The envE gene of Salmonella enterica serovar Typhimurium is encoded within Salmonella Pathogenicity Island-11 (SPI-11) and is located immediately upstream of the virulence gene msgA (macrophage survival gene A) in the same transcriptional orientation. To date, the characteristics and roles of envE remain largely unexplored. In this study, we show that EnvE, a predicted lipoprotein, is localized on the outer membrane using sucrose gradient ultracentrifugation. Under oxidative stress conditions, envE transcription is suppressed, while msgA transcription is induced, indicating an inverse correlation between the mRNA levels of the two neighboring genes. Importantly, inactivation of envE leads to constitutive transcription of msgA regardless of the presence of oxidative stress. Moreover, trans-complementation of the envE mutant with a plasmid-borne envE fails to prevent the induction of msgA transcription, suggesting that envE functions as a cis-regulatory element rather than a trans-acting factor. We further show that both inactivation and complementation of envE confer wild-type levels of resistance to oxidative stress by ensuring the expression of msgA. Our data suggest that the S. enterica envE gene encodes an outer membrane lipoprotein, and its transcription represses msgA expression in a cis-acting manner, probably by transcriptional interference, although the exact molecular details are yet unclear.
Salmonella Typhimurium is an invasive gastrointestinal pathogen for both humans and animals. To investigate the genetic framework and diversity of S.
Typhimurium, a total of 194 S. Typhimurium isolates were collected from patients in a tertiary hospital between 2020 and 2021. Antimicrobial susceptibility testing was used to confirm the resistance phenotype. Whole-genome sequencing and bioinformatics analysis were performed to determine the sequence type, phylogenetic relationships, resistance gene profiles, Salmonella pathogenicity island (SPI) and the diversity of the core and pan genome. The result showed that 57.22% of S. Typhimurium isolates were multidrug resistant and resistance of total isolates to the first-line drug ciprofloxacin was identified in 60.82%. The population structure of S. Typhimurium was categorized into three lineages: ST19 (20.10%, 39/194), ST34-1 (47.42%, 92/194) and ST34-2 (40.65%, 63/194), with the population size exhibiting increasing trends. All lineages harbored variety of fimbrial operons, prophages, SPIs and effectors that contributed to the virulence and long-term infections of S. Typhimurium. Importantly, ST34-1 lineage might potentially be more invasive due to the possession of SPI1-effector gene sopE which was essential for the proliferation, internalization and intracellular presence of S. Typhimurium in hosts. Multiple antimicrobial resistance genes were characteristically distributed across three lineages, especially carbapenem genes only detected in ST34-1&2 lineages. The distinct functional categories of pan genome among three lineages were observed in metabolism, signaling and gene information processing. This study provides a theoretical foundation for the evolved adaptation and genetic diversity of S. Typhimurium ST19 and ST34, among which ST34 lineages with multidrug resistance and potential hypervirulence need to pay more attention to epidemiological surveillance.
This study aimed to develop synthetic Claudin18.2 (CLDN18.2) chimeric antigen receptor (CAR)-T (CAR-T) cells as a treatment for advanced gastric cancer using lentiviral vector genetic engineering technology that targets the CLDN18.2 antigen and simultaneously overcomes the immunosuppressive environment caused by programmed cell death protein 1 (PD-1). Synthetic CAR T cells are a promising approach in cancer immunotherapy but face many challenges in solid tumors. One of the major problems is immunosuppression caused by PD-1. CLDN18.2, a gastric-specific membrane protein, is considered a potential therapeutic target for gastric and other cancers. In our study, CLDN18.2 CAR was a second-generation CAR with inducible T-cell costimulatory (CD278), and CLDN18.2-PD1/CD28 CAR was a third-generation CAR, wherein the synthetic PD1/CD28 chimeric-switch receptor (CSR) was added to the second-generation CAR. In vitro, we detected the secretion levels of different cytokines and the killing ability of CAR-T cells. We found that the secretion of cytokines such as interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) secreted by three types of CAR-T cells was increased, and the killing ability against CLDN18.2-positive GC cells was enhanced. In vivo, we established a xenograft GC model and observed the antitumor effects and off-target toxicity of CAR-T cells. These results support that synthetic anti-CLDN18.2 CAR-T cells have antitumor effect and anti-CLDN18.2-PD1/CD28 CAR could provide a promising design strategy to improve the efficacy of CAR-T cells in advanced gastric cancer.
