scholarly journals Characterization of the Three New Kayviruses and Their Lytic Activity Against Multidrug-Resistant Staphylococcus aureus

2019 ◽  
Vol 7 (10) ◽  
pp. 471 ◽  
Author(s):  
Natalia Łubowska ◽  
Bartłomiej Grygorcewicz ◽  
Katarzyna Kosznik-Kwaśnicka ◽  
Agata Zauszkiewicz-Pawlak ◽  
Alicja Węgrzyn ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Burst Size ◽  
Genomic Analysis ◽  
Multidrug Resistant ◽  
Biological Properties ◽  
Lytic Activity ◽  
Broad Host Range ◽  
Transmission Electron ◽  
Short Latent Period

The development of antimicrobial resistance has become a global concern. One approach to overcome the problem of drug resistance is the application of bacteriophages. This study aimed at characterizing three phages isolated from sewage, which show lytic activity against clinical isolates of multidrug-resistant Staphylococcus aureus. Morphology, genetics and biological properties, including host range, adsorption rate, latent time, phage burst size and lysis profiles, were studied in all three phages. As analyzed by transmission electron microscopy (TEM), phages vB_SauM-A, vB_SauM-C, vB_SauM-D have a myovirion morphology. One of the tested phages, vB_SauM-A, has relatively rapid adsorption (86% in 17.5 min), short latent period (25 min) and extremely large burst size (~500 plaque-forming units (PFU) per infected cell). The genomic analysis revealed that vB_SauM-A, vB_SauM-C, vB_SauM-D possess large genomes (vB_SauM-A 139,031 bp, vB_SauM-C 140,086 bp, vB_SauM-D 139,088 bp) with low G+C content (~30.4%) and are very closely related to the phage K (95–97% similarity). The isolated bacteriophages demonstrate broad host range against MDR S. aureus strains, high lytic activity corresponding to strictly virulent life cycle, suggesting their potential to treat S. aureus infections.

scholarly journals JD419, a Staphylococcus aureus Phage With a Unique Morphology and Broad Host Range

2021 ◽  
Vol 12 ◽  
Author(s):  
Tingting Feng ◽  
Sebastian Leptihn ◽  
Ke Dong ◽  
Belinda Loh ◽  
Yan Zhang ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Complete Characterization ◽  
Resistant Bacteria ◽  
Broad Host Range ◽  
Gene Engineering ◽  
Mrsa Strains

Phage therapy represents a possible treatment option to cure infections caused by multidrug-resistant bacteria, including methicillin and vancomycin-resistant Staphylococcus aureus, to which most antibiotics have become ineffective. In the present study, we report the isolation and complete characterization of a novel phage named JD219 exhibiting a broad host range able to infect 61 of 138 clinical strains of S. aureus tested, which included MRSA strains as well. The phage JD419 exhibits a unique morphology with an elongated capsid and a flexible tail. To evaluate the potential of JD419 to be used as a therapeutic phage, we tested the ability of the phage particles to remain infectious after treatment exceeding physiological pH or temperature. The activity was retained at pH values of 6.0–8.0 and below 50°C. As phages can contain virulence genes, JD419’s complete genome was sequenced. The 45509 bp genome is predicted to contain 65 ORFs, none of which show homology to any known virulence or antibiotic resistance genes. Genome analysis indicates that JD419 is a temperate phage, despite observing rapid replication and lysis of host strains. Following the recent advances in synthetic biology, JD419 can be modified by gene engineering to remove prophage-related genes, preventing potential lysogeny, in order to be deployed as a therapeutic phage.

scholarly journals Morphological, biological, and genomic characterization of a newly isolated lytic phage Sfk20 infecting Shigella flexneri, Shigella sonnei, and Shigella dysenteriae1

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Bani Mallick ◽  
Payel Mondal ◽  
Moumita Dutta
Keyword(s):  
Burst Size ◽  
Shigella Flexneri ◽  
Alternative Therapy ◽  
Genomic Analysis ◽  
Multidrug Resistant ◽  
Lytic Activity ◽  
Lytic Phage ◽  
Tem Analysis ◽  
Host Interaction ◽  
Outbreak Area

AbstractShigellosis, caused by Shigella bacterial spp., is one of the leading causes of diarrheal morbidity and mortality. An increasing prevalence of multidrug-resistant Shigella species has revived the importance of bacteriophages as an alternative therapy to antibiotics. In this study, a novel bacteriophage, Sfk20, has been isolated from water bodies of a diarrheal outbreak area in Kolkata (India) with lytic activity against many Shigella spp. Phage Sfk20 showed a latent period of 20 min and a large burst size of 123 pfu per infected cell in a one-step growth analysis. Phage-host interaction and lytic activity confirmed by phage attachment, intracellular phage development, and bacterial cell burst using ultrathin sectioning and TEM analysis. The genomic analysis revealed that the double-stranded DNA genome of Sfk20 contains 164,878 bp with 35.62% G + C content and 241 ORFs. Results suggested phage Sfk20 to include as a member of the T4 myoviridae bacteriophage group. Phage Sfk20 has shown anti-biofilm potential against Shigella species. The results of this study imply that Sfk20 has good possibilities to be used as a biocontrol agent.

