5
2.0
1.5
1.3
1.0
0.7
0.5
100 nm
Fig. 1. Electron micrograph of FSP1 negatively stained with 2.5 %
samarium triacetate. Scale bar indicates 100 nm
FEMS Microbiol Lett 359 (2014) 166–172
Fig. 3. Digestion patterns of the FSP1 genome with restriction
endonucleases. Lane 1: Gene ladder wide I (Nippon gene, Tokyo,
Japan), lane 2: FSP1 DNA, lanes 3, 4, and 5: FSP1 DNA digested with
EcoR I, Hind III, and Xba I (Nippon gene, Tokyo, Japan), lane 6: 2.5 kb
DNA ladder (Takara Bio, Tokyo, Japan)
ª 2014 Federation of European Microbiological Societies.
Published by John Wiley & Sons Ltd. All rights reserved
170
S. Yamaki et al.
(a) 8
(a)
Viable cell count
(log CFU mL–1)
Log PFU mL–1
6
5
4
3
2
7
6
5
4
0
10
20
30
40
50
0
60
(b)
Viable cell count
(log CFU mL–1)
7
6
5
4
3
6
9
12
15
Incubation time (h)
(b) 8
Log PFU mL–1
8
3
Incubation time (min)
3
2
1
Phage infection
9
7
1
10
10
9
8
7
6
5
4
3
2
1
Phage infection
0
3
6
9
12
15
Incubation time (h)
1
3
5
7
9
11
pH
Fig. 4. Effect of thermal (A) and pH treatment (B) on the viability of
FSP1 in TSB. Closed circle: heated at 50C, open circle: heated at
55C, closed square: heated at 60C, open square: heated at 65C,
closed diamond: heated at 70C. The results are shown as the mean
± standard deviation from three independent experiments
55 and 60 °C caused a decrease of c. 3.5 log PFU mL 1
of FSP1 virions, and no FSP1 virions were detected after
heating at 65 and 70 °C (Fig. 4a). In addition, FSP1 was
stable frozen at 40 °C for 24 h (data not shown). A pH
stability analysis showed that FSP1 was stable at various
pH levels, except strong acidic (lower than pH 3.0) and
alkaline (pH 12.0) levels (Fig. 4b). This thermal and pH
stability is beneficial for the use of FSP1 as a biocontrol
agent against M. morganii.
Antimicrobial action of FSP1 against
M. morganii
A bactericidal effect of FSP1 (MOI = 1.0 or 10) was
examined by changes in the viable cell count of
M. morganii NBRC3848T. At a MOI = 1.0, the viable
cell count of M. morganii decreased from 7.2 log
CFU mL 1 to 3.7 log CFU mL 1 (Fig. 5a). At a
MOI = 10, the bacterial count was not detectable (< 1
log CFU mL 1) after 1 h of FSP1 inoculation (Fig. 5b).
These results suggest that FSP1 is expected to be an
antimicrobial agent for M. morganii control and that its
ª 2014 Federation of European Microbiological Societies.
Published by John Wiley & Sons Ltd. All rights reserved
Fig. 5. Effect of FSP1 on viable cell count of M. morganii 19 at MOI
= 1.0 (A) and MOI = 10 (B) in TSB at 30C. FSP1 was added when
OD600 was reached at 0.1 which correspond to approximately 107
CFU mL 1 of viable cell count. Open diamond: non-treated; closed
diamond: FSP1 treated. The results are shown as the mean standard deviation from three independent experiments
bactericidal effect is related to the MOI. However, the
bacterial count of M. morganii began to recover after
an additional incubation for 2 h at MOI = 1.0 and 5 h
at MOI = 10.
Discussion
Phages, which possess a different mode of action compared antibiotics, are expected to be an alternative to
antibiotics (O’Flaherty et al., 2009). Controlling pathogens through the use of phages has been investigated for
various applications such as therapeutic drugs for human,
domestic animal, plants, hatchery fish, and food additives
(Carvalho et al., 2010b; Gupta & Prasad, 2011; Addy
et al., 2012; Chibeu et al., 2013; Madsen et al., 2013).
However, studies on the growth inhibition of M. morganii by lytic phages have not been reported. The aim of
this study was to isolate and characterize a phage to control M. morganii.