Candida albicans (C. albicans) is one of the most common opportunistic fungi worldwide, which is associated with a high
mortality rate. Despite treatment, C. albicans remains the leading cause of life-threatening invasive infections. Consequently,
antimicrobial peptides (AMPs) are potential alternatives as antifungal agents with excellent antifungal activity. We previously
reported that Css54, found in the venom of Centrurodies suffusus suffusus (C. s. suffusus) showed antibacterial activity
against zoonotic bacteria. However, the antifungal activity of Css54 has not yet been elucidated. The obj!ective of this study
was to identify the antifungal activity of Css54 against C. albicans and analyze its mechanism. Css54 showed high antifungal
activity against C. albicans. Css54 also inhibited biofilm formation in fluconazole-resistant fungi. The antifungal mechanism
of action of Css54 was investigated using membrane-related assays, including the membrane depolarization assay and
analysis of the membrane integrity of C. albicans after treatment with Css54. Css54 induced reactive oxygen species (ROS)
production in C. albicans, which affected its antifungal activity. Our results indicate that Css54 causes membrane damage
in C. albicans, highlighting its value as a potential therapeutic agent against C. albicans infection.
Synthetic Short Cryptic Antimicrobial Peptides as Templates for the Development of Novel Biotherapeutics Against WHO Priority Pathogen Manjul Lata, Vrushti Telang, Pooja Gupta, Garima Pant, Mitra Kalyan, Jesu Arockiaraj, Mukesh Pasupuleti International Journal of Peptide Research and Therapeutics.2024;[Epub] CrossRef
Atopic dermatitis (AD) is a chronic inflammatory skin disease with repeated exacerbations of eczema and pruritus. Probiotics
can prevent or treat AD appropriately via modulation of immune responses and gut microbiota. In this study, we evaluated
effects of Lactobacillus acidophilus (L. acidophilus) KBL409 using a house dust mite (Dermatophagoides farinae)-induced
in vivo AD model. Oral administration of L. acidophilus KBL409 significantly reduced dermatitis scores and decreased
infiltration of immune cells in skin tissues. L. acidophilus KBL409 reduced in serum immunoglobulin E and mRNA levels
of T helper (Th)1 (Interferon-γ), Th2 (Interleukin [IL]-4, IL-5, IL-13, and IL-31), and Th17 (IL-17A) cytokines in skin tissues.
The anti-inflammatory cytokine IL-10 was increased and Foxp3 expression was up-regulated in AD-induced mice with
L. acidophilus KBL409. Furthermore, L. acidophilus KBL409 significantly modulated gut microbiota and concentrations
of short-chain fatty acids and amino acids, which could explain its effects on AD. Our results suggest that L. acidophilus
KBL409 is the potential probiotic for AD treatment by modulating of immune responses and gut microbiota of host.
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The Skin Histopathology of Pro- and Parabiotics in a Mouse Model of Atopic Dermatitis Hun Hwan Kim, Se Hyo Jeong, Min Yeong Park, Pritam Bhagwan Bhosale, Abuyaseer Abusaliya, Jeong Doo Heo, Hyun Wook Kim, Je Kyung Seong, Tae Yang Kim, Jeong Woo Park, Byeong Soo Kim, Gon Sup Kim Nutrients.2024; 16(17): 2903. CrossRef
Limosilactobacillus fermentum KBL674 Alleviates Vaginal Candidiasis Sung Jae Jang, Eun Jung Jo, Cheonghoon Lee, Bo-Ram Cho, Yun Jeong Shin, Jun Soo Song, Woon-Ki Kim, Nanhee Lee, Hyungjin Lee, SungJun Park, GwangPyo Ko Probiotics and Antimicrobial Proteins.2024;[Epub] CrossRef
Salmonella enterica serovar Typhimurium (S. Typhimurium) is a globally recognized foodborne pathogen that affects both
animals and humans. Endoribonucleases mediate RNA processing and degradation in the adaptation of bacteria to environmental
changes and have been linked to the pathogenicity of S. Typhimurium. Not much is known about the specific regulatory
mechanisms of these enzymes in S. Typhimurium, particularly in the context of environmental adaptation. Thus, this
study carried out a comparative transcriptomic analysis of wild-type S. Typhimurium SL1344 and its mutant (Δrnc), which
lacks the rnc gene encoding RNase III, thereby elucidating the detailed regulatory characteristics that can be attributed to the
rnc gene. Global gene expression analysis revealed that the Δrnc strain exhibited 410 upregulated and 301 downregulated
genes (fold-change > 1.5 and p < 0.05), as compared to the wild-type strain. Subsequent bioinformatics analysis indicated
that these differentially expressed genes are involved in various physiological functions, in both the wild-type and Δrnc
strains. This study provides evidence for the critical role of RNase III as a general positive regulator of flagellar-associated
genes and its involvement in the pathogenicity of S. Typhimurium.
