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  • Open AccessReview Article

    Filippov et al., J Bioterr Biodef 2013, S3 DOI: 10.4172/2157-2526.S3-010

    ISSN:2157-2526 JBTBD, an open access journal Advances in Biosciences: BioterrorismJ Bioterr Biodef

    therapy of these infections. Many bacterial viruses (bacteriophages or phages) active against Y. pestis, B. anthracis and Brucella species have been described. Characteristics and practical applications of such phages are the subject of this review. Due to the lack of data on lytic phages of F. tularensis and very limited information on practical importance of phages capable of lysing B. pseudomallei and B. mallei, this review does not cover the literature on these bacteria.

    General Characteristics of Phages Active against Biothreat Bacteria

    Keywords: Yersinia pestis; Bacillus anthracis; Brucella; Bacteriophages;Phage diagnostics; Phage typing; Phage decontamination; Phage therapy

    Abbreviations: ATP: Adenosine Triphosphate; CDC: Centers forDisease Control and Prevention; CFU: Colony-forming Unit; EOP: Efficiency of Plating; FDA: Food and Drug Administration; FSU: Former Soviet Union; h: hour; LD50: 50% Lethal Dose; LPS: Lipopolysaccharide; PCR: Polymerase Chain Reaction; qPCR: Quantitative Real-time Polymerase Chain Reaction; PFU: Plaque-forming Unit; RTD: Routine Test Dilution; sbsp.: subspecies; UA: Unavailable

    IntroductionBioterrorism or biothreat agents are highly infectious and pathogenic

    microorganisms (bacteria, viruses, and fungi) and their toxins that can be used by individuals or groups of terrorists or as biowarfare agents in military operations. The Centers for Disease Control and Prevention (CDC) has established two groups of the most dangerous biothreatagents, categories A and B [1,2]. The highest-priority category A, inaddition to botulinum toxin and several especially dangerous viruses,includes three biothreat bacteria, the causative agents of bubonicand pneumonic plague (Yersinia pestis), anthrax (Bacillus anthracis)and tularemia (Francisella tularensis). Category B comprises severaltoxins, viruses and bacterial pathogens including the causative agentsof brucellosis (Brucella species), glanders (Burkholderia mallei), andmelioidosis (Burkholderia pseudomallei). In this review, we focus onplague, anthrax and brucellosis. Plague [3] and anthrax [4] are severefulminant primarily zoonotic infections that can be transmitted tohumans and easily disseminated in human populations, mainly vianatural or artificial aerosolization of the bacteria, resulting in epidemicswith high mortality rates. Brucellosis is a globally spread zoonosis whichis also prevalent in humans in the form of severe systemic disease. Thereare four species of Brucella pathogenic for humans, Brucella melitensis,Brucella suis, Brucella abortus, and Brucella canis [5]. The serious publichealth concern about these biothreat infections is aggravated by theemergence of multidrug-resistant strains of Y. pestis [6,7] and isolatesof Brucella resistant to some clinically relevant antibiotics [8,9], as wellas by easy induction of antibiotic resistance in B. anthracis [10], anda potential for the application of genetically engineered multidrug-resistant strains of biothreat bacteria in biological attacks [11-13].The severity of these biothreat bacterial infections requires efficientbiosurveillance and biodefense, including availability of a rich arsenalof up-to-date methods of rapid detection and identification of thebacteria, strain characterization, diagnostics, efficient prophylaxis, and

    *Corresponding author: Andrey A Filippov, Department of Emerging Bacterial Infections, Bacterial Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA, E-mail:

    Received December 27, 2012; Accepted January 18, 2013; Published January 21, 2013

    Citation: Filippov AA, Sergueev KV, Nikolich MP (2013) Bacteriophages against Biothreat Bacteria: Diagnostic, Environmental and Therapeutic Applications. J Bioterr Biodef S3: 010. doi:10.4172/2157-2526.S3-010

    Copyright: 2013 Filippov AA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits un-restricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

    Bacteriophages against Biothreat Bacteria: Diagnostic, Environmental and Therapeutic ApplicationsAndrey A Filippov*, Kirill V Sergueev and Mikeljon P NikolichDepartment of Emerging Bacterial Infections, Bacterial Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA

    AbstractPlague, anthrax and brucellosis are severe bacterial infections presenting a serious threat to public health. Their

    causative agents can be weaponized and a number of drug-resistant strains have been described. This requires improvement of existing and development of new methods of diagnostics, strain characterization, prophylaxis and therapy of these infections. This review article focuses on lytic bacteriophages (phages) active against Yersinia pestis, Bacillus anthracis and Brucella including the uses of phages for diagnostics, strain typing, specific decontamination, and antibacterial therapy.