scholarly journals Phage JS02, A Putative Temperate Phage, A Novel Biofilm-Degrading Agent for Staphylococcus Aureus

2020 ◽  
Author(s):  
Lili Zhang ◽  
Huiyan Ding ◽  
Khashayar Shahin ◽  
Abbas Soleimani-Delfan ◽  
Heye Wang ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Burst Size ◽  
Multidrug Resistant ◽  
Temperate Phage ◽  
Natural Resistance ◽  
Broad Host Range ◽  
Planktonic Cells ◽  
Livestock Wastewater ◽  
Promising Alternative ◽  
Packaging Structure

Abstract Background: Staphylococcus aureus is a biofilm-producing organism that is frequently isolated from various environments worldwide. Because of the natural resistance of S. aureus biofilm to antibiotics, bacteriophages are considered as a promising alternative for its removal. Results: The bacteriophage vB_SauS_JS02 was isolated from livestock wastewater and showed activity against multidrug-resistant (MDR) S. aureus. The phage vB_SauS_JS02 was morphologically classified as Siphoviridae; it had a broad host range (45 out of 81 strains, 55.6%) and high burst size (52 plaque-forming unit (PFU)/infected cell) and could survive in a pH range of 4 to 11 and a temperature range of 40 ºC to 50 ºC. Bioinformatics analysis showed that the phage genome contained a long double-stranded linear DNA genome of 46,435 base pairs with a G＋C content of 33.1% and had 66 putative open reading frames (ORFs). The predicted protein products of the ORFs were clustered functionally into five groups as follows: replication/regulation, DNA packaging, structure/morphogenesis, lysis, and lysogeny. The phage vB_SauS_JS02 was a temperate phage with a higher inhibiting and degrading activity against planktonic cells (~86% reduction) and S. aureus biofilm (∼68% reduction in biofilm formation). Moreover, the removal activity of the phage vB_SauS_JS02 against both planktonic cells and S. aureus biofilms was even better than that of the antibiotic (ceftazidime). Conclusion: In summary, the present study introduced the phage vB_SauS_JS02 as a potential biocontrol agent against biofilm-producing S. aureus.

scholarly journals TSP, a virulent Podovirus can control the growth of Staphylococcus aureus till 12 hours

2021 ◽  
Author(s):  
Rabia Tabassum ◽  
Iqbal Ahmed Alvi ◽  
Muhammad Asif ◽  
Abdul Basit ◽  
Shafiq ur Rehman
Keyword(s):  
Staphylococcus Aureus ◽  
Latent Period ◽  
Genome Sequence ◽  
Burst Size ◽  
Gc Content ◽  
Comparative Genomic ◽  
Broad Host Range ◽  
Lytic Bacteriophage ◽  
Short Latent Period ◽  
Promising Therapeutic Approach

Abstract Methicillin-resistant Staphylococcus aureus (MRSA) is a prevailing nosocomial pathogen that causes a large number of diseases in healthcare and community settings. The MRSA causes infections in different tissues of immunocompromised individuals leading to increased morbidity and mortality. It possess various virulence mechanisms to show resistance against to a lot of beta-lactam antibiotics. To tackle this emerging issue of MRSA, there is an urgent need of antibiotic alternatives and utilizing lytic bacteriophages is one of the best promising therapeutic approach. In the present study, a lytic bacteriophage TSP was isolated from hospital wastewater against MRSA. Its morphology, physiology, host specificity, burst size and lytic spectrum were determined and complete genome sequence was analyzed. TSP phage efficiently inhibit bacterial growth for up to 12 hours. TSP phage showed broad lytic spectrum against clinical isolates of MRSA (78%) and MSSA (37%). It showed stability at varying temperatures (25ºC, 37ºC) and pH (5–9), while its maximum storage stability was observed at 4ºC. It had short latent period (20min) and high burst size (103 PFU/ infected cell). TSP genome sequence and restriction analysis revealed that its genome is linear having 17,987 bp in length with an average GC content of 29.7%. The TSP genome showed 98% similarity to S aureus phages SCH1, SCH11 and vB SauP-436A1. According to comparative genomic analysis and phylogenetic tree analysis, TSP phage can be considered as a member of genus “P68viruses”. The strong lytic activity, broad host range and short latent period along with absence of any lysogenic and toxic genes make TSP a very good candidate for phage therapy against MRSA infections if prove safe during in vivo studies.

