Molecular Characterization of Ahp2, a Lytic Bacteriophage of Aeromonas hydrophila

Aeromonas hydrophila is an opportunistic pathogen that infects fish, amphibians, mammals, and humans. This study isolated a myophage, vB_AhyM_Ahp2 (Ahp2), that lytically infects A. hydrophila. We observed that 96% of the Ahp2 particles adsorbed to A. hydrophila within 18 min. Ahp2 also showed a latent period of 15 min with a burst size of 142 PFU/cell. This phage has a linear double-stranded DNA genome of 47,331 bp with a GC content of 57%. At least 20 Ahp2 proteins were detected by SDS-polyacrylamide gel electrophoresis; among them, a 40-kDa protein was predicted as the major capsid protein. Sequence analysis showed that Ahp2 has a genome organization closely related to a group of Aeromonas phages (13AhydR10RR, 14AhydR10RR, 85AhydR10RR, phage 3, 32 Asp37, 59.1), which infect Aeromonas hydrophila and Aeromonas salmonicida. The tail module encompassing ORF27-29 in the Ahp2 genome was present in all Aeromonas phages analyzed in this study and likely determines the host range of the virus. This study found that Ahp2 completely lyses A. hydrophila AH300206 in 3.5 h at a MOI of 0.0001 and does not lysogenize its host. Altogether, these findings show that Ahp2 is a lytic Aeromonas phage and could be a candidate for therapeutic phage cocktails.

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A moderately halophilic Vibrio from a Spanish saltern and its lytic bacteriophage

Canadian Journal of Microbiology ◽

10.1139/m96-130 ◽

1996 ◽

Vol 42 (10) ◽

pp. 1015-1023 ◽

Cited By ~ 11

Author(s):

Usha Goel ◽

Tiiu Kauri ◽

Donn J. Kushner ◽

Hans-W. Ackermann

Keyword(s):

Burst Size ◽

Lytic Phage ◽

Host Bacterium ◽

Lytic Bacteriophage ◽

Moderate Halophile ◽

Double Stranded Dna ◽

Halophilic Archaebacteria ◽

Water Salt ◽

Inhibitory Agents ◽

Number Of Bacteria

A number of bacteria and their phages were isolated from a saltern near Alicante, Spain. One isolate, Vibrio B1, a moderate halophile that is probably a strain of Vibrio costicola, was host to a lytic phage, UTAK. Studies of the host bacterium included the effects of salt concentrations on the action of a number of inhibitory agents. Phage UTAK has a head, a tail, and a baseplate. It contains 80 kbp of double-stranded DNA with no unusual bases. It was stable for long periods in the absence of high salt concentrations and even in distilled water. Salt concentrations had little effect on adsorption of UTAK to its host but resulted in considerable changes in burst size. It appears that phages of halophilic and salt-tolerant eubacteria, and also of some marine bacteria, have much lower salt requirements for stability than the phages of halophilic archaebacteria. Our results suggest that ionic controls of phage replication in these eubacteria may differ from those of growth.Key words: halophiles, Vibrio sp., bacteriophage, salt responses.

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Complete Genomic Sequence of the Lytic Bacteriophage φYeO3-12 of Yersinia enterocolitica Serotype O:3

Journal of Bacteriology ◽

10.1128/jb.183.6.1928-1937.2001 ◽

2001 ◽

Vol 183 (6) ◽

pp. 1928-1937 ◽

Cited By ~ 71

Author(s):

Maria I. Pajunen ◽

Saija J. Kiljunen ◽

M. E.-Lotta Söderholm ◽

Mikael Skurnik

Keyword(s):

