Isolation and genomic characterization of P.A-5, a novel virulent bacteriophage against Enterobacter hormaechei

Chen, Cao-wei; Yuan, Lei; Zhou, Wenyuan; Mgomi, Fedrick C.; Zhang, Yuan‐song; Wang, Yang; Zheng, Xizhong; Rao, Shengqi; Yang, Zhenquan; Jiao, Xinan

doi:10.1016/j.micpath.2021.104767

Cited by 15 publications

(5 citation statements)

References 41 publications

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“…Interestingly, studies using the new phage against Enterococcus faecalis producing biofilm in Foley catheters initially reported more efficacy of phage lytic activity at a higher MOI (MOI = 10). On the other hand, after 6 h and 24 h of exposing the bacterial biofilm to the phage, the preparations with a lower phage-to-bacteria ratio (MOI = 0.01 and MOI = 0.0001) were more effective [ 80 ]. However, we describe the MOI as the ratio of the number of phages to the number of bacteria at the start of the experiment, and it is suggested to also consider, inter alia, the changing densities of bacteria during the course of the experiment [ 81 ].…”

Section: Discussionmentioning

confidence: 99%

Two Newly Isolated Enterobacter-Specific Bacteriophages: Biological Properties and Stability Studies

Cieślik

Harhala

Orwat

et al. 2022

Viruses

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In an era of antibiotic therapy crisis caused by spreading antimicrobial resistance, and when recurrent urinary tract infections constitute a serious social and medical problem, the isolation and complex characterization of phages with a potential therapeutic application represents a promising solution. It is an inevitable, and even a necessary direction in the development of current phage research. In this paper, we present two newly isolated myoviruses that show lytic activity against multidrug-resistant clinical isolates of Enterobacter spp. (E. cloacae, E. hormaechei, and E. kobei), the genomes of which belong to a poorly represented phage group. Both phages were classified as part of the Tevenvirinae subfamily (Entb_43 was recognized as Karamvirus and Entb_45 as Kanagawavirus). Phage lytic spectra ranging from 40 to 60% were obtained. The most effective phage-to-bacteria ratios (MOI = 0.01 and MOI = 0.001) for both the phage amplification and their lytic activity against planktonic bacteria were also estimated. Complete adsorption to host cells were obtained after about 20 min for Entb_43 and 10 min for Entb_45. The phage lysates retained their initial titers even during six months of storage at both −70 °C and 4 °C, whereas storage at 37 °C caused a complete loss in their activity. We showed that phages retained their activity after incubation with solutions of silver and copper nanoparticles, which may indicate possible synergistic antibacterial activity. Moreover, a significant reduction in phage titers was observed after incubation with a disinfectant containing octenidinum dihydrochloridum and phenoxyethanol, as well as with 70% ethanol. The observed maintenance of phage activity during incubation in a urine sample, along with other described properties, may suggest a therapeutic potential of phages at the infection site after intravesical administration.

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Section: Discussionmentioning

confidence: 99%

Two Newly Isolated Enterobacter-Specific Bacteriophages: Biological Properties and Stability Studies

Cieślik

Harhala

Orwat

et al. 2022

Viruses

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“…Phage titres were determined using the double agar plaque assay as described by Chen et al . (2021). In brief, 100 μL of the filtrate and 100 μL of an overnight culture of host C. sakazakii were incubated at 25°C for 10 min, mixed with 5 mL of nutrient broth (contained 0.5% of agar powder), followed by pouring onto solid nutritional agar plates (contained 1.5% of agar powder), and then incubated for 10 h at 37°C until clear plaques appeared.…”

Section: Methodsmentioning

confidence: 99%

“…Afterwards, 5 mL of phage stock and 100 μL overnight culture of host C. sakazakii were added to 5 mL of fresh nutrient broth; the mixtures were incubated at 37°C at a constant rate (120 rpm) for 12 h. After incubation, the cultures were vortexed and centrifuged as previously described, and the supernatant was filtered by passing through a 0.45-μm filter. Phage titres were determined using the double agar plaque assay as described by Chen et al (2021). In brief, 100 μL of the filtrate and 100 μL of an overnight culture of host C. sakazakii were incubated at 25°C for 10 min, mixed with 5 mL of nutrient broth (contained 0.5% of agar powder), followed by pouring onto solid nutritional agar plates (contained 1.5% of agar powder), and then incubated for 10 h at 37°C until clear plaques appeared.…”