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CspA regulates stress resistance, flagellar motility and biofilm formation in Salmonella Enteritidis Xiang Li, Yan Cui, Xiaohui Sun, Chunlei Shi, Shoukui He, Xianming Shi Food Bioscience.2025; 66: 106237. CrossRef
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Bacteria employ a diverse array of cellular regulatory
mechanisms to successfully adapt and thrive in ever-changing
environments, including but not limited to temperature
changes, fluctuations in nutrient availability, the presence
or absence of electron acceptors such as oxygen, the availability
of metal ions crucial for enzyme activity, and the
existence of antibiotics. Bacteria can virtually modulate
any step of gene expression from transcr!ptional initiation
to posttranslational modification of a protein for the control
of cellular processes. Furthermore, one gene regulator
often controls another in a complex gene regulatory network.
Thus, it is not easy to fully understand the intricacies of
bacterial regulatory mechanisms in various environments. In
this special issue, while acknowledging the challenge of covering
all aspects of bacterial regulatory mechanisms across
diverse environments, seven review articles are included to
provide insight into the recent progress in understanding
such mechanisms from different perspectives: positive regulatory
mechanisms by secondary messenger (cAMP receptor
protein), two-component signal transduction mechanisms
(Rcs and Cpx), diverse regulatory mechanisms by a specific
environmental factor in specific bacteria (oxygen availability
in Mycobacterium and manganese ion availability in Salmonella),
diverse regulatory mechanisms by a specific environmental
factor (temperature and antibiotics), and regulatory
mechanisms by antibiotics in cell wall synthesis.
Bacteria, as ubiquitous organisms that can be found in
almost every environment, carry out complex cellular processes
that allow them to survive and thrive in a variety of
different conditions despite their small size and relative simplicity.
One of the key factors that allows bacteria to carry
out these complex processes is their ability to regulate gene
expression through various mechanisms. Gene expression
is a fundamental biological process by which the genetic
information encoded in a gene is transcribed into an RNA
molecule and subsequently translated into a functional gene
product, often a protein. Furthermore, the activity levels of
proteins may further be altered by posttranslational modification.
Regulation of gene expression refers to the control
of the amount and timing of gene expression, and thus it
can be divided into transcr!ptional, translational, and posttranslational
levels.