Yersinia Enterocolitica ◽

Genomic Sequence ◽

Gc Content ◽

Lytic Bacteriophage ◽

Gene Products ◽

Serotype O ◽

Double Stranded Dna ◽

Promoter Specificity ◽

Dna Strand ◽

Complete Genomic

ABSTRACT φYeO3-12 is a T3-related lytic bacteriophage of Yersinia enterocolitica serotype O:3. The nucleotide sequence of the 39,600-bp linear double-stranded DNA (dsDNA) genome was determined. The phage genome has direct terminal repeats of 232 bp, a GC content of 50.6%, and 54 putative genes, which are all transcribed from the same DNA strand. Functions were assigned to 30 genes based on the similarity of the predicted products to known proteins. A striking feature of the φYeO3-12 genome is its extensive similarity to the coliphage T3 and T7 genomes; most of the predicted φYeO3-12 gene products were >70% identical to those of T3, and the overall organizations of the genomes were similar. In addition to an identical promoter specificity, φYeO3-12 shares several common features with T3, nonsubjectibility to F exclusion and growth on Shigella sonneiD2371-48 (M. Pajunen, S. Kiljunen, and M. Skurnik, J. Bacteriol. 182:5114–5120, 2000). These findings indicate that φYeO3-12 is a T3-like phage that has adapted to Y. enterocolitica O:3 or vice versa. This is the first dsDNA yersiniophage genome sequence to be reported.

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Biological Characteristics and Genomic Analysis of Aeromonas Hydrophila Phage BUCT551 Isolated From Aquatic Sewage

10.21203/rs.3.rs-409219/v1 ◽

2021 ◽

Author(s):

Hongbo Qin ◽

Shiting He ◽

Xuling Xu ◽

Xiaoping An ◽

Ke Liu ◽

...

Keyword(s):

Aeromonas Hydrophila ◽

Burst Size ◽

Genomic Analysis ◽

Opportunistic Pathogen ◽

Range Analysis ◽

Transmission Electron ◽

Blastn Analysis ◽

One Step ◽

Ph Range ◽

The One

Abstract Aeromonas hydrophila is a common opportunistic pathogen in aquaculture and is ubiquitous in aquatic environment. Whereby its accessibility, variety and host specificity, phage is increasingly considered as a promising complementary medicine for antibiotics. However, a small amount of A. hydrophila phages have been characterized, which suggests the significance to isolate and characterize novel A. hydrophila phages. In this study, we isolated a novel Aeromonas hydrophila phage using A. hydrophila strain A18 as an indicator and designated it as BUCT551, and it was identified as Myoviridae phage by transmission electron microscopy (TEM). The whole genome sequencing of the phage BUCT551 revealed that it has a linear DNA genome of 613,82 bp. BLASTn analysis showed that phage BUCT551 shared 86.75% homology with A. hydrophila phage LAh_7 (Genebank ID: MK838113.1). The one-step growth curve demonstrated that phage BUCT551 had a latent period of 20 min and the burst size of 32 pfu/cell at its optimal MOI of 0.1. The phage BUCT551 had a survival pH range from 5 to 10 and tolerant temperature from 0℃ to 40℃. Host range analysis shown that the phage was able to lyse not only A. hydrophila, but also A. veronii.

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Biological characteristics and whole-genome analysis of the Enterococcus faecalis phage PEf771

Canadian Journal of Microbiology ◽

10.1139/cjm-2019-0336 ◽

2020 ◽

Vol 66 (9) ◽

pp. 505-520 ◽

Cited By ~ 1

Author(s):

Yingying Xiang ◽

Wenyu Li ◽

Fei Song ◽

Xianghong Yang ◽

Jing Zhou ◽

...

Keyword(s):