According to the method of Carvalho et al. (2010a) with
minor modification, we isolated a M. morganii phage,
named FSP1, from river water in Hakodate, Hokkaido,
Japan. Next, we examined the host range of FSP1 because
FEMS Microbiol Lett 359 (2014) 166–172
171
Characterization of Morganella morganii bacteriophage
the infection specificity of phages is generally species- or
strain dependent. FSP1 formed plaques for all of the strains
of M. morganii tested in this study, but FSP1 could not
form plaques against other gram-positive or gram-negative
bacteria, suggesting that FSP1 was a highly specific phage
to M. morganii and these M. morganii strains might have a
common and conservative phage receptor among this species. Restriction analyses of FSP1 DNA and transmission
electron microscopic observation showed that FSP1 had a
45.6–49.4 kb genome and belonged to the Myoviridae family from the virion tail structure (Figs 1 and 3). Because
agarose electrophoresis showed bands digested by restriction endonucleases, FSP1 DNA was estimated to be linear,
double-strand DNA. Another M. morganii phage, named
Mmp1, is in the Podoviridae family and has a genome size
of c. 38 kb (Zhu et al., 2010).
To examine the potential of FSP1 as an antimicrobial
agent for M. morganii, we evaluated the adsorption rate,
latent period, rise period, and burst size of FSP1 against
M. morganii NBRC3848T. The adsorption rate, latent period, eclipse period, rise period, and burst size were determined to be 40% for 5, 30, 20, 15 min, and 42 PFU
infected cell 1, respectively (Fig. 2). Other Myoviridae
phages generally have a latent period, eclipse period, and
burst size of 15–50, 10–40 min, and 50–150 PFU infected
cell 1, respectively (Raya et al., 2006; Carey-Smith et al.,
2006; Uchiyama et al., 2008; Hsieh et al., 2011; Park
et al., 2012). Therefore, the growth parameters of FSP1
were similar to those of other Myoviridae phages. FSP1
was stable at pH 4.0–10.0, freezing at 40 °C for 24 h,
and heating at 50 °C for 1 h (Fig. 4a and b). These
results suggest that the growth parameters and stability of
FSP1 will be suitable for use as a biocontrol agent against
M. morganii in food, especially in ready-to-eat food.
The bactericidal action of phages is one of the most
important factors to evaluate for potential use as an antimicrobial agent. The viable cell count of M. morganii
NBRC3848T was substantially reduced to below the detection limit at a MOI = 10 in a challenge test (Fig. 5a and b).
The extremely high lytic activity indicates that FSP1 can be
used for the efficient control of M. morganii although the
challenge test was performed in media. The in vivo challenge test will be the next step on our research. The phage
therapy on the fishery products should be carried out by
spraying or dipping because M. morganii is often found on
the gill and skin of fishes (Kim et al., 2003).
Despite the marked prevention of M. morganii,
regrowth of host strain began after 5 h of FSP1 inoculation. This phenomenon was observed not only in FSP1 but
also in other phages, such as E. coli phage PP01, SFP10,
and Salmonella spp. phage FGCSSa1 (Morita et al., 2002;
Carey-Smith et al., 2006; Park et al., 2012). The regrowth
of the host strain is one of the most crucial points to
FEMS Microbiol Lett 359 (2014) 166–172
recognize in phage therapy because they might harbor the
phage resistance. Like the antibiotics resistance, the phage
resistance might be public health concern with renaissance
of the phage therapy. Bacteria acquire the phage resistance
in various ways such as prevention of phage adsorption
and phage DNA injection, cleavage of phage DNA, and
abortive infection system (Labrie et al., 2010). The utilization of a phage cocktail will be an effective way to solve
this problem because bacteria obtained resistance to one
phage can be inhibited by another phages have different
infection mechanisms. Indeed, the regrowth of E. coli
O157:H7 was substantially inhibited by the use of a phage
cocktail containing E. coli phages SP21 and SP22 (Tanji
et al., 2004). The combination of phages and chemical
compounds was found to be an efficient regrowth control
strategy for L. monocytogenes (Chibeu et al., 2013). To
suppress the development of the phage resistance and
ensure the effect of phage therapy, we will have to isolate
and characterize more phages in order to make a phage
cocktail. In addition to the phage resistance, interaction of
phages with food components and environmental conditions (e.g. protein, lipid, carbohydrate, pH, temperature) is
important issues, as attachment of phages to a host cell
might be blocked by these factors.
In this study, we isolated and characterized a phage,
named FSP1, that inhibits histamine-producing M. morganii. Our results showed that the host specificity, thermal and pH stability, growth parameters, and
antimicrobial activity of FSP1 were useful for controlling
M. morganii as a food additive and therapeutic agent. To
our knowledge, this is the first report investigating the
growth and antimicrobial activity of a Myoviridae phage
inhibiting M. morganii.
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