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The PhoBR two-component system upregulates virulence in Aeromonas dhakensis C4–1 Wei Feng, Xuesong Li, Nuo Yang, Lixia Fan, Guiying Guo, Jun Xie, Xiuqing Cai, Yuqi Meng, Jifeng Zeng, Yu Han, Jiping Zheng Aquaculture.2025; 595: 741665. CrossRef
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PhoPQ-mediated lipopolysaccharide modification governs intrinsic resistance to tetracycline and glycylcycline antibiotics in
Escherichia coli
Byoung Jun Choi, Umji Choi, Dae-Beom Ryu, Chang-Ro Lee, Mehrad Hamidian, You-Hee Cho mSystems.2024;[Epub] CrossRef
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Antimicrobial agents targeting peptidoglycan have shown
successful results in eliminating bacteria with high selective
toxicity. Bacteriophage encoded endolysin as an alternative
antibiotics is a peptidoglycan degrading enzyme with a low
rate of resistance. Here, the engineered endolysin was developed
to defeat multiple drug-resistant (MDR) Acinetobacter
baumannii. First, putative endolysin PA90 was predicted by
genome analysis of isolated Pseudomonas phage PBPA. The
His-tagged PA90 was purified from BL21(DE3) pLysS and
tested for the enzymatic activity using Gram-negative pathogens
known for having a high antibiotic resistance rate including
A. baumannii. Since the measured activity of PA90
was low, probably due to the outer membrane, cell-penetrating
peptide (CPP) DS4.3 was introduced at the N-terminus
of PA90 to aid access to its substrate. This engineered endolysin,
DS-PA90, completely killed A. baumannii at 0.25 μM,
at which concentration PA90 could only eliminate less than
one log in CFU/ml. Additionally, DS-PA90 has tolerance to
NaCl, where the ~50% of activity could be maintained in the
presence of 150 mM NaCl, and stable activity was also observed
with changes in pH or temperature. Even MDR A. baumannii
strains were highly susceptible to DS-PA90 treatment:
five out of nine strains were entirely killed and four strains
were reduced by 3–4 log in CFU/ml. Consequently, DS-PA90
could protect waxworm from A. baumannii-induced death
by ~70% for ATCC 17978 or ~44% for MDR strain 1656-2
infection. Collectively, our data suggest that CPP-fused endolysin
can be an effective antibacterial agent against Gramnegative
pathogens regardless of antibiotics resistance mechanisms.
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Endolysins: a new antimicrobial agent against antimicrobial resistance. Strategies and opportunities in overcoming the challenges of endolysins against Gram-negative bacteria Fazal Mehmood Khan, Fazal Rasheed, Yunlan Yang, Bin Liu, Rui Zhang Frontiers in Pharmacology.2024;[Epub] CrossRef
Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria Tae-Hwan Jeong, Hye-Won Hong, Min Soo Kim, Miryoung Song, Heejoon Myung Viruses.2023; 15(3): 679. CrossRef
Design strategies for positively charged endolysins: Insights into Artilysin development Jose Vicente Carratalá, Anna Arís, Elena Garcia-Fruitós, Neus Ferrer-Miralles Biotechnology Advances.2023; 69: 108250. CrossRef
An efficient and eco-friendly bioefficacy of potent Trichofusant
(Fu21) and its green nanosilver formulation against
stem rot (Sclerotium rolfsii) in groundnut was established.
Fu21 demonstrated higher in-vitro growth inhibition of pathogen
with better fungicide tolerance than the parental strains.
The green nanosilver particles were synthesized from the extracellular
metabolites of Fu21 and characterized for shape
(spherical, 59.34 nm in scanning electron microscope), purity
(3.00 KeV, energy dispersive X-ray analysis), size (54.3 nm
in particle size analyzer), and stability (53.7 mv, zeta). The field
efficacy study exhibited that the seedling emergence was high
in seeds treated with green nanosilver (minimum inhibitory
concentration-[MIC] 20 μg Ag/ml), and a low disease severity
index of stem rot during the crop growth was followed by the
live antagonist (Fu21) in addition to seed treatment with a
fungicide mix under pathogen infestation. The seed quality
analysis of harvested pods revealed a high oil content with
balanced fatty acid composition (3.10 oleic/linoleic acid ratio)
in green nanosilver treatment under pathogen infestation.
The residual analysis suggested that green nanosilver applied
at the MIC level as seed treatment yielded similar effects as the
control for silver residue in the harvested groundnut seeds.
The green nanosilver at MIC has a high pod-yield under S.
rolfsii infestation, demonstrating green chemistry and sustainability
of the nanoproduct.
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Exploring conserved and novel MicroRNA-like small RNAs from stress tolerant Trichoderma fusants and parental strains during interaction with fungal phytopathogen Sclerotium rolfsii Sacc. Darshna G. Hirpara, H.P. Gajera, Disha D. Savaliya, M.V. Parakhia Pesticide Biochemistry and Physiology.2023; 191: 105368. CrossRef
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There are lots of seamounts globally whose primary production
is disproportionally greater than the surrounding areas.