Enterococcus Faecalis ◽

Burst Size ◽

Gc Content ◽

Lytic Cycle ◽

Proteinase K ◽

Host Bacterium ◽

Whole Genome ◽

Strain Specificity ◽

Whole Genome Analysis ◽

A Genome

Enterococcus faecalis is a common pathogen causing refractory periapical periodontitis and secondary intraradicular infections. In this study, E. faecalis YN771 isolated from a re-treated root canal at a stomatology department was used as the host bacterium and was co-cultured with wastewater from the same department and patient samples to isolate a phage that lyses E. faecalis. We studied the biological and genomic characteristics of this phage. Transmission electron microscopy showed that this phage’s head is icosahedral in structure, with a head diameter of around 98.4 nm, and a contractile tail of around 228.5 nm in length and a diameter of 17.3 nm. The phage was identified as a member of the Myoviridae family and named PEf771. It is sensitive to proteinase K but resistant to chloroform and Triton X-100. Its lytic cycle is 45 min, burst size is 78, optimal multiplicity of infection is 0.1, lysis spectrum is narrow, and host strain specificity is strong. Its optimal growth temperature is 37 °C, most suitable pH is 6.0, and is sensitive to ultraviolet radiation. Whole-genome sequencing of PEf771 indicated it has a genome size of 151 052 bp, with a GC content of 36.97%, and encodes 197 proteins plus 26 tRNAs. PEf771 is most closely related to E. faecalis phage EFDG1. Phage PEf771 has strong host specificity and lytic ability, so it is important to further characterize this phage and its interaction with E. faecalis.

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

10.21203/rs.3.rs-170860/v1 ◽

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.

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Characterization of a Lytic Bacteriophage as an Antimicrobial Agent for Biocontrol of Shiga Toxin-Producing Escherichia coli O145 Strains

Antibiotics ◽

10.3390/antibiotics8020074 ◽

2019 ◽

Vol 8 (2) ◽

pp. 74 ◽

Cited By ~ 3

Author(s):

Yen-Te Liao ◽

Alexandra Salvador ◽

Leslie A. Harden ◽

Fang Liu ◽

Valerie M. Lavenburg ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Agent ◽

Shiga Toxin ◽

Burst Size ◽

Lytic Bacteriophage ◽

E Coli ◽

A Genome ◽

Wide Range ◽

Foodborne Outbreaks

Shiga toxin-producing Escherichia coli (STEC) O145 is one of the most prevalent non-O157 serogroups associated with foodborne outbreaks. Lytic phages are a potential alternative to antibiotics in combatting bacterial pathogens. In this study, we characterized a Siphoviridae phage lytic against STEC O145 strains as a novel antimicrobial agent. Escherichia phage vB_EcoS-Ro145clw (Ro145clw) was isolated and purified prior to physiological and genomic characterization. Then, in vitro antimicrobial activity against an outbreak strain, E. coli O145:H28, was evaluated. Ro145clw is a double-stranded DNA phage with a genome 42,031 bp in length. Of the 67 genes identified in the genome, 21 were annotated with functional proteins, none of which were stx genes. Ro145clw had a latent period of 21 min and a burst size of 192 phages per infected cell. The phage could sustain a wide range of pH (pH 3 to pH 10) and temperatures (−80 °C to −73 °C). Ro145clw was able to reduce E. coli O145:H28 in lysogeny broth by approximately 5 log at 37 °C in four hours. These findings indicate that the Ro145clw phage is a promising antimicrobial agent that can be used to control E. coli O145 in adverse pH and temperature conditions.

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SGP-C: A Broad Host Range Temperate Bacteriophage; Against Salmonella gallinarum

Frontiers in Microbiology ◽

10.3389/fmicb.2021.768931 ◽

2022 ◽

Vol 12 ◽

Author(s):

Syeda Zainab Ilyas ◽

Hafsa Tariq ◽

Abdul Basit ◽

Hamza Tahir ◽

Zulquernain Haider ◽

...

Keyword(s):