scholarly journals Characterization of a Broad-Host-Range Lytic Phage SHWT1 Against Multidrug-Resistant Salmonella and Evaluation of Its Therapeutic Efficacy in vitro and in vivo

2021 ◽  
Vol 8 ◽  
Author(s):  
Chenglin Tao ◽  
Zhengfei Yi ◽  
Yaodong Zhang ◽  
Yao Wang ◽  
Hong Zhu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Host Range ◽  
Therapeutic Efficacy ◽  
Multidrug Resistant ◽  
Lytic Activity ◽  
Resistant Bacteria ◽  
Lytic Phage ◽  
Broad Host Range

Inappropriate use of antibiotics has accelerated to the emergence of multidrug-resistant bacteria, becoming a major health threat. Moreover, bacterial biofilms contribute to antibiotic resistance and prolonged infections. Bacteriophage (phage) therapy may provide an alternative strategy for controlling multidrug-resistant bacterial infections. In this study, a broad-host-range phage, SHWT1, with lytic activity against multidrug-resistant Salmonella was isolated, characterized and evaluated for the therapeutic efficacy in vitro and in vivo. Phage SHWT1 exhibited specific lytic activity against the prevalent Salmonella serovars, such as Salmonella Pullorum, Salmonella Gallinarum, Salmonella Enteritidis, and Salmonella Typhimurium. Morphological analysis showed that phage SHWT1 was a member of the family Siphoviridae and the order Caudovirales. Phage SHWT1 had a latent period of 5 min and burst size of ~150 plaque-forming units (PFUs)/cell. The phage was stable from pH 3-12 and 4–65°C. Phage SHWT1 also showed capacity to lyse Salmonella planktonic cells and inhibit the biofilm formation at optimal multiplicity of infection (MOI) of 0.001, 0.01, 0.1, and 100, respectively. In addition, phage SHWT1 was able to lyse intracellular Salmonella within macrophages. Genome sequencing and phylogenetic analyses revealed that SHWT1 was a lytic phage without toxin genes, virulence genes, antibiotic resistance genes, or significant genomic rearrangements. We found that phage SHWT1 could successfully protect mice against S. enteritidis and S. typhimurium infection. Elucidation of the characteristics and genome sequence of phage SHWT1 demonstrates that this phage is a potential therapeutic agent against the salmonellosis caused by multidrug-resistant Salmonella.

scholarly journals Characterization and Application of a Lytic Phage D10 against Multidrug-Resistant Salmonella

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1626
Author(s):  
Zhiwei Li ◽  
Wanning Li ◽  
Wenjuan Ma ◽  
Yifeng Ding ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Burst Size ◽  
Foodborne Pathogen ◽  
Food Contamination ◽  
Genomic Analysis ◽  
Multidrug Resistant ◽  
Lytic Phage ◽  
Promising Tool ◽  
Short Latent Period ◽  
Dynamic Growth ◽  
Natural Foods

Salmonella is a widely distributed foodborne pathogen that is a serious threat to human health. The accelerated development of drug resistance and the increased demand for natural foods invoke new biocontrol agents to limit contamination by multidrug-resistant (MDR) Salmonella strains. In this study, a lytic Salmonella phage named D10 was characterized at the biological and genomic levels. D10 possesses a short latent period (10 min) and a large burst size (163 PFU/cell), as well as adequate stability under a range of pH conditions and moderate thermal tolerance. D10 effectively lysed different MDR Salmonella serovars and repressed their dynamic growth in the medium. Genomic analysis disclosed that D10 is a new member of the Siphoviridae family and lacks the genes implicated in lysogeny, pathogenicity, or antibiotic resistance. A three-ingredient phage cocktail was then developed by mixing D10 with previously identified myovirus D1-2 and podovirus Pu20. The cocktail significantly reduced the count of MDR strains in liquid eggs, regardless of the temperature applied (4 and 25 °C). These results suggest that phage D10 is a promising tool to prevent food contamination by MDR Salmonella.

scholarly journals Evaluation of the Efficacy of a Bacteriophage in the Treatment of Pneumonia Induced by Multidrug ResistanceKlebsiella pneumoniaein Mice

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Fang Cao ◽  
Xitao Wang ◽  
Linhui Wang ◽  
Zhen Li ◽  
Jian Che ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Burst Size ◽  
Multidrug Resistant ◽  
Lung Lesion ◽  
Lytic Bacteriophage ◽  
Short Latent Period ◽  
Dose Dependent ◽  
Antibiotic Control