Compared to other deep-sea environments, however, the
seamounts environment is relatively less explored for fungal
diversity. In the present study, we explored the fungal community
structure in deep-sea sediments from four different
stations of the Magellan seamounts environment by using
high-throughput sequencing of the ITS1 region. A total of
1,897,618 ITS1 sequences were obtained. Among these sequences,
fungal ITS1 sequences could be clustered into 1,662
OTUs. The majority of these sequences belonged to Ascomycota.
In the genera level, the most abundant genus was Mortierella
(4.79%), which was reported as a common fungal genus
in soil and marine sediments, followed by Umbelopsis
(3.80%), Cladosporium (2.98%), Saccharomycopsis (2.53%),
Aspergillus (2.42%), Hortaea (2.36%), Saitozyma (2.20%), Trichoderma
(2.12%), Penicillium (2.11%), Russula (1.86%), and
Verticillium (1.40%). Most of these recovered genera belong
to Ascomycota. The Bray-Curtis analysis showed that there
was 37 to 85% dissimilarity of fungal communities between
each two sediment samples. The Principal coordinates analysis
clearly showed variations in the fungal community among
different sediment samples. These results suggested that there
was a difference in fungal community structures not only
among four different sampling stations but also for different
layers at the same station. The depth and geographical distance
significantly affect the fungal community, and the effect of
depth and geographical distance on the structure of the fungal
community in the Magellan seamounts is basically same.
Most of the fungi were more or less related to plants, these
plant parasitic/symbiotic/endophytic fungi constitute a unique
type of seamounts environmental fungal ecology, different
from other marine ecosystems.
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A grey pink colored bacterium, strain t3-1-3T, was isolated
from the air at the foot of the Xiangshan Mountain in Beijing,
China. The cells are aerobic, Gram-stain-negative, non-sporeforming,
motile and coccoid-rod shaped (0.9–1.2 × 1.9–2.1
μm). Strain t3-1-3T was catalase-positive and oxidase-negative
and this strain grew at 4–42°C (optimum 28°C), a pH
of 4.0–9.0 (optimum pH 7.0) and under 0–2% (w/v) NaCl
(optimum 0–1% NaCl). A phylogenetic analysis based on 16S
rRNA gene sequences revealed that strain t3-1-3T was closely
related to Azohydromonas riparia UCM-11T (97.4% similarity),
followed by Azohydromonas australica G1-2T (96.8%)
and Azohydromonas ureilytica UCM-80T (96.7%). The genome
of strain t3-1-3T contains 6,895 predicted protein-encoding
genes, 8 rRNA genes, 62 tRNA genes and one sRNA
gene, as well as five potential biosynthetic gene clusters, including
clusters of genes coding for non-ribosomal peptide
synthetase (NRPS), bacteriocin and arylpolyene and two clusters
of genes for terpene. The predominant cellular fatty acids
(> 10.0% of the total) in strain t3-1-3T were summed feature
3 (C16:1ω7c and/or C16:1ω6c, 37.8%), summed feature 8
(C18:1ω7c and/or C18:1ω6c, 29.7%) and C16:0 (17.3%). Strain
t3-1-3T contained ubiquinone-8 (Q-8) as the predominant
respiratory quinone. The polar lipids of strain t3-1-3T comprised
phosphatidyl ethanolamine (PE), phosphatidyl glycerol
(PG), diphosphatidyl glycerol (DPG), an unidentified
glycolipid (GL), an unidentified aminophospholipid (APL),
two unidentified phospholipid (PL1-2) and five unidentified
lipid (L1-5). The DNA G + C content of the type strain
is 70.3%. The broader range of growth temperature, assimilation
of malic acid and trisodium citrate, presence of C18:3ω6c
and an unidentified glycolipid and absence of C12:0 2-OH and
C16:0iso differentiate strain t3-1-3T from related species. Based
on the taxonomic data presented in this study, we suggest
that strain t3-1-3T represents a novel species within the genus
Azohydromonas, for which the name Azohydromonas
aeria sp. nov. is proposed. The type strain of Azohydromonas
aeria is t3-1-3T (= CFCC 13393T = LMG 30135T).