Host Range ◽

Burst Size ◽

Gc Content ◽

Broad Host Range ◽

Temperate Bacteriophage ◽

Salmonella Gallinarum ◽

Double Stranded Dna ◽

C Genome ◽

Super Infection ◽

Lysogenic Phage

Salmonella gallinarum is a poultry restricted-pathogen causing fowl-typhoid disease in adult birds with mortality rates up-to 80% and exhibit resistance against commonly used antibiotics. In this current study, a temperate broad host range bacteriophage SGP-C was isolated against S. gallinarum from poultry digesta. It showed infection ability in all the 15 tested field strains of S. gallinarum. The SGP-C phage produced circular, turbid plaques with alternate rings. Its optimum activity was observed at pH 7.0 and 37–42°C, with a latent period of 45 min and burst size of 187 virions/bacterial cell. The SGP-C lysogens, SGPC-L5 and SGPC-L6 exhibited super-infection immunity against the same phage, an already reported feature of lysogens. A virulence index of 0.5 and 0.001 as MV50 of SGP-C suggests its moderate virulence. The genome of SGP-C found circular double stranded DNA of 42 Kbp with 50.04% GC content, which encodes 63 ORFs. The presence of repressor gene at ORF49, and absence of tRNA sequence in SGP-C genome indicates its lysogenic nature. Furthermore, from NGS analysis of lysogens we propose that SGP-C genome might exist either as an episome, or both as integrated and temporary episome in the host cell and warrants further studies. Phylogenetic analysis revealed its similarity with Salmonella temperate phages belonging to family Siphoviridae. The encoded proteins by SGP-C genome have not showed homology with any known toxin and virulence factor. Although plenty of lytic bacteriophages against this pathogen are already reported, to our knowledge SGP-C is the first lysogenic phage against S. gallinarum reported so far.

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Characterization and genome analysis of two novel Aeromonas hydrophila phages PZL-Ah1and PZL-Ah8

10.21203/rs.3.rs-933679/v1 ◽

2021 ◽

Author(s):

Huabo Yu ◽

Chao Feng ◽

Liang Zhang ◽

Teng Chi ◽

Yanling Qi ◽

...

Keyword(s):

Aeromonas Hydrophila ◽

Bacterial Infections ◽

Genomic Sequence ◽

Gene Annotation ◽

Gc Content ◽

Opportunistic Pathogen ◽

Open Reading Frames ◽

Genome Database ◽

Narrow Host Range ◽

Phage T7

Abstract Aeromonas hydrophila (A.hydrophila) is an opportunistic pathogen of fish-human-livestock, which poses a seriously affects to the development of aquaculture. Phage therapy is considered as a process to alternatively control bacterial infections and contaminations. In this study, the genomes of two Aeromonas hydrophila- specific phages PZL-Ah1 and PZL-Ah8 were isolated, characterized and genomic sequence analyzed. Transmission electron microscopy showed that the two phages had been classified as the member of the Podoviridae family. Both the two phages in this study had relatively narrow host range with lytic activity against Aeromonas spp. strains. However, they could lyse 3 common A.hydrophila strain. As revealed from the whole genomic sequence analysis, PZL-Ah1 and PZL-Ah8 coverd the double-stranded genome of 38,641 bp and 40,855 bp in length, with the GC content of 53.68% and 51.89%, respectively. Through gene comparison in NCBI database revealed that PZL-Ah1 and PZL-Ah8 were 97.67% − 95.51% identical to Stenotrophomonas phage IME15 and Aeromonas Phage T7-Ah. Phylogenetic analysis showed that PZL-Ah8, PZL-Ah1 and other two phages belonged to the same genus. A total of 44 and 52 open reading frames (ORFs) were predicted in the PZL-Ah1 and PZL-Ah8 genome, respectively. In the process of gene annotation, 28 (63.6%) ORFs in PZL-Ah1 and 29 (55.8%) ORFs in PZL-Ah8 were known to functional proteins in NCBI database, while the remaining ORFs were classified as “hypothetical proteins”, whose functions were yet unknown. By comparing, ORF 02, ORF 29 and ORF 04 in PZL-Ah1, ORF24 in PZL-Ah8 were responsible for the host cell lysis. In conclusion, genomic studies of these two novel phages would lay the foundation for expanding the phage genome database and providing good candidates for phage typing applications.

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Characterization of a Unique Bordetella bronchiseptica vB_BbrP_BB8 Bacteriophage and Its Application as an Antibacterial Agent

International Journal of Molecular Sciences ◽

10.3390/ijms21041403 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1403 ◽

Cited By ~ 3

Author(s):

Mateusz Szymczak ◽

Bartłomiej Grygorcewicz ◽

Joanna Karczewska-Golec ◽

Przemysław Decewicz ◽

Jarosław Adam Pankowski ◽

...