Section: Isolation and Preparation Of Phage Espyzu12mentioning

confidence: 99%

Specific detection of viable Cronobacter sakazakii in powdered infant formula by phage amplification combined with qPCR (PAA‐qPCR) assay

Zhang

Yuan

Chen

et al. 2022

Int J of Dairy Tech

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Cronobacter sakazakii is a foodborne pathogen that has become a significant challenge to public health and poses a serious risk to food safety, especially in powdered infant formula. This study applied a novel approach combining the Phage Amplification Assay with qPCR (PAA‐qPCR), amplifying the C. sakazakii phage EspYZU12 coupled with phage‐specific qPCR to detect viable C. sakazakii in milk samples. Results showed that the limit of detection of 6.58 × 102 CFU/mL of C. sakazakii could be assayed in less than 5.5 h with phage EspYZU12 (103 PFU/mL). Additionally, compared with plate counting, the accuracy factor value (Af), the bias factor value (Bf) and the root mean square error (RMSE) of the PAA‐qPCR method were 1.029, 1.026 and 1.808 respectively. The PAA‐qPCR method exhibited high sensitivity, rapidity and specificity for detecting live C. sakazakii in dairy products.

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“…Chinese soft‐shell turtle farming is associated with many infectious diseases during superintensive aquaculture growth (Zhou et al., 2008), including viruses, bacteria (Yuan et al., 2020) and fungi (Takuma et al., 2011), causing severe losses in this industry. These diseases are mainly caused by bacteria, such as Aeromonas hydrophila (Lin et al., 2014; Wu, 2004), Aeromonas caviae (Wu, 2004; Yu & Ma, 1997), Aeromonas sobria (Wu, 2004), Edwardsiella tarda (Pan et al., 2010), Lactococcus garvieae (Hu et al., 2010), Citrobacter freundii (Tang et al., 2023), Morganella morganii (Wei et al., 2023), Bacillus thuringiensis (Chen et al., 2014) and Bacillus cereus (Yuan et al., 2020).…”

Section: Introductionmentioning

confidence: 99%

Isolation, identification and characterization of Aeromonas jandaei from diseased Chinese soft‐shell turtles

Chen,

Xue,

Chen

et al. 2024

Journal of Fish Diseases

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Aeromonas jandaei is a gram‐negative bacterium commonly found in aquatic environments and can induce illnesses in amphibians, reptiles and aquatic animals. In this study, a strain of bacteria was isolated from the diseased Chinese soft‐shell turtle (Pelodiscus sinensis), then named strain JDP‐FX. This isolate was identified as A. jandaei after analysis of morphological, physiological and biochemical characteristics, as well as 16S rRNA and gyrB gene sequences. Virulence genetic testing further detected temperature‐sensitive protease (eprCAI), type III secretion system (TTSS) (ascv), nuclease (nuc), cytotonic enterotoxin (alt) and serine proteinase (ser) in JDP‐FX. Compared with healthy Chinese soft‐shell turtle, the serum levels of total protein (TP), albumin (ALB) and globulin (GLB) were significantly decreased in the diseased Chinese soft‐shell turtle, while, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were significantly increased. Histopathological observations showed that multiple tissues, including intestinal mucosa, liver and kidney, were severely damaged in the diseased Chinese soft‐shell turtle. Moreover, the diseased Chinese soft‐shell turtle had significant cell degeneration, necrosis, sloughing and interstitial inflammatory cell infiltration. The pathogenicity of JDP‐FX was tested via artificial infection. The median lethal dosage (LD50) of the strain was 1.05 × 105 colony forming units (CFU/g) per weight of Chinese soft‐shell turtle. Drug susceptibility analysis revealed that JDP‐FX was susceptible to ceftazidime, minocycline, cefoperazone, ceftriaxone and piperacillin. In addition, JDP‐FX was resistant to doxycycline, florfenicol, sulfonamides, gentamicin, ampicillin and neomycin. Therefore, this study may provide guidance for further research into the diagnosis, prevention and treatment of JDP‐FX infection.

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Isolation and genomic characterization of P.A-5, a novel virulent bacteriophage against Enterobacter hormaechei

Cited by 15 publications

References 41 publications

Two Newly Isolated Enterobacter-Specific Bacteriophages: Biological Properties and Stability Studies

Two Newly Isolated Enterobacter-Specific Bacteriophages: Biological Properties and Stability Studies

Specific detection of viable Cronobacter sakazakii in powdered infant formula by phage amplification combined with qPCR (PAA‐qPCR) assay

Isolation, identification and characterization of Aeromonas jandaei from diseased Chinese soft‐shell turtles

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