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In situ injectable nano-complexed hydrogel based on chitosan/dextran for combining tumor therapy via hypoxia alleviation and TAMs polarity regulation Wenxue Zhang, Yan Shi, Hu Li, Miao Yu, Jiaxuan Zhao, Hao Chen, Ming Kong Carbohydrate Polymers.2022; 288: 119418. CrossRef
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The bacterial flagellum is an appendage structure that provides
a means for motility to promote survival in fluctuating
environments. For the intracellular pathogen Salmonella enterica
serovar Typhimurium to survive within macrophages,
flagellar gene expression must be tightly regulated, and thus,
is controlled at multiple levels, including DNA recombination,
transcription, post-transcription, protein synthesis, and
assembly within host cells. To understand the contribution of
flagella to Salmonella pathogenesis within the host, it is critical
to detect flagella production within macrophages via
microscopy. In this paper, we describe two methods for detecting
bacterial flagella by microscopy both in vitro and in
vivo infection models.
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Multi-drug resistant (MDR) non-typhoidal Salmonella (NTS)
is increasingly common worldwide. While food animals are
thought to contribute to the growing antimicrobial resistance
(AMR) problem, limited data is documenting this relationship,
especially in low and middle-income countries (LMIC).
Herein, we aimed to assess the role of non-clinical NTS of bovine
origin as reservoirs of AMR genes of human clinical significance.
We evaluated the phenotypic and genotypic AMR
profiles in a set of 44 bovine-associated NTS. For comparative
purposes, we also included genotypic AMR data of additional
isolates from Mexico (n = 1,067) that are publicly available.
The most frequent AMR phenotypes in our isolates involved
tetracycline (40/44), trimethoprim-sulfamethoxazole (26/44),
chloramphenicol (19/44), ampicillin (18/44), streptomycin
(16/44), and carbenicillin (13/44), while nearly 70% of the
strains were MDR. These phenotypes were correlated with
a widespread distribution of AMR genes (i.e. tetA, aadA,
dfrA12, dfrA17, sul1, sul2, bla-TEM-1, blaCARB-2) against
multiple antibiotic classes, with some of them contributed by
plasmids and/or class-1 integrons. We observed different
AMR genotypes for betalactams and tetracycline resistance,
providing evidence of convergent evolution and adaptive AMR.
The probability of MDR genotype occurrence was higher in
meat-associated isolates than in those from other sources (odds
ratio 11.2, 95% confidence interval 4.5–27.9, P < 0.0001). The
study shows that beef cattle are a significant source of MDR NTS in Mexico, highlighting the role of animal production
on the emergence and spread of MDR Salmonella in LMIC.
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pathogens, on-site applicable rapid detection methods have
been required for its control. The purpose of this study was
to isolate and purify S. Enteritidis-specific phage (KFS-SE2
phage) from an eel farm and to investigate its feasibility as a
novel, efficient, and reliable bio-receptor for its employment.
KFS-SE2 phage was successfully isolated at a high concentration
of (2.31 ± 0.43) × 1011 PFU/ml, and consisted of an
icosahedral head of 65.44 ± 10.08 nm with a non-contractile
tail of 135.21 ± 12.41 nm. The morphological and phylogenetic
analysis confirmed that it belongs to the Pis4avirus genus
in the family of Siphoviridae. KFS-SE2 genome consisted
of 48,608 bp with 45.7% of GC content. Genome analysis
represented KFS-SE2 to have distinctive characteristics as a
novel phage. Comparative analysis of KFS-SE2 phage with
closely related strains confirmed its novelty by the presence
of unique proteins. KFS-SE2 phage exhibited excellent specificity
to S. Enteritidis and was stable under the temperature
range of 4 to 50°C and pH of 3 to 11 (P < 0.05). The latent time
was determined to be 20 min. Overall, a new lytic KFS-SE2
phage was successfully isolated from the environment at a
high concentration and the excellent feasibility of KFS-SE2
phage was demonstrated as a new bio-receptor for S. Enteritidis
detection.
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In this study, we sought to isolate Salmonella Enteritidis-specific
lytic bacteriophages (phages), and we found a lytic phage
that could lyse not only S. Enteritidis but also other Gramnegative
foodborne pathogens. This lytic phage, SS3e, could
lyse almost all tested Salmonella enterica serovars as well as
other enteric pathogenic bacteria including Escherichia coli,
Shigella sonnei, Enterobacter cloacae, and Serratia marcescens.
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