Keyword(s):

Galleria Mellonella ◽

Therapeutic Potential ◽

Burst Size ◽

Genome Structure ◽

Bacterial Biofilm ◽

Bordetella Bronchiseptica ◽

Atrophic Rhinitis ◽

Lytic Bacteriophage ◽

Safe Alternative ◽

A Genome

Bordetella bronchiseptica, an emerging zoonotic pathogen, infects a broad range of mammalian hosts. B. bronchiseptica-associated atrophic rhinitis incurs substantial losses to the pig breeding industry. The true burden of human disease caused by B. bronchiseptica is unknown, but it has been postulated that some hypervirulent B. bronchiseptica isolates may be responsible for undiagnosed respiratory infections in humans. B. bronchiseptica was shown to acquire antibiotic resistance genes from other bacterial genera, especially Escherichia coli. Here, we present a new B. bronchiseptica lytic bacteriophage—vB_BbrP_BB8—of the Podoviridae family, which offers a safe alternative to antibiotic treatment of B. bronchiseptica infections. We explored the phage at the level of genome, physiology, morphology, and infection kinetics. Its therapeutic potential was investigated in biofilms and in an in vivo Galleria mellonella model, both of which mimic the natural environment of infection. The BB8 is a unique phage with a genome structure resembling that of T7-like phages. Its latent period is 75 ± 5 min and its burst size is 88 ± 10 phages. The BB8 infection causes complete lysis of B. bronchiseptica cultures irrespective of the MOI used. The phage efficiently removes bacterial biofilm and prevents the lethality induced by B. bronchiseptica in G. mellonella honeycomb moth larvae.

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vB-Ea-5: a lytic bacteriophage against multi-drug-resistant Enterobacter aerogenes

Iranian Journal of Microbiology ◽

10.18502/ijm.v13i2.5984 ◽

2021 ◽

Author(s):

Fatemeh Habibinava ◽

Mohammad Reza Zolfaghari ◽

Mohsen Zargar ◽

Salehe Sabouri Shahrbabak ◽

Mohammad Soleimani

Keyword(s):

Burst Size ◽

Gc Content ◽

Enterobacter Aerogenes ◽

Resistant Bacteria ◽

Potential Candidate ◽

Lytic Bacteriophage ◽

Drug Resistant ◽

Drug Resistant Bacteria ◽

Transmission Electron ◽

Functional Classes

Background and Objectives: Multi-drug-resistant Enterobacter aerogenes is associated with various infectious diseases that cannot be easily treated by antibiotics. However, bacteriophages have potential therapeutic applications in the control of multi-drug-resistant bacteria. In this study, we aimed to isolate and characterize of a lytic bacteriophage that can lyse specifically the multi-drug-resistant (MDR) E. aerogenes. Materials and Methods: Lytic bacteriophage was isolated from Qaem hospital wastewater and characterized morphologically and genetically. Next-generation sequencing was used to complete genome analysis of the isolated bacteriophage. Results: Based on the transmission electron microscopy feature, the isolated bacteriophage (vB-Ea-5) belongs to the family Myoviridae. vB-Ea-5 had a latent period of 25 minutes, a burst size of 13 PFU/ml, and a burst time of 40 min. Genome sequencing revealed that vB-Ea-5 has a 135324 bp genome with 41.41% GC content. The vB-Ea-5 genome codes 212 ORFs 90 of which were categorized into several functional classes such as DNA replication and modification, transcriptional regulation, packaging, structural proteins, and a host lysis protein (Holin). No antibiotic resistance and toxin genes were detected in the genome. SDS-PAGE of vB-Ea-5 proteins exhibited three major and four minor bands with a molecular weight ranging from 18 to 50 kD. Conclusion: Our study suggests vB-Ea-5 as a potential candidate for phage therapy against MDR E. aerogenes infections.

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