    The Most important bacteriophages lytic for Y. pestis, B. anthracis and Brucella species are listed in table 1. The listed bacteriophages are members of the same order, Caudovirales (tailed viruses) but belong to five different families (Myoviridae, Podoviridae, Siphoviridae, Tectiviridae, and Inoviridae) and different groups. The first three families have different tail structures: long contractile, short noncontractile, or long noncontractile tails, respectively. The features of the Tectiviridae phages are a double coat and a tail equivalent, and the Inoviridae family representatives have a filamentous structure. Phages of these families contain linear double-stranded DNA, except the Inoviridae members which carry circular single-stranded DNA [83]. The genomes of many phages active against biothreat bacteria were sequenced (Table 1). The bacterial cell surface receptors were identified for Y. pestis-specific phages A1122 [16,84], Pokrovskaya, L-413C, Y, JA1, R, and PST [16], mainly in different sugar residues of the LPS outer and inner core. It was found that A1122 is specific for Y. pestis when grown at 26-28C but can efficiently lyse both Y. pestis and Yersinia pseudotuberculosis (the closest phylogenetic relative of Y. pestis) at 37C [17,20-22,84], due to repressed production of O-antigen which blocks the Y. pseudotuberculosis phage receptor in the LPS inner core when grown at lower temperatures [84].



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    ISSN: 2157-2526

    Journal of Bioterrorism & Biodefense

  • Citation: Filippov AA, Sergueev KV, Nikolich MP (2013) Bacteriophages against Biothreat Bacteria: Diagnostic, Environmental and Therapeutic Applications. J Bioterr Biodef S3: 010. doi:10.4172/2157-2526.S3-010

    Page 2 of 8

    ISSN:2157-2526 JBTBD, an open access journal J Bioterr Biodef Advances in Biosciences: Bioterrorism

    The receptor for B. anthracis-specific phage gamma was localized in a cell surface-anchored protein, GamR [85]. The AP50 phage of B. anthracis was recently shown to require for its adsorption the intact structure of another cell surface-anchored protein, CsaB, important for S-layer assembly [53]. Since the main difference between smooth and rough strains of Brucella is the presence or absence of the lipopolysaccharide (LPS) O antigen, receptors of brucellaphages should at least partially involve LPS; the receptors for S-specific phages of Brucella were roughly associated with a protein-phospholipid-LPS complex [79,86].

    Lytic Y. pestis phages were isolated from animals infected with plague [87] or their carcasses [14], blood of patients [19], rodent burrow soil [88], and from sewage [16,17]. Several groups of temperate phages were isolated from Y. pestis cultures with subsequent selection of virulent (lytic) mutants from some of them [35,89-91]. Phages capable of lysing Y. pestis are similar to T7, T4, T1 or P2 phages that typically grow on enteric bacteria such as Escherichia coli and Shigella, with a prevalence of highly lytic T7-like phages (Table 1). Some of plague phages can be considered host range mutants of coliphages. For example, we isolated a Y. pestis-specific T4-like phage JA1 from Maryland sewage [16,17].


    Bacteriophage Family Group Sequence No. Reference(s)

    Y. pestis Pokrovskaya(YepE2, YpP-G)

    Podoviridae T7 NC_011038;JQ9657021


    Y. pestis A1122 Podoviridae T7 NC_004777 [16,17,19-23]Y. pestis Y Podoviridae T7 JQ9657001 [16-18,24,25]Y. pestis2 R Podoviridae T7 JQ9657011 [16-18,24,26-28]Y. pestis2 dHerelle-m

    (YpsP-G)Podoviridae T7 JQ9657031 [17,18,27,29,30]

    Y. pestis Yep-phi Podoviridae T7 HQ333270 [31]Y. pestis2 PST Myoviridae T4 UA1 [16,17,24,32,33]Y. pestis JA1 Myoviridae T4 UA1 [16,17]Y. pestis3 PY100 Siphoviridae T1 AM076770 [34]Y. pestis L-413C Myoviridae P2 NC_004745 [15-17,22,23,35,36]B. anthracis4 (W; beta) Siphoviridae DQ289555 [37-40]B. anthracis4 (W; gamma) Siphoviridae DQ221100



    B. anthracis Fah Siphoviridae NC_007814 [46-48]B. anthracis AP50 Tectiviridae ? NC_011523 [49-53]B. anthracis5 8a Myoviridae ? UA [54]B. anthracis Nk Myoviridae ? UA [55]B. anthracis DB Myoviridae ? UA [55]B. anthracis MH Podoviridae ? UA [55]B. anthracis JRB7 Inoviridae ? UA [56,57]B. anthracis K ? ? UA [43,47,58,59]B. anthracis VA-9 ? ? UA [38,47]B. abortus6 Tb (Tbilisi) Podoviridae Tb NC_019446 [60-67]B. abortus7 Np (Nepean) Podoviridae Tb UA [65]B. abortus8 Fz (Firenze) Podoviridae Tb UA9 [63,65,68-70]B. abortus10 Pr (Perote) Podoviridae Tb NC_019447 [67]


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