Multidrug-resistantKlebsiella pneumoniae(MRKP) has steadily grown beyond antibiotic control. However, a bacteriophage is considered to be a potential antibiotic alternative for treating bacterial infections. In this study, a lytic bacteriophage, phage 1513, was isolated using a clinical MRKP isolate KP 1513 as the host and was characterized. It produced a clear plaque with a halo and was classified as Siphoviridae. It had a short latent period of 30 min, a burst size of 264 and could inhibit KP 1513 growthin vitrowith a dose-dependent pattern. Intranasal administration of a single dose of 2 × 109 PFU/mouse 2 h after KP 1513 inoculation was able to protect mice against lethal pneumonia. In a sublethal pneumonia model, phage-treated mice exhibited a lower level ofK. pneumoniaeburden in the lungs as compared to the untreated control. These mice lost less body weight and exhibited lower levels of inflammatory cytokines in their lungs. Lung lesion conditions were obviously improved by phage therapy. Therefore, phage 1513 has a great effectin vitroandin vivo, which has potential to be used as an alternative to an antibiotic treatment of pneumonia that is caused by the multidrug-resistantK. pneumoniae.

Molecular Characterization and Genomic Analysis of Novel Phage vB_ ShiP-A7 Infecting Multidrug-Resistant Shiglla Fexneri and Escherichia Coli

2020 ◽  
Author(s):  
Jing Xu ◽  
Yu Gu ◽  
Xinyan Yu ◽  
Ruiyang Zhang ◽  
Xuesen Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Case Reports ◽  
Burst Size ◽  
Shigella Flexneri ◽  
Genomic Analysis ◽  
Multidrug Resistant ◽  
Open Reading Frames ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Spherical Head

Abstract BackgroundPhage therapy has regained more attention due to the rise of multidrug-resistant (MDR) bacteria. Several case reports demonstrated clinical application of phage in resolving infections caused by MDR bacteria in recent years. ResultsWe isolated a new phage, vB_ShiP-A7, and then investigated its characteristics. Phage vB_ShiP-A7 is a member of Podoviridae that has an icosahedral spherical head and a short tail. vB_ShiP-A7 has large burst size and short replication time. vB_ShiP-A7’s genome is linear double stranded DNA composed of 40058 bp, encoding forty-three putative open reading frames. Comparative genome analysis demonstrated vB_ShiP-A7’s genome sequence is closely related to fifteen different phages (coverage 74-88%, identity 86-93%). Mass Spectrometry analysis revealed that twelve known proteins and six hypothetical proteins exist in particles of vB_ShiP-A7. Genome and proteome analyses confirmed the absence of lysogen-related proteins and toxic proteins in this phage. In addition, phage vB_ShiP-A7 can significantly reduce the growth of clinical MDR stains of Shigella flexneri and Escherichia coli in liquid culture. Furthermore, vB_ShiP-A7 can disrupt biofilms formed by Shigella flexneri or Escherichia coli in vitro. ConclusionPhage vB_ShiP-A7 is a stable novel phage, which has a strong application potential to inhibit MDR stains of Shigella flexneri and Escherichia coli. Comparing the genomes between vB_ShiP-A7 and other closely-related phages will help us better understand the evolutionary mechanism of phages.

scholarly journals Characterization and genomic analysis of two Staphylococcus aureus bacteriophages isolated from poultry/livestock farms

2013 ◽  
Vol 94 (11) ◽  
pp. 2569-2576 ◽  
Author(s):  
Hyunjin Yoon ◽  
Jiae Yun ◽  
Jeong-A Lim ◽  
Eunjung Roh ◽  
Kyu-Seok Jung ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Genomic Analysis ◽  
Enzyme Treatment ◽  
Comparative Genomic ◽  
The Family ◽  
Phage Dna ◽  
Host Range Determination ◽  
Range Determination ◽  
Livestock Farms

Staphylococcus aureus is one of the most important pathogens, causing various diseases in humans and animals. As methicillin-resistant S. aureus (MRSA) has become increasingly prevalent, controlling this pathogen with standard antibiotic treatment has become challenging. Bacteriophages (phages) have attracted interest as alternative antibacterial agents to control MRSA. In this study, we isolated six S. aureus phages from soils of poultry/livestock farms. Based on the results of host range determination with 150 S. aureus strains and restriction enzyme treatment of phage DNA, two phages, designated SP5 and SP6, were selected for further characterization and genome sequencing. Both SP5 and SP6 were classified as members of the family Siphoviridae. The genome of SP5 comprises 43 305 bp and contains 63 ORFs, while the SP6 genome comprises 42 902 bp and contains 61 ORFs. Although they have different host spectra, the phage genomes exhibit high nucleotide similarity to each other. Adsorption assay results suggested that the host range determinants of the two phages are involved in both adsorption and infection. Comparative genomic analyses of the two phages provided evidence that the lysogenic/lytic control module and tail proteins may be important for host specificity.

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